Antigen binding molecule specifically binding to tfr1 and aβ, and medical use thereof

Abstract

The present invention relates to an antigen binding molecule specifically binding to TfR1 and Aβ, and a medical use thereof. Specifically, the present invention relates to the use of the antigen-binding molecule specifically binding to TfR1 and Aβ in the preparation of a medicament for preventing or treating a disease or disorder.

Description

Antigen-binding molecules that specifically bind to TfR1 and Aβ and their pharmaceutical uses

[0001] This application claims priority to Chinese patent application CN202510366281.0 filed on March 26, 2025, Chinese patent application CN202610267205.9 filed on March 5, 2026, and Chinese patent application CN202610288920.0 filed on March 10, 2026. Technical Field

[0002] This disclosure pertains to the field of biotechnology, and more specifically, to antigen-binding molecules that specifically bind to TfR1 and Aβ and their pharmaceutical uses. Background Technology

[0003] The statements herein are provided only as background information in connection with this disclosure and do not necessarily constitute prior art.

[0004] Alzheimer's disease (AD), also known as senile dementia, is a serious neurodegenerative disease. Its main clinical manifestations are cognitive decline, mental symptoms, and behavioral disorders, with severe cases resulting in the loss of basic living abilities. It is generally believed that the deposition of β-amyloid protein (Aβ) triggers neurofibrillary tangles, leading to a reduction in synapses between nerve cells, cell death, and cognitive decline (Corbett A, et al. Nature reviews. Drug discovery, 2012, 11(11):833–846). In recent years, the FDA has approved two Aβ antibody drugs, Aducanumab and Lecanemab, for the early treatment of Alzheimer's disease, indicating a dose-response relationship between Aβ clearance and clinical benefits. However, their application is still limited by insufficient central clearance efficiency of Aβ and adverse reactions such as cerebral hemorrhage and cerebral edema (Karran E, et al. Nature reviews. Drug discovery, 2022, 21(4):306–318). WO2007108756A1, WO2008081008A1, WO2020015637A1, and other publications disclose a variety of anti-Aβ antibodies.

[0005] Transferrin receptor protein 1 (TfR1), also known as CD71 or TFRC, is a type II transmembrane protein and the most important membrane protein regulating intracellular iron transport (Pierre VC, et al. Frontiers in immunology, 2021, 12:607692). Under normal physiological conditions, the primary function of TfR1 is to bind to transferrin (TF) and mediate cellular iron uptake through endocytosis. Therefore, the TF-TfR1 system is considered an important pathway for the body to acquire iron ions. TfR1 is a membrane protein widely expressed in almost all cell and tissue types in the human body, and is widely distributed in various tissues such as the immune system, hematopoietic system, nervous system, liver, and kidneys (Kawabata H. Free radical biology & medicine, 2019, 133:46–54). The blood-brain barrier (BBB) ​​in the circulatory and central nervous systems makes drug delivery to the brain difficult, posing a significant challenge to the treatment of central nervous system diseases (Neuwelt, E., et al. The Lancet Neurology, 2008, 7(1):84–96). Studies have shown that the brain uptake of traditional IgG antibodies is only 0.1%–0.2% of the injected dose (Yu, YJ, et al. Neurootherapeutics, 10(3):459–472). To improve the efficiency of intracranial drug delivery, endocytosis of certain receptors or transporters highly expressed on brain capillary endothelial cells (BCECs) is utilized to cross the blood-brain barrier. TfR1 is a receptor highly expressed on BCECs and can serve as a good cross-BBB transport vehicle for targeted delivery of therapeutic drugs to the brain (Johnsen, KB, et al. Progress in neurobiology, 2019, 181:101665). Bispecific antibody drugs targeting TfR1 have been shown to significantly increase antibody exposure in the brain of non-human primates, by 4-18 times compared to monoclonal antibody drugs (Grimm, HP, et al. mAbs, 2023, 15(1):2261509). WO1993010819A1 discloses a murine anti-TfR1 antibody ALK 128.1, while WO2021092482A1, WO2023287238A1, and WO2024155066A1 disclose various humanized anti-TfR1 antibodies based on ALK 128.1.

[0006] WO2017055540A1, WO2024200267A1, and WO2024206161A1 have disclosed several antigen-binding molecules that specifically bind to TfR1 and Aβ. New therapeutic agents for the central nervous system (CNS) are still needed clinically. Summary of the Invention

[0007] This disclosure provides a novel antigen-binding molecule that specifically binds to TfR1 and Aβ to overcome the BBB barrier, thereby establishing a central nervous system (CNS) drug delivery platform and improving the efficacy of drugs for treating CNS diseases.

[0008] This disclosure provides antigen-binding molecules that specifically bind to TfR1, antigen-binding molecules that specifically bind to Aβ, and antigen-binding molecules that specifically bind to both TfR1 and Aβ.

[0009] <Antigen-binding molecules that specifically bind to TfR1>

[0010] This disclosure provides an antigen-binding molecule that specifically binds to TfR1, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0011] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 3, HCDR2 contains the amino acid sequence of SEQ ID NO: 11, and HCDR3 contains the amino acid sequence of SEQ ID NO: 36; the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 9, LCDR2 contains the amino acid sequence of SEQ ID NO: 10, and LCDR3 contains the amino acid sequence of SEQ ID NO: 8, wherein:

[0012] X1 can be S or Y, with S being preferred; X2 can be T or S, with T being preferred.

[0013] This disclosure provides an antigen-binding molecule that specifically binds to TfR1, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0014] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain the amino acid sequences of HCDR1, HCDR2, and HCDR3 in SEQ ID NO: 27, 12, or 26, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain the amino acid sequences of LCDR1, LCDR2, and LCDR3 in SEQ ID NO: 13, 14, 15, or 16.

[0015] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein:

[0016] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain the amino acid sequences of HCDR1, HCDR2, and HCDR3 in SEQ ID NO: 27, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain the amino acid sequences of LCDR1, LCDR2, and LCDR3 in SEQ ID NO: 13.

[0017] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments, wherein the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to a numbering rule selected from Kabat, IMGT, Chothia, AbM, and Contact. In some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to the Kabat numbering rule. In some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to the IMGT numbering rule. In some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to the Chothia numbering rule. In some embodiments, the heavy chain variable regions HCDR1, HCDR2, and HCDR3, and the light chain variable regions LCDR1, LCDR2, and LCDR3, are defined according to the AbM numbering rules. In some embodiments, the heavy chain variable regions HCDR1, HCDR2, and HCDR3, and the light chain variable regions LCDR1, LCDR2, and LCDR3, are defined according to the Contact numbering rules.

[0018] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein:

[0019] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 3, HCDR2 contains the amino acid sequence of SEQ ID NO: 11, and HCDR3 contains the amino acid sequence of SEQ ID NO: 25, 5, or 24. The light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 9, LCDR2 contains the amino acid sequence of SEQ ID NO: 10, and LCDR3 contains the amino acid sequence of SEQ ID NO: 8.

[0020] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein:

[0021] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 3, HCDR2 contains the amino acid sequence of SEQ ID NO: 11, and HCDR3 contains the amino acid sequence of SEQ ID NO: 25; and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 9, LCDR2 contains the amino acid sequence of SEQ ID NO: 10, and LCDR3 contains the amino acid sequence of SEQ ID NO: 8; or

[0022] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 3, HCDR2 contains the amino acid sequence of SEQ ID NO: 11, and HCDR3 contains the amino acid sequence of SEQ ID NO: 5; and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 9, LCDR2 contains the amino acid sequence of SEQ ID NO: 10, and LCDR3 contains the amino acid sequence of SEQ ID NO: 8; or

[0023] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 3, HCDR2 contains the amino acid sequence of SEQ ID NO: 11, and HCDR3 contains the amino acid sequence of SEQ ID NO: 24, and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 9, LCDR2 contains the amino acid sequence of SEQ ID NO: 10, and LCDR3 contains the amino acid sequence of SEQ ID NO: 8.

[0024] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments includes a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region includes HCDR1, HCDR2, and HCDR3, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, wherein:

[0025] The amino acid sequences of the heavy chain variable regions HCDR1 are shown in SEQ ID NO: 3, HCDR2 in SEQ ID NO: 11, and HCDR3 in SEQ ID NO: 25; the amino acid sequences of the light chain variable regions LCDR1 in SEQ ID NO: 9, LCDR2 in SEQ ID NO: 10, and LCDR3 in SEQ ID NO: 8; or

[0026] The amino acid sequences of the heavy chain variable regions HCDR1 are shown in SEQ ID NO: 3, HCDR2 in SEQ ID NO: 11, and HCDR3 in SEQ ID NO: 5; the amino acid sequences of the light chain variable regions LCDR1 in SEQ ID NO: 9, LCDR2 in SEQ ID NO: 10, and LCDR3 in SEQ ID NO: 8; or

[0027] The amino acid sequences of the heavy chain variable regions HCDR1 are shown in SEQ ID NO: 3, HCDR2 are shown in SEQ ID NO: 11, and HCDR3 are shown in SEQ ID NO: 24. The amino acid sequences of the light chain variable regions LCDR1 are shown in SEQ ID NO: 9, LCDR2 are shown in SEQ ID NO: 10, and LCDR3 are shown in SEQ ID NO: 8.

[0028] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the Kabat numbering rules.

[0029] In some embodiments, the antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, is an antibody. In some embodiments, the antigen-binding molecule that specifically binds to TfR1 is a monoclonal antibody, a monospecific antibody, a bispecific antibody, a multispecific antibody, or a fusion protein. In some embodiments, the antigen-binding molecule that specifically binds to TfR1 is a monoclonal antibody. In some embodiments, the antigen-binding molecule that specifically binds to TfR1 is a monospecific antibody. In some embodiments, the antigen-binding molecule that specifically binds to TfR1 is a murine antibody, a chimeric antibody, a humanized antibody, or a fully human antibody. In some embodiments, the antigen-binding molecule that specifically binds to TfR1 is a humanized antibody.

[0030] In some implementations, the antigen-binding molecule that specifically binds to TfR1, as described in the preceding one, includes the frame region (FR) of the human antibody.

[0031] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments, wherein the heavy chain variable region has FR1, FR2, FR3 fragments derived from IGHV1-2*06 and FR4 fragments derived from IGHJ6*01, and is unsubstituted or contains one or more amino acid substitutions selected from the group consisting of 1E, 48I, 69L, 71V and / or 73K; and / or the light chain variable region has FR1, FR2, FR3 fragments derived from IGKV3-11*01 and FR4 fragments derived from IGKJ4*01, and is unsubstituted or contains one or more amino acid substitutions selected from the group consisting of 46R, 47W and / or 58V. In some embodiments, the antigen-binding molecule that specifically binds to TfR1, wherein the heavy chain variable region HCDR1 comprises the amino acid sequence of SEQ ID NO: 3, HCDR2 comprises the amino acid sequence of SEQ ID NO: 11, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 25, 5, or 24, and the FR of the heavy chain variable region is unsubstituted or comprises one or more amino acid substitutions selected from the group consisting of 1E, 48I, 69L, 71V, and / or 73K; and the light chain variable region LCDR1 comprises the amino acid sequence of SEQ ID NO: 9, LCDR2 comprises the amino acid sequence of SEQ ID NO: 10, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 8, and the FR of the light chain variable region is unsubstituted or comprises one or more amino acid substitutions selected from the group consisting of 46R, 47W, and / or 58V. In some embodiments, the above-mentioned variable regions and CDRs are defined according to the Kabat numbering rules.

[0032] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein:

[0033] The heavy chain variable region comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 27, 12, or 26, and the light chain variable region comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 13, 14, 15, or 16.

[0034] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein:

[0035] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 27, 12 or 26, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 13, 14, 15 or 16.

[0036] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein:

[0037] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 27, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 13, 14, 15, or 16; or

[0038] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 12, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 13, 14, 15, or 16; or

[0039] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 26, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 13, 14, 15 or 16.

[0040] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein:

[0041] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 27, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 13; or

[0042] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 27, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 14; or

[0043] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 27, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 15; or

[0044] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 27, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 16; or

[0045] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 12, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 13; or

[0046] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 12, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 14; or

[0047] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 12, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 15; or

[0048] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 12, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 16; or

[0049] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 26, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 13; or

[0050] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 26, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 14; or

[0051] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 26, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 15; or

[0052] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 26, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 16.

[0053] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein:

[0054] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13; or

[0055] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 14; or

[0056] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 15; or

[0057] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 16; or

[0058] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 12, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13; or

[0059] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 12, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 14; or

[0060] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 12, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 15; or

[0061] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 12, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 16; or

[0062] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 26, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13; or

[0063] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 26, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 14; or

[0064] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 26, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 15; or

[0065] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 26, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 16.

[0066] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein the antigen-binding molecule that specifically binds to TfR1 is humanized, reversed mutation, affinity maturation, T-cell epitope removal, antibody deamidation reduction, and / or antibody isomerization reduction modification.

[0067] In some embodiments, the antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, is an antigen-binding fragment. In some embodiments, the antigen-binding fragment is an antibody fragment. In some embodiments, the antigen-binding fragment is selected from Fab, Fab′, F(ab′)2, Fd, Fv, scFv, dsFv, and dAb.

[0068] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises a heavy chain constant region and a light chain constant region. In some embodiments, the heavy chain constant region is a heavy chain constant region of human IgG1, IgG2, IgG3, IgG4, or variants thereof, and the light chain constant region is a light chain constant region of human κ chain, λ chain, or variants thereof. In some embodiments, the heavy chain constant region is a human IgG1 heavy chain constant region or a variant thereof, and the light chain constant region is a human κ light chain constant region or a variant thereof. In some embodiments, the heavy chain constant region is a human IgG1 heavy chain constant region, and the light chain constant region is a human κ light chain constant region. In some embodiments, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 17, and the light chain constant region comprises the amino acid sequence of SEQ ID NO: 18.

[0069] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, comprises a heavy chain and a light chain, wherein:

[0070] The heavy chain comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 29, 19, or 28, and the light chain comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 20, 21, 22, or 23.

[0071] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein:

[0072] The heavy chain comprises the amino acid sequence of SEQ ID NO: 29, 19 or 28, and the light chain comprises the amino acid sequence of SEQ ID NO: 20, 21, 22 or 23.

[0073] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein:

[0074] The heavy chain comprises the amino acid sequence of SEQ ID NO: 29, and the light chain comprises the amino acid sequence of SEQ ID NO: 20, 21, 22, or 23; or

[0075] The heavy chain comprises the amino acid sequence of SEQ ID NO: 19, and the light chain comprises the amino acid sequence of SEQ ID NO: 20, 21, 22, or 23; or

[0076] The heavy chain contains the amino acid sequence of SEQ ID NO: 28, and the light chain contains the amino acid sequence of SEQ ID NO: 20, 21, 22 or 23.

[0077] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein:

[0078] The heavy chain comprises the amino acid sequence of SEQ ID NO: 29, and the light chain comprises the amino acid sequence of SEQ ID NO: 20; or

[0079] The heavy chain comprises the amino acid sequence of SEQ ID NO: 29, and the light chain comprises the amino acid sequence of SEQ ID NO: 21; or

[0080] The heavy chain comprises the amino acid sequence of SEQ ID NO: 29, and the light chain comprises the amino acid sequence of SEQ ID NO: 22; or

[0081] The heavy chain comprises the amino acid sequence of SEQ ID NO: 29, and the light chain comprises the amino acid sequence of SEQ ID NO: 23; or

[0082] The heavy chain comprises the amino acid sequence of SEQ ID NO: 19, and the light chain comprises the amino acid sequence of SEQ ID NO: 20; or

[0083] The heavy chain comprises the amino acid sequence of SEQ ID NO: 19, and the light chain comprises the amino acid sequence of SEQ ID NO: 21; or

[0084] The heavy chain comprises the amino acid sequence of SEQ ID NO: 19, and the light chain comprises the amino acid sequence of SEQ ID NO: 22; or

[0085] The heavy chain comprises the amino acid sequence of SEQ ID NO: 19, and the light chain comprises the amino acid sequence of SEQ ID NO: 23; or

[0086] The heavy chain comprises the amino acid sequence of SEQ ID NO: 28, and the light chain comprises the amino acid sequence of SEQ ID NO: 20; or

[0087] The heavy chain comprises the amino acid sequence of SEQ ID NO: 28, and the light chain comprises the amino acid sequence of SEQ ID NO: 21; or

[0088] The heavy chain comprises the amino acid sequence of SEQ ID NO: 28, and the light chain comprises the amino acid sequence of SEQ ID NO: 22; or

[0089] The heavy chain contains the amino acid sequence of SEQ ID NO: 28, and the light chain contains the amino acid sequence of SEQ ID NO: 23.

[0090] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein:

[0091] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 29, and the amino acid sequence of the light chain is shown in SEQ ID NO: 20; or

[0092] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 29, and the amino acid sequence of the light chain is shown in SEQ ID NO: 21; or

[0093] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 29, and the amino acid sequence of the light chain is shown in SEQ ID NO: 22; or

[0094] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 29, and the amino acid sequence of the light chain is shown in SEQ ID NO: 23; or

[0095] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 19, and the amino acid sequence of the light chain is shown in SEQ ID NO: 20; or

[0096] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 19, and the amino acid sequence of the light chain is shown in SEQ ID NO: 21; or

[0097] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 19, and the amino acid sequence of the light chain is shown in SEQ ID NO: 22; or

[0098] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 19, and the amino acid sequence of the light chain is shown in SEQ ID NO: 23; or

[0099] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 28, and the amino acid sequence of the light chain is shown in SEQ ID NO: 20; or

[0100] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 28, and the amino acid sequence of the light chain is shown in SEQ ID NO: 21; or

[0101] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 28, and the amino acid sequence of the light chain is shown in SEQ ID NO: 22; or

[0102] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 28, and the amino acid sequence of the light chain is shown in SEQ ID NO: 23.

[0103] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises a heavy chain constant region CH1 and a light chain constant region CL, wherein the heavy chain constant region CH1 is a heavy chain constant region CH1 of human IgG1, IgG2, IgG3, IgG4, or variants thereof, and the light chain constant region CL is a light chain constant region CL of human κ chain, λ chain, or variants thereof. In some embodiments, the heavy chain constant region CH1 is a heavy chain constant region CH1 of human IgG1 or variants thereof, and the light chain constant region CL is a light chain constant region CL of human κ chain or variants thereof. In some embodiments, the heavy chain constant region CH1 comprises the amino acid sequence of SEQ ID NO: 82, and the light chain constant region CL comprises the amino acid sequence of SEQ ID NO: 18.

[0104] In some embodiments, the antigen-binding molecule that specifically binds to TfR1, as described in the preceding one, includes an Fc region.

[0105] In some implementations, the Fc region is the Fc region of human IgG1, IgG2, IgG3, IgG4 or variants thereof.

[0106] In some embodiments, the Fc region is either unsubstituted or contains amino acid substitutions of 351I and 356K, numbered according to the EU index.

[0107] In some implementations, the Fc region contains amino acid substitutions of 351I and 356K, numbered according to the EU index.

[0108] In some embodiments, the Fc region is the Fc region of a human IgG1 variant, wherein the Fc1 is an unsubstituted amino acid or contains L351I and D356K substituted amino acids, numbered according to the EU index.

[0109] In some embodiments, the Fc region is the Fc region of a human IgG1 variant, wherein the Fc1 contains amino acid substitutions of L351I and D356K, numbered according to the EU index.

[0110] In some embodiments, the Fc region contains the amino acid sequence of SEQ ID NO: 80 or 79.

[0111] In some embodiments, the Fc region contains the amino acid sequence of SEQ ID NO: 80.

[0112] In some embodiments, the amino acid sequence of the Fc region is shown in SEQ ID NO: 80.

[0113] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments includes an Fc region comprising two Fc1 subunits capable of associating with each other; and includes two identical haptens comprising Fab and Fc1 regions that bind to TfR1.

[0114] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments includes an Fc region comprising two Fc1 subunits capable of associating with each other; and includes two identical haptens comprising at least one (e.g., 1, 2, 3, 4, 5, or 6) TfR1-binding Fab and at least one (e.g., 1, 2, 3, 4, 5, or 6) Fc1 region.

[0115] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments includes an Fc region comprising two Fc1 subunits capable of associating with each other; and includes two identical haptens that include at least one (e.g., 1, 2, 3, 4, 5 or 6) TfR1-binding Fab and one Fc1 region in the direction from the N-terminus to the C-terminus.

[0116] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments includes an Fc region comprising two Fc1 subunits capable of associating with each other; and includes two identical haptens comprising a TfR1-binding Fab and an Fc1 region from the N-terminus to the C-terminus.

[0117] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments includes an Fc region comprising two Fc1 subunits capable of associating with each other; and includes two identical haptens comprising, from the N-terminus to the C-terminus, a TfR1-binding Fab and an Fc1 region, wherein the C-terminus of the TfR1-binding Fab is operatively linked to the N-terminus of the Fc1 region.

[0118] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments includes an Fc region comprising two mutually associative subunits Fc1; and includes two identical haptens comprising, from the N-terminus to the C-terminus, a TfR1-binding Fab and an Fc1 region, wherein the C-terminus of the CH1 of the TfR1-binding Fab is operatively linked to the N-terminus of the Fc1 region.

[0119] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments includes an Fc region comprising two Fc1 subunits capable of associating with each other; and includes two identical haptens comprising, from the N-terminus to the C-terminus, a TfR1-binding Fab and an Fc1 region, wherein the C-terminus of the TfR1-binding Fab is fused directly or via a linker to the N-terminus of the Fc1 region.

[0120] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments includes an Fc region comprising two Fc1 subunits capable of associating with each other; and includes two identical haptens comprising, from the N-terminus to the C-terminus, a TfR1-binding Fab and an Fc1 region, wherein the C-terminus of the TfR1-binding Fab is fused to the N-terminus of the Fc1 region via a linker.

[0121] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments includes an Fc region comprising two Fc1 subunits capable of associating with each other; and includes two identical haptens comprising, from the N-terminus to the C-terminus, a TfR1-binding Fab and an Fc1 region, wherein the C-terminus of the CH1 of the TfR1-binding Fab is fused directly or via a linker to the N-terminus of the Fc1 region.

[0122] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments includes an Fc region comprising two mutually associative subunits Fc1; and includes two identical haptens comprising, from the N-terminus to the C-terminus, a TfR1-binding Fab and an Fc1 region, wherein the C-terminus of the CH1 of the TfR1-binding Fab is fused to the N-terminus of the Fc1 region via a linker.

[0123] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises a first chain with the structure shown in formula (a) and a second chain with the structure shown in formula (b), wherein the structures shown in formula (a) and formula (b) are arranged from the N-terminus to the C-terminus:

[0124] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[0125] (b)[TfR1-VL]-[CL].

[0126] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) first strand with the structure shown in formula (a), and at least one (e.g., 1, 2, 3, 4, 5, or 6) second strand with the structure shown in formula (b), wherein the structures shown in formulas (a) and (b) are arranged from the N-terminus to the C-terminus.

[0127] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[0128] (b)[TfR1-VL]-[CL].

[0129] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises two first strands with the structure shown in formula (a) and two second strands with the structure shown in formula (b), wherein the structures shown in formula (a) and formula (b) are arranged from the N-terminus to the C-terminus:

[0130] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[0131] (b)[TfR1-VL]-[CL].

[0132] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein the linker 1 is a peptide linker, or the linker 1 is absent.

[0133] In some embodiments, an antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, wherein the first and second chains are linked together to form an antigen-binding molecule that specifically binds to TfR1.

[0134] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises two first strands with the structure shown in formula (a) and two second strands with the structure shown in formula (b), wherein the structures shown in formula (a) and formula (b) are arranged from the N-terminus to the C-terminus:

[0135] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[0136] (b)[TfR1-VL]-[CL];

[0137] The linker 1 is a peptide linker, or the linker 1 does not exist;

[0138] The first and second chains combine to form an antigen-binding molecule that specifically binds to TfR1.

[0139] In some embodiments, the structure of the antigen-binding molecule that specifically binds to TfR1, as described in any of the preceding embodiments, is shown in Format 1 of Figure 1A.

[0140] In some embodiments, as described in any of the preceding embodiments, the linker is a peptide linker in an antigen-binding molecule that specifically binds to TfR1. In some embodiments, the linker 1 is a peptide linker in an antigen-binding molecule that specifically binds to TfR1. In some embodiments, the peptide linker may be a flexible peptide containing 1-50 or 1-20 amino acid residues. In some embodiments, each of the peptide linkers independently has L1-(GGGGS). n The structure is -L2, wherein L1 is a bond, A, G, GS, GGG, GGS, GGGS (SEQ ID NO: 98) or GGGG (SEQ ID NO: 99), n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, L2 is a bond, G, GG, GGG or GGGG, and the peptide linker is not a bond. In some embodiments, the peptide linker is 2-20 amino acid residues in length. In some embodiments, the peptide linker is as follows: (GS) a (GGS) b (GGGS) c (GGGGS) d (GGGGG) ea, b, c, d, and e are independent integers greater than or equal to 0; or the peptide linker is selected from: (EAAAK)3 (SEQ ID NO: 100), (EAAAR)3 (SEQ ID NO: 101), (EGGGK)3 (SEQ ID NO: 102), (EGGGR)3 (SEQ ID NO: 103), (DAAAR)3 (SEQ ID NO: 104), (DAAAK)3 (SEQ ID NO: 105), (DGGGR)3 (SEQ ID NO: 106) or (DGGGK)3 (SEQ ID NO: 107), SS, GGSEGKSSGSGSESKSTGGS (SEQ ID NO: 108), EPKSS (SEQ ID NO: 109), RTVAA (SEQ ID NO: 110), ASEPKSS (SEQ ID NO: 111); or the peptide linker is (G x S) y Where x is an integer selected from 1 to 5, and y is an integer selected from 1 to 6, including but not limited to GGGS, GGGGS (SEQ ID NO: 112), GGGGSGGGGS (SEQ ID NO: 113), GGGGSGGGGSGGGGS (SEQ ID NO: 84), GGGGSGGGGSGGGGSGGGS (SEQ ID NO: 83); or the peptide linker is GGGGG (SEQ ID NO: 114). In some embodiments, the peptide linker comprises the amino acid sequence of SEQ ID NO: 83. In some embodiments, the amino acid sequence of the peptide linker is as shown in SEQ ID NO: 83.

[0141] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises:

[0142] A first strand comprising at least one (e.g., 1, 2, 3, 4, 5, or 6) an amino acid sequence containing SEQ ID NO: 87, 85, or 86, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the first strand, and at least one (e.g., 1, 2, 3, 4, 5, or 6) an amino acid sequence containing SEQ ID NO: 20, 21, 22, or 23, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the second strand.

[0143] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises:

[0144] Two first strands containing amino acid sequences of SEQ ID NO: 87, 85, or 86, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with them, and two second strands containing amino acid sequences of SEQ ID NO: 20, 21, 22, or 23, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with them.

[0145] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises:

[0146] Two first strands containing the amino acid sequence SEQ ID NO: 87, 85 or 86, and two second strands containing the amino acid sequence SEQ ID NO: 20, 21, 22 or 23.

[0147] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises:

[0148] Two first strands containing the amino acid sequence SEQ ID NO: 87, 85, or 86, and two second strands containing the amino acid sequence SEQ ID NO: 20; or

[0149] Two first strands containing the amino acid sequence SEQ ID NO: 85, 86, or 87, and two second strands containing the amino acid sequence SEQ ID NO: 21; or

[0150] Two first strands containing the amino acid sequence SEQ ID NO: 85, 86, or 87, and two second strands containing the amino acid sequence SEQ ID NO: 22; or

[0151] Two first strands containing the amino acid sequence of SEQ ID NO: 85, 86 or 87, and two second strands containing the amino acid sequence of SEQ ID NO: 23.

[0152] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises:

[0153] Two first strands containing the amino acid sequence SEQ ID NO: 87, and two second strands containing the amino acid sequence SEQ ID NO: 20; or

[0154] Two first strands containing the amino acid sequence SEQ ID NO: 85, and two second strands containing the amino acid sequence SEQ ID NO: 20; or

[0155] Two first strands containing the amino acid sequence SEQ ID NO: 86, and two second strands containing the amino acid sequence SEQ ID NO: 20; or

[0156] Two first strands containing the amino acid sequence SEQ ID NO: 85, and two second strands containing the amino acid sequence SEQ ID NO: 21; or

[0157] Two first strands containing the amino acid sequence SEQ ID NO: 86, and two second strands containing the amino acid sequence SEQ ID NO: 21; or

[0158] Two first strands containing the amino acid sequence SEQ ID NO: 87, and two second strands containing the amino acid sequence SEQ ID NO: 21; or

[0159] Two first strands containing the amino acid sequence SEQ ID NO: 85, and two second strands containing the amino acid sequence SEQ ID NO: 22; or

[0160] Two first strands containing the amino acid sequence SEQ ID NO: 86, and two second strands containing the amino acid sequence SEQ ID NO: 22; or

[0161] Two first strands containing the amino acid sequence SEQ ID NO: 87, and two second strands containing the amino acid sequence SEQ ID NO: 22; or

[0162] Two first strands containing the amino acid sequence SEQ ID NO: 85, and two second strands containing the amino acid sequence SEQ ID NO: 23; or

[0163] Two first strands containing the amino acid sequence SEQ ID NO: 86, and two second strands containing the amino acid sequence SEQ ID NO: 23; or

[0164] Two first strands containing the amino acid sequence SEQ ID NO: 87, and two second strands containing the amino acid sequence SEQ ID NO: 23.

[0165] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises:

[0166] Two first strands containing the amino acid sequence of SEQ ID NO: 87, and two second strands containing the amino acid sequence of SEQ ID NO: 20.

[0167] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments comprises:

[0168] Two first strands of amino acid sequences as shown in SEQ ID NO: 87, and two second strands of amino acid sequences as shown in SEQ ID NO: 20; or

[0169] Two first strands of amino acid sequences as shown in SEQ ID NO: 85, and two second strands of amino acid sequences as shown in SEQ ID NO: 20; or

[0170] Two first strands of amino acid sequences as shown in SEQ ID NO: 86, and two second strands of amino acid sequences as shown in SEQ ID NO: 20; or

[0171] Two first strands of amino acid sequences as shown in SEQ ID NO: 85, and two second strands of amino acid sequences as shown in SEQ ID NO: 21; or

[0172] Two first strands of amino acid sequences as shown in SEQ ID NO: 86, and two second strands of amino acid sequences as shown in SEQ ID NO: 21; or

[0173] Two first strands of amino acid sequences as shown in SEQ ID NO: 87, and two second strands of amino acid sequences as shown in SEQ ID NO: 21; or

[0174] Two first strands of amino acid sequences as shown in SEQ ID NO: 85, and two second strands of amino acid sequences as shown in SEQ ID NO: 22; or

[0175] Two first strands of amino acid sequences as shown in SEQ ID NO: 86, and two second strands of amino acid sequences as shown in SEQ ID NO: 22; or

[0176] Two first strands of amino acid sequences as shown in SEQ ID NO: 87, and two second strands of amino acid sequences as shown in SEQ ID NO: 22; or

[0177] Two first strands of amino acid sequences as shown in SEQ ID NO: 85, and two second strands of amino acid sequences as shown in SEQ ID NO: 23; or

[0178] Two first strands of amino acid sequences as shown in SEQ ID NO: 86, and two second strands of amino acid sequences as shown in SEQ ID NO: 23; or

[0179] The first strand consists of two amino acid sequences as shown in SEQ ID NO: 87, and the second strand consists of two amino acid sequences as shown in SEQ ID NO: 23.

[0180] In some embodiments, such as the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments, the first and second chains are interlinked to form an antigen-binding molecule that specifically binds to TfR1.

[0181] In some embodiments, the antigen-binding molecules that specifically bind to TfR1 provided in this disclosure are in EC5 concentrations of less than 100 nM (e.g., less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, less than 0.05 nM, less than 0.01 nM, less than 0.001 nM). 50 The value is combined with human TfR1, the EC 50 The value was measured using ELISA.

[0182] In some embodiments, the antigen-binding molecules that specifically bind to TfR1 provided in this disclosure do not block or have an IC50 value greater than 3.5 nM (e.g., greater than 4 nM, greater than 5 nM, greater than 10 nM, greater than 100 nM, greater than 1000 nM). 50 The IC blocks the binding of TF and TfR1. 50 The value was measured using ELISA.

[0183] <Antigen-binding molecules that specifically bind to Aβ>

[0184] This disclosure provides an antigen-binding molecule that specifically binds to Aβ, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0185] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain the amino acid sequences of HCDR1, HCDR2, and HCDR3 in SEQ ID NO: 54 or 55, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain the amino acid sequences of LCDR1, LCDR2, and LCDR3 in SEQ ID NO: 56, 57, 58, 59, or 116.

[0186] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0187] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain the amino acid sequences of HCDR1, HCDR2, and HCDR3 in SEQ ID NO: 54, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain the amino acid sequences of LCDR1, LCDR2, and LCDR3 in SEQ ID NO: 56, 57, 58, 59, or 116; or

[0188] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain the amino acid sequences of HCDR1, HCDR2, and HCDR3 in SEQ ID NO: 55, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain the amino acid sequences of LCDR1, LCDR2, and LCDR3 in SEQ ID NO: 56, 57, 58, 59, or 116.

[0189] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0190] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain the amino acid sequences of HCDR1, HCDR2, and HCDR3 in SEQ ID NO: 54, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain the amino acid sequences of LCDR1, LCDR2, and LCDR3 in SEQ ID NO: 56.

[0191] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments, wherein the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to a numbering rule selected from Kabat, IMGT, Chothia, AbM, and Contact. In some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to the Kabat numbering rule. In some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to the IMGT numbering rule. In some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to the Chothia numbering rule. In some embodiments, the heavy chain variable regions HCDR1, HCDR2, and HCDR3, and the light chain variable regions LCDR1, LCDR2, and LCDR3, are defined according to the AbM numbering rules. In some embodiments, the heavy chain variable regions HCDR1, HCDR2, and HCDR3, and the light chain variable regions LCDR1, LCDR2, and LCDR3, are defined according to the Contact numbering rules.

[0192] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0193] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50. The light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 60 or 115, LCDR2 contains the amino acid sequence of SEQ ID NO: 61 or 52, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0194] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0195] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50; and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 60, LCDR2 contains the amino acid sequence of SEQ ID NO: 61, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53; or

[0196] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50. The light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 115, LCDR2 contains the amino acid sequence of SEQ ID NO: 52, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0197] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0198] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50, and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 60, LCDR2 contains the amino acid sequence of SEQ ID NO: 61, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0199] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments includes a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region includes HCDR1, HCDR2, and HCDR3, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, wherein:

[0200] The amino acid sequences of the heavy chain variable regions HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO: 62, SEQ ID NO: 63, and SEQ ID NO: 50, respectively. The amino acid sequences of the light chain variable regions LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO: 60, SEQ ID NO: 61, and SEQ ID NO: 53, respectively.

[0201] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0202] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50. The light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 115, LCDR2 contains the amino acid sequence of SEQ ID NO: 52, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0203] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments includes a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region includes HCDR1, HCDR2, and HCDR3, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, wherein:

[0204] The amino acid sequences of the heavy chain variable region HCDR1 are shown in SEQ ID NO: 62, HCDR2 in SEQ ID NO: 63, and HCDR3 in SEQ ID NO: 50. The amino acid sequences of the light chain variable region LCDR1 are shown in SEQ ID NO: 115, LCDR2 in SEQ ID NO: 52, and LCDR3 in SEQ ID NO: 53.

[0205] In some embodiments, the antigen-binding molecules that specifically bind to Aβ as described in any of the preceding embodiments, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the Kabat numbering rules.

[0206] In some embodiments, the antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, is an antibody. In some embodiments, the antigen-binding molecule that specifically binds to Aβ is a monoclonal antibody, a monospecific antibody, a bispecific antibody, or a multispecific antibody. In some embodiments, the antigen-binding molecule that specifically binds to Aβ is a monoclonal antibody. In some embodiments, the antigen-binding molecule that specifically binds to Aβ is a monospecific antibody. In some embodiments, the antigen-binding molecule that specifically binds to Aβ is a murine antibody, a chimeric antibody, a humanized antibody, or a fully human antibody. In some embodiments, the antigen-binding molecule that specifically binds to Aβ is a humanized antibody.

[0207] In some implementations, the antigen-binding molecule that specifically binds to Aβ, as described in the preceding one, contains the frame region (FR) of the human antibody.

[0208] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments, the heavy chain variable region has FR1, FR2, FR3 fragments derived from IGHV2-5*09 and FR4 fragments derived from IGHJ6*01, and is unsubstituted or contains one or more amino acid substitutions selected from the group consisting of 1E, 40A, 44G, 94R and / or 105E; and / or the light chain variable region has FR1, FR2, FR3 fragments derived from IGKV2-30*02 and FR4 fragments derived from IGKJ4*01, and is unsubstituted or contains one or more amino acid substitutions selected from the group consisting of 17E, 36Y, 42E, 46L and / or 100E. In some embodiments, the antigen-binding molecule that specifically binds to Aβ, wherein the heavy chain variable region HCDR1 comprises the amino acid sequence of SEQ ID NO: 62, HCDR2 comprises the amino acid sequence of SEQ ID NO: 63, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 50, and the FR of the heavy chain variable region is unsubstituted or comprises one or more amino acid substitutions selected from the group consisting of 1E, 40A, 44G, 94R, and / or 105E; and the light chain variable region LCDR1 comprises the amino acid sequence of SEQ ID NO: 60 or 115, LCDR2 comprises the amino acid sequence of SEQ ID NO: 61 or 52, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 53, and the FR of the light chain variable region is unsubstituted or comprises one or more amino acid substitutions selected from the group consisting of 17E, 36Y, 42E, 46L, and / or 100E. In some embodiments, the above-mentioned variable regions and CDRs are defined according to the Kabat numbering rules.

[0209] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0210] The heavy chain variable region comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 54 or 55, and the light chain variable region comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 56, 57, 58, 59, or 116.

[0211] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0212] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 54 or 55, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 56, 57, 58, 59 or 116.

[0213] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0214] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 56, 57, 58, 59, or 116; or

[0215] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 55, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 56, 57, 58, 59 or 116.

[0216] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0217] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 54, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 56, 57, 58, 59 or 116.

[0218] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0219] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 56; or

[0220] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 57; or

[0221] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 58; or

[0222] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 59; or

[0223] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 116; or

[0224] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 56; or

[0225] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 57; or

[0226] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 58; or

[0227] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 55, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 59; or

[0228] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 55, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 116.

[0229] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0230] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 56; or

[0231] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 57; or

[0232] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 58; or

[0233] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 59; or

[0234] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 116; or

[0235] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 55, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 56; or

[0236] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 55, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 57; or

[0237] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 55, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 58; or

[0238] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 55, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 59; or

[0239] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 55, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 116.

[0240] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0241] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 56; or

[0242] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 57; or

[0243] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 58; or

[0244] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 59; or

[0245] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 54, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 116.

[0246] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0247] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 56; or

[0248] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 57; or

[0249] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 58; or

[0250] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 59; or

[0251] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 116.

[0252] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein the antigen-binding molecule that specifically binds to Aβ is humanized, reversed mutation, affinity maturation, reduced hydrophobicity, removed T-cell epitopes, reduced antibody deamidation, and / or reduced antibody isomerization.

[0253] In some embodiments, the antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, is an antigen-binding fragment. In some embodiments, the antigen-binding fragment is an antibody fragment. In some embodiments, the antigen-binding fragment is selected from Fab, Fab′, F(ab′)2, Fd, Fv, scFv, dsFv, and dAb.

[0254] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises a heavy chain constant region and a light chain constant region. In some embodiments, the heavy chain constant region is a heavy chain constant region of human IgG1, IgG2, IgG3, IgG4, or variants thereof, and the light chain constant region is a light chain constant region of human κ chain, λ chain, or variants thereof. In some embodiments, the heavy chain constant region is a human IgG1 heavy chain constant region or a variant thereof, and the light chain constant region is a human κ light chain constant region or a variant thereof. In some embodiments, the heavy chain constant region is a human IgG1 heavy chain constant region, and the light chain constant region is a human κ light chain constant region. In some embodiments, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 17, and the light chain constant region comprises the amino acid sequence of SEQ ID NO: 18.

[0255] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein the antigen-binding molecule that specifically binds to Aβ comprises a heavy chain and a light chain, wherein:

[0256] The heavy chain comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 64 or 65, and the light chain comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 66, 67, 68, 69, or 117.

[0257] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0258] The heavy chain comprises the amino acid sequence of SEQ ID NO: 64 or 65, and the light chain comprises the amino acid sequence of SEQ ID NO: 66, 67, 68, 69 or 117.

[0259] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0260] The heavy chain comprises the amino acid sequence of SEQ ID NO: 64, and the light chain comprises the amino acid sequence of SEQ ID NO: 66, 67, 68, 69, or 117; or

[0261] The heavy chain comprises the amino acid sequence of SEQ ID NO: 65, and the light chain comprises the amino acid sequence of SEQ ID NO: 66, 67, 68, 69 or 117.

[0262] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0263] The heavy chain comprises the amino acid sequence of SEQ ID NO: 64, and the light chain comprises the amino acid sequence of SEQ ID NO: 66, 67, 68, 69 or 117.

[0264] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0265] The heavy chain comprises the amino acid sequence of SEQ ID NO: 64, and the light chain comprises the amino acid sequence of SEQ ID NO: 66; or

[0266] The heavy chain comprises the amino acid sequence of SEQ ID NO: 64, and the light chain comprises the amino acid sequence of SEQ ID NO: 67; or

[0267] The heavy chain comprises the amino acid sequence of SEQ ID NO: 64, and the light chain comprises the amino acid sequence of SEQ ID NO: 68; or

[0268] The heavy chain comprises the amino acid sequence of SEQ ID NO: 64, and the light chain comprises the amino acid sequence of SEQ ID NO: 69; or

[0269] The heavy chain comprises the amino acid sequence of SEQ ID NO: 64, and the light chain comprises the amino acid sequence of SEQ ID NO: 117; or

[0270] The heavy chain comprises the amino acid sequence of SEQ ID NO: 65, and the light chain comprises the amino acid sequence of SEQ ID NO: 66; or

[0271] The heavy chain comprises the amino acid sequence of SEQ ID NO: 65, and the light chain comprises the amino acid sequence of SEQ ID NO: 67; or

[0272] The heavy chain comprises the amino acid sequence of SEQ ID NO: 65, and the light chain comprises the amino acid sequence of SEQ ID NO: 68; or

[0273] The heavy chain comprises the amino acid sequence of SEQ ID NO: 65, and the light chain comprises the amino acid sequence of SEQ ID NO: 69; or

[0274] The heavy chain contains the amino acid sequence of SEQ ID NO: 65, and the light chain contains the amino acid sequence of SEQ ID NO: 117.

[0275] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0276] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 66; or

[0277] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 67; or

[0278] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 68; or

[0279] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 69; or

[0280] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 117; or

[0281] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 65, and the amino acid sequence of the light chain is shown in SEQ ID NO: 66; or

[0282] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 65, and the amino acid sequence of the light chain is shown in SEQ ID NO: 67; or

[0283] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 65, and the amino acid sequence of the light chain is shown in SEQ ID NO: 68; or

[0284] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 65, and the amino acid sequence of the light chain is shown in SEQ ID NO: 69; or

[0285] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 65, and the amino acid sequence of the light chain is shown in SEQ ID NO: 117.

[0286] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0287] The heavy chain comprises the amino acid sequence of SEQ ID NO: 64, and the light chain comprises the amino acid sequence of SEQ ID NO: 66; or

[0288] The heavy chain comprises the amino acid sequence of SEQ ID NO: 64, and the light chain comprises the amino acid sequence of SEQ ID NO: 67; or

[0289] The heavy chain comprises the amino acid sequence of SEQ ID NO: 64, and the light chain comprises the amino acid sequence of SEQ ID NO: 68; or

[0290] The heavy chain comprises the amino acid sequence of SEQ ID NO: 64, and the light chain comprises the amino acid sequence of SEQ ID NO: 69; or

[0291] The heavy chain contains the amino acid sequence of SEQ ID NO: 64, and the light chain contains the amino acid sequence of SEQ ID NO: 117.

[0292] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein:

[0293] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 66; or

[0294] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 67; or

[0295] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 68; or

[0296] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 69; or

[0297] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 117.

[0298] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises a heavy chain constant region CH1 and a light chain constant region CL, wherein the heavy chain constant region CH1 is a heavy chain constant region CH1 of human IgG1, IgG2, IgG3, IgG4, or variants thereof, and the light chain constant region CL is a light chain constant region CL of human κ chain, λ chain, or variants thereof. In some embodiments, the heavy chain constant region CH1 is a heavy chain constant region CH1 of human IgG1 or variants thereof, and the light chain constant region CL is a light chain constant region CL of human κ chain or variants thereof. In some embodiments, the heavy chain constant region CH1 comprises the amino acid sequence of SEQ ID NO: 82, and the light chain constant region CL comprises the amino acid sequence of SEQ ID NO: 18.

[0299] In some embodiments, the antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, includes an Fc region.

[0300] In some implementations, the Fc region is the Fc region of human IgG1, IgG2, IgG3, IgG4 or variants thereof.

[0301] In some embodiments, the Fc region is either unsubstituted or contains amino acid substitutions of 351I and 439E, numbered according to the EU index.

[0302] In some implementations, the Fc region contains amino acid substitutions of 351I and 439E, numbered according to the EU index.

[0303] In some embodiments, the Fc region is the Fc region of a human IgG1 variant, wherein the Fc1 is unsubstituted or contains amino acid substitutions of 351I and 439E, numbered according to the EU index.

[0304] In some embodiments, the Fc region is the Fc region of a human IgG1 variant, wherein the Fc1 contains amino acid substitutions of 351I and 439E, numbered according to the EU index.

[0305] In some embodiments, the Fc region contains the amino acid sequence of SEQ ID NO: 81 or 79.

[0306] In some embodiments, the Fc region contains the amino acid sequence of SEQ ID NO: 81.

[0307] In some embodiments, the amino acid sequence of the Fc region is shown in SEQ ID NO: 81.

[0308] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments includes an Fc region comprising two mutually associative subunits Fc2; and includes two identical haptens comprising Fab and Fc2 regions that bind to Aβ.

[0309] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments includes an Fc region comprising two Fc2 subunits capable of associating with each other; and includes two identical haptens comprising at least one (e.g., 1, 2, 3, 4, 5, or 6) Aβ-binding Fab and at least one (e.g., 1, 2, 3, 4, 5, or 6) Fc2 region.

[0310] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments includes an Fc region comprising two mutually associative subunits Fc2; and comprises two identical haptens that include at least one (e.g., 1, 2, 3, 4, 5, or 6) Aβ-binding Fab and one Fc2 region in the direction from the N-terminus to the C-terminus.

[0311] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments includes an Fc region comprising two mutually associative subunits Fc2; and includes two identical haptens that comprise two identical or different Aβ-binding Fab regions and one Fc2 region from the N-terminus to the C-terminus.

[0312] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments includes an Fc region comprising two mutually associative subunits Fc2; and includes two identical haptens, each hapten comprising two identical or different Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus, wherein the C-terminus of the first Aβ-binding Fab is operatively linked to the N-terminus of the second Aβ-binding Fab, and the C-terminus of the second Aβ-binding Fab is operatively linked to the N-terminus of the Fc2 region.

[0313] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments includes an Fc region comprising two mutually associative subunits Fc2; and includes two identical haptens, each hapten comprising two identical or different Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus, wherein the C-terminus of the CH1 of the first Aβ-binding Fab is operatively linked to the N-terminus of the heavy chain variable region of the second Aβ-binding Fab, and the C-terminus of the CH1 of the second Aβ-binding Fab is operatively linked to the N-terminus of the Fc2 region.

[0314] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments includes an Fc region comprising two mutually associative subunits Fc2; and includes two identical haptens, each hapten comprising two identical or different Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus, wherein the C-terminus of the first Aβ-binding Fab is fused directly or via a linker to the N-terminus of the second Aβ-binding Fab, and the C-terminus of the second Aβ-binding Fab is fused directly or via a linker to the N-terminus of the Fc2 region.

[0315] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments includes an Fc region containing two mutually associative subunits Fc2; and includes two identical haptens, each hapten containing two identical or different Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus, wherein the C-terminus of the first Aβ-binding Fab is fused to the N-terminus of the second Aβ-binding Fab via a linker, and the C-terminus of the second Aβ-binding Fab is directly fused to the N-terminus of the Fc2 region.

[0316] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments includes an Fc region comprising two mutually associative subunits Fc2; and includes two identical haptens, each hapten comprising two identical or different Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus, wherein the C-terminus of the CH1 of the first Aβ-binding Fab is fused directly or via a linker to the N-terminus of the heavy chain variable region of the second Aβ-binding Fab, and the C-terminus of the CH1 of the second Aβ-binding Fab is fused directly or via a linker to the N-terminus of the Fc2 region.

[0317] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments includes an Fc region comprising two mutually associative subunits Fc2; and includes two identical haptens, each hapten comprising two identical or different Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus, wherein the C-terminus of the CH1 of the first Aβ-binding Fab is fused to the N-terminus of the heavy chain variable region of the second Aβ-binding Fab via a linker, and the C-terminus of the CH1 of the second Aβ-binding Fab is directly fused to the N-terminus of the Fc2 region.

[0318] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises a first chain with the structure shown in formula (c) and a second chain with the structure shown in formula (d), wherein the structures shown in formulas (c) and (d) are arranged from the N-terminus to the C-terminus:

[0319] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[0320] (d)[Aβ-VL]-[CL].

[0321] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) first strand with the structure shown in formula (c), and at least one (e.g., 1, 2, 3, 4, 5, or 6) second strand with the structure shown in formula (d), wherein the structures shown in formulas (c) and (d) are arranged from the N-terminus to the C-terminus.

[0322] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[0323] (d)[Aβ-VL]-[CL].

[0324] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises two first strands with the structure shown in formula (c) and four second strands with the structure shown in formula (d), wherein the structures shown in formulas (c) and (d) are arranged from the N-terminus to the C-terminus:

[0325] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[0326] (d)[Aβ-VL]-[CL].

[0327] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein the linker 2 is a peptide linker, or the linker 2 is absent.

[0328] In some embodiments, an antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, wherein the first and second chains bind to each other to form an antigen-binding molecule that specifically binds to Aβ.

[0329] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises two first strands with the structure shown in formula (c) and four second strands with the structure shown in formula (d), wherein the structures shown in formulas (c) and (d) are arranged from the N-terminus to the C-terminus:

[0330] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[0331] (d)[Aβ-VL]-[CL];

[0332] The linker 2 is a peptide linker, or the linker 2 is absent;

[0333] The first and second chains combine to form an antigen-binding molecule that specifically binds to Aβ.

[0334] In some embodiments, the structure of the antigen-binding molecule that specifically binds to Aβ, as described in any of the preceding embodiments, is shown in Format 2 of Figure 1B.

[0335] In some embodiments, as described in any of the preceding embodiments, the linker is a peptide linker in an antigen-binding molecule that specifically binds to Aβ. In some embodiments, as described in any of the preceding embodiments, the linker 2 is a peptide linker. In some embodiments, the peptide linker may be a flexible peptide containing 1-50 or 1-20 amino acid residues. In some embodiments, each of the peptide linkers independently has L1-(GGGGS). n The structure is -L2, wherein L1 is a bond, A, G, GS, GGG, GGS, GGGS (SEQ ID NO: 98) or GGGG (SEQ ID NO: 99), n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, L2 is a bond, G, GG, GGG or GGGG, and the peptide linker is not a bond. In some embodiments, the peptide linker is 2-20 amino acid residues in length. In some embodiments, the peptide linker is as follows: (GS) a (GGS) b (GGGS) c (GGGGS) d (GGGGG) ea, b, c, d, and e are independent integers greater than or equal to 0; or the peptide linker is selected from: (EAAAK)3 (SEQ ID NO: 100), (EAAAR)3 (SEQ ID NO: 101), (EGGGK)3 (SEQ ID NO: 102), (EGGGR)3 (SEQ ID NO: 103), (DAAAR)3 (SEQ ID NO: 104), (DAAAK)3 (SEQ ID NO: 105), (DGGGR)3 (SEQ ID NO: 106) or (DGGGK)3 (SEQ ID NO: 107), SS, GGSEGKSSGSGSESKSTGGS (SEQ ID NO: 108), EPKSS (SEQ ID NO: 109), RTVAA (SEQ ID NO: 110), ASEPKSS (SEQ ID NO: 111); or the peptide linker is (G x S) y Where x is an integer selected from 1 to 5, and y is an integer selected from 1 to 6, including but not limited to GGGS, GGGGS (SEQ ID NO: 112), GGGGSGGGGS (SEQ ID NO: 113), GGGGSGGGGSGGGGS (SEQ ID NO: 84), GGGGSGGGGSGGGGSGGGS (SEQ ID NO: 83); or the peptide linker is GGGGG (SEQ ID NO: 114). In some embodiments, the peptide linker comprises the amino acid sequence of SEQ ID NO: 84. In some embodiments, the amino acid sequence of the peptide linker is as shown in SEQ ID NO: 84.

[0336] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises:

[0337] A first strand comprising at least one (e.g., 1, 2, 3, 4, 5, or 6) an amino acid sequence containing SEQ ID NO: 88 or 89, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with it, and a second strand comprising at least one (e.g., 1, 2, 3, 4, 5, or 6) an amino acid sequence containing SEQ ID NO: 66, 67, 68, 69, or 117, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with it.

[0338] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises:

[0339] Two first strands containing amino acid sequences of SEQ ID NO: 88 or 89, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with them, and four second strands containing amino acid sequences of SEQ ID NO: 66, 67, 68, 69, or 117, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with them.

[0340] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises:

[0341] Two first strands containing the amino acid sequence SEQ ID NO: 88 or 89, and four second strands containing the amino acid sequence SEQ ID NO: 66, 67, 68, 69 or 117.

[0342] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises:

[0343] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 66, 67, 68, 69, or 117; or

[0344] Two first strands containing the amino acid sequence SEQ ID NO: 89, and four second strands containing the amino acid sequences SEQ ID NO: 66, 67, 68, 69, or 117.

[0345] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises:

[0346] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequences SEQ ID NO: 66, 67, 68, 69, or 117.

[0347] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises:

[0348] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 66; or

[0349] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 67; or

[0350] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 68; or

[0351] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 69; or

[0352] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 117; or

[0353] Two first strands containing the amino acid sequence SEQ ID NO: 89, and four second strands containing the amino acid sequence SEQ ID NO: 66; or

[0354] Two first strands containing the amino acid sequence SEQ ID NO: 89, and four second strands containing the amino acid sequence SEQ ID NO: 67; or

[0355] Two first strands containing the amino acid sequence SEQ ID NO: 89, and four second strands containing the amino acid sequence SEQ ID NO: 68; or

[0356] Two first strands containing the amino acid sequence SEQ ID NO: 89, and four second strands containing the amino acid sequence SEQ ID NO: 69; or

[0357] Two first strands containing the amino acid sequence SEQ ID NO: 89, and four second strands containing the amino acid sequence SEQ ID NO: 117.

[0358] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises:

[0359] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 66; or

[0360] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 67; or

[0361] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 68; or

[0362] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 69; or

[0363] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 117; or

[0364] Two first strands of amino acid sequences as shown in SEQ ID NO: 89, and four second strands of amino acid sequences as shown in SEQ ID NO: 66; or

[0365] Two first strands of amino acid sequences as shown in SEQ ID NO: 89, and four second strands of amino acid sequences as shown in SEQ ID NO: 67; or

[0366] Two first strands of amino acid sequences as shown in SEQ ID NO: 89, and four second strands of amino acid sequences as shown in SEQ ID NO: 68; or

[0367] Two first strands of amino acid sequences as shown in SEQ ID NO: 89, and four second strands of amino acid sequences as shown in SEQ ID NO: 69; or

[0368] The first strand consists of two amino acid sequences as shown in SEQ ID NO: 89, and the second strand consists of four amino acid sequences as shown in SEQ ID NO: 117.

[0369] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises:

[0370] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 66; or

[0371] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 67; or

[0372] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 68; or

[0373] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 69; or

[0374] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 117.

[0375] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises:

[0376] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 66.

[0377] In some embodiments, the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments comprises:

[0378] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 66; or

[0379] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 67; or

[0380] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 68; or

[0381] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 69; or

[0382] The first strand consists of two amino acid sequences as shown in SEQ ID NO: 88, and the second strand consists of four amino acid sequences as shown in SEQ ID NO: 117.

[0383] In some embodiments, such as the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments, the first and second chains are interlinked to form an antigen-binding molecule that specifically binds to Aβ.

[0384] In some embodiments, the antigen-binding molecules that specifically bind to Aβ provided in this disclosure are in EC5 concentrations of less than 10 nM (e.g., less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, less than 0.05 nM, less than 0.01 nM, less than 0.001 nM).50 The value binds to human Aβ1-42fibril, the EC 50 The value was measured using ELISA.

[0385] In some embodiments, the antigen-binding molecules that specifically bind to Aβ provided in this disclosure have a PBS pH 7.4 solubility of greater than 10 mg / mL (e.g., greater than 50 mg / mL, greater than 100 mg / mL, greater than 150 mg / mL, greater than 200 mg / mL, greater than 250 mg / mL, greater than 300 mg / mL, greater than 350 mg / mL, greater than 400 mg / mL, greater than 450 mg / mL, greater than 500 mg / mL).

[0386] <Antigen-binding molecules that specifically bind to TfR1 and Aβ>

[0387] This disclosure provides an antigen-binding molecule that specifically binds to TfR1 and Aβ, comprising a first antigen-binding domain that specifically binds to TfR1 and a second antigen-binding domain that specifically binds to Aβ.

[0388] In some embodiments, the antigen-binding molecule that specifically binds TfR1 and Aβ as described in any of the preceding embodiments comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) first antigen-binding domain that specifically binds TfR1 and at least one (e.g., 1, 2, 3, 4, 5, or 6) second antigen-binding domain that specifically binds Aβ.

[0389] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0390] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0391] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain one, two, or three HCDR amino acid sequences from SEQ ID NO: 27, 12, or 26, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain one, two, or three LCDR amino acid sequences from SEQ ID NO: 13, 14, 15, or 16.

[0392] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0393] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0394] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain the amino acid sequences of HCDR1, HCDR2, and HCDR3 in SEQ ID NO: 27, 12, or 26, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain the amino acid sequences of LCDR1, LCDR2, and LCDR3 in SEQ ID NO: 13, 14, 15, or 16.

[0395] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0396] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0397] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain the amino acid sequences of HCDR1, HCDR2, and HCDR3 in SEQ ID NO: 27, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain the amino acid sequences of LCDR1, LCDR2, and LCDR3 in SEQ ID NO: 13.

[0398] In some embodiments, the antigen-binding molecules that specifically bind TfR1 and Aβ as described in any of the preceding embodiments, wherein the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to a numbering rule selected from Kabat, IMGT, Chothia, AbM, and Contact. In some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to the Kabat numbering rule. In some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to the IMGT numbering rule. In some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to the Chothia numbering rule. In some embodiments, the heavy chain variable regions HCDR1, HCDR2, and HCDR3, and the light chain variable regions LCDR1, LCDR2, and LCDR3, are defined according to the AbM numbering rules. In some embodiments, the heavy chain variable regions HCDR1, HCDR2, and HCDR3, and the light chain variable regions LCDR1, LCDR2, and LCDR3, are defined according to the Contact numbering rules.

[0399] In some embodiments, the antigen-binding molecule that specifically binds TfR1 and Aβ as described in any of the preceding claims comprises at least one (e.g., 1, 2, 3, 4, 5 or 6) first antigen-binding domain that specifically binds TfR1 and at least one (e.g., 1, 2, 3, 4, 5 or 6) second antigen-binding domain that specifically binds Aβ.

[0400] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0401] The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, wherein:

[0402] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 3, HCDR2 contains the amino acid sequence of SEQ ID NO: 11, and HCDR3 contains the amino acid sequence of SEQ ID NO: 25, 5, or 24. The light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 9, LCDR2 contains the amino acid sequence of SEQ ID NO: 10, and LCDR3 contains the amino acid sequence of SEQ ID NO: 8.

[0403] In some embodiments, the antigen-binding molecule that specifically binds TfR1 and Aβ as described in any of the preceding claims comprises at least one (e.g., 1, 2, 3, 4, 5 or 6) first antigen-binding domain that specifically binds TfR1 and at least one (e.g., 1, 2, 3, 4, 5 or 6) second antigen-binding domain that specifically binds Aβ.

[0404] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0405] The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, wherein:

[0406] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 3, HCDR2 contains the amino acid sequence of SEQ ID NO: 11, and HCDR3 contains the amino acid sequence of SEQ ID NO: 25, and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 9, LCDR2 contains the amino acid sequence of SEQ ID NO: 10, and LCDR3 contains the amino acid sequence of SEQ ID NO: 8.

[0407] In some embodiments, the antigen-binding molecule that specifically binds TfR1 and Aβ as described in any of the preceding claims comprises one first antigen-binding domain that specifically binds TfR1 and two second antigen-binding domains that specifically bind Aβ.

[0408] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0409] The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, wherein:

[0410] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 3, HCDR2 contains the amino acid sequence of SEQ ID NO: 11, and HCDR3 contains the amino acid sequence of SEQ ID NO: 25, and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 9, LCDR2 contains the amino acid sequence of SEQ ID NO: 10, and LCDR3 contains the amino acid sequence of SEQ ID NO: 8.

[0411] In some embodiments, antigen-binding molecules that specifically bind TfR1 and Aβ, as described in any of the preceding embodiments, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the Kabat numbering rules.

[0412] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0413] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0414] The heavy chain variable region comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 27, 12, or 26, and the light chain variable region comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 13, 14, 15, or 16.

[0415] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0416] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0417] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 27, 12 or 26, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 13, 14, 15 or 16.

[0418] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0419] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0420] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 27, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 13, 14, 15, or 16; or

[0421] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 12, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 13, 14, 15, or 16; or

[0422] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 26, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 13, 14, 15 or 16.

[0423] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0424] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0425] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 27, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 13.

[0426] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0427] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0428] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain one, two, or three amino acid sequences of HCDR from SEQ ID NO: 54 or 55, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain one, two, or three amino acid sequences of LCDR from SEQ ID NO: 56, 57, 58, 59, or 116.

[0429] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0430] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0431] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain the amino acid sequences of HCDR1, HCDR2, and HCDR3 in SEQ ID NO: 54 or 55, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain the amino acid sequences of LCDR1, LCDR2, and LCDR3 in SEQ ID NO: 56, 57, 58, 59, or 116.

[0432] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0433] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0434] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain the amino acid sequences of HCDR1, HCDR2, and HCDR3 in SEQ ID NO: 54, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain the amino acid sequences of LCDR1, LCDR2, and LCDR3 in SEQ ID NO: 56, 57, 58, 59, or 116.

[0435] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0436] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0437] The heavy chain variable regions HCDR1, HCDR2, and HCDR3 respectively contain the amino acid sequences of HCDR1, HCDR2, and HCDR3 in SEQ ID NO: 54, and the light chain variable regions LCDR1, LCDR2, and LCDR3 respectively contain the amino acid sequences of LCDR1, LCDR2, and LCDR3 in SEQ ID NO: 56.

[0438] In some embodiments, the antigen-binding molecules that specifically bind TfR1 and Aβ as described in any of the preceding embodiments, wherein the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to a numbering rule selected from Kabat, IMGT, Chothia, AbM, and Contact. In some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to the Kabat numbering rule. In some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to the IMGT numbering rule. In some embodiments, the HCDR1, HCDR2, and HCDR3 of the heavy chain variable regions and the LCDR1, LCDR2, and LCDR3 of the light chain variable regions are defined according to the Chothia numbering rule. In some embodiments, the heavy chain variable regions HCDR1, HCDR2, and HCDR3, and the light chain variable regions LCDR1, LCDR2, and LCDR3, are defined according to the AbM numbering rules. In some embodiments, the heavy chain variable regions HCDR1, HCDR2, and HCDR3, and the light chain variable regions LCDR1, LCDR2, and LCDR3, are defined according to the Contact numbering rules.

[0439] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0440] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0441] The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, wherein:

[0442] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50. The light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 60 or 115, LCDR2 contains the amino acid sequence of SEQ ID NO: 61 or 52, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0443] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0444] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0445] The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, wherein:

[0446] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50; and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 60, LCDR2 contains the amino acid sequence of SEQ ID NO: 61, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53; or

[0447] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50. The light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 115, LCDR2 contains the amino acid sequence of SEQ ID NO: 52, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0448] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0449] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0450] The heavy chain variable region includes HCDR1, HCDR2, and HCDR3, and the light chain variable region includes LCDR1, LCDR2, and LCDR3, wherein:

[0451] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50, and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 60, LCDR2 contains the amino acid sequence of SEQ ID NO: 61, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0452] In some embodiments, antigen-binding molecules that specifically bind TfR1 and Aβ, as described in any of the preceding embodiments, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the Kabat numbering rules.

[0453] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0454] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0455] The heavy chain variable region comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 54 or 55, and the light chain variable region comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 56, 57, 58, 59, or 116.

[0456] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0457] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0458] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 54 or 55, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 56, 57, 58, 59 or 116.

[0459] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0460] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0461] The heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 54, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 56, 57, 58, 59, or 116; or

[0462] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 55, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 56, 57, 58, 59 or 116.

[0463] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0464] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0465] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 54, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 56, 57, 58, 59 or 116.

[0466] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0467] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0468] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 54, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 56.

[0469] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0470] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0471] The TfR1-VH comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 3, HCDR2 containing the amino acid sequence of SEQ ID NO: 11, and HCDR3 containing the amino acid sequence of SEQ ID NO: 25, 5, or 24; and the TfR1-VL comprises LCDR1 containing the amino acid sequence of SEQ ID NO: 9, LCDR2 containing the amino acid sequence of SEQ ID NO: 10, and LCDR3 containing the amino acid sequence of SEQ ID NO: 8; and

[0472] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0473] The Aβ-VH's HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50; the Aβ-VL's LCDR1 contains the amino acid sequence of SEQ ID NO: 60 or 115, LCDR2 contains the amino acid sequence of SEQ ID NO: 61 or 52, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0474] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0475] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0476] The TfR1-VH comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 3, HCDR2 containing the amino acid sequence of SEQ ID NO: 11, and HCDR3 containing the amino acid sequence of SEQ ID NO: 25, 5, or 24; and the TfR1-VL comprises LCDR1 containing the amino acid sequence of SEQ ID NO: 9, LCDR2 containing the amino acid sequence of SEQ ID NO: 10, and LCDR3 containing the amino acid sequence of SEQ ID NO: 8; and

[0477] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0478] The Aβ-VH's HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50; and the Aβ-VL's LCDR1 contains the amino acid sequence of SEQ ID NO: 60, LCDR2 contains the amino acid sequence of SEQ ID NO: 61, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53; or

[0479] The Aβ-VH has HCDR1 containing the amino acid sequence of SEQ ID NO: 62, HCDR2 containing the amino acid sequence of SEQ ID NO: 63, and HCDR3 containing the amino acid sequence of SEQ ID NO: 50. The Aβ-VL has LCDR1 containing the amino acid sequence of SEQ ID NO: 115, LCDR2 containing the amino acid sequence of SEQ ID NO: 52, and LCDR3 containing the amino acid sequence of SEQ ID NO: 53.

[0480] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0481] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0482] The TfR1-VH comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 3, HCDR2 containing the amino acid sequence of SEQ ID NO: 11, and HCDR3 containing the amino acid sequence of SEQ ID NO: 25; and the TfR1-VL comprises LCDR1 containing the amino acid sequence of SEQ ID NO: 9, LCDR2 containing the amino acid sequence of SEQ ID NO: 10, and LCDR3 containing the amino acid sequence of SEQ ID NO: 8; and

[0483] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0484] The Aβ-VH has HCDR1 containing the amino acid sequence of SEQ ID NO: 62, HCDR2 containing the amino acid sequence of SEQ ID NO: 63, and HCDR3 containing the amino acid sequence of SEQ ID NO: 50; and the Aβ-VL has LCDR1 containing the amino acid sequence of SEQ ID NO: 60, LCDR2 containing the amino acid sequence of SEQ ID NO: 61, and LCDR3 containing the amino acid sequence of SEQ ID NO: 53.

[0485] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0486] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0487] The TfR1-VH comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 27, 12, or 26, and the TfR1-VL comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 13, 14, 15, or 16; and

[0488] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0489] The Aβ-VH comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 54 or 55, and the Aβ-VL comprises an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with SEQ ID NO: 56, 57, 58, 59, or 116.

[0490] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0491] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0492] The TfR1-VH comprises the amino acid sequence of SEQ ID NO: 27, 12, or 26, and the TfR1-VL comprises the amino acid sequence of SEQ ID NO: 13, 14, 15, or 16; and

[0493] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0494] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54 or 55, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 56, 57, 58, 59 or 116.

[0495] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0496] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0497] The TfR1-VH contains the amino acid sequence of SEQ ID NO: 27, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13, 14, 15, or 16; or

[0498] The TfR1-VH contains the amino acid sequence of SEQ ID NO: 12, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13, 14, 15, or 16; or

[0499] The TfR1-VH contains the amino acid sequence of SEQ ID NO: 26, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13, 14, 15, or 16; and

[0500] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0501] The Aβ-VH comprises the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL comprises the amino acid sequence of SEQ ID NO: 56, 57, 58, 59, or 116; or

[0502] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 55, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 56, 57, 58, 59 or 116.

[0503] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0504] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0505] The TfR1-VH contains the amino acid sequence of SEQ ID NO: 27, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13; and

[0506] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0507] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 56, 57, 58, 59 or 116.

[0508] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0509] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0510] The TfR1-VH contains the amino acid sequence of SEQ ID NO: 27, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13; and

[0511] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0512] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 56; or

[0513] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 57; or

[0514] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 58; or

[0515] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 59; or

[0516] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 116.

[0517] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0518] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0519] The TfR1-VH contains the amino acid sequence of SEQ ID NO: 27, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13; and

[0520] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0521] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 56.

[0522] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0523] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0524] The TfR1-VH contains the amino acid sequence of SEQ ID NO: 27, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13; and

[0525] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0526] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 57.

[0527] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0528] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0529] The TfR1-VH contains the amino acid sequence of SEQ ID NO: 27, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13; and

[0530] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0531] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 58.

[0532] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0533] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0534] The TfR1-VH contains the amino acid sequence of SEQ ID NO: 27, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13; and

[0535] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0536] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 59.

[0537] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0538] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein:

[0539] The TfR1-VH contains the amino acid sequence of SEQ ID NO: 27, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13; and

[0540] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein:

[0541] The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 116.

[0542] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0543] The first antigen-binding domain that specifically binds to TfR1 and the second antigen-binding domain that specifically binds to Aβ are selected from Fab, Fab′, F(ab′)2, Fd, Fv, scFv, dsFv and dAb.

[0544] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0545] The first antigen-binding domain that specifically binds to TfR1 and the second antigen-binding domain that specifically binds to Aβ are selected from Fab, Fab′, F(ab′)2, Fv, scFv, dsFv and dAb.

[0546] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0547] Both the first antigen-binding domain that specifically binds to TfR1 and the second antigen-binding domain that specifically binds to Aβ are Fab.

[0548] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ, as described in any of the preceding embodiments, comprises a heavy chain constant region CH1 and a light chain constant region CL.

[0549] In some embodiments, the antigen-binding molecule that specifically binds TfR1 and Aβ as described in any of the preceding embodiments comprises a heavy chain constant region CH1 and a light chain constant region CL, wherein the heavy chain constant region CH1 is the heavy chain constant region CH1 of human IgG1, IgG2, IgG3, IgG4 or variants thereof, and the light chain constant region CL is the light chain constant region CL of human κ chain, λ chain or variants thereof.

[0550] In some embodiments, the heavy chain constant region CH1 is the heavy chain constant region CH1 of human IgG1 or a variant thereof, and the light chain constant region CL is the light chain constant region CL of human κ chain or a variant thereof.

[0551] In some embodiments, the heavy chain constant region CH1 contains the amino acid sequence of SEQ ID NO: 82, and the light chain constant region CL contains the amino acid sequence of SEQ ID NO: 18.

[0552] In some embodiments, the amino acid sequence of the heavy chain constant region CH1 is shown in SEQ ID NO: 82, and the amino acid sequence of the light chain constant region CL is shown in SEQ ID NO: 18.

[0553] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2 capable of associating with each other.

[0554] In some implementations, the Fc region is the Fc region of human IgG1, IgG2, IgG3, IgG4 or variants thereof.

[0555] In some embodiments, the Fc region contains the amino acid sequence of SEQ ID NO: 79.

[0556] In some embodiments, the amino acid sequence of the Fc region is shown in SEQ ID NO: 79.

[0557] In some embodiments, Fc1 is an unsubstituted amino acid or contains 351I and 356K, and Fc2 is an unsubstituted amino acid or contains 351I and 439E, numbered according to the EU index.

[0558] In some embodiments, Fc1 contains amino acid substitutions of 351I and 356K, and Fc2 contains amino acid substitutions of 351I and 439E, numbered according to the EU index.

[0559] In some embodiments, the Fc region is the Fc region of a human IgG1 variant, wherein Fc1 is an unsubstituted amino acid or contains L351I and D356K substitutions, and Fc2 is an unsubstituted amino acid or contains L351I and K439E substitutions, numbered according to the EU index.

[0560] In some embodiments, the Fc region is the Fc region of a human IgG1 variant, wherein Fc1 contains amino acid substitutions of L351I and D356K, and Fc2 contains amino acid substitutions of L351I and K439E, numbered according to the EU index.

[0561] In some embodiments, Fc1 contains the amino acid sequence of SEQ ID NO: 80 or 79; and Fc2 contains the amino acid sequence of SEQ ID NO: 81 or 79.

[0562] In some embodiments, Fc1 comprises the amino acid sequence of SEQ ID NO: 80; and Fc2 comprises the amino acid sequence of SEQ ID NO: 81.

[0563] In some embodiments, the amino acid sequence of Fc1 is shown in SEQ ID NO: 80; and the amino acid sequence of Fc2 is shown in SEQ ID NO: 81.

[0564] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ, as described in any of the preceding embodiments, is an antibody. In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ is a multispecific antibody. In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ is a bispecific antibody. In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ is a trivalent bispecific antibody. In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ is a bivalent bispecific antibody. In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ is a murine antibody, a chimeric antibody, a humanized antibody, or a fully human antibody. In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ is a humanized antibody.

[0565] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0566] The antigen-binding molecule that specifically binds to TfR1 and Aβ is a bispecific antibody, and comprises a first half-antibody and a second half-antibody, wherein:

[0567] The first half-antibody includes Fab and Fc1 regions that bind TfR1; and the second half-antibody includes Fab and Fc2 regions that bind Aβ; and the first half-antibody and the second half-antibody are associated with each other through the Fc1 and Fc2 regions.

[0568] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein:

[0569] The antigen-binding molecule that specifically binds to TfR1 and Aβ is a bispecific antibody, and comprises a first half-antibody and a second half-antibody, wherein:

[0570] The first half-antibody includes one TfR1 binding Fab and one Fc1 region from the N-terminus to the C-terminus; and the second half-antibody includes two identical or different Aβ binding Fabs and one Fc2 region from the N-terminus to the C-terminus; and the first half-antibody and the second half-antibody associate with each other through the Fc1 region and the Fc2 region.

[0571] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2; and includes a first half-antibody and a second half-antibody.

[0572] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2; and includes a first hapten and a second hapten, wherein:

[0573] The first half-antibody includes Fab and Fc1 regions that bind TfR1; and the second half-antibody includes Fab and Fc2 regions that bind Aβ; and the first half-antibody and the second half-antibody are associated with each other through the Fc1 and Fc2 regions.

[0574] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2; and includes a first hapten and a second hapten, wherein:

[0575] The first hapten comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) Fab region that binds to TfR1 and at least one (e.g., 1, 2, 3, 4, 5, or 6) Fc1 region; and the second hapten comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) Fab region that binds to Aβ and at least one (e.g., 1, 2, 3, 4, 5, or 6) Fc2 region; and the first hapten and the second hapten associate with each other through the Fc1 and Fc2 regions.

[0576] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2; and includes a first hapten and a second hapten, wherein:

[0577] The first hapten contains at least one (e.g., 1, 2, 3, 4, 5, or 6) Fab region binding to TfR1 and one Fc1 region from the N-terminus to the C-terminus; and the second hapten contains at least one (e.g., 1, 2, 3, 4, 5, or 6) Fab region binding to Aβ and one Fc2 region from the N-terminus to the C-terminus; and the first hapten and the second hapten associate with each other through the Fc1 and Fc2 regions.

[0578] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2; and includes a first hapten and a second hapten, wherein:

[0579] The first half-antibody includes one TfR1 binding Fab and one Fc1 region from the N-terminus to the C-terminus; and the second half-antibody includes two identical or different Aβ binding Fabs and one Fc2 region from the N-terminus to the C-terminus; and the first half-antibody and the second half-antibody associate with each other through the Fc1 region and the Fc2 region.

[0580] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2; and includes a first hapten and a second hapten, wherein:

[0581] The first hapten comprises, from N-terminus to C-terminus, one TfR1-binding Fab and one Fc1 region, wherein the C-terminus of the TfR1-binding Fab is operatively connected to the N-terminus of the Fc1 region; and the second hapten comprises, from N-terminus to C-terminus, two identical or different Aβ-binding Fabs and one Fc2 region, wherein the C-terminus of the first Aβ-binding Fab is operatively connected to the N-terminus of the second Aβ-binding Fab, and the C-terminus of the second Aβ-binding Fab is operatively connected to the N-terminus of the Fc2 region; and the first hapten and the second hapten associate with each other through the Fc1 and Fc2 regions.

[0582] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2; and includes a first hapten and a second hapten, wherein:

[0583] The first hapten comprises, from N-terminus to C-terminus, one TfR1-binding Fab and one Fc1 region, wherein the C-terminus of the CH1 region of the TfR1-binding Fab is operatively connected to the N-terminus of the Fc1 region; and the second hapten comprises, from N-terminus to C-terminus, two identical or different Aβ-binding Fabs and one Fc2 region, wherein the C-terminus of the CH1 region of the first Aβ-binding Fab is operatively connected to the N-terminus of the heavy chain variable region of the second Aβ-binding Fab, and the C-terminus of the CH1 region of the second Aβ-binding Fab is operatively connected to the N-terminus of the Fc2 region; and the first hapten and the second hapten associate with each other through the Fc1 and Fc2 regions.

[0584] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2; and includes a first hapten and a second hapten, wherein:

[0585] The first hapten comprises one TfR1-binding Fab and one Fc1 region from the N-terminus to the C-terminus, wherein the C-terminus of the TfR1-binding Fab is fused directly or via a linker to the N-terminus of the Fc1 region; and the second hapten comprises two identical or different Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus, wherein the C-terminus of the first Aβ-binding Fab is fused directly or via a linker to the N-terminus of the second Aβ-binding Fab, and the C-terminus of the second Aβ-binding Fab is fused directly or via a linker to the N-terminus of the Fc2 region; and the first hapten and the second hapten associate with each other through the Fc1 and Fc2 regions.

[0586] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2; and includes a first hapten and a second hapten, wherein:

[0587] The first hapten includes one TfR1-binding Fab and one Fc1 region from the N-terminus to the C-terminus, with the C-terminus of the TfR1-binding Fab fused to the N-terminus of the Fc1 region via a linker; and the second hapten includes two identical or different Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus, with the C-terminus of the first Aβ-binding Fab fused to the N-terminus of the second Aβ-binding Fab via a linker, and the C-terminus of the second Aβ-binding Fab directly fused to the N-terminus of the Fc2 region; and the first and second haptens associate with each other through the Fc1 and Fc2 regions.

[0588] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2; and includes a first hapten and a second hapten, wherein:

[0589] The first hapten comprises one TfR1-binding Fab and one Fc1 region from the N-terminus to the C-terminus, wherein the C-terminus of the CH1 region of the TfR1-binding Fab is fused directly or via a linker to the N-terminus of the Fc1 region; and the second hapten comprises two identical or different Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus, wherein the C-terminus of the CH1 region of the first Aβ-binding Fab is fused directly or via a linker to the N-terminus of the heavy chain variable region of the second Aβ-binding Fab, and the C-terminus of the CH1 region of the second Aβ-binding Fab is fused directly or via a linker to the N-terminus of the Fc2 region; and the first and second haptens associate with each other through the Fc1 and Fc2 regions.

[0590] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2; and includes a first hapten and a second hapten, wherein:

[0591] The first hapten includes one TfR1-binding Fab and one Fc1 region from the N-terminus to the C-terminus. The C-terminus of the CH1 region of the TfR1-binding Fab is fused to the N-terminus of the Fc1 region via a linker. The second hapten includes two identical or different Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus. The C-terminus of the CH1 region of the first Aβ-binding Fab is fused to the N-terminus of the heavy chain variable region of the second Aβ-binding Fab via a linker. The C-terminus of the CH1 region of the second Aβ-binding Fab is directly fused to the N-terminus of the Fc2 region. The first and second haptens are associated with each other through the Fc1 and Fc2 regions.

[0592] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments comprises:

[0593] The first chain of the structure shown in equation (a), the second chain of the structure shown in equation (b), the third chain of the structure shown in equation (c), and the fourth chain of the structure shown in equation (d) are arranged from the N end to the C end.

[0594] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[0595] (b)[TfR1-VL]-[CL];

[0596] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[0597] (d)[Aβ-VL]-[CL].

[0598] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments comprises:

[0599] At least one (e.g., 1, 2, 3, 4, 5, or 6) of a first chain with the structure shown in Equation (a), at least one (e.g., 1, 2, 3, 4, 5, or 6) of a second chain with the structure shown in Equation (b), at least one (e.g., 1, 2, 3, 4, 5, or 6) of a third chain with the structure shown in Equation (c), and at least one (e.g., 1, 2, 3, 4, 5, or 6) of a fourth chain with the structure shown in Equation (d), wherein the structures shown in Equations (a), (b), (c), and (d) are arranged from the N-end to the C-end:

[0600] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[0601] (b)[TfR1-VL]-[CL];

[0602] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[0603] (d)[Aβ-VL]-[CL].

[0604] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments comprises:

[0605] One first chain as shown in equation (a), one second chain as shown in equation (b), one third chain as shown in equation (c), and two fourth chains as shown in equation (d), wherein the structures shown in equations (a), (b), (c), and (d) are arranged from the N end to the C end:

[0606] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[0607] (b)[TfR1-VL]-[CL];

[0608] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[0609] (d)[Aβ-VL]-[CL].

[0610] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein linker 1 and linker 2 are the same or different peptide linkers, or wherein linker 1 and / or linker 2 are absent.

[0611] In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding embodiments, wherein Fc1 and Fc2 are interchangeable.

[0612] In some embodiments, an antigen-binding molecule that specifically binds to TfR1 and Aβ, as described in any of the preceding embodiments, wherein the first and second chains bind to each other, the first and third chains bind to each other, and the third and fourth chains bind to each other to form an antigen-binding molecule that specifically binds to TfR1 and Aβ.

[0613] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments comprises:

[0614] The first chain of the structure shown in equation (a), the second chain of the structure shown in equation (b), the third chain of the structure shown in equation (c), and the fourth chain of the structure shown in equation (d) are arranged from the N end to the C end.

[0615] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[0616] (b)[TfR1-VL]-[CL];

[0617] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[0618] (d)[Aβ-VL]-[CL];

[0619] The linker 1 and linker 2 may be the same or different peptide linkers, or the linker 1 and / or linker 2 may not exist;

[0620] Fc1 and Fc2 are interchangeable;

[0621] The first and second chains combine with each other, the first and third chains combine with each other, and the third and fourth chains combine with each other to form antigen-binding molecules that specifically bind to TfR1 and Aβ.

[0622] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments comprises:

[0623] One first chain as shown in equation (a), one second chain as shown in equation (b), one third chain as shown in equation (c), and two fourth chains as shown in equation (d), wherein the structures shown in equations (a), (b), (c), and (d) are arranged from the N end to the C end:

[0624] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[0625] (b)[TfR1-VL]-[CL];

[0626] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[0627] (d)[Aβ-VL]-[CL];

[0628] The linker 1 and linker 2 may be the same or different peptide linkers, or the linker 1 and / or linker 2 may not exist;

[0629] Fc1 and Fc2 are interchangeable;

[0630] The first and second chains combine with each other, the first and third chains combine with each other, and the third and fourth chains combine with each other to form antigen-binding molecules that specifically bind to TfR1 and Aβ.

[0631] In some embodiments, the structure of the antigen-binding molecule that specifically binds to TfR1 and Aβ, as described in any of the preceding embodiments, is shown in Format 3 of Figure 2.

[0632] In some embodiments, as described in the preceding claim, the linker is a peptide linker in the antigen-binding molecule that specifically binds to TfR1 and Aβ. In some embodiments, as described in the preceding claim, linker 1 and linker 2 are peptide linkers. In this disclosure, linker 1 and linker 2 are used only to distinguish linker positions and are not intended to limit the linker sequence. In some embodiments, the peptide linker may be a flexible peptide containing 1-50 or 1-20 amino acid residues. In some embodiments, each of the peptide linkers independently has L1-(GGGGS). nThe structure is -L2, wherein L1 is a bond, A, G, GS, GGG, GGS, GGGS (SEQ ID NO: 98) or GGGG (SEQ ID NO: 99), n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, L2 is a bond, G, GG, GGG or GGGG, and the peptide linker is not a bond. In some embodiments, the peptide linker is 2-20 amino acid residues in length. In some embodiments, the peptide linker is as follows: (GS) a (GGS) b (GGGS) c (GGGGS) d (GGGGG) e a, b, c, d, and e are independent integers greater than or equal to 0; or the peptide linker is selected from: (EAAAK)3 (SEQ ID NO: 100), (EAAAR)3 (SEQ ID NO: 101), (EGGGK)3 (SEQ ID NO: 102), (EGGGR)3 (SEQ ID NO: 103), (DAAAR)3 (SEQ ID NO: 104), (DAAAK)3 (SEQ ID NO: 105), (DGGGR)3 (SEQ ID NO: 106) or (DGGGK)3 (SEQ ID NO: 107), SS, GGSEGKSSGSGSESKSTGGS (SEQ ID NO: 108), EPKSS (SEQ ID NO: 109), RTVAA (SEQ ID NO: 110), ASEPKSS (SEQ ID NO: 111); or the peptide linker is (G x S) y Where x is an integer selected from 1 to 5, and y is an integer selected from 1 to 6, including but not limited to GGGS, GGGGS (SEQ ID NO: 112), GGGGSGGGGS (SEQ ID NO: 113), GGGGSGGGGSGGGGS (SEQ ID NO: 84), GGGGSGGGGSGGGGSGGGS (SEQ ID NO: 83); or the peptide linker is GGGGG (SEQ ID NO: 114). In some embodiments, the peptide linker comprises the amino acid sequence of SEQ ID NO: 83 or 84. In some embodiments, the amino acid sequence of the peptide linker is as shown in SEQ ID NO: 83 or 84.

[0633] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments comprises:

[0634] The first strand contains at least one amino acid sequence (e.g., 1, 2, 3, 4, 5, or 6) containing SEQ ID NO: 87, 85, or 86, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity; at least one amino acid sequence (e.g., 1, 2, 3, 4, 5, or 6) containing SEQ ID NO: 20, 21, 22, or 23, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity; and at least one amino acid sequence (e.g., 1, 2, 3, 4, 5, or 6) containing SEQ ID NO: 20, 21, 22, or 23, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) ... sequence identity; and at least one amino acid sequence (e.g., 1, 2, 3, 4, 5, or 6) containing SEQ ID NO: 20, 21, 22, or 23. The third strand comprising SEQ ID NO: 88 or 89, or an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity, and at least one (e.g., 1, 2, 3, 4, 5, or 6) amino acid sequence containing SEQ ID NO: 66, 67, 68, 69, or 117, or an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity.

[0635] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments comprises:

[0636] One first strand containing SEQ ID NO: 87, 85, or 86, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with it; one second strand containing SEQ ID NO: 20, 21, 22, or 23, or having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with it; and one second strand containing SEQ ID NO: 87, 85, or 86. The third strand comprising SEQ ID NO: 88 or 89, or an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the amino acid sequence, and two fourth strands comprising SEQ ID NO: 66, 67, 68, 69, or 117, or an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the amino acid sequence.

[0637] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments comprises:

[0638] One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequences SEQ ID NO: 66, 67, 68, 69, or 117.

[0639] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments comprises:

[0640] One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 66; or

[0641] One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 67; or

[0642] One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 68; or

[0643] One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 69; or

[0644] One first strand containing the amino acid sequence of SEQ ID NO: 87, one second strand containing the amino acid sequence of SEQ ID NO: 20, one third strand containing the amino acid sequence of SEQ ID NO: 88, and two fourth strands containing the amino acid sequence of SEQ ID NO: 117.

[0645] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments comprises:

[0646] One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 66.

[0647] In some embodiments, as described in any of the preceding embodiments, an antigen-binding molecule that specifically binds to TfR1 and Aβ has the first and second chains interlinked, the first and third chains interlinked, and the third and fourth chains interlinked to form an antigen-binding molecule that specifically binds to TfR1 and Aβ.

[0648] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments comprises:

[0649] The antigen-binding molecule that specifically binds to TfR1 and Aβ comprises:

[0650] One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 66; or

[0651] One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 67; or

[0652] One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 68; or

[0653] One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 69; or

[0654] One first strand containing the amino acid sequence of SEQ ID NO: 87, one second strand containing the amino acid sequence of SEQ ID NO: 20, one third strand containing the amino acid sequence of SEQ ID NO: 88, and two fourth strands containing the amino acid sequence of SEQ ID NO: 117;

[0655] The first and second chains combine with each other, the first and third chains combine with each other, and the third and fourth chains combine with each other to form antigen-binding molecules that specifically bind to TfR1 and Aβ.

[0656] In some embodiments, the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding embodiments comprises:

[0657] The antigen-binding molecule that specifically binds to TfR1 and Aβ comprises:

[0658] One first strand containing the amino acid sequence of SEQ ID NO: 87, one second strand containing the amino acid sequence of SEQ ID NO: 20, one third strand containing the amino acid sequence of SEQ ID NO: 88, and two fourth strands containing the amino acid sequence of SEQ ID NO: 66;

[0659] The first and second chains combine with each other, the first and third chains combine with each other, and the third and fourth chains combine with each other to form antigen-binding molecules that specifically bind to TfR1 and Aβ.

[0660] In this disclosure, the first antigen-binding domain and the second antigen-binding domain are used only to distinguish the position of the binding domains and are not intended to limit specific sequences. In some embodiments, such as the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding claims, the first antigen-binding domain and the second antigen-binding domain are interchangeable.

[0661] In this disclosure, the terms "first hapten" and "second hapten" are used only to distinguish the position of the hapten and do not limit specific sequences. In some embodiments, as described in any of the preceding claims, antigen-binding molecules specifically bind to TfR1 and Aβ, wherein the first hapten and the second hapten are interchangeable.

[0662] In this disclosure, the first and second Aβ-binding Fabs are used only to distinguish the positions of the Aβ-binding Fabs and are not intended to limit specific sequences. In some embodiments, such as the antigen-binding molecule that specifically binds TfR1 and Aβ as described in any of the preceding claims, the first and second Aβ-binding Fabs are interchangeable.

[0663] In this disclosure, the first subunit Fc1 and the second subunit Fc2 are used only to distinguish the position of Fc and do not limit the specific sequence. In some embodiments, such as the antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding claims, wherein Fc1 and Fc2 are interchangeable.

[0664] In this disclosure, the terms "first strand," "second strand," "third strand," and "fourth strand" are used only to distinguish the positions of the strands and do not limit specific sequences. In some embodiments, antigen-binding molecules that specifically bind to TfR1 and Aβ, as described in any of the preceding claims, wherein the first, second, third, and fourth strands are interchangeable.

[0665] In some embodiments, the antigen-binding molecules that specifically bind TfR1 and Aβ provided in this disclosure are in EC5 concentrations of less than 100 nM (e.g., less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, less than 0.05 nM, less than 0.01 nM, less than 0.001 nM). 50 The value is combined with human TfR1, the EC 50 The value was measured using ELISA.

[0666] In some embodiments, the antigen-binding molecules that specifically bind TfR1 and Aβ provided in this disclosure are in EC5 concentrations of less than 10 nM (e.g., less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, less than 0.05 nM, less than 0.01 nM, less than 0.001 nM). 50 The value binds to human Aβ1-42fibril, the EC 50 The value was measured using ELISA.

[0667] In some embodiments, the antigen-binding molecules that specifically bind to TfR1 and Aβ provided in this disclosure have higher exposure levels to the hTfR1 transgenic mouse brain than positive antibodies (e.g., Trotinemab).

[0668] In some embodiments, the antigen-binding molecules that specifically bind to TfR1 and Aβ provided in this disclosure have a longer serum half-life in cynomolgus monkeys than positive antibodies (e.g., Trotinemab).

[0669] On the other hand, this disclosure provides an antigen-binding molecule that specifically binds to TfR1, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding claims, which competes with the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding claims for binding to human TfR1.

[0670] On the other hand, this disclosure provides an antigen-binding molecule that specifically binds to TfR1, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding claims, which, together with the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding claims, binds to the same human TfR1 antigenic epitope.

[0671] On the other hand, this disclosure provides an antigen-binding molecule that specifically binds to Aβ, or an antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding claims, which competes with the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding claims for binding to human Aβ.

[0672] On the other hand, this disclosure provides an antigen-binding molecule that specifically binds to Aβ, or an antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any of the preceding claims, which, together with the antigen-binding molecule that specifically binds to Aβ as described in any of the preceding claims, binds to the same human Aβ antigenic epitope.

[0673] On the other hand, this disclosure provides a pharmaceutical composition comprising an antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to Aβ as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding claims, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0674] In some embodiments, based on the total weight of the composition, the pharmaceutical composition contains 0.01-99.99% of an antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments, or an antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding embodiments. In some embodiments, the pharmaceutical composition contains 0.1-99.9% of an antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments, or an antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding embodiments. In some embodiments, the pharmaceutical composition contains 0.5%-99.5% of an antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding embodiments. In some embodiments, the pharmaceutical composition contains 1%-99% of an antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments, or an antigen-binding molecule that specifically binds to Aβ as described in any of the preceding embodiments, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding embodiments. In some embodiments, the pharmaceutical composition contains 2%-98% of an antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding embodiments, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding embodiments.

[0675] In some embodiments, the pharmaceutical composition contains 0.01% to 99.99% pharmaceutically acceptable carriers, diluents, or excipients based on the total weight of the composition. In some embodiments, the pharmaceutical composition contains 0.1% to 99.9% pharmaceutically acceptable carriers, diluents, or excipients. In some embodiments, the pharmaceutical composition contains 0.5% to 99.5% pharmaceutically acceptable carriers, diluents, or excipients. In some embodiments, the pharmaceutical composition contains 1% to 99% pharmaceutically acceptable carriers, diluents, or excipients. In some embodiments, the pharmaceutical composition contains 2% to 98% pharmaceutically acceptable carriers, diluents, or excipients.

[0676] On the other hand, this disclosure provides a nucleic acid that encodes an antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to Aβ as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding claims.

[0677] On the other hand, this disclosure provides a carrier containing the nucleic acid as described above.

[0678] On the other hand, this disclosure provides a host cell containing the nucleic acid as described above, or the vector as described above.

[0679] On the other hand, this disclosure provides a method for producing an antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to Aβ as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding claims, the method comprising culturing host cells as described in the preceding claims in a culture medium to form and accumulate the antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to Aβ as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding claims, and recovering the antigen-binding molecule from the culture.

[0680] On the other hand, this disclosure provides the use of an antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to Aβ as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding claims, or a pharmaceutical composition as described in any of the preceding claims, in the preparation of a medicament for the prevention or treatment of a disease or condition.

[0681] On the other hand, this disclosure provides a method for preventing or treating a disease or condition, the method comprising administering to a subject a preventive or therapeutically effective amount of an antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to Aβ as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding claims, or a pharmaceutical composition as described in any of the preceding claims.

[0682] On the other hand, this disclosure provides an antigen-binding molecule that specifically binds to TfR1 as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to Aβ as described in any of the preceding claims, or an antigen-binding molecule that specifically binds to both TfR1 and Aβ as described in any of the preceding claims, or a pharmaceutical composition as described in any of the preceding claims. In some embodiments, the pharmaceutical molecule is used to prevent or treat a disease or condition.

[0683] In some implementations, the disease or condition described in the preceding one is a neurological disease.

[0684] In some implementations, the disease or condition described in the preceding one is a central nervous system disease.

[0685] In some implementations, the disease or condition described in the preceding one is a neurodegenerative disease.

[0686] In some implementations, the disease or condition described in the preceding one is Alzheimer's disease. Attached Figure Description

[0687] Figure 1A: Schematic diagram of the structure of the maternal monoclonal antibody that specifically binds to TfR1.

[0688] Figure 1B: Schematic diagram of the structure of the maternal monoclonal antibody that specifically binds to Aβ.

[0689] Figure 2: Schematic diagram of the structure of a bispecific antibody that specifically binds to TfR1 and Aβ. Detailed Implementation

[0690] the term

[0691] To facilitate understanding of this disclosure, certain technical and scientific terms are described below. Unless otherwise expressly defined herein, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0692] The singular forms “a,” “an,” and “the” used in the specification and claims include plural references unless the context clearly indicates otherwise.

[0693] Unless the context clearly requires otherwise, the words “comprising,” “having,” “including,” etc., in the patent specification and claims should be understood as “including but not limited to,” rather than as exclusive or exhaustive.

[0694] The term "and / or" implies both "and" and "or". For example, the phrase "A, B and / or C" is intended to cover each of the following: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0695] The terms “TfR1,” “CD71,” and “TFRC” are used interchangeably to include variants, isotypes, homologues, orthologs, and paralogs of TfR1. Human TfR1 has the amino acid sequence Uniprot: P02786. Although specific database accession numbers are given, those skilled in the art will understand that TfR1 as referred to herein also encompasses the corresponding sequences reported in other databases or literature.

[0696] The terms "Aβ", "Abeta", "amyloid β", and "β-amyloid" are used interchangeably to refer to the fragments generated from amyloid precursor protein (APP) after cleavage by β-secretase 1 (BACE1), as well as their modified fragments and any functional equivalents, including but not limited to N3pGlu Aβ, Aβ1-40, and Aβ1-42. Aβ monomers can combine to form oligomers, protofibrils, fibrils, or plaques. The term "Aβ" includes variants, isotypes, homologues, orthologs, and paralogs of Aβ.

[0697] The three-letter and single-letter codes for amino acids used in this disclosure are as described in J. Biol. Chem., 243, p3558 (1968).

[0698] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimics that function in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those that are subsequently modified, such as hydroxyproline, γ-carboxyglutamic acid, and O-phosphoserine. Amino acid analogs are compounds that have the same basic chemical structure as naturally occurring amino acids (i.e., the α-carbon bound to hydrogen, carboxyl, amino, and R groups), such as homoserine, ortholeucine, methionine sulfoxide, and methionine methylsulfonium. These analogs have modified R groups (e.g., ortholeucine) or modified peptide backbones but retain the same basic chemical structure as naturally occurring amino acids. Amino acid mimics are chemical compounds that have a structure different from the general chemical structure of amino acids but function in a manner similar to naturally occurring amino acids.

[0699] The term "amino acid mutation" includes amino acid substitution (also known as amino acid replacement), deletion, insertion, and modification. Any combination of substitution, deletion, insertion, and modification can be performed to achieve the final construct, provided that the final construct possesses the desired properties, such as reduced or absent binding to Fc receptors. Amino acid sequence deletions and insertions include deletions and insertions at the amino and / or carboxyl ends of the polypeptide chain. A specific amino acid mutation can be an amino acid substitution. In some embodiments, an amino acid mutation is a non-conservative amino acid substitution, i.e., replacing one amino acid with another amino acid that has a different structure and / or chemical properties. Amino acid substitution includes substitution by non-naturally occurring amino acids or by derivatives of 20 naturally occurring amino acids (e.g., 4-hydroxyproline, 3-methylhistidine, ornithine, homoserine, 5-hydroxylysine). Amino acid mutations can be generated using genetic or chemical methods known in the art. Genetic methods can include site-directed mutagenesis, PCR, gene synthesis, etc. Methods other than genetic engineering that alter amino acid side chain groups, such as chemical modification, are also expected to be available. Various names may be used herein to refer to the same amino acid mutation. In this document, the amino acid residue at a specific site can be represented by the format "position + amino acid residue". For example, 351I indicates that the amino acid residue at position 351 is I. L351I indicates that the amino acid residue at position 351 has mutated from leucine (L) to isoleucine (I). It should be understood that when the amino acid sequence is defined by the format "position + residue" in the claims, the amino acid before the mutation at that site does not constitute a limitation on the technical solution.

[0700] The term "blood-brain barrier" (BBB) ​​refers to the physiological barrier between the peripheral circulation and the brain and spinal cord. It consists of tight junctions of the brain capillary endothelial plasma membranes, forming an extremely tight barrier that restricts the transport of molecules into the brain, even very small molecules such as urea (60 Da). The blood-brain barrier within the brain, the blood-spinal cord barrier within the spinal cord, and the blood-retinal barrier within the retina form a continuous capillary barrier within the central nervous system, collectively known as the blood-brain barrier. The blood-brain barrier also includes the blood-cerebrospinal fluid barrier (choroid plexus), which is composed of ependymal cells rather than capillary endothelial cells.

[0701] The term "antigen-binding molecule" is used in the broadest sense to encompass molecules that specifically bind antigens, including but not limited to antibodies, oligonucleotides, other peptides with antigen-binding activity, and antibody fusion proteins formed by the fusion of the two, as well as any molecule containing the aforementioned antibodies, oligonucleotides, peptides, or antibody fusion proteins, provided they exhibit the desired antigen-binding activity. Exemplarily, the antigen-binding molecules described herein are monoclonal antibodies, monospecific antibodies, polyclonal antibodies, and multispecific antibodies (e.g., bispecific antibodies, trispecific antibodies).

[0702] The term “antibody” is used in the broadest sense and covers a wide range of antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies), full-length antibodies, antibody fragments, and antigen-binding fragments (or antigen-binding portions), as long as they exhibit the desired antigen-binding activity.

[0703] The term "antigen-binding fragment" encompasses full-length antibodies, Fab, modified Fab, Fab', Fab'-SH, modified Fab', F(ab')2, Fv, dsFv, Fab-Fv, Fab-dsFv, Fd, single-domain antibodies (sdAb, e.g., VH, VL, or VHH), single-chain Fab (scFab), single-chain antibodies (e.g., scFv, sc(Fv)2), biantibodies, linear antibodies, bivalent, trivalent, or tetravalent antibodies, Bis-scFv, diabetic, tribody, triabody, tetrabody, and epitope-binding fragments of any of the above. Methods for generating and preparing these antigen-binding fragments are well known in the art.

[0704] The term "antibody fragment" refers to a molecule that is distinct from the intact antibody but contains a portion of the intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, dsFv, Fab, Fab′, Fab′-SH, F(ab′)2, Fd, single-domain antibodies (sdAb, such as VH, VL, or VHH), single-chain Fab (scFab), biantibodies, linear antibodies, single-chain antibodies (such as scFv, sc(Fv)2); and multispecific antibodies formed from antibody fragments.

[0705] The term "natural antibody" refers to naturally occurring immunoglobulin molecules. For example, natural IgG antibodies are heterotetraglycoproteins of approximately 150,000 Daltons, composed of two light chains and two heavy chains linked by disulfide bonds. From the N-terminus to the C-terminus, each heavy chain has a variable region (VH), also called a variable heavy domain or heavy chain variable region, followed by a heavy chain constant region. The natural IgG heavy chain constant region typically contains three constant domains (CH1, CH2, and CH3). Similarly, from the N-terminus to the C-terminus, each light chain has a variable region (VL), also called a variable light domain or light chain variable domain, followed by a constant light domain (light chain constant region, CL).

[0706] The terms "full-length antibody," "intact antibody," and "complete antibody" are used interchangeably in this document, referring to antibodies with a structure substantially similar to that of natural antibodies or with a heavy chain containing the Fc region as defined herein. The light chain of a natural intact antibody includes a variable region (VL) and a constant region (CL), with VL located at the amino terminus of the light chain. The constant region includes the κ and λ chains. The heavy chain includes a variable region (VH) and constant regions (CH1, CH2, and CH3), with VH located at the amino terminus of the heavy chain and the constant region located at the carboxyl terminus. CH3 is closest to the carboxyl terminus of the polypeptide. The heavy chain can belong to any isotype, including IgG (including IgG1, IgG2, IgG3, and IgG4 subtypes), IgA (including IgA1 and IgA2 subtypes), IgM, and IgE.

[0707] The term "multispecific antibody" refers to an antibody (including the antibody or its antigen-binding fragment) that can specifically bind to multiple different antigens or at least two different antigenic epitopes of the same antigen. In this disclosure, "anti-TfR1 and Aβ antibodies are multispecific antibodies, optionally bispecific antibodies, comprising one first antigen-binding domain specifically binding to TfR1 and two second antigen-binding domains specifically binding to Aβ. The term 'multispecific antibody' is used in the broadest sense in this disclosure and specifically covers antibodies having multi-epitope specificity. Multispecific antibodies include, but are not limited to, antibodies comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH and VL together constitute an antigen-binding domain (where VH / VL have multi-epitope specificity), antibodies having two or more VH and VL domains (each VH / VL unit binds to a different epitope), antibodies having two or more single variable domains (each single variable domain binds to a different epitope), full-length antibodies, and antibodies comprising one or more antibody fragments, as well as antibodies comprising antibody fragments covalently or non-covalently linked. Multispecific antibodies can be bispecific antibodies, trispecific antibodies, biantibodies, or similar molecules."

[0708] The term "operably linked" refers to a functional relationship between two or more peptide or polypeptide domains or nucleic acid (e.g., DNA) segments. In this disclosure, the term "operably linked" means linking two or more amino acid segments to produce a functional polypeptide. For example, in the context of antigen-binding molecules of this disclosure, a single antigen-binding domain can be linked by a peptide linker sequence. In the context of nucleic acids encoding fusion proteins, such as polypeptide chains of antigen-binding molecules of this disclosure, "operably linked" means linking two nucleic acids such that the amino acid sequences encoded by said two nucleic acids remain within the frame.

[0709] The term "association" in this disclosure refers to a functional relationship between two or more chains. Specifically, the term "association" means that two or more polypeptides associate with each other, for example, through non-covalent association via molecular interactions or through covalent association via one or more disulfide bridges or chemical crosslinks, thereby generating a functional antigen-binding molecule (e.g., a bispecific antibody), wherein a first antigen-binding domain and a second antigen-binding domain can bind their respective targets. Examples of association that may exist in the antigen-binding molecules of this disclosure include, but are not limited to, association between Fc regions in the Fc domain, association between the VH and VL regions in the Fab or Fv domain, and association between CH1 and CL in the Fab domain.

[0710] In the context of antigen-binding molecules (e.g., bispecific antibodies), the term "bivalent" refers to an antigen-binding molecule that has two antigen-binding domains.

[0711] In the context of antigen-binding molecules (e.g., bispecific antibodies), the term "trivalent" refers to an antigen-binding molecule that has three antigen-binding domains.

[0712] The term "half-antibody" refers to a molecule that contains at least one antigen-binding domain and can associate with another molecule containing an antigen-binding domain via, for example, a disulfide bridge or molecular interactions (e.g., club-and-mortar interactions between Fc heterodimers, hydrogen bonds, electrostatic interactions, or salt bridges). A half-antibody may consist of one or more polypeptide chains (e.g., two polypeptide chains of Fab). In some embodiments, the half-antibody further comprises an Fc region.

[0713] Examples of haptens are molecules comprising a heavy chain and a light chain of an antibody (e.g., an IgG antibody). In some embodiments, the hapten is a molecule comprising a first polypeptide and a second polypeptide, the first polypeptide comprising a VH domain, a CH1 domain, a hinge domain, a CH2 domain, and a CH3 domain from the N-terminus to the C-terminus, and the second polypeptide comprising a VL domain and a CL domain, wherein the VH and VL domains form an antigen-binding domain. In some embodiments, the hapten may comprise more than one antigen-binding domain; for example, the hapten may be a molecule comprising a first polypeptide and a second polypeptide, the first polypeptide comprising a VH domain, a CH1 domain, a hinge domain, a CH2 domain, and a CH3 domain from the N-terminus to the C-terminus, and the second polypeptide comprising a VL domain and a CL domain.

[0714] Therefore, an antigen-binding molecule may contain at least one, more typically one, two, or even more than two half antibodies, and a half antibody may contain one or more antigen-binding domains.

[0715] In some embodiments, the first hapten and the second hapten are associated, for example, through heterodimerization. In some embodiments, the first hapten is covalently linked to the second hapten, for example, via disulfide bridging or chemical cross-linking. In some embodiments, the first hapten and the second hapten are associated through covalent attachment and non-covalent interactions, such as disulfide bridging and mortise-and-tenon interactions.

[0716] The term "half-antibody" is intended for descriptive purposes only and does not indicate a specific configuration or method of manufacture. The descriptions of half-antibodies as "first" half-antibody, "second" half-antibody, "left" half-antibody, "right" half-antibody, etc., are merely for convenience and descriptive purposes.

[0717] The term "C-terminus," also known as, but not limited to, carboxyl-terminus, C-terminus, C-tail, C-terminus, or COOH-terminus, is the terminal end of an amino acid chain (protein or polypeptide) terminated by a free carboxyl group (-COOH). When a protein is translated from messenger RNA, it is produced from the N-terminus to the C-terminus. The convention for writing peptide sequences is to place the C-terminus on the right and write the sequence from the N-terminus to the C-terminus. In some embodiments, the C-terminus of a polypeptide includes the last amino acid residue of the polypeptide, which contributes its amino group to form a peptide bond with the carboxyl group of its adjacent amino acid residue.

[0718] The term "N-terminus," also known as, but not limited to, amino-terminus, amine-terminus, N-terminus, N-tail, N-terminus, or NH2-terminus, is the start of an amino acid chain (protein or polypeptide) beginning with a free amino group (-NH2). Normally, the amino group bonds to another carboxyl group in the protein to form a chain, but since only one of the two ends of a protein is chained, the free amino group refers to the N-terminus. As mentioned above, by convention, peptide sequences in LTR language are written from N-terminus to C-terminus, from left to right. This associates the translation direction with the text direction (because when a protein is translated from messenger RNA, it is produced from N-terminus to C-terminus—an amino acid is added to the carbonyl terminus). In some embodiments, the N-terminus of a polypeptide contains the first amino acid of the polypeptide, which contributes its carboxyl group to form a peptide bond with the amino group of its adjacent amino acid residue.

[0719] The term "linker" refers to a connecting unit that links two polypeptide fragments. In this disclosure, linkers appearing in the same structural formula may be the same or different. A linker may be a "peptide linker" containing one or more amino acids, typically about 1-30, 2-24, or 3-15 amino acids. Linkers used in this disclosure may be the same or different. When a "-" appears in a structural formula, it indicates that the units on either side are directly connected by a covalent bond.

[0720] The term "peptide linker" can be any suitable peptide chain, as long as the antigen-binding molecule can exhibit the desired antigen-binding activity.

[0721] The term "Fc region" or "fragment crystallizable region" is used to define the C-terminal region of an antibody heavy chain, including both native and engineered Fc regions. In some embodiments, the Fc region comprises two identical or different subunits. Suitable Fc regions for the antibodies described herein include the Fc regions of human IgG1, IgG2 (IgG2A, IgG2B), IgG3, and IgG4. In some embodiments, the boundaries of the Fc region may also vary, for example, by deleting the C-terminal lysine (residue 447 according to the EU numbering system) or by deleting both the C-terminal glycine and lysine (residues 446 and 447 according to the EU numbering system). Unless otherwise stated, the Fc region is numbered according to the EU numbering system, also known as the EU index.

[0722] The Fc region can be appropriately obtained by partially digesting IgG monoclonal antibodies with proteolytic enzymes such as pepsin, followed by eluting the components adsorbed on the protein A or protein G column. As the proteolytic enzyme, any enzyme capable of restrictively digesting full-length antibodies to produce Fab and F(ab')2 by appropriately setting the enzyme reaction conditions such as pH is acceptable; there is no particular limitation, and examples include pepsin and papain.

[0723] The term "variable region" or "variable domain" in an antibody refers to the domain in the antibody heavy or light chain involved in antibody binding to the antigen. In this paper, the antibody heavy chain variable region (VH) and light chain variable region (VL) each contain four conserved frame regions (FRs) and three complementarity-determining regions (CDRs). The term "complementarity-determining region" or "CDR" refers to the region within the variable domain that primarily facilitates antigen binding; "frame" or "FR" refers to the variable domain residues other than the CDR residues. The VH contains three CDR regions: HCDR1, HCDR2, and HCDR3; the VL contains three CDR regions: LCDR1, LCDR2, and LCDR3. Each VH and VL consists of three CDRs and four FRs arranged in the following order from the amino terminus (also called the N-terminus) to the carboxyl terminus (also called the C-terminus): FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0724] The amino acid sequence boundaries of CDRs can be determined using various well-known schemes, such as the "Kabat" numbering rule, the "Chothia" numbering rule, the "ABM" numbering rule, the "contact" numbering rule, and the ImMunoGenTics (IMGT) numbering rule. The correspondence between various numbering systems is well known to those skilled in the art, and is exemplarily shown in Table A below.

[0725] Table A. Relationships between CDR numbering systems

[0726] Unless otherwise stated, the variable areas and CDRs in this disclosure embodiment are subject to the "Kabat" numbering rule.

[0727] The term "chimeric" antibody refers to an antibody in which a portion of the heavy and / or light chain is derived from a specific source or species, while the remaining portion of the heavy and / or light chain is derived from another different source or species.

[0728] The term "humanized" antibody refers to an antibody that retains the reactivity of a non-human antibody while exhibiting lower immunogenicity in humans. For example, this can be achieved by retaining the non-human CDR region and replacing the rest of the antibody with its human counterpart (i.e., the frame region portion of the constant region and the variable region).

[0729] The terms "human antibody," "humanized antibody," "fully human antibody," and "completely human antibody" are used interchangeably, referring to antibodies whose variable and constant regions are human sequences. This term encompasses antibodies derived from human genes but with sequence alterations, such as reduced potential immunogenicity, increased affinity, or the elimination of cysteine ​​or glycosylation sites that might cause undesirable folding. This term also covers antibodies recombined in non-human cells (which may confer glycosylations not characteristic of human cells). The term also includes antibodies produced in transgenic mice containing some or all human immunoglobulin heavy and light chain loci. The term "human antibody" explicitly excludes humanized antibodies.

[0730] The term "affinity" refers to the overall strength of the non-covalent interaction between a single binding site of a molecule (e.g., an antibody) and its binding ligand (e.g., an antigen). Unless otherwise specified, as used herein, binding "affinity" refers to internal binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of molecule X for its ligand Y can typically be represented by the dissociation constant (KD). Affinity can be measured using conventional methods known in the art, including those described herein.

[0731] As used herein, the term "kassoc" or "ka" refers to the association rate of a specific antibody-antigen interaction, and the term "kdis" or "kd" refers to the dissociation rate of a specific antibody-antigen interaction. The term "KD" refers to the dissociation constant, which is derived from the ratio of kd to ka (i.e., kd / ka) and expressed as a molar concentration (M). The KD value of an antibody can be determined using methods known in the art. For example, it can be measured using a biosensing system such as a system for measuring surface plasmon resonance (e.g., Biacore), or by measuring affinity in solution using solution equilibrium titration (SET).

[0732] The term “surface plasmon resonance” refers to the optical phenomenon of analyzing real-time interactions by detecting changes in protein concentration within a biosensor matrix, for example, using the BIAcore system (Biacore LifeSciences division of GE Healthcare, Piscataway, NJ).

[0733] The term "effector function" refers to biological activities attributable to the antibody's Fc region (either the native Fc region or the Fc region with amino acid sequence mutations) and that vary across antibody isotypes. Examples of antibody effector functions include, but are not limited to: C1q binding and complement-dependent cytotoxicity, Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, downregulation of cell surface receptors (e.g., B cell receptors), and B cell activation.

[0734] The term "monoclonal antibody" refers to a group of antibodies that are essentially homogeneous, meaning that the antibody molecules contained in this group have the same amino acid sequence, except for the possible small number of naturally occurring mutations. In contrast, polyclonal antibodies typically comprise a variety of different antibodies with varying amino acid sequences in their variable domains, and they generally target different epitopes specifically. "Monoclonal" should not be interpreted as requiring the production of the antibody through any particular method.

[0735] The term "bispecific antibody" refers to an antibody (including the antibody or its antigen-binding fragment, such as a single-chain antibody) capable of specifically binding to two different antigens or at least two different antigenic epitopes of the same antigen. Various structures of bispecific antibodies have been disclosed in the prior art. Based on the integrity of the IgG molecule, they can be classified into IgG-like bispecific antibodies and antibody fragment-based bispecific antibodies. Based on the number of antigen-binding regions, they can be classified into bivalent, trivalent, tetravalent, or more bispecific antibodies. Based on structural symmetry, they can be classified into symmetrical and asymmetrical bispecific antibodies. Among these, bispecific antibodies based on antibody fragments, such as Fab fragments lacking the Fc fragment, form bispecific antibodies by combining two or more Fab fragments into one molecule. They exhibit lower immunogenicity, smaller molecular weight, and higher tumor tissue penetration. IgG-like bispecific antibodies (e.g., those with an Fc fragment) have a relatively larger molecular weight. The Fc fragment facilitates antibody purification and improves its solubility and stability. The Fc portion may also bind to the receptor FcRn, increasing the antibody's serum half-life. Typical bispecific antibody structural models include KiH, CrossMAb, Triomab quadroma, FcΔAdp, ART-Ig, BiMAb, Biclonics, BEAT, DuoBody, Azymetric, XmAb, 2:1 TCBs, 1Fab-IgG TDB, FynomAb, two-in-one / DAF, scFv-Fab-IgG, DART-Fc, LP-DART, CODV-Fab-TL, HLE-BiTE, F(ab)2-CrossMAb, IgG-(scFv)2, Bs4Ab, DVD-Ig, Tetravalent-DART-Fc, (scFv)4-Fc, CODV-Ig, mAb2, and F(ab)4-CrossMAb, among other bispecific antibodies.

[0736] The term "antigen" refers to a molecule or molecular part that can be selectively bound by antigen-binding proteins, including, for example, antibodies. An antigen may have one or more epitopes that can interact with different antigen-binding proteins, such as antibodies.

[0737] The term "epitope" refers to a region on an antigen that is capable of specifically binding to an antibody or its antigen-binding fragment. Epitopes can be formed from a continuous string of amino acids (linear epitopes) or contain non-continuous amino acids (conformal epitopes), such as those spatially close due to antigen folding. The difference between conformational and linear epitopes is that antibody binding to a conformational epitope is lost in the presence of a denaturing solvent. An epitope contains at least 3, at least 4, at least 5, at least 6, at least 7, or 8-10 amino acids in a unique spatial conformation. Screening for antibodies that bind to a specific epitope (i.e., those that bind the same epitope) can be performed using methods routine in the art, such as, but not limited to, alanine scanning, Western blotting, peptide cleavage analysis, epitope excision, epitope extraction, chemical modification of the antigen (see Prot. Sci. 9 (2000) 487-496), and cross-blocking.

[0738] The terms "specific binding," "specific binding," or "binding" refer to the ability of an antibody to bind to a specific antigen or epitope with a higher affinity than other antigens or epitopes. Typically, antibodies bind at an affinity of approximately 1 × 10⁻⁶. -6 M or smaller (e.g., about 1×10⁻⁶) -7 M, 1×10 -8 M, 1×10 -9 M, 1×10 -10 M, 1×10 -11 The equilibrium dissociation constant (KD) of an antibody (M or less) binds to an antigen or an epitope within the antigen. In some embodiments, the KD of the antibody binding to an antigen is 10% or less (e.g., 1%) of the KD of the antibody binding to a nonspecific antigen (e.g., BSA, casein). KD can be measured using known methods, such as by... Surface plasmon resonance assays are used to measure this. However, antibodies that specifically bind to antigens or epitopes within antigens may be cross-reactive to other related antigens, for example, to corresponding antigens from other species (homologous) (such as humans or monkeys, such as the cynomolgus (cyno), chimpanzee (chimp), or common marmoset (marmoset)).

[0739] The term "non-binding" means that the antibody cannot bind to an antigen or its epitope in the manner described above for specific binding. For example, when the antibody binds at approximately 1 × 10⁻⁶... -6 M or a larger equilibrium dissociation constant (KD) binds to the antigen or its epitope.

[0740] The term "antigen-binding molecule that specifically binds to TfR1 and Aβ" refers to an antigen-binding molecule capable of binding to TfR1 or its epitope, or Aβ or its epitope, with sufficient affinity. The terms "anti-TfR1 and Aβ bispecific antibody" and "TfR1×Aβ bispecific antibody" refer to antibodies capable of binding to TfR1 or its epitope, or Aβ or its epitope, with sufficient affinity. In some embodiments, the term "first antigen-binding domain that specifically binds to TfR1" refers to an antigen-binding domain capable of binding to TfR1 with a certain affinity, such that a molecule containing this antigen-binding domain can be used as a diagnostic and / or therapeutic agent targeting TfR1. In some embodiments, the term "second antigen-binding domain that specifically binds to Aβ" refers to an antigen-binding domain capable of binding to Aβ with a certain affinity, such that a molecule containing this antigen-binding domain can be used as a diagnostic and / or therapeutic agent targeting Aβ. In this disclosure, antigen-binding domains include "antibody," "antigen-binding fragment," and "antibody fragment" (e.g., Fab) as defined in this disclosure.

[0741] The terms “antibody-dependent cell cytotoxicity,” “antibody-dependent cell-mediated cytotoxicity,” or “ADCC” refer to mechanisms that induce cell death that rely on the interaction between antibody-coated target cells and lytic effector cells (such as natural killer (NK) cells, monocytes, macrophages, and neutrophils) via Fcγ receptors (FcγR) expressed on the effector cells. For example, NK cells express FcγRIIIa, while monocytes express FcγRI, FcγRII, and FcγRIIIa. The ADCC activity of the antibodies described herein can be assessed in vitro using cells expressing the antigen as target cells and NK cells as effector cells. Cell lysis is detected based on the release of markers (e.g., radioactive substrates, fluorescent dyes, or native intracellular proteins) from lysed cells.

[0742] The term "antibody-dependent phagocytosis (ADCP)" refers to the mechanism by which antibody-coated target cells are eliminated through internalization by phagocytes (such as macrophages or dendritic cells).

[0743] The term "complement-dependent cytotoxicity" or "CDC" refers to a mechanism that induces cell death in which the Fc effector domain of a target-binding antibody binds to and activates the complement component C1q. C1q then activates the complement cascade, leading to target cell death. Activation of complement can also result in the deposition of complement components on the surface of target cells, which promote CDC by binding to complement receptors (e.g., CR3) on leukocytes.

[0744] The term "nucleic acid" is used interchangeably with the term "polynucleotide" herein and refers to deoxyribonucleotides or ribonucleotides and their polymers in single-stranded or double-stranded form. The term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, or non-natural, have similar binding properties to a reference nucleic acid, and are metabolized in a manner similar to that of a reference nucleotide. Examples of such analogs include, but are not limited to, phosphate thioesters, aminophosphate esters, methylphosphonates, chiral methylphosphonates, 2-O-methylribonucleotides, and peptide-nucleic acids (PNAs).

[0745] Nucleic acid encoding a polypeptide refers to one or more nucleic acid molecules encoding a polypeptide, including one or more such nucleic acid molecules in a single vector or separate vectors, and one or more such nucleic acid molecules present at one or more locations in the host cell. Unless otherwise stated, a specific nucleic acid sequence also implicitly encompasses variants of its conserved modifications (e.g., degenerate codon substitutions) and complementary sequences, as well as explicitly specified sequences.

[0746] The terms “peptide” and “protein” are used interchangeably herein to refer to polymers of amino acid residues. The term applies to amino acid polymers, where one or more amino acid residues are artificial chemical mimics of the corresponding naturally occurring amino acids, as well as to both naturally occurring and non-naturally occurring amino acid polymers. Unless otherwise stated, a particular peptide sequence also implicitly encompasses variants with conserved modifications.

[0747] The term "sequence identity" refers to the degree (percentage) to which two sequences share the same amino acids / nucleic acids at equivalent positions when optimally aligned; gaps may be introduced, where necessary, to obtain the maximum percentage of sequence identity, without considering any conserved substitutions as part of sequence identity. To determine the percentage of sequence identity, alignment can be performed using techniques known in the art, such as publicly available computer software like BLAST, BLAST-2, ALIGN, ALIGN-2, or Megalign (DNASTAR) software. Those skilled in the art can determine the parameters suitable for measuring alignment, including any algorithms required to achieve maximum alignment across the full length of the sequences being compared.

[0748] The term "vector" refers to a polynucleotide molecule capable of transporting another polynucleotide linked to it. One type of vector is a "plasmid," which is a circular double-stranded DNA loop in which an additional DNA segment can be attached. Another type of vector is a viral vector, such as an adeno-associated virus vector (AAV or AAV2), in which an additional DNA segment can be attached to the viral genome. Some vectors are capable of autonomous replication in the host cells to which they are introduced (e.g., bacterial vectors with bacterial origins of replication and attachable mammalian vectors). Other vectors (e.g., non-attached mammalian vectors) can integrate into the host cell's genome after introduction into the host cell, thereby replicating along with the host genome. The term "expression vector" or "expression construct" refers to a vector capable of transforming host cells and containing a nucleic acid sequence that directs and / or controls (alongside the host cell) the expression of one or more heterologous coding regions operatively linked to it. Expression constructs can include, but are not limited to, sequences that affect or control transcription, translation, and, in the presence of introns, influence RNA splicing of coding regions operatively linked to them.

[0749] The terms “host cell,” “host cell line,” and “host cell culture” are used interchangeably and refer to cells into which exogenous nucleic acids have been introduced, including the progeny of such cells. Host cells include “transformers” and “transformed cells,” which include primary transformed cells and their derived progeny, regardless of the number of passages. Progeny may not be identical to parental cells in their nucleic acid contents and may contain mutations. Mutant progeny are included herein, which have the same function or biological activity as cells screened or selected in the initial transformed cells. Host cells include prokaryotic and eukaryotic host cells, wherein eukaryotic host cells include, but are not limited to, mammalian cells, insect cell lines, plant cells, and fungal cells. Mammalian host cells include human, mouse, rat, dog, monkey, pig, goat, cow, horse, and hamster cells, including but not limited to Chinese hamster ovary (CHO) cells, NSO, SP2 cells, HeLa cells, young hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), A549 cells, 3T3 cells, and HEK-293 cells.Fungal cells include yeast and filamentous fungal cells, including, for example, *Pichia pastoris*, *Pichia finlandica*, *Pichia trehalophila*, *Pichia koclamae*, *Pichia membranaefaciens*, *Pichia minuta* (Ogataea minuta, *Pichia lindneri*), *Pichia xiaopuntiae*, *Pichia thermotolerans*, *Pichia salictaria*, *Pichia guercuum*, *Pichia pijperi*, *Pichia stiptis*, *Pichia methanolica*, *Pichia* genus, *Saccharomyces cerevisiae*, *Saccharomyces* genus, and *Hansenula*. The fungi include *C. polymorpha*, *Kluyveromyces lactis*, *Candida albicans*, *Aspergillus*, *Aspergillus nidulans*, *Aspergillus niger*, *Aspergillus oryzae*, *Trichoderma reesei*, *Chrysosporium lucknowense*, *Fusarium sp.*, *Fusarium gramineum*, *Fusarium venenatum*, *Physcomitrella patens*, *Neurospora crassa*, and *Yarrowia lipolytica*.

[0750] The terms “cell,” “cell line,” and “cell culture” are used interchangeably, and all such names include progeny. Therefore, the terms “transformation” and “transformed cell” include primary subject cells and cultures derived from them, regardless of the number of passages. It should also be understood that due to intentional or unintentional mutations, not all progeny will have exactly the same DNA contents. This includes mutant progeny that have the same function or biological activity as the original transformed cells.

[0751] "Optional" or "optionally" means that the event or circumstances described below may, but do not have to, occur, and the description includes the circumstances under which the event or circumstances may or may not occur.

[0752] The term "pharmaceutical composition" means a mixture containing one or more antigen-binding molecules or conjugates disclosed herein, along with other chemical components, such as physiological / pharmaceutical carriers, diluents, or excipients.

[0753] The term "pharmaceutically acceptable carrier, diluent, or excipient" refers to a component in a pharmaceutical formulation that is different from the active ingredient and is non-toxic to the subject. Pharmaceutically acceptable carriers, diluents, or excipients include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0754] The terms "subject" or "individual" include both humans and non-human animals. Non-human animals include all vertebrates (e.g., mammals and non-mammals) such as non-human primates, sheep, dogs, cattle, chickens, amphibians, and reptiles. Unless otherwise specified, the terms "patient" or "subject" are used interchangeably herein. In some embodiments, the individual or subject is a human being.

[0755] "Administration" or "giving," when applied to animals, humans, experimental subjects, cells, tissues, organs, or biological fluids, refers to the contact between an exogenous drug, therapeutic agent, diagnostic agent, or composition and the animal, human, subject, cell, tissue, organ, or biological fluid.

[0756] The term "sample" refers to a collection (such as fluid, cells, or tissue) separated from a subject, as well as fluids, cells, or tissues present within a subject. Exemplary samples include biological fluids such as blood, serum and serous fluid, plasma, lymph, urine, saliva, cystic fluid, tears, excretions, sputum, mucosal secretions of secretory tissues or organs, vaginal secretions, ascites, pleura, pericardium, peritoneum, fluids in the abdominal cavity and other body cavities, fluids collected by bronchoalveolar lavage fluid, synovial fluid, liquid solutions in contact with the subject or biological sources, such as culture media (including conditioned media), lavage fluids, tissue biopsy samples, fine-needle aspiration, surgically removed tissue, organ cultures, or cell cultures.

[0757] "Treatment" and "treatment" (and their grammatical variations) refer to a clinical intervention on the individual being treated, and can be implemented for prevention or during a clinicopathological process. The expected effects of treatment include, but are not limited to, preventing the onset or recurrence of disease, alleviating symptoms, reducing / decreasing any direct or indirect pathological consequences of the disease, preventing metastasis, slowing the rate of disease progression, improving or alleviating the disease state, and resolving or improving the prognosis.

[0758] An "effective dose" is generally an amount sufficient to reduce the severity and / or frequency of symptoms, eliminate these symptoms and / or underlying causes, prevent the occurrence of symptoms and / or underlying causes, and / or improve or mitigate damage caused by or associated with a disease state (e.g., lung disease). In some implementations, an effective dose is a therapeutically effective dose or a preventatively effective dose. A "therapeuticly effective dose" is an amount sufficient to treat a disease state or symptom, especially a state or symptom associated with that disease state, or otherwise prevent, inhibit, delay, or reverse the progression of the disease state or any other undesirable symptom associated with that disease. A "preventatively effective dose" is an amount that, when given to a subject, will have a predetermined preventative effect, such as preventing or delaying the onset (or recurrence) of the disease state, or reducing the likelihood of the onset (or recurrence) of the disease state or related symptoms. Complete treatment or prevention may not occur after a single dose, but may occur after a series of doses. Therefore, a therapeutically or preventatively effective dose may be administered in a single or multiple-dose manner. "Therapeutic effective dose" and "preventive effective dose" can vary depending on a number of factors, such as an individual's disease state, age, sex, and weight, as well as the ability of the treatment or combination of treatments to elicit the desired response in the individual. Exemplary indicators of an effective treatment or combination of treatments include, for example, improved health status in the patient.

[0759] This disclosure discloses antigen-binding molecules that specifically bind to TfR1.

[0760] This disclosure provides antigen-binding molecules that specifically bind to TfR1 and possess numerous advantageous properties, such as good affinity, therapeutic activity, safety, pharmacokinetic properties, and drugability (e.g., solubility, viscosity, purity, and stability).

[0761] Exemplary antigen-binding molecules that specifically bind to TfR1

[0762] For example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0763] The amino acid sequences of the heavy chain variable region HCDR1 are shown in SEQ ID NO: 3, HCDR2 in SEQ ID NO: 11, and HCDR3 in SEQ ID NO: 36; the amino acid sequences of the light chain variable region LCDR1 are shown in SEQ ID NO: 9, LCDR2 in SEQ ID NO: 10, and LCDR3 in SEQ ID NO: 8, wherein:

[0764] X1 can be S or Y, with S being preferred; X2 can be T or S, with T being preferred.

[0765] For example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0766] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 3, HCDR2 contains the amino acid sequence of SEQ ID NO: 11, and HCDR3 contains the amino acid sequence of SEQ ID NO: 25, and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 9, LCDR2 contains the amino acid sequence of SEQ ID NO: 10, and LCDR3 contains the amino acid sequence of SEQ ID NO: 8.

[0767] For example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0768] The amino acid sequences of the heavy chain variable regions HCDR1 are shown in SEQ ID NO: 3, HCDR2 are shown in SEQ ID NO: 11, and HCDR3 are shown in SEQ ID NO: 25. The amino acid sequences of the light chain variable regions LCDR1 are shown in SEQ ID NO: 9, LCDR2 are shown in SEQ ID NO: 10, and LCDR3 are shown in SEQ ID NO: 8.

[0769] For example, the antigen-binding molecules that specifically bind to TfR1 disclosed herein, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the Kabat numbering rules.

[0770] For example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0771] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 37, HCDR2 contains the amino acid sequence of SEQ ID NO: 38, and HCDR3 contains the amino acid sequence of SEQ ID NO: 39, and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 40, LCDR2 contains the amino acid sequence of SEQ ID NO: 41, and LCDR3 contains the amino acid sequence of SEQ ID NO: 8.

[0772] For example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0773] The amino acid sequences of the heavy chain variable region HCDR1 are shown in SEQ ID NO: 37, HCDR2 in SEQ ID NO: 38, and HCDR3 in SEQ ID NO: 39. The amino acid sequences of the light chain variable region LCDR1 are shown in SEQ ID NO: 40, LCDR2 in SEQ ID NO: 41, and LCDR3 in SEQ ID NO: 8.

[0774] For example, the antigen-binding molecules that specifically bind to TfR1 disclosed herein, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the IMGT numbering rules.

[0775] For example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0776] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 42, HCDR2 contains the amino acid sequence of SEQ ID NO: 43, and HCDR3 contains the amino acid sequence of SEQ ID NO: 25. The light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 9, LCDR2 contains the amino acid sequence of SEQ ID NO: 10, and LCDR3 contains the amino acid sequence of SEQ ID NO: 8.

[0777] For example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0778] The amino acid sequences of the heavy chain variable region HCDR1 are shown in SEQ ID NO: 42, HCDR2 in SEQ ID NO: 43, and HCDR3 in SEQ ID NO: 25. The amino acid sequences of the light chain variable region LCDR1 are shown in SEQ ID NO: 9, LCDR2 in SEQ ID NO: 10, and LCDR3 in SEQ ID NO: 8.

[0779] For example, the antigen-binding molecules that specifically bind to TfR1 disclosed herein, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the Chothia numbering rules.

[0780] By way of example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein:

[0781] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 27, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 13.

[0782] By way of example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein:

[0783] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13.

[0784] By way of example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises a heavy chain and a light chain, wherein:

[0785] The heavy chain contains the amino acid sequence of SEQ ID NO: 29, and the light chain contains the amino acid sequence of SEQ ID NO: 20.

[0786] By way of example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises a heavy chain and a light chain, wherein:

[0787] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 29, and the amino acid sequence of the light chain is shown in SEQ ID NO: 20; or

[0788] The amino acid sequence of the heavy chain is shown as amino acid residues 1 to 447 of SEQ ID NO: 29, and the amino acid sequence of the light chain is shown as amino acid residues 1 to 213 of SEQ ID NO: 20.

[0789] By way of example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises a heavy chain and a light chain, wherein:

[0790] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 29, and the amino acid sequence of the light chain is shown in SEQ ID NO: 20.

[0791] For example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises:

[0792] Two first strands containing the amino acid sequence of SEQ ID NO: 87, and two second strands containing the amino acid sequence of SEQ ID NO: 20.

[0793] For example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises:

[0794] Two first strands of amino acid sequences as shown in SEQ ID NO: 87, and two second strands of amino acid sequences as shown in SEQ ID NO: 20; or

[0795] The first strand consists of two amino acid sequences as shown in amino acid residues 1 to 467 of SEQ ID NO: 87, and the second strand consists of two amino acid sequences as shown in amino acid residues 1 to 213 of SEQ ID NO: 20.

[0796] For example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein comprises:

[0797] The first strand consists of two amino acid sequences as shown in SEQ ID NO: 87, and the second strand consists of two amino acid sequences as shown in SEQ ID NO: 20.

[0798] For example, the antigen-binding molecule that specifically binds to TfR1 disclosed herein has the first and second chains linked together to form an antigen-binding molecule that specifically binds to TfR1.

[0799] The antigen-binding molecule provided in this disclosure may also contain an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the above-described amino acid sequence, such as a CDR sequence, a variable region sequence, a heavy chain, a light chain, a first chain, or a second chain sequence.

[0800] This disclosure discloses antigen-binding molecules that specifically bind to Aβ.

[0801] This disclosure provides antigen-binding molecules that specifically bind to Aβ, possessing numerous advantageous properties such as good affinity, therapeutic activity, safety, pharmacokinetic properties, and drugability (e.g., solubility, viscosity, purity, and stability).

[0802] Exemplary antigen-binding molecules that specifically bind to Aβ

[0803] Exemplarily, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0804] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50, and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 60, LCDR2 contains the amino acid sequence of SEQ ID NO: 61, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0805] Exemplarily, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0806] The amino acid sequences of the heavy chain variable regions HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO: 62, SEQ ID NO: 63, and SEQ ID NO: 50, respectively. The amino acid sequences of the light chain variable regions LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO: 60, SEQ ID NO: 61, and SEQ ID NO: 53, respectively.

[0807] For example, the antigen-binding molecules that specifically bind to Aβ disclosed herein, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the Kabat numbering rules.

[0808] Exemplarily, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0809] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 70, HCDR2 contains the amino acid sequence of SEQ ID NO: 71, and HCDR3 contains the amino acid sequence of SEQ ID NO: 72, and the light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 73, LCDR2 contains the amino acid sequence of SEQ ID NO: 74, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0810] Exemplarily, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0811] The amino acid sequences of the heavy chain variable regions HCDR1 are shown in SEQ ID NO: 70, HCDR2 in SEQ ID NO: 71, and HCDR3 in SEQ ID NO: 72; the amino acid sequences of the light chain variable regions LCDR1 in SEQ ID NO: 73, LCDR2 in SEQ ID NO: 74, and LCDR3 in SEQ ID NO: 53.

[0812] For example, the antigen-binding molecules that specifically bind to Aβ disclosed herein, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the IMGT numbering rules.

[0813] Exemplarily, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0814] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 75, HCDR2 contains the amino acid sequence of SEQ ID NO: 76, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50. The light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 60, LCDR2 contains the amino acid sequence of SEQ ID NO: 61, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0815] Exemplarily, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0816] The amino acid sequences of the heavy chain variable regions HCDR1 are shown in SEQ ID NO: 75, HCDR2 in SEQ ID NO: 76, and HCDR3 in SEQ ID NO: 50; the amino acid sequences of the light chain variable regions LCDR1 in SEQ ID NO: 60, LCDR2 in SEQ ID NO: 61, and LCDR3 in SEQ ID NO: 53.

[0817] For example, the antigen-binding molecules that specifically bind to Aβ disclosed herein, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the Chothia numbering rules.

[0818] Exemplarily, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0819] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50. The light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 115, LCDR2 contains the amino acid sequence of SEQ ID NO: 52, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0820] Exemplarily, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0821] The amino acid sequences of the heavy chain variable region HCDR1 are shown in SEQ ID NO: 62, HCDR2 in SEQ ID NO: 63, and HCDR3 in SEQ ID NO: 50. The amino acid sequences of the light chain variable region LCDR1 are shown in SEQ ID NO: 115, LCDR2 in SEQ ID NO: 52, and LCDR3 in SEQ ID NO: 53.

[0822] For example, the antigen-binding molecules that specifically bind to Aβ disclosed herein, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the Kabat numbering rules.

[0823] Exemplarily, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0824] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 70, HCDR2 contains the amino acid sequence of SEQ ID NO: 71, and HCDR3 contains the amino acid sequence of SEQ ID NO: 72. The light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 118, LCDR2 contains the amino acid sequence of SEQ ID NO: 74, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0825]

[0826] Exemplarily, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0827] The amino acid sequences of the heavy chain variable region HCDR1 are shown in SEQ ID NO: 70, HCDR2 in SEQ ID NO: 71, and HCDR3 in SEQ ID NO: 72. The amino acid sequences of the light chain variable region LCDR1 are shown in SEQ ID NO: 118, LCDR2 in SEQ ID NO: 74, and LCDR3 in SEQ ID NO: 53.

[0828] For example, the antigen-binding molecules that specifically bind to Aβ disclosed herein, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the IMGT numbering rules.

[0829] Exemplarily, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0830] The heavy chain variable region HCDR1 contains the amino acid sequence of SEQ ID NO: 75, HCDR2 contains the amino acid sequence of SEQ ID NO: 76, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50. The light chain variable region LCDR1 contains the amino acid sequence of SEQ ID NO: 115, LCDR2 contains the amino acid sequence of SEQ ID NO: 52, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53.

[0831] Exemplarily, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3, wherein:

[0832] The amino acid sequences of the heavy chain variable region HCDR1 are shown in SEQ ID NO: 75, HCDR2 in SEQ ID NO: 76, and HCDR3 in SEQ ID NO: 50. The amino acid sequences of the light chain variable region LCDR1 are shown in SEQ ID NO: 115, LCDR2 in SEQ ID NO: 52, and LCDR3 in SEQ ID NO: 53.

[0833] For example, the antigen-binding molecules that specifically bind to Aβ disclosed herein, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the Chothia numbering rules.

[0834] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein:

[0835] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 54, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 56.

[0836] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein:

[0837] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 54, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 57.

[0838] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein:

[0839] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 54, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 58.

[0840] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein:

[0841] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 54, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 59.

[0842] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein:

[0843] The heavy chain variable region contains the amino acid sequence of SEQ ID NO: 54, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 116.

[0844] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein:

[0845] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 56.

[0846] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein:

[0847] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 57.

[0848] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein:

[0849] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 58.

[0850] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein:

[0851] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 59.

[0852] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain variable region and a light chain variable region, wherein:

[0853] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 116.

[0854] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0855] The heavy chain contains the amino acid sequence of SEQ ID NO: 64, and the light chain contains the amino acid sequence of SEQ ID NO: 66.

[0856] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0857] The heavy chain contains the amino acid sequence of SEQ ID NO: 64, and the light chain contains the amino acid sequence of SEQ ID NO: 67.

[0858] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0859] The heavy chain contains the amino acid sequence of SEQ ID NO: 64, and the light chain contains the amino acid sequence of SEQ ID NO: 68.

[0860] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0861] The heavy chain contains the amino acid sequence of SEQ ID NO: 64, and the light chain contains the amino acid sequence of SEQ ID NO: 69.

[0862] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0863] The heavy chain contains the amino acid sequence of SEQ ID NO: 64, and the light chain contains the amino acid sequence of SEQ ID NO: 117.

[0864] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0865] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 66; or

[0866] The amino acid sequence of the heavy chain is shown as amino acid residues 1 to 453 of SEQ ID NO: 64, and the amino acid sequence of the light chain is shown as amino acid residues 1 to 219 of SEQ ID NO: 66.

[0867] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0868] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 67; or

[0869] The amino acid sequence of the heavy chain is shown as amino acid residues 1 to 453 of SEQ ID NO: 64, and the amino acid sequence of the light chain is shown as amino acid residues 1 to 219 of SEQ ID NO: 67.

[0870] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0871] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 68; or

[0872] The amino acid sequence of the heavy chain is shown as amino acid residues 1 to 453 of SEQ ID NO: 64, and the amino acid sequence of the light chain is shown as amino acid residues 1 to 219 of SEQ ID NO: 68.

[0873] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0874] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 69; or

[0875] The amino acid sequence of the heavy chain is shown as amino acid residues 1 to 453 of SEQ ID NO: 64, and the amino acid sequence of the light chain is shown as amino acid residues 1 to 219 of SEQ ID NO: 69.

[0876] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0877] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 69; or

[0878] The amino acid sequence of the heavy chain is shown as amino acid residues 1 to 453 of SEQ ID NO: 64, and the amino acid sequence of the light chain is shown as amino acid residues 1 to 219 of SEQ ID NO: 117.

[0879] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0880] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 66.

[0881] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0882] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 67.

[0883] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0884] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 68.

[0885] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0886] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 69.

[0887] By way of example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises a heavy chain and a light chain, wherein:

[0888] The amino acid sequence of the heavy chain is shown in SEQ ID NO: 64, and the amino acid sequence of the light chain is shown in SEQ ID NO: 117.

[0889] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0890] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 66.

[0891] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0892] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 67.

[0893] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0894] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 68.

[0895] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0896] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 69.

[0897] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0898] Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 117.

[0899] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0900] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 66; or

[0901] The first strand consists of two amino acid sequences, such as those of amino acid residues 1 to 695 in SEQ ID NO: 88, and a second strand consists of four amino acid sequences, such as those of amino acid residues 1 to 219 in SEQ ID NO: 66.

[0902] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0903] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 67; or

[0904] The first strand consists of two amino acid sequences, such as those of amino acid residues 1 to 695 in SEQ ID NO: 88, and a second strand consists of four amino acid sequences, such as those of amino acid residues 1 to 219 in SEQ ID NO: 67.

[0905] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0906] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 68; or

[0907] The first strand consists of two amino acid sequences as shown in amino acid residues 1 to 695 of SEQ ID NO: 88, and the second strand consists of four amino acid sequences as shown in amino acid residues 1 to 219 of SEQ ID NO: 68.

[0908] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0909] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 69; or

[0910] The first strand consists of two amino acid sequences as shown in amino acid residues 1 to 695 of SEQ ID NO: 88, and the second strand consists of four amino acid sequences as shown in amino acid residues 1 to 219 of SEQ ID NO: 69.

[0911] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0912] Two first strands of amino acid sequences as shown in SEQ ID NO: 88, and four second strands of amino acid sequences as shown in SEQ ID NO: 69; or

[0913] The first strand consists of two amino acid sequences, such as those of amino acid residues 1 to 695 in SEQ ID NO: 88, and a second strand consists of four amino acid sequences, such as those of amino acid residues 1 to 219 in SEQ ID NO: 117.

[0914] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0915] The first strand consists of two amino acid sequences as shown in SEQ ID NO: 88, and the second strand consists of four amino acid sequences as shown in SEQ ID NO: 66.

[0916] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0917] The first strand consists of two amino acid sequences as shown in SEQ ID NO: 88, and the second strand consists of four amino acid sequences as shown in SEQ ID NO: 67.

[0918] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0919] The first strand consists of two amino acid sequences as shown in SEQ ID NO: 88, and the second strand consists of four amino acid sequences as shown in SEQ ID NO: 68.

[0920] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0921] The first strand consists of two amino acid sequences as shown in SEQ ID NO: 88, and the second strand consists of four amino acid sequences as shown in SEQ ID NO: 69.

[0922] For example, the antigen-binding molecule that specifically binds to Aβ disclosed herein comprises:

[0923] The first strand consists of two amino acid sequences as shown in SEQ ID NO: 88, and the second strand consists of four amino acid sequences as shown in SEQ ID NO: 117.

[0924] The antigen-binding molecule provided in this disclosure may also contain an amino acid sequence having at least 80% (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with the above-described amino acid sequence, such as a CDR sequence, a variable region sequence, a heavy chain, a light chain, a first chain, or a second chain sequence.

[0925] This disclosure discloses antigen-binding molecules that specifically bind to TfR1 and Aβ.

[0926] This disclosure provides antigen-binding molecules that specifically bind to TfR1 and Aβ, possessing numerous advantageous properties such as good affinity, therapeutic activity, safety, pharmacokinetic properties, and drugability (e.g., solubility, viscosity, purity, and stability).

[0927] Exemplary antigen-binding molecules that specifically bind to TfR1 and Aβ

[0928] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[0929] A first antigen-binding domain that specifically binds to TfR1, wherein the first antigen-binding domain that specifically binds to TfR1 is Fab; and a second antigen-binding domain that specifically binds to Aβ, wherein the second antigen-binding domain that specifically binds to Aβ is Fab; and

[0930] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0931] The amino acid sequences of the heavy chain variable regions HCDR1 are shown in SEQ ID NO: 3, HCDR2 in SEQ ID NO: 11, and HCDR3 in SEQ ID NO: 25; the amino acid sequences of the light chain variable regions LCDR1 in SEQ ID NO: 9, LCDR2 in SEQ ID NO: 10, and LCDR3 in SEQ ID NO: 8; and

[0932] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0933] The amino acid sequences of the heavy chain variable regions HCDR1, HCDR2, and HCDR3 are shown in SEQ ID NO: 62, SEQ ID NO: 63, and SEQ ID NO: 50, respectively. The amino acid sequences of the light chain variable regions LCDR1, LCDR2, and LCDR3 are shown in SEQ ID NO: 60, SEQ ID NO: 61, and SEQ ID NO: 53, respectively.

[0934] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[0935] A first antigen-binding domain that specifically binds to TfR1, wherein the first antigen-binding domain that specifically binds to TfR1 is Fab; and a second antigen-binding domain that specifically binds to Aβ, wherein the second antigen-binding domain that specifically binds to Aβ is Fab; and

[0936] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0937] The amino acid sequences of the heavy chain variable regions HCDR1 are shown in SEQ ID NO: 3, HCDR2 in SEQ ID NO: 11, and HCDR3 in SEQ ID NO: 25; the amino acid sequences of the light chain variable regions LCDR1 in SEQ ID NO: 9, LCDR2 in SEQ ID NO: 10, and LCDR3 in SEQ ID NO: 8; and

[0938] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0939] The amino acid sequences of the heavy chain variable region HCDR1 are shown in SEQ ID NO: 62, HCDR2 in SEQ ID NO: 63, and HCDR3 in SEQ ID NO: 50. The amino acid sequences of the light chain variable region LCDR1 are shown in SEQ ID NO: 115, LCDR2 in SEQ ID NO: 52, and LCDR3 in SEQ ID NO: 53.

[0940] For example, the antigen-binding molecules that specifically bind TfR1 and Aβ disclosed herein, wherein the heavy chain variable regions HCDR1, HCDR2, and HCDR3 and the light chain variable regions LCDR1, LCDR2, and LCDR3 are defined according to the Kabat numbering rules.

[0941] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[0942] A first antigen-binding domain that specifically binds to TfR1, wherein the first antigen-binding domain that specifically binds to TfR1 is Fab; and a second antigen-binding domain that specifically binds to Aβ, wherein the second antigen-binding domain that specifically binds to Aβ is Fab; and

[0943] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0944] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13; and

[0945] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0946] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 56.

[0947] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[0948] A first antigen-binding domain that specifically binds to TfR1, wherein the first antigen-binding domain that specifically binds to TfR1 is Fab; and a second antigen-binding domain that specifically binds to Aβ, wherein the second antigen-binding domain that specifically binds to Aβ is Fab; and

[0949] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0950] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13; and

[0951] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0952] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 57.

[0953] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[0954] A first antigen-binding domain that specifically binds to TfR1, wherein the first antigen-binding domain that specifically binds to TfR1 is Fab; and a second antigen-binding domain that specifically binds to Aβ, wherein the second antigen-binding domain that specifically binds to Aβ is Fab; and

[0955] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0956] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13; and

[0957] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0958] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 58.

[0959] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[0960] A first antigen-binding domain that specifically binds to TfR1, wherein the first antigen-binding domain that specifically binds to TfR1 is Fab; and a second antigen-binding domain that specifically binds to Aβ, wherein the second antigen-binding domain that specifically binds to Aβ is Fab; and

[0961] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0962] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13; and

[0963] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0964] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 59.

[0965] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[0966] A first antigen-binding domain that specifically binds to TfR1, wherein the first antigen-binding domain that specifically binds to TfR1 is Fab; and a second antigen-binding domain that specifically binds to Aβ, wherein the second antigen-binding domain that specifically binds to Aβ is Fab; and

[0967] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0968] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13; and

[0969] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0970] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 54, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 116.

[0971] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein is composed of a first chain, a second chain, a third chain, and a fourth chain, wherein:

[0972] The amino acid sequence of the first chain is shown in SEQ ID NO: 87, the amino acid sequence of the second chain is shown in SEQ ID NO: 20, the amino acid sequence of the third chain is shown in SEQ ID NO: 88, and the amino acid sequence of the fourth chain is shown in SEQ ID NO: 66.

[0973] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein is composed of a first chain, a second chain, a third chain, and a fourth chain, wherein:

[0974] The amino acid sequence of the first chain is shown as amino acid residues 1 to 467 of SEQ ID NO: 87, the amino acid sequence of the second chain is shown as amino acid residues 1 to 213 of SEQ ID NO: 20, the amino acid sequence of the third chain is shown as amino acid residues 1 to 695 of SEQ ID NO: 88, and the amino acid sequence of the fourth chain is shown as amino acid residues 1 to 219 of SEQ ID NO: 66.

[0975] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein is composed of a first chain, a second chain, a third chain, and a fourth chain, wherein:

[0976] The amino acid sequence of the first chain is shown in SEQ ID NO: 87, the amino acid sequence of the second chain is shown in SEQ ID NO: 20, the amino acid sequence of the third chain is shown in SEQ ID NO: 88, and the amino acid sequence of the fourth chain is shown in SEQ ID NO: 67.

[0977] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein is composed of a first chain, a second chain, a third chain, and a fourth chain, wherein:

[0978] The amino acid sequence of the first chain is shown as amino acid residues 1 to 467 of SEQ ID NO: 87, the amino acid sequence of the second chain is shown as amino acid residues 1 to 213 of SEQ ID NO: 20, the amino acid sequence of the third chain is shown as amino acid residues 1 to 695 of SEQ ID NO: 88, and the amino acid sequence of the fourth chain is shown as amino acid residues 1 to 219 of SEQ ID NO: 67.

[0979] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein is composed of a first chain, a second chain, a third chain, and a fourth chain, wherein:

[0980] The amino acid sequence of the first chain is shown in SEQ ID NO: 87, the amino acid sequence of the second chain is shown in SEQ ID NO: 20, the amino acid sequence of the third chain is shown in SEQ ID NO: 88, and the amino acid sequence of the fourth chain is shown in SEQ ID NO: 68.

[0981] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein is composed of a first chain, a second chain, a third chain, and a fourth chain, wherein:

[0982] The amino acid sequence of the first chain is shown as amino acid residues 1 to 467 of SEQ ID NO: 87, the amino acid sequence of the second chain is shown as amino acid residues 1 to 213 of SEQ ID NO: 20, the amino acid sequence of the third chain is shown as amino acid residues 1 to 695 of SEQ ID NO: 88, and the amino acid sequence of the fourth chain is shown as amino acid residues 1 to 219 of SEQ ID NO: 68.

[0983] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein is composed of a first chain, a second chain, a third chain, and a fourth chain, wherein:

[0984] The amino acid sequence of the first chain is shown in SEQ ID NO: 87, the amino acid sequence of the second chain is shown in SEQ ID NO: 20, the amino acid sequence of the third chain is shown in SEQ ID NO: 88, and the amino acid sequence of the fourth chain is shown in SEQ ID NO: 69.

[0985] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein is composed of a first chain, a second chain, a third chain, and a fourth chain, wherein:

[0986] The amino acid sequence of the first chain is shown as amino acid residues 1 to 467 of SEQ ID NO: 87, the amino acid sequence of the second chain is shown as amino acid residues 1 to 213 of SEQ ID NO: 20, the amino acid sequence of the third chain is shown as amino acid residues 1 to 695 of SEQ ID NO: 88, and the amino acid sequence of the fourth chain is shown as amino acid residues 1 to 219 of SEQ ID NO: 69.

[0987] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein is composed of a first chain, a second chain, a third chain, and a fourth chain, wherein:

[0988] The amino acid sequence of the first chain is shown in SEQ ID NO: 87, the amino acid sequence of the second chain is shown in SEQ ID NO: 20, the amino acid sequence of the third chain is shown in SEQ ID NO: 88, and the amino acid sequence of the fourth chain is shown in SEQ ID NO: 117.

[0989] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein is composed of a first chain, a second chain, a third chain, and a fourth chain, wherein:

[0990] The amino acid sequence of the first chain is shown as amino acid residues 1 to 467 of SEQ ID NO: 87, the amino acid sequence of the second chain is shown as amino acid residues 1 to 213 of SEQ ID NO: 20, the amino acid sequence of the third chain is shown as amino acid residues 1 to 695 of SEQ ID NO: 88, and the amino acid sequence of the fourth chain is shown as amino acid residues 1 to 219 of SEQ ID NO: 117.

[0991] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[0992] A first antigen-binding domain that specifically binds to TfR1, wherein the first antigen-binding domain that specifically binds to TfR1 is Fab; and a second antigen-binding domain that specifically binds to Aβ, wherein the second antigen-binding domain that specifically binds to Aβ is Fab; and

[0993] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[0994] The amino acid sequences of the heavy chain variable regions HCDR1 are shown in SEQ ID NO: 3, HCDR2 in SEQ ID NO: 11, and HCDR3 in SEQ ID NO: 25, 5, or 24; the amino acid sequences of the light chain variable regions LCDR1 are shown in SEQ ID NO: 9, LCDR2 in SEQ ID NO: 10, and LCDR3 in SEQ ID NO: 8; and

[0995] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[0996] The amino acid sequences of the heavy chain variable regions HCDR1 are shown in SEQ ID NO: 48, HCDR2 in SEQ ID NO: 49, and HCDR3 in SEQ ID NO: 50; the amino acid sequences of the light chain variable regions LCDR1 in SEQ ID NO: 51, LCDR2 in SEQ ID NO: 52, and LCDR3 in SEQ ID NO: 53.

[0997] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[0998] A first antigen-binding domain that specifically binds to TfR1, wherein the first antigen-binding domain that specifically binds to TfR1 is Fab; and a second antigen-binding domain that specifically binds to Aβ, wherein the second antigen-binding domain that specifically binds to Aβ is Fab; and

[0999] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[1000] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 27, 12 or 26, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13, 14, 15 or 16; and

[1001] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[1002] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 46, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 47.

[1003] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[1004] A first antigen-binding domain that specifically binds to TfR1, wherein the first antigen-binding domain that specifically binds to TfR1 is Fab; and a second antigen-binding domain that specifically binds to Aβ, wherein the second antigen-binding domain that specifically binds to Aβ is Fab; and

[1005] The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region and a light chain variable region, wherein:

[1006] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13, 14, 15 or 16.

[1007] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 12, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13, 14, 15 or 16; or

[1008] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 26, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 13, 14, 15 or 16; and

[1009] The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region and a light chain variable region, wherein:

[1010] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 46, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 47.

[1011] Antigen-binding molecular structure

[1012] The bispecific antibodies disclosed herein are not limited to a specific molecular structure, as long as they possess the desired antigen-binding function. For example, the bispecific antibodies disclosed herein can be trivalent (1+2). The antigen-binding domain in the bispecific antibody can be any antibody or antibody fragment with antigen-binding activity, either directly or through linker fusion.

[1013] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein includes a first antigen-binding domain that specifically binds to TfR1 and a second antigen-binding domain that specifically binds to Aβ.

[1014] For example, the antigen-binding molecule that specifically binds TfR1 and Aβ disclosed herein comprises a first antigen-binding domain that specifically binds TfR1 and at least one (e.g., 1, 2, 3, 4, 5 or 6) second antigen-binding domain that specifically binds Aβ.

[1015] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises one first antigen-binding domain that specifically binds to TfR1 and two second antigen-binding domains that specifically bind to Aβ.

[1016] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein includes a first antigen-binding domain that specifically binds to TfR1, wherein the first antigen-binding domain that specifically binds to TfR1 is Fab; and a second antigen-binding domain that specifically binds to Aβ, wherein the second antigen-binding domain that specifically binds to Aβ is Fab.

[1017] For example, the antigen-binding molecule that specifically binds TfR1 and Aβ disclosed herein comprises a first antigen-binding domain that specifically binds TfR1, wherein the first antigen-binding domain that specifically binds TfR1 is Fab; and at least one (e.g., 1, 2, 3, 4, 5 or 6) second antigen-binding domain that specifically binds Aβ, wherein the second antigen-binding domain that specifically binds Aβ is Fab.

[1018] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises one first antigen-binding domain that specifically binds to TfR1, wherein the first antigen-binding domain that specifically binds to TfR1 is Fab; and two second antigen-binding domains that specifically bind to Aβ, wherein the second antigen-binding domains that specifically bind to Aβ are Fab.

[1019] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein includes an Fc region comprising a first subunit Fc1 and a second subunit Fc2; and includes a first half-antibody and a second half-antibody.

[1020] Exemplarily, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises an Fc region, the Fc region comprising a first subunit Fc1 and a second subunit Fc2; and comprises a first half-antibody and a second half-antibody, wherein:

[1021] The first half-antibody includes Fab and Fc1 regions that bind TfR1; and the second half-antibody includes Fab and Fc2 regions that bind Aβ; and the first half-antibody and the second half-antibody are associated with each other through the Fc1 and Fc2 regions.

[1022] Exemplarily, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises an Fc region, the Fc region comprising a first subunit Fc1 and a second subunit Fc2; and comprises a first half-antibody and a second half-antibody, wherein:

[1023] The first half-antibody includes one TfR1-binding Fab and one Fc1 region from the N-terminus to the C-terminus; and the second half-antibody includes at least one (e.g., 1, 2, 3, 4, 5 or 6) Aβ-binding Fab and one Fc2 region from the N-terminus to the C-terminus; and the first half-antibody and the second half-antibody are associated with each other through the Fc1 region and the Fc2 region.

[1024] Exemplarily, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises an Fc region, the Fc region comprising a first subunit Fc1 and a second subunit Fc2; and comprises a first half-antibody and a second half-antibody, wherein:

[1025] The first half-antibody contains one TfR1 binding Fab and one Fc1 region from the N-terminus to the C-terminus; and the second half-antibody contains two identical Aβ binding Fab and one Fc2 region from the N-terminus to the C-terminus; and the first half-antibody and the second half-antibody associate with each other through the Fc1 region and the Fc2 region.

[1026] Exemplarily, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises an Fc region, the Fc region comprising a first subunit Fc1 and a second subunit Fc2; and comprises a first half-antibody and a second half-antibody, wherein:

[1027] The first hapten comprises, from N-terminus to C-terminus, one TfR1-binding Fab and one Fc1 region, wherein the C-terminus of the TfR1-binding Fab is operatively connected to the N-terminus of the Fc1 region; and the second hapten comprises, from N-terminus to C-terminus, two identical Aβ-binding Fabs and one Fc2 region, wherein the C-terminus of the first Aβ-binding Fab is operatively connected to the N-terminus of the second Aβ-binding Fab, and the C-terminus of the second Aβ-binding Fab is operatively connected to the N-terminus of the Fc2 region; and the first hapten and the second hapten associate with each other through the Fc1 and Fc2 regions.

[1028] Exemplarily, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises an Fc region, the Fc region comprising a first subunit Fc1 and a second subunit Fc2; and comprises a first half-antibody and a second half-antibody, wherein:

[1029] The first hapten comprises, from N-terminus to C-terminus, one TfR1-binding Fab and one Fc1 region, wherein the C-terminus of the CH1 region of the TfR1-binding Fab is operatively connected to the N-terminus of the Fc1 region; and the second hapten comprises, from N-terminus to C-terminus, two identical Aβ-binding Fabs and one Fc2 region, wherein the C-terminus of the CH1 region of the first Aβ-binding Fab is operatively connected to the N-terminus of the heavy chain variable region of the second Aβ-binding Fab, and the C-terminus of the CH1 region of the second Aβ-binding Fab is operatively connected to the N-terminus of the Fc2 region; and the first hapten and the second hapten associate with each other through the Fc1 and Fc2 regions.

[1030] Exemplarily, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises an Fc region, the Fc region comprising a first subunit Fc1 and a second subunit Fc2; and comprises a first half-antibody and a second half-antibody, wherein:

[1031] The first hapten comprises one TfR1-binding Fab and one Fc1 region from the N-terminus to the C-terminus, wherein the C-terminus of the TfR1-binding Fab is fused directly or via a linker to the N-terminus of the Fc1 region; and the second hapten comprises two identical Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus, wherein the C-terminus of the first Aβ-binding Fab is fused directly or via a linker to the N-terminus of the second Aβ-binding Fab, and the C-terminus of the second Aβ-binding Fab is fused directly or via a linker to the N-terminus of the Fc2 region; and the first and second haptens associate with each other through the Fc1 and Fc2 regions.

[1032] Exemplarily, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises an Fc region, the Fc region comprising a first subunit Fc1 and a second subunit Fc2; and comprises a first half-antibody and a second half-antibody, wherein:

[1033] The first hapten includes one TfR1-binding Fab and one Fc1 region from the N-terminus to the C-terminus, with the C-terminus of the TfR1-binding Fab fused to the N-terminus of the Fc1 region via a linker; and the second hapten includes two identical Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus, with the C-terminus of the first Aβ-binding Fab fused to the N-terminus of the second Aβ-binding Fab via a linker, and the C-terminus of the second Aβ-binding Fab directly fused to the N-terminus of the Fc2 region; and the first and second haptens associate with each other through the Fc1 and Fc2 regions.

[1034] Exemplarily, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises an Fc region, the Fc region comprising a first subunit Fc1 and a second subunit Fc2; and comprises a first half-antibody and a second half-antibody, wherein:

[1035] The first hapten comprises one TfR1-binding Fab and one Fc1 region from the N-terminus to the C-terminus. The C-terminus of the CH1 region of the TfR1-binding Fab is fused directly or via a linker to the N-terminus of the Fc1 region. The second hapten comprises two identical Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus. The C-terminus of the CH1 region of the first Aβ-binding Fab is fused directly or via a linker to the N-terminus of the heavy chain variable region of the second Aβ-binding Fab. The C-terminus of the CH1 region of the second Aβ-binding Fab is fused directly or via a linker to the N-terminus of the Fc2 region. The first and second haptens associate with each other through the Fc1 and Fc2 regions.

[1036] Exemplarily, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises an Fc region, the Fc region comprising a first subunit Fc1 and a second subunit Fc2; and comprises a first half-antibody and a second half-antibody, wherein:

[1037] The first hapten includes one TfR1-binding Fab and one Fc1 region from the N-terminus to the C-terminus. The C-terminus of the CH1 region of the TfR1-binding Fab is fused to the N-terminus of the Fc1 region via a linker. The second hapten includes two identical Aβ-binding Fabs and one Fc2 region from the N-terminus to the C-terminus. The C-terminus of the CH1 region of the first Aβ-binding Fab is fused to the N-terminus of the heavy chain variable region of the second Aβ-binding Fab via a linker. The C-terminus of the CH1 region of the second Aβ-binding Fab is directly fused to the N-terminus of the Fc2 region. The first and second haptens are associated with each other through the Fc1 and Fc2 regions.

[1038] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[1039] The first chain of the structure shown in equation (a), the second chain of the structure shown in equation (b), the third chain of the structure shown in equation (c), and the fourth chain of the structure shown in equation (d) are arranged from the N end to the C end.

[1040] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[1041] (b)[TfR1-VL]-[CL];

[1042] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[1043] (d)[Aβ-VL]-[CL].

[1044] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[1045] One first chain as shown in equation (a), one second chain as shown in equation (b), one third chain as shown in equation (c), and two fourth chains as shown in equation (d), wherein the structures shown in equations (a), (b), (c), and (d) are arranged from the N end to the C end:

[1046] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[1047] (b)[TfR1-VL]-[CL];

[1048] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[1049] (d)[Aβ-VL]-[CL].

[1050] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[1051] The first chain of the structure shown in equation (a), the second chain of the structure shown in equation (b), the third chain of the structure shown in equation (c), and the fourth chain of the structure shown in equation (d) are arranged from the N end to the C end.

[1052] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[1053] (b)[TfR1-VL]-[CL];

[1054] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[1055] (d)[Aβ-VL]-[CL];

[1056] The linker 1 and linker 2 may be the same or different peptide linkers, or the linker 1 and / or linker 2 may not exist;

[1057] Fc1 and Fc2 are interchangeable;

[1058] The first and second chains combine with each other, the first and third chains combine with each other, and the third and fourth chains combine with each other to form antigen-binding molecules that specifically bind to TfR1 and Aβ.

[1059] For example, the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein comprises:

[1060] One first chain as shown in equation (a), one second chain as shown in equation (b), one third chain as shown in equation (c), and two fourth chains as shown in equation (d), wherein the structures shown in equations (a), (b), (c), and (d) are arranged from the N end to the C end:

[1061] (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1];

[1062] (b)[TfR1-VL]-[CL];

[1063] (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2];

[1064] (d)[Aβ-VL]-[CL];

[1065] The linker 1 and linker 2 may be the same or different peptide linkers, or the linker 1 and / or linker 2 may not exist;

[1066] Fc1 and Fc2 are interchangeable;

[1067] The first and second chains combine with each other, the first and third chains combine with each other, and the third and fourth chains combine with each other to form antigen-binding molecules that specifically bind to TfR1 and Aβ.

[1068] For example, the structure of the antigen-binding molecule that specifically binds to TfR1 and Aβ disclosed herein is shown in Format 3 of Figure 2.

[1069] antibody structure

[1070] In some implementations, the antigen-binding molecule provided herein is a full-length antibody.

[1071] In some implementations, the antigen-binding molecule provided herein is an antigen-binding fragment.

[1072] In some embodiments, the antigen-binding molecule provided herein is an antibody fragment. In some embodiments, the antigen-binding fragment is a Fab, Fab′, Fab′-SH, or F(ab′)2 fragment, particularly a Fab fragment. “Fab” is a monovalent fragment consisting of VL, VH, CL, and CH1 domains. A “Fab fragment” can be generated by cleavage of an antigen-binding molecule via papain. “Fab′” contains VL, CL, VH, and CH1, and also contains a region between the CH1 and CH2 domains such that interchain disulfide bonds can form between the two heavy chains of two Fab′ fragments to form an F(ab′)2 molecule. “Fab′-SH” is a Fab′ fragment in which the cysteine ​​residues in the constant region have free thiol groups. “F(ab′)2” is a divalent fragment comprising two Fab fragments linked by disulfide bonds in the hinge region.

[1073] In some implementations, the antigen-binding fragment is a biantibody, triantibody, or tetraantibody. A biantibody is an antibody fragment with two antigen-binding sites, containing linked VH and VL domains within the same polypeptide chain (VH-VL). By using a short linker that prevents pairing between two domains on the same chain, these domains are forced to pair with complementary domains on another chain, thereby creating two antigen-binding sites. The two antigens can be the same or different.

[1074] In some embodiments, the antigen-binding fragment is a single-chain Fab fragment. A “single-chain Fab fragment” or “scFab” is a polypeptide consisting of VH, CH1, VL, CL, and a linker, wherein the antigen-binding domain and the linker have one of the following sequences in the N-terminal to C-terminal direction: a) VH-CH1-linker-VL-CL, b) VL-CL-linker-VH-CH1, c) VH-CL-linker-VL-CH1, or d) VL-CH1-linker-VH-CL. In some embodiments, the linker is a polypeptide having at least 30 amino acids. In some embodiments, the linker is a polypeptide having between 32 and 50 amino acids. The single-chain Fab fragment is stabilized via a native disulfide bond between CL and CH1. Additionally, these single-chain Fab molecules can be further stabilized by inserting cysteine ​​residues (e.g., at position 44 in the heavy chain variable region and position 100 in the light chain variable region, according to Kabat numbering) to create interchain disulfide bonds.

[1075] In some implementations, the antigen-binding fragment is an Fv fragment consisting of the VH and VL domains of a single arm of the antibody.

[1076] In some embodiments, the antigen-binding fragment is a single-chain variable fragment (scFv). An “scFv” is a fusion protein comprising at least one antigen-binding fragment containing a light chain variable region and at least one antigen-binding fragment containing a heavy chain variable region, wherein the light and heavy chain variable regions are sequentially linked by a short, flexible peptide linker, capable of being expressed as a single-chain polypeptide, and wherein the scFv retains the specificity of the intact antigen-binding molecule from which it originates. Unless otherwise specified, the scFv may have VL and VH variable regions in any order herein; for example, relative to the N-terminus and C-terminus of the polypeptide, the scFv may comprise a VL-linker-VH or may comprise a VH-linker-VL.

[1077] In some embodiments, the antigen-binding fragment is dsFv, which is obtained by linking polypeptides in which one amino acid residue in each of the VH and VL is replaced by a cysteine ​​residue via disulfide bonds between the cysteine ​​residues. The amino acid residues to be replaced by cysteine ​​residues can be selected based on predictions of the three-dimensional structure of the antigen-binding molecule using known methods.

[1078] In some implementations, the antigen-binding fragment is a single-domain antibody (dAb). A single-domain antibody is an antibody fragment containing all or part of the heavy chain variable domain or all or part of the light chain variable domain.

[1079] In some embodiments, the antigen-binding molecules provided herein are chimeric antibodies. In some embodiments, the chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate, such as a monkey) and a human constant region. In some embodiments, the chimeric antibody is a "class-switched" antibody, wherein the class or subclass has been changed from the class or subclass of the parent antibody.

[1080] In some embodiments, the antigen-binding molecule is a humanized antibody. Typically, a non-human antibody is humanized to reduce its immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. Generally, a humanized antibody contains one or more variable regions, wherein the CDR or a portion thereof is derived from the non-human antibody, and the FR or a portion thereof is derived from the human antibody. Optionally, the humanized antibody may also contain a portion of a human constant region. In some embodiments, some FR residues in the humanized antibody may be replaced with corresponding residues from a non-human antibody (e.g., an antibody providing the CDR sequence).

[1081] Variants of antigen-binding molecules

[1082] In some embodiments, amino acid sequence variants of the antigen-binding molecules provided in this disclosure are included. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antigen-binding molecule. Amino acid sequence variants of the antigen-binding molecule can be prepared by introducing suitable modifications into the nucleotide sequence encoding the antigen-binding molecule, or by peptide synthesis. Such modifications include, for example, deletion, and / or insertion, and / or substitution of residues within the amino acid sequence of the antigen-binding molecule. Any combination of deletions, insertions, and substitutions can be performed to obtain the final construct, provided that the final construct possesses the desired characteristics, such as antigen-binding properties.

[1083] Replace, insert, and delete variants

[1084] In some embodiments, antigen-binding molecule variants with one or more amino acid substitutions are provided. Substitution mutagenesis sites of interest include CDR and FR. Conserved substitutions are shown in Table B under the heading “Preferred Substitutions.” More substantial variations are provided in Table B under the heading “Exemplary Substitutions” and are further described below with reference to the amino acid side chain categories. Amino acid substitutions can be introduced into the antigen-binding molecule of interest, and the product can be screened for desired activities, such as retained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC.

[1085] Table B. Substitution of Amino Acids

[1086] Based on common side-chain characteristics, amino acids can be grouped as follows:

[1087] (1) Hydrophobic: Leucine, Met, Ala, Val, Leu, Ile;

[1088] (2) Neutral and hydrophilic: Cys, Ser, Thr, Asn, Gln;

[1089] (3) Acidic: Asp, Glu;

[1090] (4) Alkaline: His, Lys, Arg;

[1091] (5) Residues that affect chain orientation: Gly, Pro;

[1092] (6) Aromatic: Trp, Tyr, Phe.

[1093] Non-conservative replacement would require replacing a member of one of these categories with a member of another category.

[1094] One class of substitution variants involves replacing one or more CDR residues in a parent antigen-binding molecule (e.g., humanized or human antibody). Generally, the resulting variants selected for further research will have alterations (e.g., improvements) to certain biological properties (e.g., increased affinity, decreased affinity, decreased immunogenicity) relative to the parent antigen-binding molecule, and / or will substantially retain some of the biological properties of the parent antigen-binding molecule. An exemplary substitution variant is an affinity-matured antigen-binding molecule, which can be conveniently generated, for example, using phage display-based affinity maturation techniques (such as those described herein). In short, one or more CDR residues are mutated, the variant antigen-binding molecule is displayed on a phage, and it is screened for specific biological activities (e.g., binding affinity). Modifications (e.g., substitutions) can be made to the CDRs, for example, to improve the affinity of the antigen-binding molecule. Such alterations can be made to CDR "hotspots"—residues encoded by codons that undergo high-frequency mutations during somatic maturation—and / or residues that come into contact with antigens, while simultaneously testing the binding affinity of the resulting variants VH or VL. In some embodiments of affinity maturation, diversity is introduced into the selected variant gene for maturation using any of a variety of methods, such as error-prone PCR, strand shuffling, or oligonucleotide-guided mutagenesis. A secondary library is then created. The library is then screened to identify any antigen-binding molecule variants with the desired affinity. Another method for introducing diversity involves CDR-directed approaches, where several CDR residues (e.g., 4-6 residues at a time) are randomized. CDR residues involved in antigen binding can be specifically identified, for example, using alanine scan mutagenesis or modeling. In particular, HCDR3 and LCDR3 are frequently targeted.

[1095] In some embodiments, substitution, insertion, or deletion can occur within one or more CDRs, provided that such changes do not materially reduce the ability of the antigen-binding molecule to bind to the antigen. For example, conserved changes (e.g., conserved substitutions, as provided herein) can be made to the CDRs that do not materially reduce binding affinity. Such changes can, for example, be external to the antigen-contacting residues in the CDR. In some embodiments of the variant VH and VL sequences provided above, each CDR is unchanged or contains no more than one, two, or three amino acid substitutions.

[1096] One method for identifying residues or regions in an antigen-binding molecule that can serve as mutagenic targets is called "alanine scanning mutagenesis." In this method, a residue or target group of residues (e.g., charged residues such as Arg, Asp, His, Lys, and Glu) is identified and replaced with a neutral or negatively charged amino acid (e.g., Ala or polyalanine) to determine whether the interaction between the antigen-binding molecule and the antigen is affected. Further substitutions can be introduced at amino acid positions that show functional sensitivity to the initial substitution. Furthermore, the contact points between the antigen-binding molecule and the antigen can be identified by studying the crystal structure of the antigen-antigen-binding molecule complex. These contact residues and adjacent residues can be targeted or eliminated as substitution candidates. Variants can be screened to determine if they contain the desired properties.

[1097] Amino acid sequence insertions include fusion of the amino and / or carboxyl ends of peptides ranging in length from 1 residue to 100 or more residues, and intra-sequence insertions of single or multiple amino acid residues. Examples of terminal insertions include antigen-binding molecules having an N-terminal methionyl residue. Other insertion variants of antigen-binding molecules include fusions of the N- or C-terminus of an antigen-binding molecule with an enzyme or a peptide that extends the serum half-life of the antigen-binding molecule.

[1098] Renovation of Fc District

[1099] The Fc region of the antigen-binding molecule disclosed herein contains one or more amino acid substitutions.

[1100] The natural IgG Fc region, specifically the IgG1 Fc region or the IgG4 Fc region, may cause the antigen-binding molecules disclosed herein to target cells expressing Fc receptors, rather than cells expressing antigens. In some embodiments, the Fc receptor is an Fcγ receptor. In some embodiments, the Fc receptor is a human Fcγ receptor, such as FcγRI, FcγRIIa, FcγRIIB, or FcγRIIIa.

[1101] When an antigen-binding molecule contains different binding modules fused to two subunits of the Fc region, undesirable homodimerization may occur. To improve yield and purity, it is advantageous to introduce modifications that promote heterodimerization into the Fc region of the antigen-binding molecule disclosed herein. In some embodiments, the Fc region of this disclosure comprises a first subunit and a second subunit, each containing a charged amino acid mutation (e.g., a positively charged amino acid or a negatively charged amino acid) in its CH3 domain, with the amino acid mutations in the CH3 domains of the first and second subunits carrying opposite charges; furthermore, each of the first and second subunits also contains an identical amino acid mutation located at the CH3 interaction interface, which can weaken the interaction between the CH3 domains of the first and / or second polypeptide homomers, making the first and / or second polypeptide homomers more readily dissociate into first and second polypeptide monomers under reducing conditions, thereby promoting the generation of a heteropolymer containing one first polypeptide and one second polypeptide. In some embodiments, the first subunit comprises amino acid substitutions of 351I and 356K, and the second subunit comprises amino acid substitutions of 351I and 439E, numbered according to the EU index. In some embodiments, the first subunit comprises amino acid substitutions of L351I and D356K, and the second subunit comprises amino acid substitutions of L351I and K439E, numbered according to the EU index. In some embodiments, the first subunit comprises the amino acid sequence of SEQ ID NO: 80; and the second subunit comprises the amino acid sequence of SEQ ID NO: 81.

[1102] In addition to the techniques mentioned above, other techniques for modifying the CH3 domain of heavy chains to achieve heterodimerization are also known in the art, such as WO1996027011A1, WO1998050431A2, EP1870459A1, WO2007110205A2, WO2009089004A1, WO2010129304A2, WO2011143545A1, WO2012058768A1, WO2013157954A1 and WO2013096291A2.

[1103] The C-terminus of the Fc region can be a complete C-terminus ending with the amino acid residue PGK; or it can be a truncated C-terminus, for example, in which one or two C-terminal amino acid residues have been removed. In some embodiments, the C-terminus of the heavy chain is a shortened C-terminus ending with PG. Therefore, in some embodiments, the composition of a complete antibody may include an antibody population with all K447 residues and / or G446+K447 residues removed. In some embodiments, the composition of a ...

Claims

1. An antigen-binding molecule that specifically binds to TfR1 and Aβ, comprising at least one first antigen-binding domain that specifically binds to TfR1 and at least one second antigen-binding domain that specifically binds to Aβ; The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein: The TfR1-VH comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 3, HCDR2 containing the amino acid sequence of SEQ ID NO: 11, and HCDR3 containing the amino acid sequence of SEQ ID NO: 25, 5, or 24; and the TfR1-VL comprises LCDR1 containing the amino acid sequence of SEQ ID NO: 9, LCDR2 containing the amino acid sequence of SEQ ID NO: 10, and LCDR3 containing the amino acid sequence of SEQ ID NO: 8; and The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein: The Aβ-VH has HCDR1 containing the amino acid sequence of SEQ ID NO: 62, HCDR2 containing the amino acid sequence of SEQ ID NO: 63, and HCDR3 containing the amino acid sequence of SEQ ID NO: 50; and the Aβ-VL has LCDR1 containing the amino acid sequence of SEQ ID NO: 60 or 115, LCDR2 containing the amino acid sequence of SEQ ID NO: 61 or 52, and LCDR3 containing the amino acid sequence of SEQ ID NO:

53. Preferably, The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein: The TfR1-VH comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 3, HCDR2 containing the amino acid sequence of SEQ ID NO: 11, and HCDR3 containing the amino acid sequence of SEQ ID NO: 25; and the TfR1-VL comprises LCDR1 containing the amino acid sequence of SEQ ID NO: 9, LCDR2 containing the amino acid sequence of SEQ ID NO: 10, and LCDR3 containing the amino acid sequence of SEQ ID NO: 8; and The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein: The Aβ-VH's HCDR1 contains the amino acid sequence of SEQ ID NO: 62, HCDR2 contains the amino acid sequence of SEQ ID NO: 63, and HCDR3 contains the amino acid sequence of SEQ ID NO: 50; and the Aβ-VL's LCDR1 contains the amino acid sequence of SEQ ID NO: 60, LCDR2 contains the amino acid sequence of SEQ ID NO: 61, and LCDR3 contains the amino acid sequence of SEQ ID NO: 53; or The Aβ-VH has HCDR1 containing the amino acid sequence of SEQ ID NO: 62, HCDR2 containing the amino acid sequence of SEQ ID NO: 63, and HCDR3 containing the amino acid sequence of SEQ ID NO: 50; and the Aβ-VL has LCDR1 containing the amino acid sequence of SEQ ID NO: 115, LCDR2 containing the amino acid sequence of SEQ ID NO: 52, and LCDR3 containing the amino acid sequence of SEQ ID NO:

53. More preferably, The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein: The TfR1-VH comprises HCDR1 containing the amino acid sequence of SEQ ID NO: 3, HCDR2 containing the amino acid sequence of SEQ ID NO: 11, and HCDR3 containing the amino acid sequence of SEQ ID NO: 25; and the TfR1-VL comprises LCDR1 containing the amino acid sequence of SEQ ID NO: 9, LCDR2 containing the amino acid sequence of SEQ ID NO: 10, and LCDR3 containing the amino acid sequence of SEQ ID NO: 8; and The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein: The Aβ-VH has HCDR1 containing the amino acid sequence of SEQ ID NO: 62, HCDR2 containing the amino acid sequence of SEQ ID NO: 63, and HCDR3 containing the amino acid sequence of SEQ ID NO: 50; and the Aβ-VL has LCDR1 containing the amino acid sequence of SEQ ID NO: 60, LCDR2 containing the amino acid sequence of SEQ ID NO: 61, and LCDR3 containing the amino acid sequence of SEQ ID NO:

53.

2. The antigen-binding molecule that specifically binds to TfR1 and Aβ according to claim 1, wherein: The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein: The TfR1-VH comprises an amino acid sequence having at least 80% sequence identity with SEQ ID NO: 27, 12, or 26, and the TfR1-VL comprises an amino acid sequence having at least 80% sequence identity with SEQ ID NO: 13, 14, 15, or 16; and The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein: The Aβ-VH comprises an amino acid sequence having at least 80% sequence identity with SEQ ID NO: 54 or 55, and the Aβ-VL comprises an amino acid sequence having at least 80% sequence identity with SEQ ID NO: 56, 57, 58, 59 or 116; Preferably, The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein: The TfR1-VH comprises the amino acid sequence of SEQ ID NO: 27, 12, or 26, and the TfR1-VL comprises the amino acid sequence of SEQ ID NO: 13, 14, 15, or 16; and The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein: The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54 or 55, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 56, 57, 58, 59 or 116; More preferably, The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein: The TfR1-VH contains the amino acid sequence of SEQ ID NO: 27, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13, 14, 15, or 16; or The TfR1-VH contains the amino acid sequence of SEQ ID NO: 12, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13, 14, 15, or 16; or The TfR1-VH contains the amino acid sequence of SEQ ID NO: 26, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13, 14, 15, or 16; and The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein: The Aβ-VH comprises the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL comprises the amino acid sequence of SEQ ID NO: 56, 57, 58, 59, or 116; or The Aβ-VH contains the amino acid sequence of SEQ ID NO: 55, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 56, 57, 58, 59 or 116; More preferably, The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein: The TfR1-VH contains the amino acid sequence of SEQ ID NO: 27, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13; and The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein: The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO: 56, 57, 58, 59 or 116; Most preferably, The first antigen-binding domain that specifically binds to TfR1 comprises a heavy chain variable region (TfR1-VH) and a light chain variable region (TfR1-VL), wherein: The TfR1-VH contains the amino acid sequence of SEQ ID NO: 27, and the TfR1-VL contains the amino acid sequence of SEQ ID NO: 13; and The second antigen-binding domain that specifically binds to Aβ comprises a heavy chain variable region (Aβ-VH) and a light chain variable region (Aβ-VL), wherein: The Aβ-VH contains the amino acid sequence of SEQ ID NO: 54, and the Aβ-VL contains the amino acid sequence of SEQ ID NO:

56.

3. The antigen-binding molecule that specifically binds to TfR1 and Aβ according to claim 1 or 2, wherein: The first antigen-binding domain that specifically binds to TfR1 and the second antigen-binding domain that specifically binds to Aβ are selected from Fab, Fab′, F(ab′)2, Fv, scFv, dsFv and dAb; Preferably, both the first antigen-binding domain that specifically binds to TfR1 and the second antigen-binding domain that specifically binds to Aβ are Fab domains.

4. The antigen-binding molecule that specifically binds to TfR1 and Aβ according to any one of claims 1 to 3, comprising a heavy chain constant region CH1 and a light chain constant region CL; Preferably, the heavy chain constant region CH1 is the heavy chain constant region CH1 of human IgG1 or its variants, and the light chain constant region CL is the light chain constant region CL of human κ chain or its variants; More preferably, the heavy chain constant region CH1 contains the amino acid sequence of SEQ ID NO: 82, and the light chain constant region CL contains the amino acid sequence of SEQ ID NO:

18.

5. The antigen-binding molecule that specifically binds to TfR1 and Aβ according to any one of claims 1 to 4, comprising an Fc region, wherein the Fc region comprises a first subunit Fc1 and a second subunit Fc2 capable of associating with each other; Preferably, the Fc region is the Fc region of a human IgG1 variant, wherein Fc1 contains amino acid substitutions of L351I and D356K, and Fc2 contains amino acid substitutions of L351I and K439E, and the numbering is based on the EU index; More preferably, Fc1 contains the amino acid sequence of SEQ ID NO: 80; and Fc2 contains the amino acid sequence of SEQ ID NO:

81.

6. The antigen-binding molecule that specifically binds to TfR1 and Aβ according to any one of claims 1 to 5, wherein the antigen-binding molecule that specifically binds to TfR1 and Aβ is an antibody; preferably a multispecific antibody; more preferably a bispecific antibody.

7. The antigen-binding molecule that specifically binds to TfR1 and Aβ according to claim 6, wherein: The antigen-binding molecule that specifically binds to TfR1 and Aβ is a bispecific antibody, and comprises a first half-antibody and a second half-antibody, wherein: The first half-antibody includes Fab and Fc1 regions that bind TfR1; and the second half-antibody includes Fab and Fc2 regions that bind Aβ; and the first half-antibody and the second half-antibody are associated with each other through the Fc1 and Fc2 regions; Preferably, The first hapten contains one TfR1 binding Fab and one Fc1 region from the N-terminus to the C-terminus; and the second hapten contains two identical or different Aβ binding Fabs and one Fc2 region from the N-terminus to the C-terminus; and the first hapten and the second hapten associate with each other through the Fc1 and Fc2 regions. More preferably, the antigen-binding molecule that specifically binds to TfR1 and Aβ comprises: One first chain as shown in equation (a), one second chain as shown in equation (b), one third chain as shown in equation (c), and two fourth chains as shown in equation (d), wherein the structures shown in equations (a), (b), (c), and (d) are arranged from the N end to the C end: (a)[TfR1-VH]-[CH1]-[connector 1]-[Fc1]; (b)[TfR1-VL]-[CL]; (c)[Aβ-VH]-[CH1]-[connector 2]-[Aβ-VH]-[CH1]-[Fc2]; (d)[Aβ-VL]-[CL]; Linker 1 and linker 2 may be the same or different peptide linkers, or linker 1 and / or linker 2 may not exist; Fc1 and Fc2 may be interchangeable; The first and second chains combine with each other, the first and third chains combine with each other, and the third and fourth chains combine with each other to form antigen-binding molecules that specifically bind to TfR1 and Aβ.

8. The antigen-binding molecule that specifically binds to TfR1 and Aβ according to any one of claims 1 to 7, comprising: One first chain containing an amino acid sequence of SEQ ID NO: 87, 85 or 86, or having at least 80% sequence identity with it; one second chain containing an amino acid sequence of SEQ ID NO: 20, 21, 22 or 23, or having at least 80% sequence identity with it; one third chain containing an amino acid sequence of SEQ ID NO: 88 or 89, or having at least 80% sequence identity with it; and two fourth chains containing an amino acid sequence of SEQ ID NO: 66, 67, 68, 69 or 117, or having at least 80% sequence identity with it. Preferably, The antigen-binding molecule that specifically binds to TfR1 and Aβ comprises: One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequences SEQ ID NO: 66, 67, 68, 69, or 117. More preferably, The antigen-binding molecule that specifically binds to TfR1 and Aβ comprises: One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 66; or One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 67; or One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 68; or One first strand containing the amino acid sequence SEQ ID NO: 87, one second strand containing the amino acid sequence SEQ ID NO: 20, one third strand containing the amino acid sequence SEQ ID NO: 88, and two fourth strands containing the amino acid sequence SEQ ID NO: 69; or One first strand containing the amino acid sequence of SEQ ID NO: 87, one second strand containing the amino acid sequence of SEQ ID NO: 20, one third strand containing the amino acid sequence of SEQ ID NO: 88, and two fourth strands containing the amino acid sequence of SEQ ID NO: 117; The first and second chains combine with each other, the first and third chains combine with each other, and the third and fourth chains combine with each other to form antigen-binding molecules that specifically bind to TfR1 and Aβ. Most preferably, The antigen-binding molecule that specifically binds to TfR1 and Aβ comprises: One first strand containing the amino acid sequence of SEQ ID NO: 87, one second strand containing the amino acid sequence of SEQ ID NO: 20, one third strand containing the amino acid sequence of SEQ ID NO: 88, and two fourth strands containing the amino acid sequence of SEQ ID NO: 66; The first and second chains combine with each other, the first and third chains combine with each other, and the third and fourth chains combine with each other to form antigen-binding molecules that specifically bind to TfR1 and Aβ.

9. An antigen-binding molecule that specifically binds to TfR1, comprising: Two first strands containing amino acid sequences of SEQ ID NO: 87, 85 or 86, or having at least 80% sequence identity with them, and two second strands containing amino acid sequences of SEQ ID NO: 20, 21, 22 or 23, or having at least 80% sequence identity with them; Preferably, The antigen-binding molecule that specifically binds to TfR1 comprises: Two first strands containing the amino acid sequence SEQ ID NO: 87, 85, or 86, and two second strands containing the amino acid sequence SEQ ID NO: 20; or Two first strands containing the amino acid sequence SEQ ID NO: 85, 86, or 87, and two second strands containing the amino acid sequence SEQ ID NO: 21; or Two first strands containing the amino acid sequence SEQ ID NO: 85, 86, or 87, and two second strands containing the amino acid sequence SEQ ID NO: 22; or Two first strands containing the amino acid sequence SEQ ID NO: 85, 86 or 87, and two second strands containing the amino acid sequence SEQ ID NO: 23; More preferably, The antigen-binding molecule that specifically binds to TfR1 comprises: Two first strands containing the amino acid sequence of SEQ ID NO: 87, and two second strands containing the amino acid sequence of SEQ ID NO:

20.

10. An antigen-binding molecule that specifically binds to Aβ, comprising: Two first strands containing amino acid sequences of SEQ ID NO: 88 or 89, or having at least 80% sequence identity with them, and four second strands containing amino acid sequences of SEQ ID NO: 66, 67, 68, 69 or 117, or having at least 80% sequence identity with them; Preferably, The antigen-binding molecule that specifically binds to Aβ comprises: Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 66, 67, 68, 69, or 117; or Two first strands containing the amino acid sequence SEQ ID NO: 89, and four second strands containing the amino acid sequence SEQ ID NO: 66, 67, 68, 69, or 117; More preferably, The antigen-binding molecule that specifically binds to Aβ comprises: Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO: 66, 67, 68, 69, or 117; Most preferably, The antigen-binding molecule that specifically binds to Aβ comprises: Two first strands containing the amino acid sequence SEQ ID NO: 88, and four second strands containing the amino acid sequence SEQ ID NO:

66.

11. A pharmaceutical composition comprising an antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any one of claims 1 to 8, or an antigen-binding molecule that specifically binds to TfR1 as described in claim 9, or an antigen-binding molecule that specifically binds to Aβ as described in claim 10, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

12. A nucleic acid encoding an antigen-binding molecule that specifically binds to TfR1 and Aβ as described in any one of claims 1 to 8, or an antigen-binding molecule that specifically binds to TfR1 as described in claim 9, or an antigen-binding molecule that specifically binds to Aβ as described in claim 10.

13. A host cell comprising the nucleic acid as described in claim 12.

14. A method for preventing or treating a disease or condition, the method comprising administering to a subject a preventive or therapeutically effective amount of an antigen-binding molecule that specifically binds to TfR1 and Aβ according to any one of claims 1 to 8, or an antigen-binding molecule that specifically binds to TfR1 according to claim 9, or an antigen-binding molecule that specifically binds to Aβ according to claim 10, or a pharmaceutical composition according to claim 11. Preferably, the disease or symptom is a neurological disease; More preferably, the disease or symptom is a neurodegenerative disease; Most preferably, the disease or symptom is Alzheimer's disease.

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