Pharmaceutical Composition of FAP / CD40 Binding Molecule and its Pharmaceutical Use

Abstract

A pharmaceutical composition of a FAP / CD40 binding molecule and the pharmaceutical use thereof. Specifically, the pharmaceutical composition contains a FAP / CD40 binding molecule and a buffer agent. The pharmaceutical composition has a good biological activity and stability.

Description

1 / 155 Pharmaceutical Composition of FAP / CD40 Binding Molecule and its Pharmaceutical Use

[001] This description claims priority to Chinese Patent Application No. 202310174524.1, filed on February 24, 2023, which is incorporated herein by reference in its entirety. TECHNICAL FIELD

[002] The present description refers to the field of pharmaceutical formulations, in particular to a pharmaceutical composition comprising a FAP / CD40 binding molecule and its pharmaceutical use. FUNDAMENTALS

[003] CD40 (TNFRSF5), a transmembrane phosphorylated glycoprotein, is a member of the tumor necrosis factor receptor (TNFRS) superfamily. CD40 is expressed on various cell types, including B cells, follicular dendritic cells (DCs), epithelial cells, monocytes, macrophages, smooth muscle cells, and tumor cells. Its ligand CD40L is primarily expressed on activated T cells, activated B cells, platelets, smooth muscle cells, etc. Binding to CD40L multimerizes CD40, generating downstream activation, growth, and differentiation signals. CD40 signaling activates a variety of downstream signaling pathways, such as NF-κB, MAPK, and STAT3 (Pype S, et al. J Biol Chem. 2000 Jun. 16; 275(24):1858-693), and Petition 870250091928, dated 08 / 10 / 2025, page 9 / 175 2 / 155 These pathways regulate gene expression by regulating the activating protein c-Jun, ATF2, and Rel transcription factors. The binding of CD40 to CD40L induces the proliferation of resting B cells, immunoglobulin switching, and antibody secretion, in addition to playing important roles in the development of germinal centers in tissues and in B cell survival. All these factors are essential for humoral immune responses (Kehry M R. J Immunol 1996; 156: 2345-2348). CD40L binds to CD40 on dendritic cells, inducing dendritic cell maturation, which manifests in increased expression of the B7 family of co-stimulatory factors (CD80 and CD86) and increased secretion of pro-inflammatory cytokines, such as interleukin 12 (IL-12). The interaction between CD40 and CD40L provides a co-stimulatory signal for T cell activation and promotes antigen presentation by dendritic cells to T cells.

[004] With the clinical success of immune checkpoint inhibitor (ICI) therapies targeting CD40, CTLA-4, and PD-L1 in the treatment of tumors, immunotherapy has become the most anticipated research field among third-generation tumor therapies. Antitumor immunotherapy, based on the mechanism by which antigen presentation by dendritic cells and T cell activation are enhanced by CD40 activation, Petition 870250091928, dated 08 / 10 / 2025, page 10 / 175 3 / 155 has been shown to be clinically effective. However, there is systemic activation of CD40, which can lead to some target-specific toxicities, such as cytokine storms, hepatotoxicity, hematotoxicity, and venous thrombosis caused by peripheral CD40 activation. These toxicities limit the therapeutic windows of CD40 agonist antibodies. Clinical studies with Pfizer's CD40 agonists CP-870 and 893 and Medimmune's CD40L-Fc fusion protein MEDI5083 (a novel fusion protein from AstraZeneca that can activate the CD40 signaling pathway, consisting of three CD40L molecules connected together and IgG4-Fc) have been terminated.Thus, in the research and development of second-generation CD40 agonist antibodies, researchers aim to mediate CD40 activation through tumor-associated antigens (TAAs), so that CD40 is specifically activated within tumors, while peripheral CD40 activation is reduced, thereby increasing the therapeutic windows of CD40 agonist antibodies.

[005] Fibroblast activating protein (FAP) α is a tumor-associated antigen. FAP is poorly expressed in normal tissues of healthy adults and is selectively expressed in 93% of tumor tissues, with 30% showing high expression, such as in colon cancer, pancreatic cancer, breast cancer, gastric cancer, prostate cancer, bladder cancer, and oral squamous cell carcinoma. The document Petition 870250091928, dated 08 / 10 / 2025, page 11 / 175 4 / 155 WO2023025194 provides an anti-FAP antibody, an anti-CD40 antibody, and a bispecific antibody for both. The bispecific antibody can mediate specific tumor activation of CD40 via FAP, has maturation-promoting and activation-promoting effects on APCs (e.g., dendritic cells [DCs]), can eliminate hepatotoxicity, peripheral hematotoxicity, and other peripheral toxicities, and has an excellent administration window and pharmacology.

[006] Due to their large molecular weights and complex structures, antibody-based drugs are prone to degradation, aggregation, unwanted chemical modifications, etc., which makes them unstable. It is particularly important to develop stable antibody-based drug formulations that make the antibodies more suitable for administration, maintain the antibodies stable during storage and subsequent use, and produce better effects. For a new FAP / CD40 binding molecule, it is still necessary to develop a suitable pharmaceutical composition. SUMMARY

[007] The present description provides a pharmaceutical composition comprising a FAP / CD40 binding molecule, and the composition exhibits excellent stability.

[008] The present description provides a pharmaceutical composition comprising a binding molecule to Petition 870250091928, dated 08 / 10 / 2025, page 12 / 175 5 / 155 FAP / CD40 and a buffer, wherein the buffer is selected from the group consisting of an acetate buffer, a succinate buffer, a histidine salt buffer, and a phosphate buffer. In some embodiments, the buffer is a histidine salt buffer. In some embodiments, the buffer is a histidine-hydrochloride buffer. In some embodiments, the buffer is a histidine-acetate buffer. In some embodiments, the buffer is a histidine-histidine hydrochloride buffer. In some embodiments, the buffer is a histidine-acetic acid buffer.

[009] In some embodiments, the FAP / CD40 binding molecule comprises a first antigen-binding domain that binds specifically to FAP and a second antigen-binding domain that binds specifically to CD40, wherein the second antigen-binding domain that binds specifically to CD40 comprises (at least one) unique immunoglobulin variable domain.

[0010] In some embodiments, the FAP / CD40 binding molecule comprises a single immunoglobulin variable domain that binds specifically to CD40. In some other embodiments, the FAP / CD40 binding molecule comprises 2, 3, 4, 5, 6, 7, 8, 9, 10 or more single immunoglobulin variable domains that bind specifically to CD40, and the single immunoglobulin variable domains may be identical or different. Petition 870250091928, dated 08 / 10 / 2025, p. 13 / 175 6 / 155

[0011] In some embodiments, the FAP / CD40 binding molecule comprises at least one (e.g., 2, 3, or 4) antigen-binding domain that binds specifically to FAP.

[0012] In some embodiments, in the FAP / CD40 binding molecule, the single variable immunoglobulin domain that specifically binds to CD40 comprises three complementarity-determining regions: CDR1, CDR2, and CDR3, in which the amino acid sequences of CDR1, CDR2, and CDR3 are presented in SEQ ID NOs: 12, 13, and 14, respectively.

[0013] In some embodiments, in the FAP / CD40 binding molecule mentioned above, the single variable immunoglobulin domain that specifically binds to CD40 comprises or is set in any of the SEQ ID NOs: 11 and 16 to 19, or has at least 90%, at least 95%, at least 98% or at least 99% sequence identity with any of the SEQ ID NOs: 23 and 35 to 38.

[0014] In some embodiments, the FAP / CD40 binding molecule comprises a first antigen-binding domain that binds specifically to FAP and a second antigen-binding domain that binds specifically to CD40, wherein the first antigen-binding domain that binds specifically to FAP comprises a variable heavy chain region and a variable light chain region; the variable heavy chain region comprises an HCDR1, a Petition 870250091928, dated 08 / 10 / 2025, p. 14 / 175 7 / 155 HCDR2 and HCDR3, and the variable region of the light chain comprises an LCDR1, an LCDR2, and an LCDR3, where: 1) The amino acid sequences of HCDR1, HCDR2, and HCDR3 are established in SEQ ID NOs 3, 4, and 5, respectively; the amino acid sequences of LCDR1, LCDR2, and LCDR3 are established in SEQ ID NOs 6, 7, and 8, respectively; or 2) The amino acid sequences of HCDR1, HCDR2, and HCDR3 are established in SEQ ID NOs: 3, 4, and 5, respectively; the amino acid sequences of LCDR1, LCDR2, and LCDR3 are established in SEQ ID NOs: 6, 32, and 8, respectively.

[0015] In some embodiments, in the FAP / CD40 binding molecule mentioned above, the variable region of the heavy chain of the first antigen-binding domain that specifically binds to FAP comprises the amino acid sequence shown in SEQ ID NO: 1 or an amino acid sequence with at least 80% identity to it, and the variable region of the light chain comprises the amino acid sequence shown in SEQ ID NO: 2 or an amino acid sequence with at least 80% identity to it.

[0016] In some embodiments, the FAP / CD40 binding molecule additionally comprises a human immunoglobulin Fc region (e.g., a human IgG1 or IgG4 Fc region).

[0017] In some embodiments, in the FAP / CD40 binding molecule mentioned earlier, the first domain of Petition 870250091928, dated 08 / 10 / 2025, p. 15 / 175 8 / 155 antigen linkage that specifically binds to FAP comprises a heavy chain (HC) and a light chain (LC);

[0018] for example, the heavy chain is of the IgG1 or IgG4 isotype, and the light chain is of the Kappa isotype.

[0019] In some specific embodiments, the heavy chain is the amino acid sequence set forth in SEQ ID NO: 24 or an amino acid sequence with at least 80% identity with it, and the light chain is the amino acid sequence set forth in SEQ ID NO: 25 or an amino acid sequence with at least 80% identity with it.

[0020] In some embodiments, in the FAP / CD40 binding molecule mentioned above, the first antigen-binding domain that specifically binds to FAP comprises a variable region of the heavy chain and a variable region of the light chain, in which: The single immunoglobulin variable domain of the second antigen-binding domain that specifically binds to CD40 is located at the N-terminus of the variable region of the heavy chain of the first antigen-binding domain that specifically binds to FAP; The single immunoglobulin variable domain of the second antigen-binding domain that specifically binds to CD40 is located at the C-terminus of the variable region of the heavy chain of the first antigen-binding domain that specifically binds to FAP; Petition 870250091928, dated 08 / 10 / 2025, page 16 / 175 9 / 155 the single immunoglobulin variable domain of the second antigen-binding domain that specifically binds to CD40 is located at the N-terminus of the variable region of the light chain of the first antigen-binding domain that specifically binds to FAP; and / or the single immunoglobulin variable domain of the second antigen-binding domain that specifically binds to CD40 is located at the C-terminus of the variable region of the light chain of the first antigen-binding domain that specifically binds to FAP.

[0021] In some embodiments, the single variable immunoglobulin domain of the second antigen-binding domain mentioned above, which specifically binds to CD40, is linked, directly or by a ligand, to the first antigen-binding domain that specifically binds to FAP; for example, the ligand has an amino acid sequence represented by (G4S)x, where x is independently selected from the group consisting of integers from 1 to 20 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10); for example, the ligand is an amino acid sequence represented by (G4S)2, (G4S)3, or (G4S)4.

[0022] In some embodiments, in the FAP / CD40 binding molecule mentioned above, the second antigen-binding domain that specifically binds to CD40 is multivalent (e.g., one of the binding molecules to Petition 870250091928, dated 08 / 10 / 2025, p. 17 / 175 10 / 155 The FAP / CD40 mentioned above comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 second antigen-binding domains that bind specifically to CD40. In some specific embodiments, the second antigen-binding domain that binds specifically to CD40 is bivalent, tetravalent, or hexavalent. In some specific embodiments, the second antigen-binding domain that binds specifically to CD40 comprises 2, 3, 4, 5, or 6 of the aforementioned single variable immunoglobulin domains.

[0023] In some embodiments, the FAP / CD40 binding molecule comprises a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain comprises the amino acid sequence set forth in any of the SEQ ID Nos: 21-23 or an amino acid sequence with at least 80% identity therewith, and the second polypeptide chain comprises the amino acid sequence set forth in SEQ ID No: 20 or an amino acid sequence with at least 80% identity therewith. In some specific embodiments, the FAP / CD40 binding molecule comprises two first polypeptide chains and two second polypeptide chains; in some specific embodiments, the two first polypeptide chains are identical and the two second polypeptide chains are identical.

[0024] In some embodiments, the binding molecule to Petition 870250091928, dated 08 / 10 / 2025, p. 18 / 175 11 / 155 The previously mentioned FAP / CD40 is capable of inhibiting tumor growth (e.g., increase in tumor volume or weight gain) and / or metastasis (e.g., metastasis to multiple organs or tissues or distant metastasis) by at least about 10%, for example, at least about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90%.

[0025] In some embodiments, the binding molecule to The FAP / CD40 mentioned above is a bispecific anti-FAP / CD40 antibody or an antigen-binding fragment thereof, wherein the antigen-binding fragment includes, but is not limited to, Fab, Fv, sFv, Fab', F(ab')2, linear antibodies, single-chain antibodies, scFv, sdAb, sdFv, nanobodies, pepticobodies, domain antibodies and multispecific antibodies (bispecific antibodies, diabodies, triabodies, tetrabodies, tandem di-scFv and tandem tri-scFv) and is, for example, an scFv, Fv, Fab or Fab' fragment.

[0026] In some embodiments, the bispecific anti-FAP / CD40 antibody comprises the single immunoglobulin variable domain in the aforementioned second antigen-binding domain, which binds specifically to CD40, and the variable heavy chain (VH) region and the variable light chain (VL) region in the aforementioned first antigen-binding domain, which bind Petition 870250091928, dated 08 / 10 / 2025, p. 19 / 175 12 / 155 specifically to FAP.

[0027] In some forms, in the bispecific anti-FAP / CD40 antibody: The first antigen-binding domain that specifically binds to FAP is a first antibody, comprising a heavy chain (HC) and a light chain (LC); and the second antigen-binding domain that specifically binds to CD40 is a second antibody, which is a VHH and has CDR1, CDR2, and CDR3 in the aforementioned CD40-binding molecule.

[0028] In some specific embodiments, VHH, as a second antibody, is located at the N-terminus and / or the C-terminus of the heavy or light chain of the first antibody.

[0029] In some specific embodiments, the bispecific anti-FAP / CD40 antibody comprises 1 first antibody and 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 (e.g., 2, 4 or 6) second VHH antibodies; the first antibody comprises two HCs and two LCs, wherein the VH of one HC of the first antibody forms an antigen-binding site with the VL of one LC, and the VH of the other HC forms an antigen-binding site with the VL of the other LC.

[0030] In some specific embodiments, the first antibody of the bispecific anti-FAP / CD40 antibody or the antigen-binding fragment thereof may be linked to 1, 2, 3, 4, 5, 6, 7 or 8 second VHH antibodies, and the Petition 870250091928, dated 08 / 10 / 2025, p. 20 / 175 13 / 155 seconds VHH antibodies may be identical or different, they may all be attached to the N-terminus of the heavy chain of the first antibody, or they may all be attached to the C-terminus of the heavy chain of the first antibody, or they may all be attached to the N-terminus of the light chain of the first antibody, or they may all be attached to the C-terminus of the light chain of the first antibody, or they may be attached to any combination of the N-terminus of the heavy chain, the C-terminus of the heavy chain, the N-terminus of the light chain and the C-terminus of the light chain.

[0031] In some specific embodiments, the bispecific anti-FAP / CD40 antibody comprises a first polypeptide chain and a second polypeptide chain, where: (i) the first polypeptide chain is, from the N-terminus to the C-terminus: [heavy chain of the first antibody]-ligand 1[second antibody]; the second polypeptide chain is the light chain of the first antibody; (ii) the first polypeptide chain is, from the N-terminus to the C-terminus: [heavy chain of the first antibody] - ligand 1 - [second antibody]1 - ligand 2 - [second antibody]2; the second polypeptide chain is the light chain of the first antibody; (iii) the first polypeptide chain is, from the N-terminus to the C-terminus: [heavy chain of the first antibody] - ligand 1 - [second antibody]1 - ligand 2 - [second antibody]2 Petition 870250091928, dated 08 / 10 / 2025, page 21 / 175 14 / 155 ligand 3-[second antibody^; the second polypeptide chain is the light chain of the first antibody; (iv) the first polypeptide chain is, from the N-terminus to the C-terminus: [second antibody]-ligand 1-[heavy chain of the first antibody]; the second polypeptide chain is the light chain of the first antibody; (v) the first polypeptide chain is, from the N-terminus to the C-terminus: [second antibody]2-ligand 2-[second antibody]1-ligand 1-[heavy chain of the first antibody]; the second polypeptide chain is the light chain of the first antibody; (vi) the first polypeptide chain is, from the N-terminus to the C-terminus: [second antibody] 3-ligand 3-[second antibody]2-ligand 2-[second antibody]1-ligand 1-[heavy chain of the first antibody]; the second polypeptide chain is the light chain of the first antibody; (vii) the first polypeptide chain is, from the N-terminus to the C-terminus: [second antibody]1-ligand 1-[heavy chain of the first antibody]-ligand 2-[second antibody]2; the second polypeptide chain is the light chain of the first antibody; (viii) the first polypeptide chain is, from the N-terminus to the C-terminus: [second antibody]1-ligand 1-[second antibody]2-ligand 2-[heavy chain of the first antibody] ligand 3-[second antibody]3; the second chain Petition 870250091928, dated 08 / 10 / 2025, page 22 / 175 15 / 155 polypeptide is the light chain of the first antibody; (ix) the first polypeptide chain is, from the N-terminus to the C-terminus: [second antibody]1-ligand 1-[heavy chain of the first antibody]-ligand 2-[second antibody]2-ligand 3-[second antibody]α; the second polypeptide chain is the light chain of the first antibody; wherein the [second antibody]1, [second antibody]2, and [second antibody]3 can be identical or different.

[0032] In some modalities, the second antibodies VHH in the bispecific anti-FAP / CD40 antibody mentioned above are linked, directly or by ligands, to the first antibody. The ligands are selected from the group consisting of amino acid sequences represented by (GmSn)x, (GGNGT)x, and (YGNGT)x, where men are each independently selected from the group consisting of integers from 1 to 8 (e.g., 1, 2, 3, 4, 5, 6, 7, or 8), ex is independently selected from the group consisting of integers from 1 to 20 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20). For example, ligands are amino acid sequences represented by G4S, (G4S)2, (G4S)3, (G4S)4, (G4S)5, or (G4S)6. In some embodiments, ligand 1, ligand 2, and ligand 3 may be identical or different.

[0033] In some embodiments, the heavy chain of the first antibody of the bispecific anti-FAP / CD40 antibody Petition 870250091928, dated 08 / 10 / 2025, page 23 / 175 16 / 155 comprises a variable region (VH) of the heavy chain and a constant region (CH) of the heavy chain, and the light chain comprises a variable region (VL) of the light chain and a constant region (CL) of the light chain. The first antibody may be a full-length antibody.

[0034] In some embodiments, the heavy chain of the first antibody of the bispecific anti-FAP / CD40 antibody is of an IgG isotype (e.g., IgG1, IgG2, IgG3, or IgG4), for example, the IgG1 isotype; and / or the light chain of the first antibody is of the Kappa isotype.

[0035] In some embodiments, the two HCs of the bispecific anti-FAP / CD40 antibody comprise identical CDRs and / or the two LCs comprise identical CDRs. In some specific embodiments, the two HCs of the first antibody comprise identical VHs and / or the two LCs comprise identical VLs. In some specific embodiments, the two HCs of the first antibody have identical amino acid sequences and / or the two LCs have identical amino acid sequences.

[0036] In some embodiments, two second VHH antibodies of the bispecific anti-FAP / CD40 antibody have identical or different amino acid sequences. For example, the two second VHH antibodies have identical amino acid sequences.

[0037] In some modalities, the bispecific antibody Petition 870250091928, dated 08 / 10 / 2025, p. 24 / 175 17 / 155 anti-FAP / CD40 comprises two first polypeptide chains and two second polypeptide chains, wherein for each polypeptide chain: a) each of the first polypeptide chains independently comprises a second VHH antibody and the heavy chain (HC) of the first antibody, and b) each of the second polypeptide chains independently comprises the light chain (LC) of the first antibody, wherein the VHH is linked to the N-terminus and / or the C-terminus of the HC of the second antibody by a ligand; or, i) each of the first polypeptide chains independently comprises the heavy chain (HC) of the first antibody, and ii) each of the second polypeptide chains independently comprises a second VHH antibody and the light chain (LC) of the first antibody, wherein the VHH is linked, directly or by a ligand, to the N-terminus and / or the C-terminus of the LC of the first antibody.

[0038] In some specific embodiments, the bispecific anti-FAP / CD40 antibody comprises two identical primary polypeptide chains and two identical secondary polypeptide chains.

[0039] In some embodiments, a mutation is introduced into the Fc region of the FAP / CD40 binding molecule or the bispecific anti-FAP / CD40 antibody. The mutation is, for example, a mutation that removes or reduces the effector function of IgG Fc, including, but not limited to, N297A or Petition 870250091928, dated 08 / 10 / 2025, p. 25 / 175 18 / 155 D265A / N297A in IgG1, or L234A / L235A, L234A / L235A / P329G, L234E, L234F, L234E / L235F or L234E / L235F / P329G in IgG1, V234A / G237A / P238S / H268A / V309L / A330S / P331S in IgG2, F234A / L235A in IgG4, S228P / F234A / L235A in IgG4, N297A in IgG2 or IgG4, V234A / G237A in IgG2, K214T / E233P / L234V / L235A / G236 / A327G / P331A / D365E / L358M deletion in IgG1, H268Q / V309L / A330S / P331S in IgG2, S267E / L328F in IgG1, L234F / L235E / D265A in IgG1, L234A / L235A / G237A / P238S / H268A / A330S / P331S in IgG1 and S228P / F234A / L235A / G237A / P238S in IgG4.

[0040] In the context of the mutations contained in the Fc region described in this document, / represents and; for example, D265A / N297A represents D265A and N297A; that is, Fc comprises the mutations D265A and N297A; the amino acid positions of the mutations are numbered according to the EU numbering scheme.

[0041] In some embodiments, the FAP / CD40 binding molecule or the bispecific anti-FAP / CD40 antibody mentioned above has one or more of the following characteristics: (a) binding to human FAP or to an epitope thereof with a KD value<10-7; (b) binding to human CD40 or to an epitope thereof with a KDd10-7 value; Petition 870250091928, dated 08 / 10 / 2025, p. 26 / 175 19 / 155 (c) induction of immunostimulation of antigen-presenting cells (APCs) expressing CD40; (d) increased activation of APCs (e.g., dendritic cells) and / or promotion of APC proliferation (e.g., dendritic cells); (e) stimulation of tumor-specific T cell responses; (f) causes or promotes apoptosis of tumor cells; and / or (g) inhibits tumor growth and / or metastasis.

[0042] In some embodiments, the FAP / CD40 binding molecule in the pharmaceutical composition is at a concentration of 0.01 mg / mL to 500 mg / mL, such as 0.05 mg / mL to 450 mg / mL, 0.05 mg / mL to 400 mg / mL, 0.05 mg / mL to 350 mg / mL, 0.05 mg / mL to 300 mg / mL, 0.05 mg / mL to 250 mg / mL, 0.05 mg / mL to 200 mg / mL, 0.05 mg / mL to 150 mg / mL, 0.05 mg / mL to 140 mg / mL, 0.05 mg / mL to 130 mg / mL, 0.05 mg / mL to 120 mg / mL, 0.05 mg / mL to 110 mg / mL, 0,05 mg / mL a 100 mg / mL, 0,1 mg / mL a 400 mg / mL, 0,1 mg / mL a 350 mg / mL, 0,1 mg / mL a 300 mg / mL, 0,1 mg / mL a 250 mg / mL, 0,1 mg / mL a 200 mg / mL, 0,1 mg / mL a 150 mg / mL, 0 ,1 mg / mL a140 mg / mL, 0,1 mg / mL a 130 mg / mL, 0,1 mg / mL a 120 mg / mL,0,1 mg / mL a 110 mg / mL, 0,1 mg / mL a 100 mg / mL, 0,5 mg / mL a350 mg / mL, 0,5 mg / mL a 300 mg / mL, 0,5 mg / mL a 250 mg / mL,0,5 mg / mL a 200 mg / mL, 0,5 mg / mL a 150 mg / mL, 0,5 mg / mL a140 mg / mL, 0,5 mg / mL a 130 mg / mL, 0,5 mg / mL a 120 mg / mL,0,5 Petição 870250091928, de 08 / 10 / 2025, pág. 27 / 175 20 / 155 mg / mL a 110 mg / mL, 0,5 mg / mL a 100 mg / mL, 1 mg / mL a 300 mg / mL, 1 mg / mL a 250 mg / mL, 1 mg / mL a 200 mg / mL, 1 mg / mL a 150 mg / mL, 1 mg / mL a 140 mg / mL, 1 mg / mL a 130 mg / mL, 1 mg / mL a 120 mg / mL, 1 mg / mL a 110 mg / mL, 1 mg / mL a 100 mg / mL, 1 mg / mL a 95 mg / mL, mg / mL a 80 mg / mL, mg / mL a 65 mg / mL, mg / mL a 50 mg / mL, mg / mL a 35 mg / mL, mg / mL a 20 mg / mL, mg / mL a 85 mg / mL, mg / mL a 70 mg / mL, mg / mL a 55 mg / mL, mg / mL a 40 mg / mL, mg / mL a 25 mg / mL, mg / mL a 90 mg / mL,1 mg / mL a 75 mg / mL,1 mg / mL a 60 mg / mL,1 mg / mL a 45 mg / mL,1 mg / mL a 30 mg / mL,1 mg / mL a 15 mg / mL,5 mg / mL a 80 mg / mL,5 mg / mL a 65 mg / mL,5 mg / mL a 50 mg / mL,5 mg / mL a 35 mg / mL,5 mg / mL a 20 mg / mL,5 mg / mL a 85 mg / mL,1 mg / mL a 70 mg / mL,1 mg / mL a 55 mg / mL,1 mg / mL a 40 mg / mL,1 mg / mL a 25 mg / mL,1 mg / mL a 90 mg / mL,5 mg / mL a 75 mg / mL,5 mg / mL a 60 mg / mL,5 mg / mL a 45 mg / mL,5 mg / mL a 30 mg / mL,5 mg / mL a 15 mg / mL,8 mg / mL a 12 mg / mL, 30 mg / mL a 250 mg / mL, mg / mL a 200 mg / mL, mg / mL a 190 mg / mL, 30 mg / mL a 180 mg / mL, 30 mg / mL a 170 mg / mL, 30 mg / mL a 160 mg / mL, 30 mg / mL a 150 mg / mL, 30 mg / mL a 140 mg / mL, 30 mg / mL a 130 mg / mL, 30 mg / mL a 120 mg / mL, 30 mg / mL a 110 mg / mL,30 mg / mL a 100 mg / mL, 50 mg / mL a 200 mg / mL, 50 mg / mL a 190 mg / mL, 50 mg / mL a 180 mg / mL, 50 mg / mL a 170 mg / mL, 50 mg / mL a 160 mg / mL, 50 mg / mL a 150 mg / mL, 50 mg / mL a 140 mg / mL, 50 mg / mL a 130 mg / mL, 50 mg / mL a 120 mg / mL, 50 mg / mL a 110 mg / mL, 50 mg / mL a 100 mg / mL, 70 mg / mL a 180 mg / mL, 70 mg / mL a 170 mg / mL, 70 mg / mL a 160 mg / mL, 70, Petição 870250091928, de 08 / 10 / 2025, pág. 28 / 175 21 / 155 mg / mL to 150 mg / mL, 70 mg / mL to 140 mg / mL, 70 mg / mL to 130 mg / mL, 70 mg / mL to 120 mg / mL, 70 mg / mL to 110 mg / mL, 70 mg / mL to 100 mg / mL, 90 mg / mL to 110 mg / mL, or 95 mg / mL to 105 mg / mL. In some embodiments, the FAP / CD40 binding molecule in the pharmaceutical composition is at a concentration of 0.1 mg / mL to 400 mg / mL. In some embodiments, the FAP / CD40 binding molecule in the pharmaceutical composition is at a concentration of 0.5 mg / mL to 200 mg / mL. In some embodiments, the FAP / CD40 binding molecule in the pharmaceutical composition is at a concentration of 1 mg / mL to 150 mg / mL. In some embodiments, the FAP / CD40 binding molecule in the pharmaceutical composition is at a concentration of 5 mg / mL to 45 mg / mL. In some embodiments, the FAP / CD40 binding molecule in the pharmaceutical composition is at a concentration of 50 mg / mL to 110 mg / mL.In some embodiments, the FAP / CD40 binding molecule in the pharmaceutical composition is at a concentration of approximately 1 mg / mL, approximately 5 mg / mL, approximately 10 mg / mL, approximately 15 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 30 mg / mL, approximately 40 mg / mL, approximately 45 mg / mL, approximately 50 mg / mL, approximately 55 mg / mL, approximately 60 mg / mL, approximately 70 mg / mL, approximately 75 mg / mL, approximately 80 mg / mL, approximately 85 mg / mL, approximately 90 mg / mL, approximately 95 mg / mL, or approximately 100 mg / mL. In some formulations, the FAP / CD40 binding molecule in the pharmaceutical composition is at a concentration of approximately 5 mg / mL, approximately 10. Petition 870250091928, dated 08 / 10 / 2025, p. 29 / 175 22 / 155 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL. In some embodiments, the FAP / CD40 binding molecule in the pharmaceutical composition is at a concentration of approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, or approximately 25 mg / mL. In some embodiments, the FAP / CD40 binding molecule in the pharmaceutical composition is at a concentration of approximately mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL. In some embodiments, the FAP / CD40 binding molecule in the pharmaceutical composition is at a concentration of at least 500 mg / mL. In some embodiments, the FAP / CD40 binding molecule in the pharmaceutical composition is at a concentration of at least 450 mg / mL, at least 400 mg / mL, at least 350 mg / mL, at least 300 mg / mL, at least 250 mg / mL, at least 200 mg / mL, at least 150 mg / mL, at least 100 mg / mL, at least 50 mg / mL, at least 10 mg / mL, at least 1 mg / mL, at least 0.1 mg / mL, or at least 0.01 mg / mL.

[0043] In some embodiments, the buffer in the pharmaceutical composition is at a concentration of 0.1 mM to 50 mM, such as 0.1 mM to 45 mM, 0.1 mM to 40 mM, 0.1 mM to 35 mM, 0.1 mM to 30 mM, 0.1 mM to 25 mM, 0.1 mM to 20 mM, 0.1 mM to 15 mM, 0.1 mM to 10 mM, 0.5 mM to 45 mM, 0.5 mM to 40 mM, 0.5 mM to 35 mM, 0.5mM to 30mM, 0.5mM to 25mM, 0.5mM to 20mM, 0.5mM to 15mM, 0.5mM to 10mM, 1mM to 40mM, 1mM to 35mM, 1mM to 30mM Petition 870250091928, dated 08 / 10 / 2025, page 30 / 175 23 / 155 mM, 1 mM to 25 mM, 1 mM to 20 mM, 1 mM to 15 mM, 1 mM to 10 mM, 5 mM to 35 mM, 5 mM to 30 mM, 5 mM to 25 mM, 5 mM to 20 mM, 5 mM to 15 mM, 5 mM to 10 mM, 8 mM to 30 mM, 8 mM to 25 mM, 8 mM to 20 mM, 8 mM to 15 mM, or 8 mM to 12 mM. In some embodiments, the buffer in the pharmaceutical composition is at a concentration of 0.5 mM to 40 mM. In some embodiments, the buffer in the pharmaceutical composition is at a concentration of 1 mM to 30 mM. In some embodiments, the buffer in the pharmaceutical composition is at a concentration of 5 mM to 20 mM.In some embodiments, the buffer in the pharmaceutical composition is at a concentration of approximately 1 mM, approximately 2 mM, approximately 3 mM, approximately 4 mM, approximately 5 mM, approximately 6 mM, approximately 7 mM, approximately 8 mM, approximately 9 mM, approximately 10 mM, approximately 11 mM, approximately 12 mM, approximately 13 mM, approximately 14 mM, approximately 15 mM, approximately 16 mM, approximately 17 mM, approximately mM, approximately 19 mM, approximately 20 mM, approximately 21 mM, approximately 22 mM, approximately 23 mM, approximately 24 mM, approximately 25 mM, approximately 26 mM, approximately 27 mM, approximately 28 mM, approximately 29 mM, or approximately 30 mM. In some embodiments, the buffer in the pharmaceutical composition is at a concentration of approximately mM. In some embodiments, the buffer in the pharmaceutical composition is at a concentration of at least 50 mM. In some embodiments, the buffer in the pharmaceutical composition is at a concentration of at least 10 mM.

[0044] In some modalities, the composition Petition 870250091928, dated 08 / 10 / 2025, p. 31 / 175 24 / 155 pharmaceutical additionally comprises a surfactant. In some embodiments, the surfactant is one or more surfactants selected from the group consisting of a polysorbate and a poloxamer. In some embodiments, the polysorbate is one or more polysorbates selected from the group consisting of polysorbate 20 and polysorbate 80. In some embodiments, the poloxamer is poloxamer 188. In some embodiments, the surfactant is a polysorbate. In some embodiments, the surfactant is polysorbate 80.

[0045] In some embodiments, the surfactant in the pharmaceutical composition is at a concentration of 0.01 mg / mL to 10 mg / mL, such as 0.01 mg / mL to 9 mg / mL, 0.01 mg / mL to 8 mg / mL, 0.01 mg / mL to 7 mg / mL, 0.01 mg / mL to 6 mg / mL, 0.01 mg / mL to 5 mg / mL, 0.01 mg / mL to 4 mg / mL, 0.01 mg / mL to 3 mg / mL, 0.01 mg / mL to 2 mg / mL, 0.01 mg / mL to 1 mg / mL, 0.01 mg / mL to 0.5 mg / mL, 0.05 mg / mL to 8 mg / mL, 0.05 mg / mL to 7 mg / mL, 0.05 mg / mL to 6 mg / mL, 0.05 mg / mL to 5 mg / mL, 0.05 mg / mL to 4 mg / mL, 0.05 mg / mL to 3 mg / mL, 0.05 mg / mL to 2 mg / mL, 0.05 mg / mL to 1 mg / mL, 0.05 mg / mL to 0.5 mg / mL, 0.1 mg / mL to 6 mg / mL, 0.1 mg / mL to 5 mg / mL, 0.1 mg / mL to 4 mg / mL, 0.1 mg / mL to 3 mg / mL, 0.1 mg / mL to 2 mg / mL, 0.1 mg / mL to 1 mg / mL, 0.1 mg / mL to 0.5 mg / mL, 0.2 mg / mL to 5 mg / mL, 0.2 mg / mL to 4.5 mg / mL, 0.2 mg / mL to 4 mg / mL, 0.2 mg / mL to 3.5 mg / mL, 0.2 mg / mL to 3 mg / mL, 0.2 mg / mL to 2.5 mg / mL, 0.2 mg / mL to 2 mg / mL, 0.2 mg / mL to 1.5 mg / mL, 0.2 mg / mL to 1 mg / mL, 0.2 mg / mL to 0.5 mg / mL, 0.3 mg / mL to 4.5 mg / mL, 0.3 Petition 870250091928, dated 08 / 10 / 2025, page 32 / 175 25 / 155 mg / mL to 4 mg / mL, 0.3 mg / mL to 3.5 mg / mL, 0.3 mg / mL to 3 mg / mL, 0.3 mg / mL to 2.5 mg / mL, 0.3 mg / mL to 2 mg / mL, 0.3 mg / mL to 1.5 mg / mL, 0.3 mg / mL to 1 mg / mL, 0.3 mg / mL to 0.5 mg / mL, 0.7 mg / mL to 3.5 mg / mL, 0.7 mg / mL to 3 mg / mL, 0.7 mg / mL to 2.5 mg / mL, 0.7 mg / mL to 2 mg / mL, 0.7 mg / mL to 1.5 mg / mL, 0.7 mg / mL to 1 mg / mL, or 0.7 mg / mL to 0.9 mg / mL. In some embodiments, the surfactant in the pharmaceutical composition is at a concentration of 0.05 mg / mL to 5 mg / mL. In some embodiments, the surfactant in the pharmaceutical composition is at a concentration of 0.1 mg / mL to 3 mg / mL. In some embodiments, the surfactant in the pharmaceutical composition is at a concentration of 0.2 mg / mL to 2 mg / mL. In some embodiments, the surfactant in the pharmaceutical composition is at a concentration of approximately 0.1 mg / mL, approximately 0.2 mg / mL, approximately 0.3 mg / mL, approximately 0.4 mg / mL, approximately 0.5 mg / mL, approximately 0.6 mg / mL, approximately 0.7 mg / mL, approximately 0.8 mg / mL, approximately 0.9 mg / mL, approximately 1 mg / mL, approximately 1.1 mg / mL, approximately 1.2 mg / mL, approximately 1.3 mg / mL, approximately 1.4 mg / mL, approximately 1.5 mg / mL, about 1.6 mg / mL, about 1.7 mg / mL, about 1.8 mg / mL, approximately 1.9 mg / mL, or approximately 2 mg / mL. In some embodiments, the surfactant in the pharmaceutical composition is at a concentration of approximately 0.2 mg / mL, approximately 0.4 mg / mL, approximately 0.6 mg / mL, or approximately 0.8 mg / mL. In some embodiments, the surfactant in the pharmaceutical composition is at a concentration of approximately 0.4 mg / mL, approximately 0.6 mg / mL, Petition 870250091928, dated 08 / 10 / 2025, p. 33 / 175 26 / 155 or approximately 0.8 mg / mL. In some embodiments, the surfactant in the pharmaceutical composition is at a concentration of approximately 0.4 mg / mL or approximately 0.6 mg / mL. In some embodiments, the surfactant in the pharmaceutical composition is at a concentration of approximately 0.4 mg / mL or approximately 0.8 mg / mL. In some embodiments, the surfactant in the pharmaceutical composition is at a concentration of at least 10 mg / mL. In some embodiments, the surfactant in the pharmaceutical composition is at a concentration of at least 0.4 mg / mL, at least 0.6 mg / mL, or at least 0.8 mg / mL.

[0046] In some embodiments, the pharmaceutical composition additionally comprises a sugar. In some embodiments, the sugar is one or more sugars selected from the group consisting of sucrose, glucose, trehalose and maltose. In some embodiments, the sugar is sucrose.

[0047] In some embodiments, the sugar in the pharmaceutical composition is at a concentration of 1 mg / mL to 200 mg / mL, such as 1 mg / mL to 150 mg / mL, 1 mg / mL to 140 mg / mL, 1 mg / mL to 130 mg / mL, 1 mg / mL to 120 mg / mL, 1 mg / mL to 110 mg / mL, 1 mg / mL to 100 mg / mL, 1 mg / mL to 95 mg / mL, 1 mg / mL to 90 mg / mL, 1 mg / mL to 85 mg / mL, 1 mg / mL to 80 mg / mL, 10 mg / mL to 150 mg / mL, 10 mg / mL to 140 mg / mL, 10 mg / mL to 130 mg / mL, 10 mg / mL to 120 mg / mL, 10 mg / mL to 110 mg / mL, 10 mg / mL to 100 mg / mL, 10 mg / mL to 95 mg / mL, 10 mg / mL to 90 mg / mL, 10 mg / mL to 85 mg / mL, 10 mg / mL to 80 mg / mL, 20 mg / mL to 130 mg / mL, 20 mg / mL to 120 mg / mL, Petition 870250091928, dated 08 / 10 / 2025, page 34 / 175 27 / 155 0 mg / mL to 110 mg / mL, 20 mg / mL to 100 mg / mL, 20 mg / mL to 95 mg / mL, 20 mg / mL to 90 mg / mL, 20 mg / mL to 85 mg / mL, 20 mg / mL to mg / mL, 30 mg / mL to 120 mg / mL, 30 mg / mL to 110 mg / mL, 30 mg / mL to 100 mg / mL, 30 mg / mL to 95 mg / mL, 30 mg / mL to 90 mg / mL, mg / mL to 85 mg / mL, 30 mg / mL to 80 mg / mL, 50 mg / mL to 110 mg / mL, 50 mg / mL to 100 mg / mL, 50 mg / mL to 95 mg / mL, 50 mg / mL to 90 mg / mL, 50 mg / mL to 85 mg / mL, 50 mg / mL to 80 mg / mL, 60 mg / mL to 100 mg / mL, 60 mg / mL to 95 mg / mL, 60 mg / mL to 90 mg / mL, 60 mg / mL to 85 mg / mL, 60 mg / mL to 80 mg / mL, 70 mg / mL to 95 mg / mL, 70 mg / mL to 90 mg / mL, 70 mg / mL to 85 mg / mL, or 70 mg / mL to 80 mg / mL. In some embodiments, the sugar in the pharmaceutical composition is at a concentration of 10 mg / mL to 150 mg / mL. In some embodiments, the sugar in the pharmaceutical composition is at a concentration of 30 mg / mL to 120 mg / mL. In some embodiments, the sugar in the pharmaceutical composition is at a concentration of 50 mg / mL to 100 mg / mL.In some embodiments, the sugar in the pharmaceutical composition is at a concentration of approximately 30 mg / mL, approximately 35 mg / mL, approximately 40 mg / mL, approximately 45 mg / mL, approximately 50 mg / mL, approximately 55 mg / mL, approximately 60 mg / mL, approximately 65 mg / mL, approximately 70 mg / mL, approximately 75 mg / mL, approximately 80 mg / mL, approximately 85 mg / mL, approximately 90 mg / mL, approximately 95 mg / mL, approximately 100 mg / mL, approximately 105 mg / mL, approximately 110 mg / mL, approximately 115 mg / mL, or approximately 120 mg / mL. In some embodiments, the sugar in the composition... Petition 870250091928, dated 08 / 10 / 2025, p. 35 / 175 28 / 155 pharmaceutical composition is at a concentration of approximately 80 mg / mL. In some embodiments, the sugar in the pharmaceutical composition is at a concentration of at least 200 mg / mL. In some embodiments, the sugar in the pharmaceutical composition is at a concentration of at least 80 mg / mL.

[0048] In some embodiments, the pharmaceutical composition also optionally comprises one or more additional auxiliary materials selected from the group consisting of a polyol and a metal chelating agent. In some embodiments, the polyol is one or more polyols selected from the group consisting of glycerol, mannitol, and sorbitol. In some embodiments, the metal chelating agent is selected from the group consisting of ethylenediaminetetraacetic acid and a pharmaceutically acceptable salt thereof. In some embodiments, the polyol is mannitol. In some embodiments, the metal chelating agent is ethylenediaminetetraacetic acid.

[0049] In some embodiments, the additional auxiliary material in the pharmaceutical composition is at a concentration of 0.001% w / v 20% w / v, such as 0.005% w / v 20% w / v, 0.005% w / v 15% w / v, 0.005% w / v 10% w / v, 0.005% w / v 9% w / v, 0.005% w / v 8% w / v, 0.005% w / v 7% w / v, 0.005% w / v 6% w / v, 0.005% w / v 5% w / v, 0.005% w / v 4% w / v, 0.005% w / v 3% w / v, 0.005% w / v 2% w / v, 0.005% w / v 1% p / v, 0.005% p / v 0.1% p / v, 0.005% p / v 0.05% p / v, 0.008% Petition 870250091928, dated 08 / 10 / 2025, p. 36 / 175 29 / 155 p / va 15% p / v, 0.008% p / va 10% p / v, 0.008% p / va 9% p / v, 0.008% p / va 8% p / v, 0.008% p / va 7% p / v, 0.008% p / va 6% p / v, 0.008% p / va 5% p / v, 0.008% p / va 4% p / v, 0.008% p / va 3% p / v, 0.008% p / va 2% p / v, 0.008% p / va 1% p / v, 0.008% p / va 0.1% p / v, 0.008% p / va 0.05% p / v, 0.01% p / va 10% p / v, 0.01% p / v 9% p / v, 0.01% p / v 8% p / v, 0.01% p / v 7% p / v, 0.01% p / v 6% p / v, 0.01% p / v 5% p / v, 0.01% p / v 4% p / v, 0.01% p / v 3% p / v, 0.01% p / v 2% p / v, 0.01% p / v 1% p / v, 0.01% p / va 0.1% p / v, 0.01% p / va 0.05% p / v, 0.1% p / va 15% p / v, 0.1% p / va 10% p / v, 0.1% p / va 10% p / v, 0.1% p / va 9% p / v, 0.1% p / va 8% p / v, 0.1% p / va 7% p / v, 0.1% p / va 6% p / v, 0.1% p / va 5% p / v, 1% p / va 15% p / v, 1% p / va 10% p / v, 1% p / va 9% p / v, 1% p / va 8% p / v, 1% p / va 7% p / v, 1% p / v 6% p / v, 1% p / va 5% w / v, 0.001% w / v 5% w / v, 0.001% w / v 4% w / v, 0.001% w / v 3% w / v, 0.001% w / v 2% w / v, 0.001% w / v 1% w / v, 0.001% w / v 0.1% w / v, or 0.001% w / v 0.05% w / v. In some embodiments, the additional auxiliary material in the pharmaceutical composition is at a concentration of 0.005% w / v 15% w / v. In some embodiments, the additional auxiliary material in the pharmaceutical composition is at a concentration of 0.008% w / v 10% w / v. In some embodiments, the additional auxiliary material in the pharmaceutical composition is at a concentration of 0.01% w / v 8% w / v.

[0050] In some embodiments, mannitol in the pharmaceutical composition is at a concentration of 0.1% w / v to 15% w / v, Petition 870250091928, dated 08 / 10 / 2025, p. 37 / 175 30 / 155 such as 0.5% w / v 12% w / v, 0.5% w / v 11% w / v, 0.5% w / v 10% p / v, 0.5% p / v 9% p / v, 0.5% p / v 8% p / v, 0.5% p / v 7% p / v, 0.5% p / v 6% p / v, 0.5% p / v 5% p / v, 1% p / v 10% w / v, 1% p / v 9% p / v, 1% p / v 8% p / v, 1% p / v 7% p / v, 1% p / v 6% w / v, or 1% w / v to 5% w / v. In some embodiments, the mannitol in the pharmaceutical composition is at a concentration of 0.5% w / v to 12% w / v. In some embodiments, the mannitol in the pharmaceutical composition is at a concentration of 1% w / v to 10% w / v. In some embodiments, the mannitol in the pharmaceutical composition is at a concentration of approximately 1% w / v, approximately 2% w / v, approximately 3% w / v, approximately 4% w / v, approximately 5% w / v, approximately 6% w / v, approximately 7% w / v, approximately 8% w / v, approximately 9% w / v, or approximately 10% w / v. In some embodiments, the mannitol in the pharmaceutical composition is at a concentration of approximately 4% w / v. In some embodiments, the mannitol in the pharmaceutical composition is at a concentration of at least 15% w / v. In some embodiments, the mannitol in the pharmaceutical composition is at a concentration of at least 4% w / v.

[0051] In some embodiments, ethylenediaminetetraacetic acid in the pharmaceutical composition is at a concentration of 0.001% w / v 5% w / v, such as 0.001% w / v 4% w / v, 0.001% w / v 3% w / v, 0.001% w / v 2% w / v, 0.001% w / v 1% w / v, 0.001% w / v 0.1% w / v, 0.001% w / v 0.05% w / v, 0.005% w / v 4% w / v, 0.005% w / v 3% w / v, 0.005% w / v 2% Petition 870250091928, dated 08 / 10 / 2025, p. 38 / 175 31 / 155 w / v, 0.005% w / va 1% w / v, 0.005% w / va 0.1% w / v, or 0.005% w / va 0.05% w / v. In some embodiments, the ethylenediaminetetraacetic acid in the pharmaceutical composition is at a concentration of 0.001% w / va 1% w / v. In some embodiments, the ethylenediaminetetraacetic acid in the pharmaceutical composition is at a concentration of 0.005% w / va 0.1% w / v. In some embodiments, the ethylenediaminetetraacetic acid in the pharmaceutical composition is at a concentration of approximately 0.005% w / v, approximately 0.006% w / v, approximately 0.007% w / v, approximately 0.008% w / v, approximately 0.009% w / v, approximately 0.01% w / v, approximately 0.02% w / v, approximately 0.03% w / v, approximately 0.04% w / v, approximately 0.05% w / v, approximately 0.06% w / v, approximately 0.07% w / v, approximately 0.08% w / v, approximately 0.09% w / v, or approximately 0.1% w / v. In some embodiments, the ethylenediaminetetraacetic acid in the pharmaceutical composition is at a concentration of approximately 0.01% w / v.In some embodiments, ethylenediaminetetraacetic acid in the pharmaceutical composition is at a concentration of at least 5% w / v. In some embodiments, ethylenediaminetetraacetic acid in the pharmaceutical composition is at a concentration of at least 0.01% w / v.

[0052] In some embodiments, the buffer in the pharmaceutical composition has a pH of 3.5 to 9, such as 3.5 to 8.5, 3.5 to 8, 3.5 to 7.5, 3.5 to 7, 3.5 to 6.9, 3.5 to 6.8, 3.5 to 6.7, 3.5 to 6.6, 3.5 to 6.5, 3.5 to 6.4, 3.5 to 6.3, 3.5 to 6.2, 3.5 Petition 870250091928, dated 08 / 10 / 2025, p. 39 / 175 32 / 155 to 6.1, 3.5 to 6, 3.5 to 5.9, 3.5 to 5.8, 3.5 to 5.7, 3.5 to 5.6, 3.5 to 5.5, 4 to 5.4, 4 to 5.3, 4 to 5.2, 4 to 5.1, 4 to 5.0.4 to 8.5, 4 to 8, 4 to 7.5, 4 to 7, 4 to 6.9, 4 to 6.8, 4 to 6,7,4 to 6,6 , 4 a 6,5, 4 a 6,4, 4 a 6,3, 4 a 6,2, 4 a 6,1, 4 a 6,4 a 5,9, 4 a 5,8, 4 a 5,7, 4 a 5,6, 4 a 5,5, 4 a 5,4, 4 a5,3, a 5,2, 4 a 5,1, 4 a 5,0, 4,2 a 8, 4,2 a 7,5, 4,2 a 7,4,2 a 6,9, 4,2 a 6,8, 4,2 a 6,7, 4,2 a 6,6, 4,2 a 6,5,4,2 a 6,4 , 4,2 a 6,3, 4,2 a 6,2, 4,2 a 6,1, 4,2 a 6, 4,2 a 5,9, 4,2 a 5,8, 4,2 a 5,7, 4,2 a 5,6, 4,2 a 5,5, 4,2 a 5,4,4,2 a 5,3, 4,2 a 5,2, 4,2 a 5,1, 4,2 a 5,0, 4,5 a 8, 4,5 a 7,5, 4,5 a 7, 4,5 a 6,9, 4,5 a 6,8, 4,5 a 6,7, 4,5 a 6,6,4,5 a 6,5 , 4,5 a 6,4, 4,5 a 6,3, 4,5 a 6,2, 4,5 a 6,1, 4,5 a 6, 4,5 a 5,9, 4,5 a 5,8, 4,5 a 5,7, 4,5 a 5,6, 4,5 a 5,5,4,5 a 5,4, 4,5 a 5,3, 4,5 a 5,2, 4,5 a 5,1, 4,5 a 5,0,4,6 a 7,5 , 4,6 a 7, 4,6 a 6,9, 4,6 a 6,8, 4,6 a 6,7, 4,6 a 6,6, 4,6 a 6,5, 4,6 a 6,4, 4,6 a 6,3, 4,6 a 6,2, 4,6 a 6,1,4,6 a 6, 4,6 a 5,9, 4,6 a 5,8, 4,6 a 5,7, 4,6 a 5,6, 4,6 a5,5, 4.6 to 5.4, 4.6 to 5.3, 4.6 to 5.2, 4.6 to 5.1, 4.6 to 5.0,4.8 to 7, 4.8 to 6.9, 4.8 to 6.8, 4.8 to 6.7, 4.8 to 6.6, 4.8 to 6.5, 4.8 to 6.4, 4.8 to 6.3, 4.8 to 6.2, 4.8 to 6.1, 4.8 to 6.4.8 to R 5.6, or 5 to 5.5. In some embodiments, the buffer in the pharmaceutical composition has a pH of 3.5 to 7. In some embodiments, the buffer in the pharmaceutical composition has Petition 870250091928, dated 08 / 10 / 2025, p. 40 / 175 33 / 155 a pH of 4.0 to 6.5. In some embodiments, the buffer in the pharmaceutical composition has a pH of 4.2 to 6.2. In some embodiments, the buffer in the pharmaceutical composition has a pH of 4.5 to 6.0. In some embodiments, the buffer in the pharmaceutical composition has a pH of approximately 4.5, approximately 4.6, approximately 4.7, approximately 4.8, approximately 4.9, approximately 5.0, approximately 5.1, approximately 5.2, approximately 5.3, approximately 5.4, approximately 5.5, approximately 5.6, approximately 5.7, approximately 5.8, approximately 5.9, approximately 6.0, approximately 6.1, approximately 6.2, approximately 6.3, approximately 6.4, or approximately 6.5. In some embodiments, the buffer in the pharmaceutical composition has a pH of approximately 5.0, approximately 5.2, approximately 5.3, approximately 5.5, approximately 5.6, approximately 5.8, or approximately 6.0. In some embodiments, the buffer in the pharmaceutical composition has a pH of approximately 5.0, approximately 5.2, approximately 5.3, or approximately 5.5.

[0053] In some embodiments, the pharmaceutical composition additionally comprises a pH adjuster, such as sodium hydroxide and / or hydrochloric acid.

[0054] In some embodiments, there is a difference of at most ±0.5 between the pH of the pharmaceutical composition and the pH of the buffer that the pharmaceutical composition comprises. In some embodiments, the pharmaceutical composition has a pH of 3.5 to 9, such as 3.5 to 8.5, 3.5 to 8, 3.5 to 7.5, 3.5 to 7, 3.5 to 6.9, 3.5 to 6.8, 3.5 to 6.7, 3.5 to 6.6, 3.5 to 6.5, 3.5 to 6.4, 3.5 to 6.3, 3.5 to 6.2, 3.5 to 6.1, 3.5 to 6, 3.5 to 5.9, 3.5 to 5.8, Petition 870250091928, dated 08 / 10 / 2025, p. 41 / 175 34 / 155 3.5 to 5.7, 3.5 to 5.6, 3.5 to 5.5, 4 to 5.4, 4 to 5.3, 4 to 5.2, 5.1, 4 to 5.0, 4 to 8.5, 4 to 8, 4 to 7.5, 4 to 7, 4 to 6.9, 6.8, 4 to 6.7, 4 to 6.6, 4 to 6.5, 4 to 6.4, 4 to 6,3,4 to 6.2, 4 to 6.1, 4 to 6, 4 to 5.9, 4 to 5.8, 4 to 5.7, 4 to 5.6, 4 to 5.5, 4 to 5.4, 4 to 5.3, 4 to 5.2, 4 to 5.1, 4 to 5.0, 4.2 to 8, 4.2 and 7.5, 4.2 and 7, 4.2 and 6.9, 4.2 and 6.8, 4.2 and 6.7, 4.2 and 6.6, 4.2 and 6.5, 4.2 and 6.4, 4.2 and 6.3, 4.2 and 6.2, 4.2 and 6.1, 4.2 and 6, 4.2 and 5.9, 4.2 and 5.8, 4.2 and 5.7, 4.2 and 5.6, 4.2 and 5.5, 4.2 and 5.4, 4.2 and 5.3, 4.2 and 5.2, 4.2 and 5.1, 4.2 and 5.0, 4.5 and 8, 4.5 and 7.5, 4.5 and 7, 4.5 and 6.9, 4.5 and 6.8, 4.5 and 6.7, 4.5 and 6.6, 4.5 and 6.5, 4.5 and 6.4, 4.5 and 6.3, 4.5 and 6.2, 4.5 and 6.1, 4.5 and 6, 4.5 and 5.9, 4.5 and 5.8, 4.5 and 5.7, 4.5a 5.6, 4.5 and 5.5, 4.5 and 5.4, 4.5 and 5.3, 4.5 and 5.2, 4.5 and 5.1, 4.5 and 5.0, 4.6 and 7.5, 4.6 and 7, 4.6 and 6.9, 4.6 and 6.8, 4.6 and 6.7, 4.6 and 6.6, 4.6 and 6.5, 4.6 and 6.4, 4.6 and 6.3, 4.6 and 6.2, 4.6 and 6.1, 4.6 and 6, 4.6 and 5.9, 4.6 and 5.8, 4.6 and 5.7, 4.6a 5.6, 4.6 and 5.5, 4.6 and 5.4, 4.6 and 5.3, 4.6 and 5.2, 4.6 and 5.1, 4.6 and 5.0, 4.8 and 7, 4.8 and 6.9, 4.8 and 6.8, 4.8 and 6.7, 4.8a 6.6, 4.8 and 6.5, 4.8 and 6.4, 4.8 and 6.3, 4.8 and 6.2, 4.8 and 6.1, 4.8 to 6, 4.8 to 5.9, 4.8 to 5.8, 4.8 to 5.7, 4.8 to 5.6,4.8 to 5.5, 5 to 6.5, 5 to 6.4, 5 to 6.3, 5 to 6.2, 5 to 6.1, 5 to 6.5 to 5.9, 5 to 5.8, 5 to 5.7, 5 to 5.6, or 5 to 5.5. In some embodiments, the pharmaceutical composition has a pH of 3.5 to 7. In some embodiments, the pharmaceutical composition has a pH of 4.0 to 6.5. In some embodiments, the composition Petition 870250091928, dated 08 / 10 / 2025, p. 42 / 175 35 / 155 pharmaceutical composition has a pH of 4.2 to 6.2. In some embodiments, the pharmaceutical composition has a pH of 4.5 to 6.0. In some embodiments, the pharmaceutical composition has a pH of approximately 4.5, approximately 4.6, approximately 4.7, approximately 4.8, approximately 4.9, approximately 5.0, approximately 5.1, approximately 5.2, approximately 5.3, approximately 5.4, approximately 5.5, approximately 5.6, approximately 5.7, approximately 5.8, approximately 5.9, approximately 5.5, approximately 6.0, approximately 6.1, approximately 6.2, approximately 6.3, approximately 6.4, or approximately 6.5. In some embodiments, the pharmaceutical composition has a pH of approximately 5.0, approximately 5.2, approximately 5.3, approximately 5.5, approximately 5.6, approximately 5.8, or approximately 6.0. In some embodiments, the buffer in the pharmaceutical composition has a pH of approximately 5.0, approximately 5.2, approximately 5.3, or approximately 5.5.

[0055] In some embodiments, the present description provides a pharmaceutical composition comprising a FAP / CD40 binding molecule (for example, Ab10-A297V3-4 comprising heavy and light chain sequences of SEQ ID NOs: 22 and 20, respectively), comprising any of the following groups 1) to 4): 1) a FAP / CD40 binding molecule; a histidine salt buffer, such as a histidine hydrochloride buffer, a histidine acetate buffer, a histidine-histidine hydrochloride buffer, or a histidine-acetic acid buffer; 2) a FAP / CD40 binding molecule; Petition 870250091928, dated 08 / 10 / 2025, page 43 / 175 36 / 155 a histidine salt buffer; a polysorbate, such as polysorbate 80; sucrose; wherein, optionally, the composition additionally includes mannitol or ethylenediaminetetraacetic acid; 3) a FAP / CD40 binding molecule; a histidine hydrochloride buffer, such as a histidine-histidine hydrochloride buffer; a polysorbate, such as polysorbate 80; sucrose; wherein, optionally, the composition additionally includes mannitol or ethylenediaminetetraacetic acid; 4) a FAP / CD40 binding molecule; a histidine acetate buffer, such as a histidine-acetic acid buffer; a polysorbate, such as polysorbate 80; sucrose; wherein, optionally, the composition additionally includes mannitol or ethylenediaminetetraacetic acid;

[0056] In some embodiments, the present description provides a pharmaceutical composition comprising a FAP / CD40 binding molecule (for example, Ab10-A297V3-4 comprising heavy and light chain sequences of SEQ ID NOs: 22 and 20, respectively), which is any of the following groups 1) to 4): Petition 870250091928, dated 08 / 10 / 2025, page 44 / 175 37 / 155 1) a FAP / CD40 binding molecule; a histidine salt buffer; a polysorbate; sucrose and water for injection; 2) a FAP / CD40 binding molecule; a histidine hydrochloride buffer; a polysorbate; sucrose and water for injection; 3) a FAP / CD40 binding molecule; a histidine-histidine hydrochloride buffer; a polysorbate; sucrose and water for injection; 4) a FAP / CD40 binding molecule; a histidine-acetic acid buffer; a polysorbate; sucrose and water for injection; 5) the pharmaceutical composition according to any one of items 1) to 4) described above, wherein the polysorbate is polysorbate 80.

[0057] In some embodiments, the present description provides a pharmaceutical composition comprising a FAP / CD40 binding molecule (for example, Ab10-A297V3-4 comprising heavy and light chain sequences of SEQ ID NOs: 22 and 20, respectively), comprising any of the following groups 1) to 6): 1) 0.01 mg / mL to 500 mg / mL of FAP / CD40 binding molecule; 0.1 mM to 50 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer, a histidine acetate buffer, a histidine-hydrochloride buffer Petition 870250091928, dated 08 / 10 / 2025, page 45 / 175 38 / 155 histidine or a histidine-acetic acid buffer; 0.01 mg / mL to 10 mg / mL polysorbate; mg / mL to 200 mg / mL of sucrose; wherein, optionally, the pharmaceutical composition additionally comprises 0.1% w / v 15% w / v of mannitol or 0.001% w / v 5% w / v of ethylenediaminetetraacetic acid; and the pharmaceutical composition has a pH of 3.5 to 7; 2) 0.1 mg / mL to 400 mg / mL of binding molecule to FAP / CD40; 0.5 mM to 40 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer, a histidine acetate buffer, a histidine-histidine hydrochloride buffer, or a histidine-acetic acid buffer; 0.05 mg / mL to 5 mg / mL polysorbate; mg / mL to 150 mg / mL of sucrose; wherein, optionally, the pharmaceutical composition additionally comprises 0.5% w / v 12% w / v of mannitol or 0.001% w / v 1% w / v of ethylenediaminetetraacetic acid; and the pharmaceutical composition has a pH of 4 to 6.5; 3) 0.5 mg / mL to 200 mg / mL of binding molecule to FAP / CD40; 30 mM to 30 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer, a histidine acetate buffer, a histidine-histidine hydrochloride buffer, or a histidine-acetic acid buffer; Petition 870250091928, dated 08 / 10 / 2025, page 46 / 175 39 / 155 0.1 mg / mL to 3 mg / mL polysorbate; mg / mL to 120 mg / mL of sucrose; wherein, optionally, the pharmaceutical composition additionally comprises 1% w / v 10% w / v of mannitol or 0.005% w / v 0.1% w / v of ethylenediaminetetraacetic acid; and the pharmaceutical composition has a pH of 4.2 to 6.2; 4) 1 mg / mL to 150 mg / mL of binding molecule to FAP / CD40; 20 mM to 20 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer, a histidine acetate buffer, a histidine-histidine hydrochloride buffer, or a histidine-acetic acid buffer; 0.2 mg / mL to 2 mg / mL polysorbate; mg / mL to 100 mg / mL of sucrose; wherein, optionally, the pharmaceutical composition additionally comprises 1% w / v 10% w / v of mannitol or 0.005% w / v 0.1% w / v of ethylenediaminetetraacetic acid; and the pharmaceutical composition has a pH of 4.5 to 6; 5) 1 mg / mL to 40 mg / mL of FAP / CD40 binding molecule; mM to 20 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer; 0.2 mg / mL to 2 mg / mL polysorbate; mg / mL to 100 mg / mL of sucrose; wherein, optionally, the pharmaceutical composition additionally comprises 1% w / v or 10% w / v of mannitol or Petition 870250091928, dated 08 / 10 / 2025, page 47 / 175 40 / 155 0.005% w / v and 0.1% w / v ethylenediaminetetraacetic acid; and the pharmaceutical composition has a pH of 4.5 to 6; 6) 50 mg / mL to 150 mg / mL of binding molecule to FAP / CD40; mM to 20 mM of histidine acetate buffer, such as a histidine-acetic acid buffer; 0.2 mg / mL to 2 mg / mL polysorbate; mg / mL to 100 mg / mL of sucrose; wherein, optionally, the pharmaceutical composition additionally comprises 1% w / v 10% w / v of mannitol or 0.005% w / v 0.1% w / v of ethylenediaminetetraacetic acid; and the pharmaceutical composition has a pH of 4.5 to 6; 7) the pharmaceutical composition according to any of items 1) to 6) described above, wherein the pharmaceutical composition additionally includes water for injection; 8) the pharmaceutical composition according to any of items 1) to 6) described above, wherein the polysorbate is polysorbate 80.

[0058] In some embodiments, the present description provides a pharmaceutical composition comprising a FAP / CD40 binding molecule (for example, Ab10-A297V3-4 comprising heavy and light chain sequences of SEQ ID NOs: 22 and 20, respectively), comprising or being any of the following groups 1) to 25): 1) 1 to 150 mg / mL of FAP / CD40 binding molecule; Petition 870250091928, dated 08 / 10 / 2025, page 48 / 175 41 / 155 approximately 10 mM of histidine hydrochloride buffer or histidine acetate buffer, such as a histidine-histidine hydrochloride buffer or a histidine-acetic acid buffer; 0.2 mg / mL to 2 mg / mL polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of 4.5 to 6; 2) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of the FAP / CD40 binding molecule, such as approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; where the pharmaceutical composition has a pH of approximately 5.5; 3) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of the FAP / CD40 binding molecule, such as approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL of the FAP / CD40 binding molecule; Petition 870250091928, dated 08 / 10 / 2025, page 49 / 175 42 / 155 approximately 10 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5.3; 4) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of the FAP / CD40 binding molecule, such as approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5.2; 5) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of the FAP / CD40 binding molecule, such as approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine hydrochloride buffer, such Petition 870250091928, dated 08 / 10 / 2025, page 50 / 175 43 / 155 as a histidine buffer - histidine hydrochloride; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5; 6) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of the FAP / CD40 binding molecule, such as approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer; approximately 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; where the pharmaceutical composition has a pH of approximately 5.5; 7) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of the FAP / CD40 binding molecule, such as approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer; Petition 870250091928, dated 08 / 10 / 2025, page 51 / 175 44 / 155 approximately 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5.3; 8) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of the FAP / CD40 binding molecule, such as approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer; approximately 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5.2; 9) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of the FAP / CD40 binding molecule, such as approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer; approximately 0.6 mg / mL of polysorbate 80; Petition 870250091928, dated 08 / 10 / 2025, page 52 / 175 45 / 155 approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5; 10) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine-acetic acid buffer; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5; 11) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine-acetic acid buffer; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5.2; 12) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL of the FAP / CD40 binding molecule; Petition 870250091928, dated 08 / 10 / 2025, page 53 / 175 46 / 155 approximately 10 mM histidine-acetic acid buffer; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5.3; 13) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine-acetic acid buffer; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; where the pharmaceutical composition has a pH of approximately 5.5; 14) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine-acetic acid buffer; approximately 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5; 15) about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, about 25 mg / mL, about 50 mg / mL, about 80 Petition 870250091928, dated 08 / 10 / 2025, page 54 / 175 47 / 155 mg / mL or approximately 100 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine-acetic acid buffer; approximately 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5.2; 16) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine-acetic acid buffer; approximately 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5.3; 17) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine-acetic acid buffer; approximately 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; where the pharmaceutical composition has a pH of approximately 5.5; Petition 870250091928, dated 08 / 10 / 2025, page 55 / 175 48 / 155 18) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine-acetic acid buffer; approximately 0.8 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5; 19) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine-acetic acid buffer; approximately 0.8 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of approximately 5.2; 20) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine-acetic acid buffer; approximately 0.8 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; Petition 870250091928, dated 08 / 10 / 2025, page 56 / 175 49 / 155 in which the pharmaceutical composition has a pH of approximately 5.3; 21) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine-acetic acid buffer; approximately 0.8 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; where the pharmaceutical composition has a pH of approximately 5.5; 22) 1 to 45 mg / mL of the FAP / CD40 binding molecule, such as approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine hydrochloride buffer, such as a histidine hydrochloride buffer; 0.2 mg / mL to 2 mg / mL polysorbate 80; approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of 4.5 to 6; 23) 50 to 120 mg / mL of the FAP / CD40 binding molecule, such as approximately 50 mg / mL, approximately 80 mg / mL, or approximately 100 mg / mL of the FAP / CD40 binding molecule; approximately 10 mM of histidine acetate buffer, such as a histidine-acetic acid buffer; 0.2 mg / mL to 2 mg / mL polysorbate 80; Petition 870250091928, dated 08 / 10 / 2025, page 57 / 175 50 / 155 approximately 80 mg / mL of sucrose; in which the pharmaceutical composition has a pH of 4.5 to 6; 24) the pharmaceutical compositions from 1) to 23), wherein the pharmaceutical compositions additionally comprise 4% w / v of mannitol or 0.01% w / v of ethylenediaminetetraacetic acid; 25) the pharmaceutical compositions from 1) to 24) with a final volume of 1 mL, the volume being completed to 1 mL with the use of water for injection, when necessary.

[0059] In another aspect, the present description provides a pharmaceutical composition comprising a FAP / CD40 binding molecule at a low concentration (for example, 1 to 45 mg / mL, about 5 mg / mL, about 10 mg / mL, about 20 mg / mL or about 25 mg / mL) or a method of preparation thereof, and the pharmaceutical composition is obtained by diluting a pharmaceutical composition comprising a FAP / CD40 binding molecule at a high concentration (for example, 50 to 120 mg / mL, about 50 mg / mL, about 80 mg / mL or about 100 mg / mL) with a diluent. In some embodiments, the diluent includes, but is not limited to, water, normal saline solution and a glucose solution.

[0060] The pharmaceutical composition described herein already has sufficient stability to be prepared as a medicament and may be stable after long-term storage.

[0061] In some embodiments, the pharmaceutical composition remains stable at 2-8 °C for at least 3 Petition 870250091928, dated 08 / 10 / 2025, p. 58 / 175 51 / 155 months, at least 6 months, at least 12 months, at least 18 months, at least 24 months or at least 36 months. In some embodiments, the pharmaceutical composition remains stable at 25 °C for at least 3 months, at least 6 months, at least 12 months, at least 18 months or at least 24 months. In some embodiments, the pharmaceutical composition remains stable at 40 °C for at least 7 days, at least 14 days, at least 28 days, at least 1 month, at least 3 months, at least 6 months, at least 12 months, at least 18 months or at least 24 months.

[0062] The present description provides a method for preparing the pharmaceutical composition mentioned above, which comprises the step of dissolving the FAP / CD40 binding molecule.

[0063] The pharmaceutical composition of the present description may be further prepared in a lyophilized formulation to facilitate drug administration.

[0064] In certain embodiments, the pharmaceutical composition is provided in accordance with any of the above items, wherein the pharmaceutical composition is a liquid formulation. In some embodiments, the solvent of the liquid formulation is water, normal saline solution or a glucose solution.

[0065] The present description also provides a freeze-dried formulation, wherein the freeze-dried formulation is capable of forming the pharmaceutical composition according to any of Petition 870250091928, dated 08 / 10 / 2025, page 59 / 175 52 / 155 of the above items after reconstitution.

[0066] The present description also provides a freeze-dried formulation, wherein the freeze-dried formulation is obtained by freeze-drying the pharmaceutical composition in accordance with any of the above items.

[0067] The present description provides a reconstituted solution, wherein the reconstituted solution is prepared by reconstituting the lyophilized formulation mentioned above. In certain embodiments, the reconstituted solution is selected from the group consisting of, but not limited to, water for injection, normal saline solution or a glucose solution.

[0068] This description also provides a manufactured article comprising a container, wherein the container contains the pharmaceutical composition mentioned above, the lyophilized formulation mentioned above, or the reconstituted solution mentioned above. In certain embodiments, the container is a tubular injection vial made of neutral borosilicate glass. In certain embodiments, the manufactured article comprises a package insert.

[0069] The present description also provides the pharmaceutical composition, the lyophilized formulation or the reconstituted solution of the lyophilized formulation as a medicament for use in the treatment or improvement of a disease or disorder. Petition 870250091928, dated 08 / 10 / 2025, page 60 / 175 53 / 155 A method for treating diseases and pharmaceutical use.

[0070] In some embodiments, the present description provides a method for preventing and / or treating a cancer or tumor, comprising administering to a patient or individual a prophylactic and / or therapeutically effective amount of the aforementioned pharmaceutical composition, lyophilized formulation or reconstituted solution of the lyophilized formulation to inhibit the growth of tumor cells in the patient or individual. In some specific embodiments, the cancer is preferably, but not limited to, a cancer responsive to immunotherapy.

[0071] In the above method or use, non-limiting examples of cancer or tumor include lung cancer, ovarian cancer, colon cancer, rectal cancer, melanoma (e.g., metastatic malignant melanoma), renal cancer, bladder cancer, breast cancer, liver cancer, lymphoma, hematologic malignancies, head and neck cancer, glioma, gastric cancer, nasopharyngeal carcinoma, laryngeal cancer, cervical cancer, uterine body tumors, and osteosarcoma. Examples of other cancers that may be treated using the method of the present description include: bone cancer, pancreatic cancer, skin cancer, prostate cancer, malignant melanoma of the skin or intraocular region, uterine cancer, anal cancer, testicular cancer, fallopian tube cancer, endometrial cancer, vaginal cancer, vulvar cancer, Petition 870250091928, dated 08 / 10 / 2025, page 61 / 175 54 / 155 Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small bowel cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia, including acute myeloid leukemia, chronic myeloid leukemia, acute lymphocytic leukemia and chronic lymphocytic leukemia, childhood solid tumors, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal pelvis cancer, central nervous system (CNS) tumor, primary CNS lymphoma, tumor angiogenesis, spinal tumor, brainstem neuroglioma, pituitary adenoma, Kaposi's sarcoma, epidermal carcinoma, squamous cell carcinoma, T-cell lymphoma and environment-induced cancers, including asbestos-induced cancers and combinations of cancers. In some cases, the tumor or cancer described above is metastatic and / or advanced.

[0072] In addition, the present description also provides a method for preventing and / or treating an infectious disease in an individual or patient, comprising administering to the individual or patient the aforementioned pharmaceutical composition, lyophilized formulation or reconstituted solution of the lyophilized formulation, so that the infectious disease in the individual is prevented and / or treated. Similar to the use for cancer or tumor described above, individual use or in combination with vaccines may be undertaken for Petition 870250091928, dated 08 / 10 / 2025, page 62 / 175 55 / 155 stimulate immune responses to pathogens, toxins, and autoantigens. Examples of pathogens to which the treatment method may be particularly applicable include pathogens for which no effective vaccine is currently available or pathogens for which conventional vaccines are not fully effective. Pathogens include, among others, HIV, hepatitis viruses (A, B, and C), influenza viruses, herpes viruses, giardia, malaria, leishmania, Staphylococcus aureus, and Pseudomonas aeruginosa.

[0073] The pharmaceutical composition comprising the FAP / CD40 binding molecule of the present description can be used to treat a patient requiring such treatment by parenteral administration. For parenteral routes of administration, subcutaneous injection, intramuscular injection or intravenous injection may be selected. BRIEF DESCRIPTION OF THE DRAWINGS

[0074] FIG. 1 shows the binding of anti-FAP antibodies Ab9, Ab10, Ab14 and Ab15 to the human FAP cell surface, with 28H1 as a positive control and the IgG1 isotype as a negative control.

[0075] FIG. 2 shows the binding of anti-FAP antibodies Ab9, Ab10, Ab14 and Ab15 to the mouse FAP cell surface, with 28H1 as a positive control and the IgG1 isotype as a negative control.

[0076] FIG. 3 shows the binding of anti-antibodies Petition 870250091928, dated 08 / 10 / 2025, page 63 / 175 56 / 155 CD40 A12 and A297 to human cell surface CD40 Raji, with 9E5-SELFNS as a positive control and IgG1 as a negative control.

[0077] FIG. 4 shows the binding of antiCD40 A12 and A297 antibodies to human cell surface CD40 HEK293, with 9E5-SELFNS as a positive control and IgG1 as a negative control; the ordinate represents the percentage of agonist activity relative to 200 nM of 9E5-SELFNS.

[0078] FIG. 5 shows the affinities of humanized anti-CD40 antibodies A297_V1, A297_V2, A297_V3 and A297_V4 for HEK293 cells overexpressing human CD40, as measured by flow cytometry, with 9E5-SELFNS as a positive control and hIgG as a negative control.

[0079] FIG. 6 shows FcyRIIb-mediated activation of HEK-Blue™CD40L cells by humanized anti-CD40 antibodies A297_V1, A297_V2, A297_V3 and A297_V4, with 9E5 SELFNS as a positive control and hIgG as a negative control.

[0080] FIG. 7 shows a schematic diagram of the structures of bispecific anti-FAP / CD40 antibodies.

[0081] FIG. 8A shows the affinities of the bispecific anti-FAP / CD40 antibodies Ab10-A12V2-2, Ab10-A12V2-4, Ab10-A297V3-2, and Ab10-A297V3-4 for a stably transfected CHOK1 / FAP strain highly expressing the human FAP antigen, as measured by flow cytometry. FIGS. 8B and 8C show the antibody affinities. Petition 870250091928, dated 08 / 10 / 2025, page 64 / 175 57 / 155 bispecific anti-FAP / CD40 antibodies mentioned above for stably transfected CHOK1 / FAP strains, strongly expressing murine FAP antigen and cynomolgus monkey FAP antigen, respectively, as measured by flow cytometry.

[0082] FIG. 9A shows the affinities of the bispecific anti-FAP / CD40 antibodies Ab10-A12V2-2, Ab10-A12V2-4, Ab10-A297V3-2, and Ab10-A297V3-4 for a stably transfected HEK-Blue™CD40L strain highly expressing the human CD40 antigen, as measured by flow cytometry. FIG. 9B shows the affinities of the aforementioned bispecific anti-FAP / CD40 antibodies for HEK293 cells strongly expressing the cynomolgus monkey CD40 antigen, as measured by flow cytometry.

[0083] FIG. 10 shows the affinities of the bispecific anti-FAP / CD40 antibodies Ab10-A297V3-2 and Ab10-A297V3-4 for CD40 on immature human dendritic cells, as measured by flow cytometry.

[0084] FIG. 11 shows the maturation-promoting effect of the bispecific antiFAP / CD40 antibodies Ab10-A12V2-2, Ab10-A12V2-4, Ab10-A297V3-2 and Ab10A297V3-4 in a concentration gradient in the absence of crosslinking of stably transfected CHOK1 / FAP cells, with LPC, hIgG1 and the vehicle as controls.

[0085] FIG. 12 shows the ripening-promoting effect Petition 870250091928, dated 08 / 10 / 2025, page 65 / 175 58 / 155 of dendritic cells of the bispecific antibodies antiFAP / CD40 Ab10-A12V2-2, Ab10-A12V2-4, Ab10-A297V3-2 and Ab10A297V3-4 from a concentration gradient in the presence of cross-linking of stably transfected CHOK1 / FAP cells, with LPC, hIgG1 and vehicle as controls.

[0086] FIG. 13A shows tumor growth curves of mice after single-dose injections of the antibody Ab10 A297V3-2 or Ab10-A297V3-4 in humanized hCD40 mice bearing mFAP-MC38 tumors. FIG. 13B shows corresponding activation of peripheral blood B cells in mice. FIG. 13C shows curves of corresponding changes in body weight in mice. FIG. 13D shows corresponding changes in platelet count in mice. FIG. 13E shows the corresponding influence on liver function in mice.

[0087] FIG. 14A shows tumor growth curves of mice after multiple dose injections of the antibody Ab10-A297V3-2 or Ab10-A297V3-4 in humanized hCD40 mice bearing mFAP-MC38 tumors. FIG. 14B shows corresponding activation of peripheral blood B cells in the mice. FIG. 14C shows the curves of the corresponding changes in body weight of the mice. FIG. 14D shows the corresponding changes in platelet count of the mice. FIG. 14E shows the corresponding influence on liver function in the mice. Petition 870250091928, dated 08 / 10 / 2025, p. 66 / 175 59 / 155

[0088] In FIGS. 13A to 14E, P values ​​were calculated relative to the hIgG1 control, unless otherwise indicated. The statistical method used was Student's t-test. ns represents no significant difference. * represents p < 0.05, ** represents p < 0.01, *** represents p < 0.001 and **** represents p < 0.0001. DETAILED DESCRIPTION I. Terminology

[0089] To facilitate understanding of the present description, certain technical and scientific terms are specifically defined below. Unless explicitly defined in this document, all other technical and scientific terms used in this document have the meanings generally understood by those skilled in the art to which the present description pertains.

[0090] The three-letter and single-letter codes for amino acids in the present description are as described in J. biol. chem., 243, p3558 (1968).

[0091] CD40 and CD40 antigen refer to a glycoprotein of approximately 48 kDa expressed on the surface of normal and neoplastic B cells, which acts as a receptor for signals involved in cell proliferation and differentiation (Ledbetter et al., 1987, J. Immunol. 138:788-785). A cell that endogenously expresses CD40 is any cell distinguished by the surface expression of Petition 870250091928, dated 08 / 10 / 2025, page 67 / 175 60 / 155 CD40, including, among others, normal and neoplastic B cells, interdigitating cells, basal epithelial cells, carcinoma cells, macrophages, endothelial cells, follicular dendritic cells, tonsil cells, and bone marrow-derived plasma cells. In the present description, CD40 refers to any native CD40 derived from any vertebrate, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise indicated. A cDNA molecule encoding CD40 was isolated from a library prepared from the Raji Burkitt lymphoma cell line (Stamenkovic et al., 1989, EMBO J. 8:1403). Sequence information can be found in Table 8 of the present description.In the present description, "CD40" encompasses unprocessed full-length CD40, as well as any form of CD40 resulting from processing in a cell, and also includes natural variants of CD40, such as splicing variants or allelic variants. In one embodiment, the CD40-binding molecule of the present description is capable of specifically binding to human, mouse, and / or cynomolgus monkey CD40.

[0092] Fibroblast activating protein (FAP)” and FAP antigen”, also known as prolyl endopeptidase FAP or seprase (EC 3.4.21), refer to any naturally occurring FAP derived from any vertebrate, including mammals, such as Petition 870250091928, dated 08 / 10 / 2025, p. 68 / 175 61 / 155 primates (e.g., humans), non-human primates (e.g., cynomolgus monkeys), and rodents (e.g., mice and rats), unless otherwise indicated. In the present description, FAP encompasses unprocessed full-length FAP as well as any form of FAP resulting from processing in a cell, and further includes natural variants of FAP, such as splicing variants or allelic variants. In one embodiment, the FAP-binding molecule of the present description is capable of specifically binding to human, mouse, and / or cynomolgus monkey FAP. The amino acid sequence of human FAP is shown under UniProt accession number Q12884 (www.uniprot.org) (version 149, SEQ ID NO: 2), or NCBI (www.ncbi.nlm.nih.gov / ) RefSeq NP_004451.2, or GeneBank accession number AAC51668. The extracellular domain (ECD) of human FAP extends from amino acid position 26 to 760.Amino acid sequences, such as the His-labeled human FAP ECD, are shown in Table 2 of this description. The amino acid sequence of murine FAP is shown under UniProt accession number P97321 (version 126, SEQ ID NO: 143), or NCBI RefSeq NP_032012.1. The mouse FAP extracellular domain (ECD) extends from amino acid position 26 to 761. In some embodiments, the FAP-binding molecule of this description binds to the FAP extracellular domain.

[0093] Antibody is used in the broadest sense and Petition 870250091928, dated 08 / 10 / 2025, p. 69 / 175 62 / 155 covers a variety of antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies), full-length antibodies, and antibody fragments (or antigen-binding fragments or antigen-binding fractions), provided they exhibit the desired antigen-binding activity. An antibody refers to an immunoglobulin, which is a four-peptide chain structure formed by the linkage of two identical heavy chains and two identical light chains by interchain disulfide bonds. The constant regions of the heavy chain of immunoglobulins differ in their amino acid composition and arrangement and therefore in their antigenicity.Consequently, immunoglobulins can be divided into five classes, also called immunoglobulin isotypes, namely IgM, IgD, IgG, IgA, and IgE, their corresponding heavy chains being μ chain, δ chain, γ chain, α chain, and ε chain, respectively. Igs of the same class can be divided into different subclasses according to differences in the amino acid composition of the hinge regions and the number and positions of the disulfide bonds of the heavy chains; for example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4. The light chains are divided into κ or λ chains according to differences in... Petition 870250091928, dated 08 / 10 / 2025, p. 70 / 175 63 / 155 constant regions. Each of the five Ig classes can have either a κ chain or a λ chain. In the antibody heavy and light chains, the sequences of approximately 110 amino acids near the N-terminus vary considerably and are therefore referred to as variable regions (V regions); the remaining amino acid sequences near the C-terminus are relatively stable and are therefore referred to as constant regions (C regions). The variable regions comprise 3 hypervariable regions (CDRs) and 4 structural regions (FRs) with relatively conservative sequences. The 3 hypervariable regions determine the antibody specificity and are therefore also known as complementarity-determining regions (CDRs). Each of the variable regions of the light chain (VLs) and variable regions of the heavy chain (VHs) consists of 3 CDRs and 4 FRs arranged from the amino terminal to the carboxyl terminal as follows: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.The 3 CDRs of the light chain refer to LCDR1, LCDR2, and LCDR3, and the 3 CDRs of the heavy chain refer to HCDR1, HCDR2, and HCDR3.

[0094] “Bispecific antibody” encompasses antibodies (including antibodies or antigen-binding fragments thereof, such as single-chain antibodies) capable of specifically binding to two different antigens or at least two different antigenic epitopes of the same antigen. Petition 870250091928, dated 08 / 10 / 2025, p. 71 / 175 64 / 155 antigen. Typical structural models of bispecific antibodies include bispecific antibodies such as KiH, CrossMAb, Triomab quadroma, FcΔAdp, ART-Ig, BiMAb, Biclonics, BEAT, DuoBody, Azymetric, XmAb, 2:1 TCBs, 1FabIgG TDB, FynomAb, two-in-one / DAF, scFv-Fab-IgG, DART-Fc, LPDART, 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 (see Aran F. Labrijn et al., Nature Reviews Drug Discovery, volume 18, pages 585-608). (2019); Chen S1 et al., J Immunol Res., February. 11, 2019; 2019: 4516041).

[0095] For the determination or definition of CDRs, the deterministic representation of CDRs and the identification of residues comprising antigen-binding sites of an antibody can be performed by solving the antibody structure and / or solving the structure of the antibody-ligand complex. This can be performed by any of a variety of techniques known to those skilled in the art, such as X-ray crystallography. A variety of analytical methods can be used to identify CDRs, including, but not limited to, the Kabat numbering scheme, the Chothia numbering scheme, the AbM numbering scheme, the IMGT numbering scheme, contact definition, and conformational definition.

[0096] Kabat's numbering scheme is a standard for Petition 870250091928, dated 08 / 10 / 2025, page 72 / 175 The 65 / 155 residue numbering scheme in antibodies is generally used to identify CDRs (see, for example, Johnson & Wu, 2000, Nucleic Acids Res., 28: 214-8). Chothia's numbering scheme is similar to Kabat's, except that it takes into account the positions of certain structural loop regions (see, for example, Chothia et al., 1986, J. Mol. Biol., 196: 901-17; Chothia et al., 1989, Nature, 342: 877- 83). The AbM numbering scheme adopts a set of integrated computer programs for modeling antibody structures manufactured by the Oxford Molecular Group (see, for example, Martin et al., 1989, Proc Natl Acad Sci (USA), 86: 9268-9272; “AbMTM, A Computer Program for Modeling Variable Regions of Antibodies”, Oxford, UK; Oxford Molecular, Ltd.). The AbM numbering scheme adopts a combination of a knowledge database and the de novo method to model the tertiary structure of antibodies from basic sequences (see those described in Samudrala et al., 1999, “Ab Initio Protein Structure”). Prediction Using a Combined Hierarchical Approach”, PROTEINS, Structure, Function and Genetics Suppl., 3: 194 - 198). The definition of contact is based on the analysis of available complex crystal structures (see, for example, MacCallum et al., 1996, J. Mol. Biol., 5: 732-45). In the conformational definition, the positions of CDRs can be identified as residues that contribute enthalpy to Petition 870250091928, dated 08 / 10 / 2025, p. 73 / 175 66 / 155 antigen binding (see, for example, Makabe et al., 2008, Journal of Biological Chemistry, 283: 1156-1166). Furthermore, other definitions of CDR limits may not strictly follow one of the methods described above, but still overlap with at least some of Kabat's CDRs, although they may be shortened or lengthened based on predictions or experimental results that a specific residue or a specific group of residues does not significantly affect antigen binding. As used in the present description, a CDR may refer to a CDR defined by any method known in the art, including combinations of methods. The correspondence between the various numbering schemes is well known to those skilled in the art, and examples are shown in Table 1 below. Table 1. The relationships between the numbering schemes of CDR CDR IMGT Kabat AbM Chothia Contact HCDR1 27-38 31-35 26-35 26-32 30-35 HCDR2 56-65 50-65 50-58 52-56 47-58 HCDR3 105-117 95-102 95-102 95-102 93-101 LCDR1 27-38 24-34 24-34 24-34 30-36 LCDR2 56-65 50-56 50-56 50-56 46-55 LCDR3 105-117 89-97 89-97 89-97 89-96

[0097] The amino acid residues of the CDRs of the regions VL and VH of the antibody described herein conform to the known Kabat numbering scheme, in terms of Petition 870250091928, dated 08 / 10 / 2025, page 74 / 175 67 / 155 number and positions.

[0098] “Domain” of a polypeptide or protein refers to a folded protein structure that is able to maintain its tertiary structure independently of the rest of the protein. In general, a domain is responsible for a single functional property of a protein and, in many cases, can be added, deleted, or transferred to other proteins without loss of function of the rest of the protein and / or the protein domain.

[0099] “Immunoglobulin domain” refers to a globular region of an antibody chain (e.g., a chain of a conventional antibody with a four-peptide chain structure or a heavy chain antibody) or a polypeptide consisting essentially of such globular regions. The immunoglobulin domain is distinguished by retaining the characteristic immunoglobulin fold of an antibody molecule and consists of a 2-layer sandwich with approximately 7 antiparallel β-strands arranged in two β-sheets, optionally stabilized by a conserved disulfide bond.

[00100] “Variable immunoglobulin domain” refers to an immunoglobulin domain consisting essentially of four “structural regions” referred to in the art and hereafter as “structural region 1” or “FR1”, “structural region 2” or “FR2”, “structural region 3” or “FR3” and Petition 870250091928, dated 08 / 10 / 2025, page 75 / 175 68 / 155 “structural region 4 or “FR4”, wherein the structural regions are spaced by three “complementarity-determining regions or “CDRs” referred to in the technique and hereafter as “complementarity-determining region 1 or “CDR1”, “complementarity-determining region 2 or “CDR2” and “complementarity-determining region 3 or “CDR3”. Thus, the overall structure or sequence of an immunoglobulin variable domain can be expressed as follows: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. Immunoglobulin variable domains have specificity for an antigen by virtue of having an antigen-binding site.

[00101] “Antibody frame (RF) refers to a portion of a variable domain that serves as a framework for the antigen-binding loops (CDRs) of the variable domain.

[00102] “Single variable immunoglobulin domain” is generally used to refer to an immunoglobulin variable domain (which may be a heavy or light chain domain, including a VH, VHH, or VL domain) that can form a functional antigen-binding site without interacting with other variable domains (e.g., without VH / VL interactions, as is required between the VH and VL domains of conventional four-chain monoclonal antibodies). Examples of “single variable immunoglobulin domains” include nanobodies (including VHHs, humanized VHHs, and / or Petition 870250091928, dated 08 / 10 / 2025, page 76 / 175 69 / 155 Camelid-like VHs (e.g., camelized human VHs), IgNARs, domains, antibodies (single-domain) such as VH domains or derived from VH domains (such as dAbs™) and antibodies (single-domain) such as VL domains or derived from VL domains (such as dAbs™). Single variable immunoglobulin domains based on and / or derived from variable heavy chain domains (such as VH or VHH domains) are generally preferred. A specific example of a single variable immunoglobulin domain is a “VHH domain” (or “VHH for short”), as defined below.

[00103] “VHH domain”, also known as single heavy chain domain antibody, VHH, VHH domain, VHH antibody fragment, VHH antibody or nanobody, is a variable domain of an antigen-binding immunoglobulin known as a “heavy chain antibody” (i.e., “an antibody devoid of light chains”) (HamersCasterman C, Atarhouch T, Muyldermans S, Robinson G, Hamers C, Songa EB, Bendahman N, Hamers R., “Naturally occurring antibodies devoid of light chains”; Nature 363, 446-448 (1993)). The term “VHH domain” is used to distinguish the variable domain from the heavy chain variable domain (which is referred to in this description as a “VH domain”) and the light chain variable domain (which is referred to in this description as a “VL domain”) present in conventional antibodies with a four-chain structure. Petition 870250091928, dated 08 / 10 / 2025, page 77 / 175 70 / 155 peptide chains. VHH domains bind specifically to an epitope without the need for an additional antigen-binding domain (unlike the VH or VL domain in conventional antibodies with a four-peptide chain structure, in which case the epitope is recognized by the VL domain along with the VH domain). A VHH domain is a small, stable, and efficient antigen recognition unit consisting of a single immunoglobulin domain. The terms “single-domain heavy chain antibody,” “VHH domain,” “VHH,” “VhH domain,” “VHH antibody fragment,” “VHH antibody,” “Nanobody®,” and “Nanobody® domain” (“Nanobody is a registered trademark of Ablynx NV, Ghent, Belgium”) are used interchangeably. “VHH domains” include, but are not limited to, naturally occurring camelid-produced antibodies, camelid-produced and subsequently humanized antibodies, or antibodies obtained by screening with phage display techniques.The total number of amino acid residues in a VHH domain will generally be in the range of 110 to 120, frequently between 112 and 115. However, it should be noted that shorter and longer sequences may also be suitable for the purposes described in this description. Methods for obtaining VHHs that bind to a specific antigen or epitope have been previously described in the following documents: R. van der Linden et al., Journal of Immunological Methods, 240 (2000). Petition 870250091928, dated 08 / 10 / 2025, p. 78 / 175 71 / 155 185-195; Li et al., J Biol Chem., 287 (2012) 13713-13721; Deffar et al., African Journal of Biotechnology Vol. 8 (12), pp.2645-2652, June 17, 2009, and WO94 / 04678.

[00104] As is well known in the art for VH and VHH domains, the total number of amino acid residues in each of the CDRs can vary and may not correspond to the total number of amino acid residues indicated by Kabat numbering (i.e., one or more positions according to Kabat numbering may not be occupied in the actual sequence, or the actual sequence may contain more amino acid residues than the number allowed by Kabat numbering). This means that, in general, Kabat numbering may or may not correspond to the actual numbering of amino acid residues in the actual sequence. Other numbering systems or schemes include Chothia, IMGT, and AbM.

[00105] “Humanized antibody, also known as CDR-grafted antibody, refers to an antibody produced by grafting non-human CDR sequences into the structure of variable regions of a human antibody. Chimeric antibodies, due to their significant non-human protein content, can induce a strong immune response. This problem can be overcome with the use of humanized antibodies. To avoid the decrease in activity caused by decreased immunogenicity, the variable regions of a fully human antibody can be subjected to a Petition 870250091928, dated 08 / 10 / 2025, page 79 / 175 72 / 155 minimum reverse mutation required to maintain activity. Examples of humanization include the humanization of camelid-derived VHH domains by replacing one or more amino acid residues in the amino acid sequence of the original VHH sequence with one or more amino acid residues present in the corresponding positions in a VH domain of a conventional human antibody with a four-peptide chain structure (also referred to in this description as sequence optimization; in addition to humanization, sequence optimization may also encompass other sequence modifications by one or more mutations that provide enhanced VHH properties, such as the removal of potential post-translational modification sites). The humanized VHH domain may contain one or more sequences of a fully human structural region.Furthermore, to avoid decreased activity caused by decreased immunogenicity, the structural sequence in the variable region of the human antibody can undergo minimal reverse mutation or retromutation to maintain activity.

[00106] A fully human antibody includes antibodies with variable and constant regions derived from human germline immunoglobulin sequences. The fully human antibody described herein may include amino acid residues not encoded by human germline immunoglobulin sequences. Petition 870250091928, dated 08 / 10 / 2025, page 80 / 175 73 / 155 (for example, mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutations in vivo). “Fully human antibody” does not include “humanized antibodies.”

[00107] Typically, the CD40-binding molecule or the FAP-binding molecule of the present description will bind to the antigen to be bound (i.e., CD40 or FAP) with a dissociation constant (Kd), preferably from 10⁻⁷ to 10⁻¹⁰ mol / L (M), more preferably from 10⁻⁸ to 10⁻¹⁰ mol / L and even more preferably from 10⁻⁹ to 10⁻¹⁰ or less, and / or with an association constant (KA) of at least 10⁻⁷ M, preferably from at least 10⁻⁸ M, more preferably from at least 10⁻⁹ M and most preferably from at least 10⁻¹⁰ M, as measured in a Biacore, KinExA or Fortibio assay. Any Kd value greater than 10⁻⁴ M is generally considered indicative of non-specific binding.The specific binding of an antigen-binding protein to an antigen or epitope can be measured by any known suitable method, including, for example, surface plasmon resonance (SPR) assay, Scatchard assay and / or competitive binding assay (e.g., radioimmunoassay (RIA), enzyme immunoassay (EIA) and sandwich competitive assay) described in this description.

[00108] “Compete, when used in a case where Petition 870250091928, dated 08 / 10 / 2025, page 81 / 175 74 / 155 antigen-binding proteins (e.g., neutralizing antigen-binding proteins or neutralizing antibodies) compete for the same epitope, refers to the competition between antigen-binding proteins, which is measured by the following assays in which a test antigen-binding protein (e.g., an antibody or an immunologically functional fragment thereof) prevents or inhibits (e.g., reduces) the specific binding of a reference antigen-binding protein (e.g., a reference ligand or antibody) to a common antigen (e.g., CD40 or a fragment thereof). Several types of competitive binding assays are available to determine whether one antigen-binding protein competes with another, such as direct or indirect solid-phase radioimmunoassay (RIA), direct or indirect solid-phase enzyme immunoassay (EIA), and sandwich competition assay (see, for example, Stahli et al.)., 1983, Methods in Enzymology 9: 242-253); direct solid-phase biotin-avidin EIA (see, for example, Kirkland et al., 1986, J. Immunol. 137: 3614-3619), direct solid-phase labeling assay and direct solid-phase sandwich labeling assay (see, for example, Harlow and Lane, 1988, Antibodies: A Laboratory Manual, Cold Spring Harbor Press); direct solid-phase labeling RIA with I-125 marker (see, for example, Morel et al., 1988, Molec. Immunol. 25: 7-15); direct solid-phase biotin-avidin EIA. Petition 870250091928, dated 08 / 10 / 2025, p. 82 / 175 75 / 155 solid (see, for example, Cheung, et al., 1990, Virology 176: 546-552); and direct labeling RIA (Moldenhauer et al., (1990, Scand. J. Immunol. 32: 77-82). Generally, the assay involves the use of a purified antigen (found on a solid or cell surface) that can bind to an unlabeled detection antigen-binding protein and a labeled reference antigen-binding protein. Competitive inhibition is measured by measuring the amount of marker bound to the solid surface or cell in the presence of the test antigen-binding protein. Usually, the test antigen-binding protein is present in excessive amounts.Antigen-binding proteins identified by competitive assays (competitive antigen-binding proteins) include: antigen-binding proteins that bind to the same epitope as the reference antigen-binding protein; and antigen-binding proteins that bind to an epitope sufficiently close to the epitope to which the reference antigen-binding protein binds, where the two epitopes spatially impede binding to each other. Generally, when the competitive antigen-binding protein exists in excessive quantity, the specific binding of the reference antigen-binding protein to the common antigen will be inhibited (i.e., reduced) by at least 40% to 45%, 45% to 50%, 50% to 55%, 55% to 60%, 60% to 65%, 65% to 70%, 70% to 75%, or 75% or more. Petition 870250091928, dated 08 / 10 / 2025, p. 83 / 175 76 / 155 In certain cases, the binding is inhibited by at least 80% to 85%, 85% to 90%, 90% to 95%, 95% to 97%, or 97% or more.

[00109] Antibodies can be competitively screened for binding to the same epitope using conventional techniques known to those skilled in the art. For example, competition and cross-competition studies can be performed to obtain antibodies that compete or cross-compete with each other for binding to an antigen. A high-throughput method for obtaining antibodies that bind to the same epitope based on their cross-competition is described in International Patent Publication No. WO03 / 48731. Therefore, antibodies that compete for binding to the same epitope on CD40 or FAP with the antibody molecules of the present description can be obtained by conventional techniques known to those skilled in the art.

[00110] “CD40-binding molecule” refers to any protein capable of specifically binding to CD40 or any molecule that composes it. The CD40-binding molecule may include antibodies against CD40, as defined in this description, or their conjugates. The CD40-binding molecule also includes antibodies of the immunoglobulin superfamily (IgSF) or CDR-grafted molecules. The “CD40-binding molecule” of the present description may comprise at least one unique variable immunoglobulin domain (such as a VHH) that binds Petition 870250091928, dated 08 / 10 / 2025, p. 84 / 175 77 / 155 to CD40. In some embodiments, the “CD40-binding molecule” may comprise 2, 3, 4 or more unique variable immunoglobulin domains (such as VHHs) that bind to CD40. The CD40-binding molecule of the present description may further comprise, in addition to the unique variable immunoglobulin domain of CD40, a ligand and / or a fraction with effector function, such as a half-life extender fraction (e.g., a unique variable immunoglobulin domain that binds to serum albumin) and / or a fusion partner (such as serum albumin) and / or a conjugated polymer (such as PEG) and / or an Fc region.In some embodiments, the "CD40-binding molecule" of the present description further encompasses bispecific / multispecific antibodies comprising immunoglobulins that bind to different antigens (e.g., a first antibody that binds to a first antigen (e.g., CD40) and a second antibody that binds to a second antigen (e.g., FAP), optionally a third antibody that binds to a third antigen and, optionally, a fourth antibody that binds to a fourth antigen).

[00111] “FAP-binding molecule” refers to any protein capable of specifically binding to FAP or any molecule that composes it. The FAP-binding molecule may include antibodies against FAP, as defined in this description, or their conjugates. Petition 870250091928, dated 08 / 10 / 2025, p. 85 / 175 78 / 155

[00112] “FAP / CD40 binding molecule” refers to any protein capable of specifically binding to CD40 and FAP, or to any molecule that comprises them. The FAP / CD40 binding molecule may include antibodies against CD40 and FAP, as defined in this description, or their conjugates.

[00113] “CD40 binding” or “CD40 binding” refers to the ability to interact with CD40 or an epitope thereof, wherein CD40 or its epitope may be human-derived. “FAP binding” or “FAP binding” refers to the ability to interact with FAP or an epitope thereof, wherein FAP or its epitope may be human-derived. “Antigen-binding site in the present description” refers to a discontinuous three-dimensional spatial site on an antigen that is recognized by the antibody of the present description.

[00114] “Antigen refers to a molecule used for immunization of an immunocompetent vertebrate to produce an antibody that recognizes the antigen or to screen an expression library (e.g., particularly a display library of phages, yeast, or ribosomes). In the present description, antigen is defined in a broader sense and includes a target molecule that is specifically recognized by the antibody and a portion or mimic of a molecule used in an immunization process to produce the antibody or in screening for Petition 870250091928, dated 08 / 10 / 2025, page 86 / 175 79 / 155 library to select the antibody. For example, for the antibody that binds to human CD40 of the present description, monomers and multimers (e.g., dimers, trimers, etc.) of human CD40, and truncated variants and other variants of human CD40 are all referred to as antigens.

[00115] An epitope refers to a site on an antigen to which an immunoglobulin or antibody binds. An epitope may be formed from contiguous or non-contiguous amino acids juxtaposed by tertiary folding of the protein. An epitope formed from contiguous amino acids is generally retained after exposure to a denaturing solvent, while an epitope formed by tertiary folding is generally lost after treatment with a denaturing solvent. An epitope generally comprises, for example, at least 3 to 15 amino acids in a distinct spatial conformation. Methods for determining which epitope is bound to a given antibody are well known in the art and include an immunoblotting assay, an immunoprecipitation assay, and the like. Methods for determining the spatial conformation of an epitope include techniques in the art and techniques described in this description, such as X-ray crystallography and two-dimensional nuclear magnetic resonance.

[00116] Specific binding or selective binding refers to the binding of an antibody to an epitope on a Petition 870250091928, dated 08 / 10 / 2025, page 87 / 175 80 / 155 predetermined antigen. For example, an antibody binds to a predetermined antigen or its epitope with an equilibrium dissociation constant (KD) of approximately less than 10-7 M or even less and with an affinity that is at least twice as high as its affinity to bind to a nonspecific antigen other than the predetermined antigen or its epitope (or nonspecific antigens other than closely related antigens, e.g., BSA, etc.), when measured by surface plasmon resonance (SPR) techniques on an instrument using human CD40 or an epitope thereof as analyte and the antibody as ligand. “Antigen-recognizing antibody” is used interchangeably in the present description with “specifically bound antibody.”

[00117] “Binding affinity” or “affinity” is used in the present description as a measure of the strength of a non-covalent interaction between two molecules (e.g., an antibody or a portion thereof and an antigen). The binding affinity between two molecules can be quantified by determining the dissociation constant (KD). KD can be determined by measuring the kinetics of complex formation and dissociation using, for example, the surface plasmon resonance (SPR) method (Biacore). The rate constants corresponding to the association and dissociation of a monovalent complex are called Petition 870250091928, dated 08 / 10 / 2025, page 88 / 175 81 / 155 association rate constant ka (or kon) and dissociation rate constant kd (or koff), respectively. Kd is related to ka and kd by the equation Kd = kd / ka. The value of the dissociation constant can be determined directly by well-known methods and can be calculated by methods such as those described by Caceci et al. (1984, Byte 9: 340-362) even for complex mixtures. For example, Kd can be determined using a double-filtration nitrocellulose filter binding assay, as described by Wong & Lohman (1993, Proc. Natl. Acad. Sci. USA 90: 5428-5432). Other standard assays for evaluating the binding capacity of an antibody to a target antigen are known in the art and include, for example, ELISA, western blot, RIA, and flow cytometry, as well as other assays exemplified elsewhere in this description.The binding kinetics and binding affinity of the antibody can also be assessed by standard assays known in the art, such as surface plasmon resonance (SPR), for example, using the Biacore™ or KinExA system. The binding affinities associated with different molecular interactions, for example, the binding affinities of different antibodies for a given antigen, can be compared by comparing the Kd values ​​of antibody / antigen complexes. Similarly, the specificity of an interaction can be assessed by determining and comparing the Kd value for the interaction. Petition 870250091928, dated 08 / 10 / 2025, page 89 / 175 82 / 155 of interest (e.g., a specific interaction between an antibody and an antigen) with the Kd value for an interaction not of interest (e.g., a control antibody known not to bind to CD40).

[00118] “Conservative substitution refers to the substitution by another amino acid residue with properties similar to the original amino acid residue. For example, lysine, arginine, and histidine have similar properties because they have basic side chains, and aspartic acid and glutamic acid have similar properties because they have acidic side chains. Additionally, glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, and tryptophan have similar properties because they have uncharged polar side chains, and alanine, valine, leucine, threonine, isoleucine, proline, phenylalanine, and methionine have similar properties because they have nonpolar side chains. Furthermore, tyrosine, phenylalanine, tryptophan, and histidine have similar properties because they have aromatic side chains.”Thus, it will be apparent to those skilled in the art that even when an amino acid residue in a group exhibiting properties similar to those described above is substituted, it will not exhibit a particular change in properties.

[00119] “Homology, “identity or “sequence identity” refers to sequence similarity between two Petition 870250091928, dated 08 / 10 / 2025, p. 90 / 175 83 / 155 polynucleotide sequences or between two polypeptides. When positions in two compared sequences are occupied by identical nucleotides or amino acid monomers, for example, if the position in each of the two DNA molecules is occupied by an identical nucleotide, the molecules are homologous at that position. The percentage of homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the number of positions compared χ² 100%. For example, if 6 out of 10 positions are matching or homologous when two sequences are optimally aligned, the two sequences are 60% homologous. In general, when two sequences are aligned, a comparison is performed to obtain the maximum percentage of homology.

[00120] Inhibition and “blocking” are used interchangeably and encompass both partial and complete inhibition / blocking. Growth inhibition (e.g., involving cells) is intended to include any measurable reduction in cell growth.

[00121] Agonistic activity, agonist activity, or agonism refers to the function of a substance as an agonist. The binding of an agonist to a cellular receptor initiates a reaction or activity that is similar to or the same as that initiated by the receptor's natural ligand. For example, a CD40 agonist or CD40 agonist antibody can induce Petition 870250091928, dated 08 / 10 / 2025, page 91 / 175 84 / 155 any or all of the following responses: cell proliferation and / or differentiation; positive regulation of intercellular adhesion through molecules such as ICAM-1, E-selectin, and VCAM; secretion of pro-inflammatory cytokines such as IL-1, IL-6, IL-8, IL-12, and TNF; signal transduction through the CD40 receptor via pathways such as TRAF (e.g., TRAF2 and / or TRAF3), MAP kinases such as NIK (NF-κB kinase inducer), 1-kB kinases (IKKα / β), NF-kB transcription factor, Ras and the MEK / ERK pathway, the PI3K / Akt pathway, and the P38 MAPK pathway; anti-apoptotic signal transduction by molecules such as XIAP, Mcl-1, and BCLx; generation of B and / or T cell memory: antibody production for B cells; B cell isotype switching; upregulation of cell surface expression of MHC class II and CD80 / 86, and similar effects.

[00122] “CD40-related disease” or “CD40-related disorder” refers to a disorder in which cells expressing CD40 are altered or eliminated. These cells include CD40-expressing cells that demonstrate abnormal proliferation or CD40-expressing cells that are associated with cancerous or malignant growth. More specific examples of cancers that demonstrate abnormal CD40 antigen expression include B-cell lymphoblastoid cancer, Burkitt lymphoma, multiple myeloma, T-cell lymphomas, Kaposi's sarcoma, osteosarcoma, epidermal and endothelial tumors, pancreatic cancer, lung cancer, cancer of Petition 870250091928, dated 08 / 10 / 2025, page 92 / 175 85 / 155 breast, ovarian cancer, colon cancer, prostate cancer, head and neck cancer, skin cancer (melanoma), bladder cancer, and kidney cancer. Such disorders include, among others, leukemias, lymphomas (including B-cell lymphoma and non-Hodgkin lymphoma), multiple myeloma, Waldenstrom's macroglobulinemia; solid tumors, including sarcomas such as osteosarcoma, Ewing's sarcoma, malignant melanoma, adenocarcinoma (including ovarian adenocarcinoma), Kaposi's sarcoma / Kaposi's tumor, and squamous cell carcinoma.

[00123] “Proliferative disease refers to a disorder associated with a certain degree of abnormal cell proliferation. In one form, the proliferative disorder is cancer.

[00124] “Cancer,” “cancerous,” “proliferative disorder,” and “tumor” are not mutually exclusive when mentioned in this description.

[00125] “Preventing cancer refers to delaying, inhibiting, or preventing the onset of cancer in an individual in whom the onset of oncogenesis or tumorigenesis has not been evidenced, but a predisposition to cancer has been identified, as determined, for example, by genetic screening or otherwise. The term also encompasses the treatment of an individual with a pre-malignant disorder to halt progression or cause regression of the pre-malignant disorder to malignancy. Petition 870250091928, dated 08 / 10 / 2025, page 93 / 175 86 / 155

[00126] Optionally or optionally means that the event or circumstance described below may occur, but not necessarily, and this description includes cases where the event or circumstance either occurs or does not occur. For example, optionally including mannitol or ethylenediaminetetraacetic acid means that it may, but not necessarily, be present in the pharmaceutical composition.

[00127] The term about and approximately as used in this document means that a numerical value is within an acceptable error range for the specific value determined by one skilled in the art, and the numerical value depends in part on how the value is measured or determined (i.e., the limits of the measurement system). For example, about may mean a standard deviation within 1 or more than 1 in each practice in the art. Alternatively, about or substantially comprise may mean a range up to ±30%; for example, a pH of about 5.5 means a pH of 5.5 ± 1.65. Furthermore, particularly for biological systems or processes, the term may mean up to an order of magnitude or up to 5 times the numerical value.Unless otherwise specified, when a specific value is provided in this application and claims, the meaning of "about" or "substantially comprises" should be assumed to be within an acceptable range of error for that specific value. Petition 870250091928, dated 08 / 10 / 2025, page 94 / 175 87 / 155

[00128] Buffer refers to a buffer that resists changes in pH through the action of its conjugate acid-base components. Examples of buffers that control pH within an appropriate range include tris(hydroxymethyl)aminomethane (Tris), acetate, succinate, gluconate, histidine salt, oxalate, lactate, phosphate, citrate, tartrate, fumarate, glycylglycine, and other organic acid buffers.

[00129] An acetate buffer is a buffer comprising acetate ions. Examples of acetate buffers include acetic acid-sodium acetate, histidine-histidine acetate, acetic acid-potassium acetate, acetic acid-calcium acetate, acetic acid-magnesium acetate, and the like. The preferred acetate buffer is acetic acid-sodium acetate.

[00130] A succinate buffer is a buffer comprising succinate ions. Examples of succinate buffers include succinic acid-sodium succinate, succinic acid-potassium succinate, succinic acid-calcium succinate, and the like. The preferred succinate buffer is succinic acid-sodium succinate. Illustratively, succinic acid-sodium succinate can be prepared from succinic acid and sodium hydroxide, or from succinic acid and sodium succinate.

[00131] A histidine salt buffer is a buffer comprising histidine ions. Examples of histidine salt buffers include a histidine hydrochloride buffer, a Petition 870250091928, dated 08 / 10 / 2025, page 95 / 175 88 / 155 histidine-acetate buffer, a histidine-phosphate buffer, a histidine-sulfate buffer and the like, histidine-hydrochloride buffer or histidine-acetate buffer being preferred. The histidine-acetate buffer is prepared from histidine and acetic acid, and the histidine-hydrochloride buffer is prepared from histidine and histidine hydrochloride, or histidine and hydrochloric acid.

[00132] “Phosphate buffer is a buffer comprising phosphate ions. Examples of phosphate buffers include disodium hydrogen phosphate-sodium dihydrogen phosphate, disodium hydrogen phosphate-potassium dihydrogen phosphate, disodium hydrogen phosphate-citric acid and the like. The preferred phosphate buffer is disodium hydrogen phosphate-sodium dihydrogen phosphate.

[00133] “Pharmaceutical composition refers to a mixture comprising one or more of the antibodies described in this document and other chemical components, and the other components are, for example, physiologically / pharmaceutically acceptable carriers and excipients. The objective of the pharmaceutical composition is to maintain the stability of the active ingredient and promote administration to an organism, which facilitates the absorption of the active ingredient, thus exerting biological activity.

[00134] In the present description, “pharmaceutical composition” and “formulation” are not mutually exclusive. Petition 870250091928, dated 08 / 10 / 2025, page 96 / 175 89 / 155

[00135] Unless otherwise specified, the solvent in the pharmaceutical composition described herein in solution form is water.

[00136] “Lyophilized formulation” refers to a pharmaceutical formulation or composition obtained by lyophilizing a pharmaceutical composition or formulation in liquid form or a vacuum solution.

[00137] The pharmaceutical composition described in this description can achieve a stable effect, i.e., the FAP / CD40 binding molecule in the pharmaceutical composition substantially retains its physical and / or chemical stability and / or biological activity after storage; for example, the pharmaceutical composition substantially retains its physical and chemical stability as well as its biological activity after storage. The storage period is generally selected based on a predetermined shelf life of the pharmaceutical composition. A variety of analytical techniques are currently available to measure protein stability, and stability after storage for a selected period of time at a selected temperature can be measured.

[00138] A stable pharmaceutical antibody formulation is one in which no significant change is observed under the following conditions: storage at refrigeration temperature (2 to 8 °C) for at least 3 Petition 870250091928, dated 08 / 10 / 2025, page 97 / 175 90 / 155 months, at least 6 months, at least 1 year, at least 2 years, or at most 2 years. In addition, stable liquid formulations include liquid formulations that exhibit desirable characteristics after storage at 25°C for periods including 1 month, 3 months, and 6 months, or storage at 40°C for periods including 1 month. Typical acceptable criteria for stability are as follows: typically, no more than about 10%, preferably no more than about 5%, of antibody monomer is degraded as measured by SEC-HPLC. The pharmaceutical antibody formulation is colorless, or transparent to slightly opalescent, by visual analysis. The concentration, pH, and osmolality of the formulation show a change not exceeding ±10%. Typically, cutoffs not exceeding about 10%, preferably not exceeding about 5%, are observed. Typically, an aggregation of no more than about 10% is formed, preferably no more than about 5%.

[00139] An antibody “maintains its physical stability in a pharmaceutical formulation if it does not show a significant increase in aggregation, precipitation, and / or denaturation after visual inspection of color and / or clarity, or as determined by UV light scattering, size exclusion chromatography (SEC), and dynamic light scattering (DLS). Changes in protein conformation may be Petition 870250091928, dated 08 / 10 / 2025, page 98 / 175 91 / 155 evaluated by fluorescence spectroscopy (which determines the tertiary structure of the protein) and by FTIR spectroscopy (which determines the secondary structure of the protein).

[00140] An antibody “maintains its chemical stability” in a pharmaceutical formulation if it does not exhibit significant chemical alteration. Chemical stability can be assessed by detecting and quantifying chemically altered proteins. Degradation processes that often alter the chemical structure of proteins include hydrolysis or cleavage (evaluated by methods such as size exclusion chromatography and SDS-PAGE), oxidation (evaluated by methods such as peptide mapping in combination with mass spectrometry or MALDI / TOF / MS), deamidation (evaluated by methods such as ion-exchange chromatography, capillary isoelectric focusing, peptide mapping, and isoaspartic acid determination), and isomerization (evaluated by isoaspartic acid content determination, peptide mapping, etc.).

[00141] An antibody “retains its biological activity” in a pharmaceutical formulation if the biological activity of the antibody at a given time is within a predetermined range of the biological activity exhibited during the preparation of the pharmaceutical formulation. The biological activity of an antibody can be determined, for example, by an antigen binding assay. Petition 870250091928, dated 08 / 10 / 2025, page 99 / 175 92 / 155

[00142] Administering, giving, and treating, when applied to animals, humans, experimental subjects, cells, tissues, organs, or biological fluids, refer to the contact of an exogenous drug, a therapeutic agent, a diagnostic agent, or a composition with animals, humans, subjects, cells, tissues, organs, or biological fluids. Administering, giving, and treating may refer, for example, to therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Cell treatment comprises the contact of a reagent with cells and the contact of the reagent with a fluid, where the fluid is in contact with the cells. Administering, giving, and treating also refer to the treatment, for example, of cells by reagents, diagnostic, or binding compositions, or by another cell in vitro and ex vivo.Treatment, when applied to humans, veterinarians, or research subjects, refers to therapeutic treatment, preventive or prophylactic measures, and research and diagnostic applications.

[00143] Treating or treatment refers to the administration of a therapeutic agent, such as a therapeutic agent comprising any of the antibodies described herein or a pharmaceutical composition thereof, internally or externally, to an individual who has had, is suspected of having, or is predisposed to having one or more diseases. Petition 870250091928, dated 08 / 10 / 2025, pages 100 / 175 93 / 155 proliferative diseases or symptoms thereof in which the therapeutic agent is known to have a therapeutic effect. Typically, the therapeutic agent is administered in an amount effective to relieve one or more disease symptoms in the individual or population being treated, either by inducing regression of such symptoms or by inhibiting the development of such symptoms to any clinically measurable degree. The amount of therapeutic agent effective to relieve any specific disease symptom (also referred to as the “therapeutically effective amount”) may vary depending on a variety of factors such as the disease state, age, and weight of the individual, and the ability of the drug to produce the desired therapeutic effect in the individual. Whether a disease symptom has been relieved or not can be assessed by any clinical testing method commonly used by physicians or other healthcare professionals to assess symptom severity or progression.Although the modalities of the present description (e.g., treatment methods or products) may be ineffective in relieving a symptom of the disease of interest in a given individual, they should relieve the symptom of the disease of interest in a statistically significant number of individuals, as determined by any statistical testing method known in the art, such as Student's t-test, chi-square test, Mann-Whitney U test, Kruskal-Wallis test (H-test), etc. Petition 870250091928, dated 08 / 10 / 2025, page 101 / 175 94 / 155 Jonckheere-Terpstra and Wilcoxon test.

[00144] “Effective amount” comprises an amount sufficient to improve or prevent a symptom or sign of a medical disorder. An effective amount also means an amount sufficient to permit or facilitate diagnosis. The effective amount for an individual may vary depending on factors such as the disorder being treated, the individual’s general health, the method, route, and dose of administration, and the severity of side effects. An effective amount may be the maximum dose or administration regimen to avoid significant side effects or toxic effects. The subject of this description may be an animal or a human being.

[00145] The “individual” or “patient” in the present description means a mammal, particularly a primate, and especially a human being.

[00146] The equipment and methods used in the detection are shown below: Appearance:

[00147] A visual method was used. The sample vial was cleaned with a cloth, and the color, clarity, and presence of visible foreign matter in the sample were observed in a clarity tester under a white background and a black background, with an illumination intensity of 1000 to 1500 lx. Petition 870250091928, dated 08 / 10 / 2025, page 102 / 175 95 / 155

[00148] Instrument for appearance analysis: Jingtuo Instrument YB-2A clarity analyzer. Molecular exclusion chromatography (SEC):

[00149] This is an analytical method that separates a solute based on the relative relationship between the pore size of the gel and the spiral size of the polymer sample molecule.

[00150] Percentage of monomer content in SEC = A monomer / A total x 100% (A monomer represents the peak area of ​​the main monomer peak in the sample and A total represents the sum of all peak areas).

[00151] Instrument for SEC determination: Agilent 1260-Bio; chromatographic column: Waters, XBrige BEH200® SEC (300 x 7.8 mm, 3.5 μm) NR-CE capillary gel electrophoresis:

[00152] This is a type of electrophoresis in which a gel is moved into a capillary as a support medium and a method that achieves separation under a given voltage based on the molecular weight of the sample.

[00153] Percentage of purity of unreduced CE = Main peak area / Total area x 100% (Main peak area represents the peak area of ​​the main peak in the sample and Total area represents the sum of all peak areas).

[00154] Instrument for determining CE: Sciex model PA800 plus. Petition 870250091928, dated 08 / 10 / 2025, page 103 / 175 96 / 155 Capillary isoelectric focusing electrophoresis with icIEF imaging:

[00155] This is a separation technique based on the difference in isoelectric points (pI) of proteins.

[00156] Percentage of content of the main peak of icIEF = area of ​​the main peak / total area χ 100% (the total area represents the sum of the areas of the acid, main and basic peaks).

[00157] Instrument manufacturer for determining icIEF: Protein Simple, model: Muarice. Determination of protein concentration:

[00158] Instrument for determining protein concentration: ultraviolet-visible spectrophotometer; Model: Nano Drop 2000; Optical path length: 1 mm. Process for preparing an exemplary antibody pharmaceutical composition (formulation).

[00159] Step 1: A FAP / CD40 binding antibody (e.g., Ab10-A297V3-4, with heavy and light chain sequences established at SEQ ID NOs: 22 and 20, respectively) was mixed with the following amounts of auxiliary materials to prepare a stock solution containing the FAP / CD40 binding molecule. The stock solution was filtered, subjected to in-process control sampling, and tested for sterility. The stock solution was filtered through a 0.22 μm filter and the filtrate was Petition 870250091928, dated 08 / 10 / 2025, page 104 / 175 97 / 155 collected.

[00160] Step 2: The filling quantity was adjusted to 1.15 mL and the filtrate was bottled in 2 mL vials. Stoppers were applied and in-process control samples were taken at the beginning, middle, and end of filling to detect differences in the amount of charge.

[00161] Step 3: The closing machine was switched on, the aluminum covers were put on, and the closing was completed.

[00162] Step 4: Visual inspection was performed to confirm that the products were free of defects, such as inaccurate filling quantity. Bottle labels were printed and placed on the bottles; box labels were printed, the boxes were folded, the packaging was carried out, and the box labels were placed on the boxes. II. Examples and Test Examples

[00163] The present description is illustrated in detail by the following examples. These examples are for illustrative purposes only and are not intended to limit the scope of the present description.

[00164] Experimental methods without the specific conditions indicated in the examples of this description were generally conducted under conventional conditions or conditions recommended by the material manufacturer. Petition 870250091928, dated 08 / 10 / 2025, pp. 105 / 175 98 / 155 batch or commercial products. Reagents without specific origins indicated were conventional, commercially available reagents. Examples

[00165] The methods for preparing and purifying the FAP / CD40 binding molecule in the present application are described in Patent Application No. WO2023025194, which is incorporated herein by reference. totality. Example 1. Screening and Preparation of Anti-FAP Monoclonal Antibodies 1.1. Sequencing and preparation of screening and detection antigens

[00166] Human fibroblast activating protein (FAP; GeneBank accession number: AAC51668) is a serine oligopeptidase with a molecular weight of approximately 170 kDa and is a homodimer. The sequences, sources, and uses of the recombinant FAP proteins used in this description are presented in Table 2. Table 2. Sources of amino acid sequences for recombinant proteins Name Start and end of amino acid sequence Sino Biological Cat. No. Use h-FAPbiotin Leu26-Asp760 (GenBank Accession No. AAC51668) 10464-H07H-B Liquid-phase magnetic bead screening with phage display h-FAP-His 10464-H07H ELISA screening for Petition 870250091928, dated 08 / 10 / 2025, pp. 106 / 175 99 / 155 h-FAP-FITC 10464-H07H-F positive clones CHO cell surface antibody display screening 1.2. Screening of specific h-FAP binding antibody fragments from the human natural Fab phage display library

[00167] With h-FAP-biotin antigen as the target molecule, antigen-specific phages were captured using avidin-coated magnetic beads, and phage enrichment was performed using a magnetic support. Antigen-specific phages were eluted with a glycine solution (pH 2.2).

[00168] After two rounds of screening, 284 clones were randomly selected and screened by ELISA for phages looking for positive clones, which included the following specific steps: the growing colonies on the plate were counted (284 colonies in total). The phages were inoculated into a 96-well plate, each well containing 400 μL of culture medium (2YT + Amp + 0.2% glucose). The plate was shaken at 37 °C at 250 rpm for 6 h. The plate was coated with h-FAP-His antigen at 100 ng / 100 μL / well and incubated overnight at 4 °C. The following day, after the 96-well plate had been washed and blocked, 100 μL of overnight-incubated bacterial fluid was added to each well, and the Petition 870250091928, dated 08 / 10 / 2025, page 107 / 175 The 100 / 155 plate was incubated at 37 °C for 1 h. After washing, a secondary antibody (an HRP-labeled anti-human IgG-Fab antibody) was added, and the plate was incubated at °C for 40 min. After washing, the substrate solution was added, and the plate was incubated in a dark place for 30 min. Then, the OD600 was measured using a microplate reader. A total of 122 positive clones obtained in two screening rounds were sequenced, and 74 distinct VHs (constituting a VH-enriched library), 48 distinct κ light chains (constituting a KLC-enriched library), and 10 distinct λ light chains (constituting an LLC-enriched library) were obtained. 1.3. Construction of the complete antibody library for display in CHO cells.

[00169] 1) A complete antibody library was constructed for display in CHO cells, which included: the VH-enriched library, the KLC-enriched library, and the LLC-enriched library, which were subjected to PCR amplification with three sets of primers, respectively. The three enriched libraries were digested with enzymes and then inserted into the corresponding component vectors to construct the corresponding component libraries. The KLC, LLC, and VH component libraries were digested with enzymes. The libraries of Petition 870250091928, dated 08 / 10 / 2025, pp. 108 / 175 101 / 155 KLC, LLC, and VH fragments, after digestion, were analyzed and purified by electrophoresis. The complete antibody vector for cell display was digested with an enzyme, and the vector fragments were purified. The purified vector fragments and the KLC, LLC, and VH fragment libraries were mixed and ligated. The ligation products were purified and transferred to Escherichia coli cells by electroporation. Cells were seeded in a plate and cultured overnight at 37 °C, and colonies were counted. The library capacity was 3.6 χ¹⁰E⁶, representing more than 800 times the theoretical diversity (74 χ¹⁸ = 429²). All colonies were collected, and vector DNA was extracted to obtain a complete library of antibody cell display genes.

[00170] 2) A full-length antibody cell display library specific for the h-FAP antigen was constructed, which included: CHO cells were transformed with the DNA of the full-length antibody cell display gene library vector to construct a full-length antibody CHO cell display library. The cell library was screened under hygromycin pressure. A stably transformed cell library was obtained, and the cell library was double-stained with a chain antibody. Petition 870250091928, dated 08 / 10 / 2025, page 109 / 175 102 / 155 light chain κ (or λ) anti-human mouse antigen labeled with PE and an h-FAP antigen labeled with FITC, and sorted by FACS for cells doubly positive for PE and FITC. Each well contained one cell. The κ light chain library sorting used 2 plates of 96 wells, and the λ light chain library sorting used 1 plate of 96 wells. Cells were cultured under hygromycin pressure.

[00171] 3) FACS analysis was performed to identify single-cell clones exhibiting specific antibodies to the h-FAP antigen, including: cells were cultured under hygromycin pressure for 14 days, and 153 single-cell κ-chain clones and 50 single-cell λ-chain clones were stably transformed. The cells were digested with a 0.5 mM EDTA-PBS buffer solution, and the single-cell clones were double-stained with a PE-labeled mouse anti-human κ (or λ) light chain antibody and a FITC-labeled h-FAP antigen. FACS analysis showed that 138 single-cell clones with double-positive fluorescence for PE and FITC were obtained.

[00172] 4) The antibody genes were cloned, including: the affinities of the positive clones were analyzed by FACS, and 45 clones were subjected to antibody gene amplification by PCR. After centrifugation, the positive clones were collected and the Petition 870250091928, dated 08 / 10 / 2025, pages 110 / 175 103 / 155 supernatant was discarded. Genomic DNA was extracted from the cells, and VH and CL cells were amplified by PCR. The fragments obtained from the amplification were separated by electrophoresis and sequenced, and 6 distinct VH cells and 6 distinct κ strands were identified. Their combinations can produce 12 clones with distinct sequences. The VH and VL sequences of one of the clones are shown in Table 3. Table 3. The amino acid sequences of VH and VL (κ of KLC) of the h-FAP antibody No. Amino acid sequence Ab10 VH QVQLQQSGVEVKKPGASVTVSCRASGYSFADHFIHWVRQAPGQGFQWMGWINPNRG VTHHAQDFQGRVAMTRDMSTDTVYMELTSLRSDDTAVYYCARDASLTARPYYFYGF DVWGQGTLVTVSS (SEQ ID NO:1) VL DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYAASSLQS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPPAFGQGTKVEIK (SEQ ID NO:2)

[00173] Note: CDRs defined by Kabat's numbering scheme are underlined.

[00174] The present description also provides the sequences of the full-length heavy and light chains of Ab10. Full-length heavy chain of Ab10 QVQLQQSGVEVKKPGASVTVSCRASGYSFADHFIHWVRQAPGQGFQWMGWINPN RGVTHHAQDFQGRVAMTRDMSTDTVYMELTSLRSDDTAVYYCARDASLTARPYYFYGFD VWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG Petition 870250091928, dated 08 / 10 / 2025, pp. 111 / 175 104 / 155 QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 24) > Ab10 full-length light chain DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYAASSL QSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPPAFGQGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDST YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVVTKSFNRGEC (SEQ ID NO: 25) Tabela 4. As CDRs do anticorpo anti-FAP N° CDRs de VH CDRs de VL Ab10 HCDR1 DHFIH (SEQ ID NO:3) LCDR1 RASQGISSWLA (SEQ ID NO:6) HCDR2 WINPNRGVTHHAQDFQG (SEQ ID NO:4) LCDR2 AASSLQ (SEQ ID NO:7) AASSLQS (SEQ ID NO:32) HCDR3 DASLTARPYYFYGFDV (SEQ ID NO:5) LCDR3 QQANSFPPA (SEQ ID NO:8) 1.4. Construção da biblioteca completa de anticorpos para exibição em células CHO

[00175] 1) A soluble antibody expression vector was constructed, which included: VH and LC positive fragments with distinct sequences were digested with enzymes and purified. The VH fragments were inserted into a soluble heavy chain (IgG1) expression vector, and the LC fragments were inserted into a soluble light chain expression vector. The colonies were sent for sequencing and the DNA was extracted.

[00176] 2) A soluble antibody was expressed and Petition 870250091928, dated 08 / 10 / 2025, page 112 / 175 105 / 155 purified, which included: Expi293 cells were expanded by suspension culture. According to the light-heavy chain pair determined above, 18 μg of the light chain expression vector and 12 μg of the heavy chain expression vector were mixed. The vector DNA and PEI were mixed in a weight ratio of 1:2.5, and the mixture was used to transform Expi293 cells. On day 6, the culture medium supernatant was collected and the antibody was purified by the Protein-A method. SDS-PAGE denaturing gel electrophoresis analysis showed that the antibody purity reached more than 90%. The antibody was stored at -80 °C. 1.5. ELISA bond strength test

[00177] A plate was directly coated with a recombinant FAP protein labeled with His. After the addition of the antibody, a secondary antibody (an anti-human IgG antibody conjugated with HRP) and the HRP substrate TMB were added to determine the antibody's binding activity to the antigen. The assay included: A 96-well microplate was coated with a 0.5 μg / mL solution of human FAP-His protein at 100 μL / well, and the plate was incubated overnight at 4 °C. After thorough washing, a blocking solution was added at 200 μL / well, and the plate was incubated at room temperature for 2 h. After thorough washing, the test anti-FAP antibody, diluted with a diluent, was added at 100 μL / well. The plate was incubated. Petition 870250091928, dated 08 / 10 / 2025, pp. 113 / 175 106 / 155 at room temperature for 1 h. After thorough washing, an HRP-labeled anti-human goat IgG secondary antibody, diluted with a diluent at a ratio of 1:20000, was added at 100 μL / well. The plate was incubated at room temperature for 1 h. After thorough washing, TMB was added at 100 μL / well, and the plate was incubated in a dark place for 15 min. 0.16 M sulfuric acid was added at 50 μL / well. The OD value at 450 nm was measured using a Thermo MultiSkanFc microplate reader, and the EC50 value of anti-FAP antibody binding was calculated. The result is shown in Table 5. Table 5. EC50 value for the binding affinity of the anti-FAP antibody to the human FAP antigen. EC50 No. (mg / mL) Ab10 0.0920 1.6. Surface plasmon resonance (SPR) binding assay

[00178] A CM5 sensor chip was used and HBS-EP+ buffer solution (10 mM HEPES, 150 mM NaCl, 3 mM EDTA, 0.05% P20 surfactant) was used as the mobile phase. A 30 μg / mL solution of an anti-human IgG (Fc) antibody was prepared in 10 mM sodium acetate buffer (pH 5.0) and amino coupling immobilization was performed. The test antibody was diluted in the HBS-EP+ buffer solution. Petition 870250091928, dated 08 / 10 / 2025, pp. 114 / 175 107 / 155 and captured by anti-human IgG (Fc) antibody in the chip channels. h-FAP-His was dissolved in HBSEP+ buffer solution as the analyte, and the resulting solution was serially diluted twice. The diluted antibody was allowed to flow through the experimental and reference channels at a flow rate of 30 μL / min, with 1 min of association and 15 min of dissociation. 10 mM glycine pH 1.5 was administered as a regeneration buffer solution at a flow rate of 10 μL / min for 30 s. The data were analyzed. The results in Table 6 show that the affinity of Ab10 for the FAP antigen was 3 to 6 times greater than that of the control anti-FAP antibody 28H1 (sequences 219 and 233 of US patent 9266938B2). Table 6. Ka, Kd, ​​and KD affinity values ​​of the anti-FAP antibody determined by SPR. No. ka (1 / Ms) kd (1 / s) Kd (M) Ratio of Kd of 28H1 to Kd of test antibody Ratio of Kd of 28H1 to Kd of test antibody Ab10 2.97E+05 7.28E05 2.45E-10 5.3 3.50

[00179] The sequences of the variable region of the heavy and light chain of the 28H1 control antibody are as follows: > VH of 28H1 EVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWAS GEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVT VSS (SEQ ID NO: 9) > VL of 28H1 Petition 870250091928, dated 08 / 10 / 2025, pages 115 / 175 108 / 155 EIVLTQSPGTLSLSPGERATLSCRASQSVSRSYLAWYQQKPGQAPRLLIIGAST RATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGQVIPPTFGQGTKVEIK (SEQ ID NO: 10) 1.7. FACS bonding test

[00180] A FACS assay was used to evaluate the binding properties of anti-FAP antibodies to cells. The assay included: A human FAP overexpressing CHO cell line was constructed and plated (1E5 / well). Test antibodies were added at 100 μL / well. The maximum concentration was 100 nM and was diluted 5 times to obtain 8 concentrations in total. The plate was incubated at 4 °C for 1 h. The anti-hIgG antibody Alexa Fluor-647 was added as a secondary antibody at a ratio of 1:500. The plate was incubated at 4 °C for 30 min and a FACS assay was performed. Antibodies capable of binding to FAP can mark cells. The relationship between the percentage of labeled cells overexpressing human FAP among all cells and the antibody concentration is shown in FIG. 1. The EC50 values ​​for antibody binding to human FAP are shown in Table 7. The relationship between the percentage of labeled cells overexpressing mouse FAP among all cells and the antibody concentration is shown in FIG. 2, where Ab9, Ab14, and Ab15 are the other 3 anti-FAP antibodies obtained by screening in this application. Table 7. EC50 values ​​for affinities of Petition 870250091928, dated 08 / 10 / 2025, pp. 116 / 175 109 / 155 binding of anti-FAP antibodies to human FAP antigen Antibody No. EC50 (pg / mL) 28H1 0.0007839 Ab10 0.0004320 Isotype (NC) - Example 2. Screening and Preparation of Anti-CD40 Nanobodies (VHHs) 2.1. Sequencing and preparation of immunizing and screening antigens

[00181] A recombinant human CD40 protein labeled with His (h-CD40-His), a recombinant human CD40 protein biotinylated at the C-end (hCD40-biotin), and a recombinant monkey CD40 protein labeled with His at the C-end (cyno-CD40-His) were selected. Their sequences and sources are shown in Table 8. The protein reagents can be used in the experiments of the following examples. Table 8. Amino acid sequences and sources of recombinant proteins Name Start and end of amino acid sequence Acrobiosystems Cat. No. h-CD40-His Glu21-Arg193 CD0-H5228 h-CD40-biotin CD0-H82E8 cyno-CD40-His CD0-C52H6 2.2. Immunization process and determination of alpaca titer Petition 870250091928, dated 08 / 10 / 2025, pp. 117 / 175 110 / 155

[00182] An alpaca was immunized with h-CD40-his once every two weeks, totaling four immunizations. In the first immunization, 0.5 mg of antigen and 1 mL of complete Freund's adjuvant (CFA) were thoroughly mixed and injected subcutaneously. In the following three immunizations, 0.25 mg of antigen and 1 mL of incomplete Freund's adjuvant (IFA) were thoroughly mixed and injected subcutaneously. Blank serum was collected before immunizations. One week after the third immunization and one week after the fourth immunization, 50 mL of peripheral blood were collected and lymphocytes were separated. The serum titer was determined. 2.3. Determining the title and constructing the phage library.

[00183] After four alpaca immunizations, the serum titer was determined. After determining the titer as acceptable, PBMCs were separated and total RNA was extracted. Purity was determined and RNA was reverse transcribed into DNA. After two rounds of nested PCR, the purified vector and the VHH fragment of interest were digested with enzymes and ligated. Transfection was performed by electroporation, clones were selected, and phage A and B libraries were obtained. 2.4. Phage library affinity selection and identification by ELISA.

[00184] With the h-CD40 antigen as the target molecule, the Petition 870250091928, dated 08 / 10 / 2025, pp. 118 / 175 Screening of 111 / 155 cells was performed using an acid elution method with Gly-HCl to elute specific phages. After two rounds of screening, 384 clones were randomly selected from the titer determination plates of the first and second rounds and screened by ELISA for phages searching for positive clones, and the optical density at 450 nm was measured. According to the sequencing results, the sequences were subjected to sequence alignment and phylogenetic tree analysis, and 16 sequences were obtained by screening. Among them, the amino acid sequences of the best-functioning anti-CD40 antibody are shown in Tables 9 and 10. Table 9. The sequence of the single-domain anti-CD40 antibody. Amino Acid Sequence No. A2 97 QVQLVESGGGMVEPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGPEWVSTINSHGDTTYYA DSVKGRFTISRDNAQNTVYLQMNSLKPEDTALYYCLRIDYSDYSSRGQGTQVTVSS (SEQ ID NO:11)

[00185] Note: CDRs defined by Kabat's numbering scheme are underlined. Table 10. CDRs of the anti-CD40 antibody CDR Sequence Numbers Ab2 97 CDR1 RYGMK (SEQ ID NO:12) CDR2 TINSHGDTTYYADSVKG (SEQ ID NO:13) CDR3 IDYSDYSS (SEQ ID NO:14) 2.5. Expression and purification of the anti-CD40 single-domain Fc antibody fusion protein Petition 870250091928, dated 08 / 10 / 2025, p. 119 / 175 112 / 155

[00186] The two VHHs above were separately linked to human IgG1-Fc containing the N297A mutation. The resulting VHH-Fc fusion protein sequence is shown in Table 11. Human IgG1-Fc is underlined and the N297A mutation is in bold.

[00187] 9E5-SELFNS was used as a positive control CD40 agonist (see sequences 58 and 59 of document WO2020108611A1). Table 11. Fusion protein sequence obtained from anti-CD40 single-domain antibody and human IgG1-Fc. No. Amino Acid Sequence A297-Fc PKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF NWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQ ID NO: 15)

[00188] Plasmids were constructed and transiently transfected into cells, and antibodies were expressed and purified. Analysis showed that the antibody of interest was obtained. 2.6. FACS bonding test

[00189] A FACS assay was used to evaluate the binding properties of anti-CD40 antibodies to cells. The assay included: Individual Raji cells were obtained and plated. Test antibodies were added to 100 Petition 870250091928, dated 08 / 10 / 2025, pp. 120 / 175 113 / 155 μL / well. The highest concentration point was 100 nM. A 5-fold dilution was performed, resulting in 8 concentration points in total. The plate was incubated at 4 °C for 1 h. The anti-hIgG antibody Alexa Fluor-647 was added as a secondary antibody at a ratio of 1:500. The plate was incubated at 4 °C for 30 min. A FACS assay was performed. A CD40 agonist was used as a positive control. The EC50 values ​​of the antibodies are shown in FIG. 3 and Table 12. A12 was another anti-CD40 antibody obtained by screening in the present application. Table 12. EC50 values ​​for the binding affinities of anti-CD40 antibodies to human CD40 antigen. No. EC50 (nM) A297 0.5293 9E5-SELFNS 0.1961

[00190] For a CD40 agonist antibody used in a bispecific FAP / CD40 antibody, its EC50 affinity for CD40 should not be too high; otherwise, the bispecific antibody will preferentially bind to CD40. It is more desirable for the bispecific antibody to preferentially bind to FAP, thus playing a role. Therefore, A297 presents advantages that are more aligned with the above features. This is also reflected in Table 13. 2.7. Measurement of anti-CD40 nanobody activity

[00191] Activation of cells with reporter genes Petition 870250091928, dated 08 / 10 / 2025, pp. 121 / 175 114 / 155 The agonist activity of CD40 antibodies was measured, and the agonist activity of the CD40 antibodies was evaluated according to EC50. The measurement included: HEK-Blue™ CD40L cells (acquired from Invivogen Cat# hkb-cd40, having been stably transfected with the human CD40 gene and the NF-κB-mediated SEAP genome; the activation level of the CD40 signaling pathway can be distinguished by measuring the amount of SEAP secreted into the supernatant using the SEAP QUANTI-Blue substrate) were prepared. The culture medium was DMEM containing 10% FBS, 100 μg / mL Normocin, 100 μg / mL Zeocin, and 30 pg / mL Blasticidin. Cells were seeded in a 96-well plate at 5E4 / well. The culture medium was DMEM containing 10% FBS and 100 μg / mL of Normocin. Cells were cultured overnight. After cell adhesion, serially diluted test antibodies were added at 100 μL / well, and the plate was incubated overnight at 37 °C. Cells were centrifuged.The cell supernatant was transferred to a new 96-well plate, and 180 μL of QUANTI-Blue substrate solution was added. The plate was incubated in the dark for 15 minutes. Absorbance at 620 nm was measured using an Envision microplate reader, and EC50 values ​​were calculated. A CD40 agonist (9E5-SELFNS) was used as a positive control. The EC50 was calculated for the ratio between the percentage of relative agonist activity (compared to 200 nM 9E5-SELFNS) and the concentration of... Petition 870250091928, dated 08 / 10 / 2025, pp. 122 / 175 115 / 155 antibodies are shown in FIG. 4 and Table 13. A12 was another anti-CD40 antibody obtained by screening in the present application. Table 13. EC50 values ​​of agonist activity of anti-CD40 antibodies and their percentages of activating activity (%) in relation to the positive control. EC50 No. (nM) Maximum response (percentage relative to 200 nM 9E5-SELFNS) A2 97 0.556 84.15 9E5-SELFNS 0.466 102.55 IgG1 No activation No activation 2.8. Humanization of anti-CD40 nanobodies

[00192] According to the results above, A297 was humanized. Based on the typical VHH structures of the obtained A297 nanobody, the sequences of the VHH variable region were compared with a database of antibody germlines to obtain highly homologous human germline templates. A preferred human germline template for the A297 antibody of the present description was IGHV3-48*03. CDRs were then grafted onto human FRs, and key amino acids affecting antibody structure and function were mutated to restore binding capacity and activity. The humanized sequences are presented in Table 14. Table 14. Sequences of humanized anti-CD40 antibodies Name Amino acid sequence A2 97 1 V1 QVQLVESGGGVVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTINSHGDTT Petition 870250091928, dated 08 / 10 / 2025, pp. 123 / 175 116 / 155 YYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCARIDYSDYSSRGQGTMVTVS S(SEQ ID NO: 16) V2 QVQLVESGGGVVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTINSHGDTT YYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCLRIDYSDYSSRGQGTMVTVS S(SEQ ID NO: 17) V3 EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTINSHGDTT YYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCLRIDYSDYSSRGQGTLVTVS S(SEQ ID NO: 18) V4 EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTINSHGDST YYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCLKIDYSDYSSRGQGTLVTVS S(SEQ ID NO: 19)

[00193] The 4 VHHs above were separately linked to human IgG1-Fc containing the S267E / L328F mutations (numbered according to EU). The resulting VHH-Fc fusion protein sequences are shown in Table 15. Human IgG1-Fc is underlined and the S267E / L328F mutations are in bold. Table 15. Fusion protein sequences obtained from humanized single-domain anti-CD40 antibodies and human IgG1-Fc (S267E / L328F). N° Sequência de aminoácidos A297V1-Fc QVQLVESGGGVVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTINSHGDT TYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCARIDYSDYSSRGQGTMVT VSSEPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVEHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSSK AFPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK(SEQ ID NO: 26) A297V2-Fc QVQLVESGGGVVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTINSHGDT TYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCLRIDYSDYSSRGQGTMVT VSSEPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVEHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSSK AFPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK(SEQ ID NO: 27) A297V3-Fc EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTINSHGDT TYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCLRIDYSDYSSRGQGTLVT VSSEPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVEHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSSK AFPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK(SEQ ID NO: 28) A297V4-Fc EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTINSHGDS Petition 870250091928, dated 08 / 10 / 2025, pp. 124 / 175 117 / 155 TYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCLKIDYSDYSSRGQGTLVT VSSEPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVEHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSSK AFPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK(SEQ ID NO: 29)

[00194] The 4 VHH-Fc above were used to transiently transfect cells, and the antibodies were expressed and purified. Analysis demonstrated that the antibody proteins of interest were obtained. 2.9. Affinities of humanized CD40 antibodies for cell strains with high CD40 expression

[00195] The binding capacity of humanized antibodies to cells with high expression of the human antigenic protein CD40 was determined by FACS. HEK293 cells were transiently transfected with CD40 plasmids (CD40 cDNA ORF Clone, Human, C-DYKDDDDK (Flag®) Tag; Sino Biological; HG10774-CF), resulting in HEK293 cells with high CD40 expression. The resulting cells were resuspended in flow cytometry staining buffer (PBS + 2% FBS) and the test antibodies, serially diluted at different concentrations, were added. After 1 h of ice incubation, the cells were washed with PBS and centrifuged at 400 g for 5 min. A goat anti-human Fc antibody labeled with the Alexa Fluor 647 fluorescent group was added as a secondary antibody. The cells were stained in an ice bath for 1 h and washed twice with Petition 870250091928, dated 08 / 10 / 2025, pages 125 / 175 118 / 155 PBS, and the fluorescent signals on the cell surface were then detected by FACS. The results are shown in FIG. 5, and the EC50 values ​​are shown in Table 16. All humanized antibodies described here exhibit relatively high affinities for the cell strain with high CD40 expression. Table 16. EC50 values ​​for the affinities of humanized anti-CD40 antibodies for the cell strain with high human CD40 expression. EC50 Antibody (nM) Emax MFI A2 97 1.12 16728 A297 V1 1.11 7300 A297 V2 1.07 12764 A297 V3 1.25 12827 A297 V4 2.70 8255 9E5-SELFNS 1.12 12120 2.10. Activation of HEK-Blue™CD40L cells by humanized CD40 antibodies

[00196] The in vitro agonist activity of CD40 antibodies was evaluated by activating HEKBlue™CD40L cells with CD40 antibodies when crosslinked with FcγRIIb. S267E / L328F mutations contained in the Fc of humanized CD40 antibodies increase the affinity of IgG1 Fc for FcγRIIb and thus increase antibody crosslinking by FcγRIIb. The agonist activity of fully crosslinked CD40 antibodies was simulated by FcγRIIb-mediated activation of HEK-Blue™CD40L cells. Petition 870250091928, dated 08 / 10 / 2025, pp. 126 / 175 119 / 155 by CD40 antibodies. HEK293 cells were transiently transfected with FcyRIIb plasmids (CD32B / Fcgr2b cDNA ORF Clone, Human, N-His tag; Sino Biological; HG10259-NH) to obtain HEK293 cells with high FcyRIIb expression. HEK-Blue™CD40L cells were seeded in a 96-well cell culture plate at 5E4 / well (culture medium was DMEM, 10% FBS, 100 μg / mL Normocin), and FcyRIIb-expressing HEK293 cells were added at 5E4 / well. Serially diluted test antibodies were added to 100 μL / well, and the plate was incubated overnight at 37°C. The cells were centrifuged. 20 μL of the cell supernatant were transferred to a new 96-well plate and 180 μL of QUANTI-Blue substrate solution were added. The plate was incubated in a dark place for 30 min. Absorbance at 620 nm was measured using an Envision microplate reader, and EC50 and Emax values ​​(relative to the fluorescence intensity of the group without antibodies) were calculated. The in vitro cellular agonist activity of the CD40 antibodies was evaluated based on the EC50 values.

[00197] FcyRIIb-mediated activation of HEK-Blue™CD40L cells by CD40 antibodies is shown in FIG. 6. EC50 and Emax (relative fluorescence intensity) values ​​are presented in Table 17.

[00198] The results show that, in the cellular reporter gene system described above, all CD40 antibodies Petition 870250091928, dated 08 / 10 / 2025, pp. 127 / 175 120 / 155 humanized cells showed relatively strong agonist activity when cross-linked with FcYRIIb. Table 17. Activation of HEK-Blue™CD40L cells by humanized anti-CD40 antibodies. Antibody Activation of HEK-Blue™CD4 0L cells mediated by FcγRIIb EC50 (nM) Emax (relative fluorescence intensity) A2 97 0.243 17.34 A297-V1 0.021 18.37 A297-V2 0.005 19.99 A297_V3 0.006 12.84 A297_V4 0.038 13.31 9E5-SELFNS 0.009 16.05 Example 3. Design and Preparation of Bispecific Anti-FAP / CD40 Antibodies 3.1. Design, expression and purification of bispecific anti-FAP / CD40 antibodies

[00199] Based on the results of the humanized anti-CD40 antibody screening, antibody A297V3 was selected. Based on the results of the anti-FAP antibody screening, antibody Ab10 was selected. The selected antibodies were used to construct bispecific anti-FAP / CD40 antibodies. Antibody Ab10 was used as the IgG scaffold for the bispecific antibodies, and each of the C-terminus of the two heavy chains of antibody Ab10 was connected to an anti-CD40 nanobody, or to two or three anti-CD40 nanobodies in sequence. The linkers used between the CD40 nanobodies and the linkers used for the Petition 870250091928, dated 08 / 10 / 2025, pages 128 / 175 121 / 155 binding to the C-terminus of the Ab10 antibody heavy chains were “GGGGSGGGGS”. Each of the bispecific antibodies contains bivalent, tetravalent, and hexavalent CD40 nanobodies, as shown in FIG. 7. In the antibody names, for example, Ab10-A297V3-2, Ab10 represents the anti-FAP antibody used, A297V3 represents the humanized anti-CD40 antibody used, and the “2” at the end represents that the CD40 was bivalent. The other antibodies are all named in this way. Transient transfection and expression and purification of the antibodies were performed using the methods in section 2.5 of Example 2. Analysis showed that the bispecific antibody molecules of interest were obtained. The amino acid sequences of the bispecific antibody molecules are as follows: Ab10-A12V2-2 light chain DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYAASSL QSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPPAFGQGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDST YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 20) > The heavy chain of Ab10-A297V3-2 QVQLQQSGVEVKKPGASVTVSCRASGYSFADHFIHWVRQAPGQGFQWMGWINPN RGVTHHAQDFQGRVAMTRDMSTDTVYMELTSLRSDDTAVYYCARDASLTARPYYFYGFD VWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGÇLVKDYFPEPVTVSWNSGALT. SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV Petition 870250091928, dated 08 / 10 / 2025, pp. 129 / 175 122 / 155 EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSEV QLVESGGGLVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTINSHGDTTYYA DSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCLRIDYSDYSSRGQGTLVTVSS (SEQ ID NO: 21) > The heavy chain of Ab10-A297V3-4 QVQLQQSGVEVKKPGASVTVSCRASGYSFADHFIHWVRQAPGQGFQWMGWINPN RGVTHHAQDFQGRVAMTRDMSTDTVYMELTSLRSDDTAVYCARDASLTARPYYFYGFD VWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLPKGGGGSGGGGSEV QLVESGGGLVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTINSHGDTTYYA DSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCLRIDYSDYSSRGQGTLVTVSS GGG GSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTIN SHGDTTYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCLRIDYSDYSSRGQGTLVTVSS (SEQ ID NO: 22) > A chain pesada de Ab10-A297V3-6 QVQLQQSGVEVKKPGASVTVSCRASGYSFADHFIHWVRQAPGQGFQWMGWINPN RGVTHHAQDFQGRVAMTRDMSTDTVYMELTSLRSDDTAVYCARDASLTARPYYFYGFD VWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT Petition 870250091928, de 08 / 10 / 2025, pág. 130 / 175 123 / 155 HTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLPKGGGGSGGGGSEV QLVESGGGLVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTINSHGDTTYYA DSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCLRIDYSDYSSRGQGTLVTVSS GGG GSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGKGLEWVSTIN SHGDTTYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCLRIDYSDYSSRGQGTLVTVSS LVTVSSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSRYGMKWVRQAPGK GLEWVSTINSHGDTTYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCLRIDYS DYSSRGQGTLVTVSS (SEQ ID NO: 23)

[00200] The light chains of Ab10-A297V3-2, Ab10A297V3-4 and Ab10-A297V3-6 are all established in SEQ ID NO: 20.

[00201] Note: The CH1 and Fc regions in the heavy chains are underlined; the CL regions in the light chains are underlined; the ligands are in italics; LALA (the L234A and L235A mutations) is in bold. 3.2. Measurement of antigen-binding affinities of bispecific anti-FAP / CD40 antibodies

[00202] The affinities of bispecific anti-FAP / CD40 antibodies for their antigens, human FAP protein (Acro, FAP-H5244) and human CD40 protein (Acro, CD0-H5228), were measured by SPR (GE Healthcare; Biacore 8K). A CM5 sensor chip was used and the HBS-EP+ buffer solution (10 mM of Petition 870250091928, dated 08 / 10 / 2025, pp. 131 / 175 124 / 155 HEPES (150 mM NaCl, 3 mM EDTA, 0.05% P20 surfactant) was used as the mobile phase. An anti-human IgG (Fc) antibody was immobilized by amino coupling. Test antibodies were separately diluted in HBS-EP+ buffer solution as ligands and captured by the anti-human IgG (Fc) antibody in the chip channels. Antigenic proteins from different species were used as analytes. The results are presented in Tables 18 and 19. The results show that the prepared bispecific antibodies can normally bind to FAP and, as their affinities for CD40 are lower, they preferentially bind to FAP, thus playing a role. Table 18. Affinities of bispecific anti-FAP / CD40 antibodies for human FAP. Antibody Ka (1 / Ms) Kd (1 / s) KD (M) Ab10 2.17E+05 7.39E-05 3.40E-10 Ab10-A297V3-2 2.03E+05 8.22E-05 4.05E-10 Ab10-A297V3-4 1.84E+05 6.96E-05 3.78E-10 Table 19. Affinities of bispecific anti-FAP / CD40 antibodies for human CD40 Ka (1 / Ms) Antibody 1.37E-01 2.62E-07 3.3. Measurement of the affinities of bispecific anti-FAP / CD40 antibodies for cell lines with high FAP expression. Petition 870250091928, dated 08 / 10 / 2025, pp. 132 / 175 125 / 155

[00203] In this experiment, the binding of bispecific anti-FAP / CD40 antibodies to transfected CHOK1 cell lines with high expression of human FAP, cynomolgus monkey FAP, and mouse FAP was measured by FACS. The bispecific antibodies demonstrated binding to cell surface FAP antigens from humans, mice, and cynomolgus monkeys, and their binding capabilities are similar to those of mAbs. See FIGS. 8A-8C. EC50 values ​​are shown in Table 20. Ab10-A12V2-2 and Ab10-A12V2-4 were two other bispecific anti-FAP / CD40 antibodies obtained by screening in this application. Table 20. EC50 values ​​for the binding of bispecific anti-FAP / CD40 antibodies to human and mouse cell surface FAP antigens. Human FAP antibody Mouse FAP Monkey FAP cynomolgus EC50 (nM) Emax (MFI) EC50 (nM) Emax (MFI) EC50 (nM) Emax (MFI) Ab10-A297V32 2.51 65204 0.79 12633 1.09 28178 Ab10-A297V34 1.57 53835 1.65 12747 1.35 32480 Ab10 1.41 71968 10.41 26381 7.34 64618 3.4. Measurement of the affinities of bispecific anti-FAP / CD40 antibodies for cell lines with high CD40 expression.

[00204] The binding of bispecific antibodies to cell surface CD40 was measured by FACS, including HEK-Blue™CD40L cells with high expression of human CD40. Petition 870250091928, dated 08 / 10 / 2025, pp. 133 / 175 126 / 155 (Invivogen, Cat# hkb-cd40) and HEK293 cells that were transiently transfected with cynomolgus monkey CD40 plasmids (Sino Biological, cat# CG90970-UT) and thus induced to highly express cynomolgus monkey CD40. The results show that bispecific antibodies can bind to CD40 on the surface of human and cynomolgus monkey cell membranes. See FIGs. 9A, 9B, and 11 and Table 21 for details. Ab10-A12V2-2 and Ab10-A12V2-4 were two other bispecific anti-FAP / CD40 antibodies obtained by screening in the present application. Table 21. EC50 values ​​for the binding of bispecific antibodies to human cell surface CD40 antigen. Human CD40 antibody Monkey CD40 cynomolgus EC50 (nM) Emax (MFI) EC50 (nM) Emax (MFI) Ab10-A297V32 1.66 6254 1.31 4927 Ab10-A297V34 0.94 5085 0.88 5124 3.5. Measurement of the maturation-promoting activity of FAP-dependent dendritic cells using humanized bispecific anti-FAP / CD40 antibodies.

[00205] To validate the effect of bispecific anti-FAP / CD40 antibodies on dendritic cell maturation, monocytes were isolated from fresh PBMCs using CD14-positive magnetic beads and cultured in Petition 870250091928, dated 08 / 10 / 2025, pp. 134 / 175 127 / 155 culture medium 1640 containing 50 ng / mL of GM-CSF and 50 ng / mL of hIL-4 was incubated for 5 days. Medium was changed every 2-3 days. On day 5, dendritic cells obtained by induction were added to a 96-well plate at a density of 1E5 cells / well, and CHOK1 cells in the logarithmic growth phase and CHOK1 cells overexpressing human FAP were added separately. Meanwhile, serially diluted test antibodies were added, with 1 μg / mL of LPS as a positive control and 100 nM of hIgG1 and a group without antibodies as negative controls. After 48 h of culture, the expression of CD83 molecules on the surface of dendritic cells was measured by FACS.

[00206] The results in FIG. 11 (in the absence of CHOK1 / FAP) and FIG. 12 (in the presence of CHOK1 / FAP) show that the bivalent molecule Ab10-A297V3-2 cannot activate DCs in the absence of CHOK1 / FAP: its CD40 agonist activity is completely dependent on FAP-mediated crosslinking. However, the tetravalent molecule Ab10-A297V3-4 can induce DC cell maturation in the absence of CHOK1 / FAP cells, and DC cell activation can be further increased in the presence of CHOK1 / FAP cells. Thus, FAP expressed on the surface of dendritic cells may provide an activation window for bispecific antibodies, whether bivalent or tetravalent. Ab10-A12V2-2 and Ab10-A12V2-4 Petition 870250091928, dated 08 / 10 / 2025, pages 135 / 175 128 / 155 were another 2 bispecific anti-FAP / CD40 antibodies obtained by screening in the present application. Example 4. In vivo efficacy of bispecific anti-FAP / CD40 antibodies Ab10-A297V3-2 / 4 in mice.

[00207] Humanized mice with B-hCD40 were from Biocytogen Jiangsu Co., Ltd. (Species: Mus musc μLuS; strain: C57BL / 6; female). Mouse colorectal carcinoma MC38 cells were transfected with full-length mouse FAP plasmids to construct a stably transfected strain. Cells from the strain were inoculated subcutaneously into the right axilla of mice at a ratio of 5 χ¹⁰⁵ cells / 0.1 mL / mouse. When the mean tumor volume reached 80–100 mm³, mice with adequate individual tumor volumes were selected and randomized into groups of 6. Mice received intraperitoneal injections twice weekly. Forty-eight hours after the first administration, peripheral blood samples were collected and analyzed for B-cell count and CD86 expression on the cell surface. On the eighth day after grouping, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) concentrations were measured. Administration was discontinued after four administrations. When there were mice with tumor volumes greater than 2.000 mm3 in the control group hIgGl, the last administration was performed. Twenty-four hours after the. Petition 870250091928, dated 08 / 10 / 2025, pp. 136 / 175 129 / 155 last administration, a complete blood count was performed. 4.1. In vivo efficacy and toxicity of a single dose of Ab10-A297V3-2 or Ab10-A297V3-4 in mice

[00208] The antitumor activity of Ab10-A297V3-2 or Ab10-A297V3-4 was compared with that of the CD40 agonist monoclonal antibody 9E5-mIgG1 under single-dose conditions. 9E5-mIgG1 and 9E5-SELFNS have the same variable region, but use the constant region of the mouse mIgG1 heavy chain and the constant region of the mouse kappa chain as their heavy and light chain constant regions. Since the activity of CD40 agonist monoclonal antibodies depends on the cross-linking of their Fc by FcyRIIb, 9E5-mIgG1, which uses the constant region of the mouse IgG1 heavy chain, may exhibit better CD40 agonist activity in mice. In the in vivo efficacy experiment in mice, 9E5-mIgG1 was a better control antibody than 9E5-SELFNS.

[00209] The antibody Ab10-A297V3-4 showed stronger antitumor activity than the monoclonal antibody CD40 9E5-mIgG1 at the same molar dose, both in terms of TGI (tumor growth inhibition) and CR (complete response). The sequences of 9E5-mIgG1 are as follows: The heavy chain of 9E5-mIgG1 QVQLVQSGAEVKKPGASVKVSCKASGYILTTYWITWVRQAPGQGLEWMGDIHPG Petition 870250091928, dated 08 / 10 / 2025, pages 137 / 175 130 / 155 SGSTKYNEKFKSRVTLTVDTSISTAYMELSRLRSEDTAVYYCARRDYWGQGTTVTVSSA KTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSD LYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFI FPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFR SVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAK DKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEA GNTFTCSVLHEGLHNHHTEKSLSHSPGK (SEQ ID NO: 30) > The 9E5-mIgG1 light chain DIVMTQSPLSLPVTPGEPASISCRSSQNIVNSQGNTYLEWYLQKPGQSPQLLIY KVTNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQASLVPWTFGGGTKVEIKR ADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQD SKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC (SEQ ID NO: 31)

[00210] The antitumor activity of Ab10-A297V3-2 at twice the molar concentration is still weaker than that of Ab10-A297V3-4, but is similar to that of 9E5-mIgG1.

[00211] Peripheral CD40 activation by bispecific antibody molecules was measured by measuring the activation of peripheral blood B lymphocytes in mice. The results are consistent with the in vitro experiment. Ab10-A297V3-4 was able to partially activate peripheral B lymphocytes due to its CD40-activating activity, partially independent of FAP, while Ab10A297V3-2 did not cause detectably significant activation of peripheral B cells (one-way ANOVA), as its Petition 870250091928, dated 08 / 10 / 2025, pages 138 / 175 131 / 155 CD40 activation is completely dependent on FAP. Information on the antibodies tested is presented in Table 22. In vivo efficacy results are presented in FIG. 13A (P < 0.0001 for all antibody groups relative to the hIgG1 control group) and FIG. 13B. Table 22. Information on the antibodies tested. Drug Concentration Molecular weight (kDa) Dose of administration hIgG1 14.17 mg / mL 150 3 mg / kg Ab10A297V3-2 2.95 mg / mL 174 6.96 mg / kg Ab10A297V3-4 3 mg / mL 200.03 4 mg / kg 9E5- mIgG1 4.4 mg / mL 150 3 mg / kg

[00212] In terms of toxicity in mice, neither Ab10-A297V3-2 nor Ab10-A297V3-4 caused changes in the body weight of the mice. See FIG. 13C.

[00213] Complete blood count results show that, compared to the hIgG1 control group at the same molar dose, 9E5-mIgG1 caused some platelet reduction, similar to hepatotoxicity; Ab10-A297V3-2 did not cause platelet reduction; Ab10-A297V3-4 caused the same platelet reduction as 9E5-mIgG1. See FIG. 13D. The dose of Ab10A297V3-2 / 4 administered to mice and statistics on tumor growth inhibition rates are detailed in Table 23.

[00214] The formula for calculating TGI is: TGI = (daily tumor volume of the blank group - volume Petition 870250091928, dated 08 / 10 / 2025, pp. 139 / 175 132 / 155 daily tumor volume of the treatment group) / (daily tumor volume of the blank group) χ 100%. Table 23. The dose of Ab10-A297V3-2 or 4 administered to mice and the rates of inhibition of tumor growth. Antibody No. Dose (mpk) TGI hIgG1 3 - Ab10-A297V3-2 6.96 (double molar dose) 54.6% Ab10-A297V3-4 4 (equimolar dose) 96.4% 9E5-mIgG1 3 59.2%

[00215] As can be seen from the ALT and AST results, 9E5-mIgG1 showed some hepatotoxicity on the eighth day after dosing, through CD32B-mediated crosslinking (i.e., FcyRIIB). Ab10-A297V3-4 shows some hepatotoxicity due to a certain level of background activation, but since its activation is independent of CD32B, the hepatotoxicity is less than that of 9E5-mIgG1. Ab10-A297V32 does not show hepatotoxicity, as its CD40 activation depends on FAP. See FIG. 13E. 4.2. In vivo efficacy and toxicity of multiple doses of Ab10-A297V3-2 or Ab10-A297V3-4 in mice

[00216] To explore the therapeutic window of Ab10A297V3-2 or 4, the dose of Ab10-A297V3-2 was increased and the dose of the Ab10-A297V3-4 molecule was reduced. See Table 24 for details. When the dose of the bivalent Ab10A297V3-2 molecule was increased, its antitumor activity was not... Petition 870250091928, dated 08 / 10 / 2025, pp. 140 / 175 133 / 155 improved, and no activation of peripheral B lymphocytes was observed. When the dose of Ab10-A297V3-4 was reduced to 1.3 mg / kg (i.e., the molar dose of 1 / 3 of 9E5-mIgG1), it still showed a GI tract of 92.6%, which was much better than that of 9E5-mIgG1.

[00217] To evaluate the role of the FAP antibody in the tetravalent bispecific CD40 antibody in in vivo experiments, the antitumor activity of Ab10-A297V3-4 was compared with that of another tetravalent bispecific CD40 antibody without FAP binding function (isotype A297V3-4). The A297V3-4 isotype is structurally identical to Ab10-A297V3-4, except that it contains another isotypic antibody that did not bind to any murine protein, instead of the anti-FAP Ab10 fraction.

[00218] The results in FIGS. 14A-14E show that the tetravalent bispecific antibody CD40 can exhibit relatively good antitumor activity even without binding to FAP; furthermore, its antitumor activity may be stronger if it binds to FAP. Regarding toxicity, the increased dose of Ab10-A297V3-2 did not cause changes in body weight, ALT / AST levels, or platelet count, and the reduced dose of Ab10-A297V3-4 did not cause the abnormal ALT / AST levels or platelet count observed previously with a higher dose, maintaining a very strong antitumor effect (P < 0.0001). This indicates that both Ab10-A297V3-2 and Ab10-A297V3-4 have windows of activity. Petition 870250091928, dated 08 / 10 / 2025, pp. 141 / 175 134 / 155 broader therapeutic ranges than mAb CD40 9E5-mIgG1. Information on antibodies is presented in Table 24. Statistics on the in vivo efficacy of multiple doses of Ab10-A297V3-2 or Ab10-A297V3-4 are presented in Table 25. Table 24. Information on the antibodies tested. Drug Concentration Molecular weight (kDa) Dose hIgG1 14.17 mg / mL 150 3 mg / kg Ab10-A297V3-2 2 mg / mL 174 6.96 mg / kg Ab10-A297V3-2 13.92 mg / kg Ab10-A297V3-4 2 mg / mL 200.03 4 mg / kg Ab10-A297V3-4 1.3 mg / kg Isotype- A297V3-4 3.44 mg / mL 200.03 1.3 mg / kg 9E5-mIgG1 4.79 mg / mL 150 3 mg / kg Table 25. Statistics on the in vivo efficacy of multiple doses of Ab10-A297V3-2 / 4 Drug Dose (mg / kg) GI Tract CR Ratio (<100 mm3) hIgG1 3 - 0% Ab10-A297V3-2 6.96 (double molar dose) 32.8% 0% Ab10-A297V3-2 13.92 (quadruple molar dose) 46.9% 0% Ab10-A297V3-4 4 (equimolar dose) 101.6% 100% Ab10-A297V3-4 1.3 (1 / 3 molar dose) 92.6% 33% Isotype- A297V3-4 1.3 (1 / 3 molar dose) 79.3% 0% 9E5-mIgG1 3 59.2% 0%

[00219] The bispecific anti-FAP / CD40 antibody used in the following examples was the Ab10-A297V3-4 mentioned. Petition 870250091928, dated 08 / 10 / 2025, pp. 142 / 175 135 / 155 previously, whose heavy chain and light chain amino acid sequences are SEQ ID NOs: 22 and 20, respectively. Example 5. Screening of Buffer Systems and pH Values ​​for Anti-FAP / CD40 Bispecific Antibody Formulations

[00220] The following buffers were prepared and 20 mg / mL bispecific anti-FAP / CD40 antibody formulations were prepared. A high-temperature (40 °C) stability study was performed. 1) 10 mM acetic acid-sodium acetate, pH 5.5 2) 10 mM histidine-histidine hydrochloride, pH 5.5 3) 10 mM histidine-histidine hydrochloride, pH 6.0 4) 10 mM histidine-histidine hydrochloride, pH 6.5 5) 10 mM succinic acid-sodium succinate, pH 5.0 6) 10 mM succinic acid-sodium succinate, pH 5.5 7) 10 mM succinic acid-sodium succinate, pH 6.0 8) 10 mM phosphate buffer, pH 6.5 9) 10 mM phosphate buffer, pH 7.0 10) 10 mM phosphate buffer, pH 7.5 Table 26. Results for the buffer system and pH-1 value screening. No. Appearance T0 40 °C-2 W 40 °C-4 W F1 Light yellow, slightly Light yellow, slightly Light yellow, slightly Petition 870250091928, dated 08 / 10 / 2025, pp. 143 / 175 136 / 155 Opalescent, free of visible particles; Opalescent, with few particles; Opalescent, with a large number of particles. F2 Light yellow, slightly opalescent, free of visible particles; Light yellow, slightly opalescent, free of visible particles; Light yellow, slightly opalescent, free of visible particles. F3 Light yellow, slightly opalescent, free of visible particles; Light yellow, slightly opalescent, free of visible particles; Light yellow, slightly opalescent, free of visible particles. F4 Light yellow, slightly opalescent, free of visible particles; Light yellow, slightly opalescent, with a large number of particles; Light yellow, slightly opalescent, with a large number of particles. F5 Light yellow, slightly opalescent, free of visible particles; Light yellow, slightly opalescent, free of visible particles; Light yellow, slightly opalescent, free of visible particles; Light yellow, slightly opalescent, free of visible particles. F6 Light yellow,Slightly opalescent, free of visible particles Light yellow, slightly opalescent, free of visible particles Light yellow, slightly opalescent, with few particles F7 Light yellow, slightly opalescent, free of visible particles Light yellow, slightly opalescent, free of visible particles Light yellow, slightly opalescent, with few particles F8 Light yellow, slightly opalescent, free of visible particles Light yellow, slightly opalescent, with few particles Light yellow, slightly opalescent, with a large number of particles F9 Light yellow, slightly opalescent, free of visible particles Light yellow, opalescent, with a large number of particles Light yellow, opalescent, with a large number of particles F10 Light yellow, slightly opalescent, free of visible particles Light yellow, opalescent, with a large number of particles Light yellow, opalescent, with a large number of particles Petition 870250091928, dated 08 / 10 / 2025, pages 144 / 175 137 / 155 Table 27. Results for the buffer system and pH-2 value screening. N° SEC-HPLC CEX NR-CE % at 40 °C 2 W 40 °C 4 W at 40 °C -2 W 40 °C -4 W at 40 °C 2 W 40 °C 4 W % Monomer % Monomer % Monomer % Principal Peak % Acid Peak % Basic Peak % Principal Peak % Acid Peak % Basic Peak % Principal Peak % Acid Peak % Basic Peak % Principal Peak % Principal Peak % Principal Peak F1 96.1 95.3 94.6 53.5 43.6 3.0 40.8 55.0 4.3 33.8 63.6 2.6 95.7 94.4 91.0 F2 96.1 96.7 95.3 53.5 43.6 2.9 38.6 55.4 6.1 29.4 67.1 3.5 96.0 94.3 90.2 F3 96.1 96.7 95.3 53.4 43.6 3.0 37.7 56.0 6.3 32.5 67.5 0.0 95.9 95.5 88.3 F4 95.9 95.9 94.2 53.2 43.9 2.9 31.9 63.6 4.5 25.8 74.2 0.09 96.0 89.3 87.0 53.7 43.3 3.0 28.7 66.9 4.4 14.6 82.0 3.4 96.5 93.1 90.5 F6 95.9 85.6 81.1 53.6 43.4 3.0 32.3 59.2 8.5 22.2 73.8 4.0 96.5 94.2 90.9 F7 95.5 85.3 80.6 53.4 43.4 3.2 36.0 57.1 7.0 27.5 69.1 3.4 96.4 94.9 87.5 F8 95.6 88.8 84.3 53.5 43.4 3.1 33.6 61.5 4.9 20.4 74.6 5.0 96.5 94.7 83.0 F9 95.3 82.5 75.8 53.0 44.0 3.0 26.2 69.2 4.6 18.3 81.7 0.0 96.4 94.1 75,6 F1 0 94.6 78.6 75.4 52.8 44.1 3.1 17.3 79.0 3.7 8.9 91.1 0.0 96.2 91.4 64.1, Petition 870250091928, dated 08 / 10 / 2025, pages 145 / 175 138 / 155 Experimental results:

[00221] The appearance results demonstrate that: after storing formulations F1 to F10 at 40 °C for 4 weeks, no significant particles were observed in formulations F2 (histidine salt buffer, pH 5.5), F3 (histidine salt buffer, pH 6.0) and F5 (succinate buffer, pH 5.0); visible particles were observed in all other formulations. The results indicate that the histidine salt buffers (pH 5.5, pH 6.0) and the succinate buffer (pH 5.0) were superior to the other buffer systems.

[00222] The SEC results show that: after storage at an elevated temperature of 40 °C for 4 weeks, all formulations showed decreases in SEC purity; the decreases in the histidine salt systems (F2-F4) and the acetate system (F1) were smaller than those in the succinate systems (F5-F7) and the phosphate buffer systems (F8-F10), and formulations F2 and F3 did not show a significant change in monomer purity.

[00223] CEX results show that: after storage at 40 °C for 4 weeks, the main peaks of formulations F1 and F3 decreased by about 20%, the main peak of formulation F2 decreased by about 24%, falling behind only the peaks of formulations F1 and F3, and the main peaks of Petition 870250091928, dated 08 / 10 / 2025, pages 146 / 175 139 / 155 other formulations decreased more significantly.

[00224] NR-CE results show that: after storage at 40 °C for 4 weeks, formulations F1, F2, F5 and F6 showed the smallest decrease in purity, about 6%, compared to T0, formulation F3 showed the second smallest decrease in purity and the other formulations showed more significant decreases in purity.

[00225] In summary, in terms of appearance, SEC, CEX, and NR-CE detection results of the formulations, formulations F2 and F3 are better than the other formulations and exhibit better stability. Therefore, the histidine salt buffer system was selected as the final buffer system. F2 (10 mM histidine salt buffer, pH 5.5) and F3 (10 mM histidine salt buffer, pH 6.0) were selected as the buffer systems used for the next round of pH screening. Example 6. Surfactant Screening in Anti-FAP / CD40 Bispecific Antibody Formulations

[00226] Histidine-histidine hydrochloride buffer systems at pH 5.0 and pH 5.5 were selected, and formulations containing 25 mg / mL of bispecific anti-FAP / CD40 antibody, 80 mg / mL of sucrose, and surfactants of different types and concentrations were prepared to investigate stability after repeated freeze-thaw cycles and shaking: Petition 870250091928, dated 08 / 10 / 2025, pages 147 / 175 140 / 155 15) 10 mM histidine-histidine hydrochloride pH 5.0, 0.2 mg / mL of polysorbate 80; 16) 10 mM histidine-histidine hydrochloride pH 5.5, 0.4 mg / mL of polysorbate 80; 17) 10 mM histidine-histidine hydrochloride pH 5.5, 0.8 mg / mL of polysorbate 80; 18) 10 mM histidine-histidine hydrochloride pH 5.5, poloxamer 188 mg / mL. Table 28. Results of screening for surfactant type and concentration. Test Item Condition F15 F16 F17 F18 Appearance to LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF Freeze-thaw - 3 rounds LY,SO,1* LY,SO,1* LY,SO,PF LY,SO,AFP Freeze-thaw - 5 rounds LY,SO,AFP LY,SO,1* LY,SO,AFP LY,SO,AFP Agitation - 1 day LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF Agitation - 3 days LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,AFP Microinsoluble Particle (MFI) to t2 μη 305 305 189 359 t1Q μη 19 19 19 27 ^25 μη 7 7 5 4 Freeze-thaw - 5 rounds ^2 μη 596 610 510 763 ^10 μη 23 12 71 61 ^25 μη 4 4 17 9 Agitation - 3 days t2 μη 303 724 700 740 t10 μη 17 50 32 41 t2 5 μη 2 5 0 4 SEC to % Aggregate 2.2 2.1 2.1 2.1 % Monomer 97.8 97.9 97.9 97.9 % Fragment 0.0 0.0 0.0 0.0 Freeze-thaw % Aggregate 2.2 2.1 2.1 2.1 Petition 870250091928, dated 08 / 10 / 2025, pages 148 / 175 141 / 155 - 5 rounds % Monomer 97.8 97.9 97.9 97.9 % Fragment 0.0 0.0 0.0 0.0 Agitation - 3 days % Aggregate 2.3 2.1 2.1 2.2 % Monomer 97.7 97.9 97.9 97.9 % Fragment 0.0 0.0 0.0 0.0 CEX to % Acid peak 37.4 37.4 37.3 37.4 % Main peak 58.2 56.9 58.4 58.4 % Basic peak 4.4 5.7 4.3 4.2 Freeze-thaw - 5 rounds % Acid peak 37.0 37.2 36.5 37.3 % Main peak 58.8 58.8 59.7 58.7 % Basic Peak 4.2 4.0 3.8 4.0 Agitation - 3 days % Acid Peak 37.2 36.6 37.3 37.5 % Main Peak 58.6 59.7 58.6 58.3 % Basic Peak 4.2 3.7 4.0 4.1 NR-CE % to 95.6 95.6 95.4 95.5 Freeze-thaw - 5 rounds 95.6 95.2 95.3 95.6 Agitation - 3 days 95.4 95.3 95.3 95.5

[00227] Note: LY = light yellow; SO = slightly opalescent; PF = free of visible particles; AFP = some particles. * : 1 denotes 1 particle. Experimental results

[00228] The appearance results show that: after 5 rounds of freeze-thaw cycles, 1 particle was observed in formulation F16 and some visible particles were observed in formulations F15, F17 and F18. After 3 days of agitation, no visible particles were observed in formulations F15-F17 and some visible particles were observed. Petition 870250091928, dated 08 / 10 / 2025, pages 149 / 175 142 / 155 observed in formulation F18.

[00229] The results of the insoluble microparticles show that: after 3 days of agitation and 5 rounds of freezing and thawing, the particle counts of formulations F15-F18 did not change significantly and the difference between the formulations was not large.

[00230] SEC results show that: after 5 rounds of freeze-thaw cycles and 3 days of agitation, the SEC monomer contents of formulations F15 and F18 did not change significantly and the difference between formulations was not large.

[00231] CEX results show that: after 3 days of agitation and 5 freeze-thaw cycles, the main peaks, acid peaks and basic peaks of formulations F15-F18 did not show significant changes, and the difference between formulations was not large.

[00232] NR-CE results show that: after 5 freeze-thaw cycles and 3 days of agitation, formulations F15-F18 did not show significant changes in purity, and the difference between formulations was not large.

[00233] In summary, in histidine salt buffer systems at pH 5.0-5.5, as can be seen from the appearance results, after 5 rounds of freezing and Petition 870250091928, dated 08 / 10 / 2025, pages 150 / 175 143 / 155 thawing, visible particles were observed in formulations F15-F18, and only 1 particle was observed in formulation F16, which is better than in the other formulations; after 3 days of agitation, some particles were observed in formulation F18, and no visible particles were observed in the other formulations. There was no significant difference in the results of insoluble microparticles and purity between the different formulations. Therefore, polysorbate 80 was selected as the surfactant with a content of 0.04% (w / v). Example 7. Screening of Auxiliary Materials in Anti-FAP / CD40 Bispecific Antibody Formulations

[00234] Histidine-histidine hydrochloride 10 mM buffer systems were selected, and bispecific anti-FAP / CD40 antibody formulations containing 0.4 mg / mL of polysorbate 80 and different auxiliary materials were prepared, with different antibody concentrations: 19) 10 mM histidine-histidine hydrochloride pH 5.0, 8% sucrose, 25 mg / mL of bispecific anti-FAP / CD40 antibody; 20) 10 mM histidine-histidine hydrochloride pH 5.5, 8% sucrose, 25 mg / mL of bispecific anti-FAP / CD40 antibody; 21) 10 mM histidine-histidine hydrochloride pH 5.5, 1% sucrose + 4% mannitol, 25 mg / mL bispecific anti-FAP / CD40 antibody; Petition 870250091928, dated 08 / 10 / 2025, pp. 151 / 175 144 / 155 22) 10 mM arginine-glutamic acid pH 5.0, 8% sucrose, 25 mg / mL bispecific anti-FAP / CD40 antibody; 23) 10 mM histidine-histidine hydrochloride pH 5.5, 8% sucrose + 0.01% EDTA, 25 mg / mL bispecific anti-FAP / CD40 antibody; 24) 10 mM histidine-histidine hydrochloride pH 5.5, 8% sucrose + 0.01% EDTA, 5 mg / mL bispecific anti-FAP / CD40 antibody; 25) 10 mM histidine-histidine hydrochloride pH 5.5, 8% sucrose, 5 mg / mL of bispecific anti-FAP / CD40 antibody.

[00235] The formulations described above were left to stand at 2-8 °C, 25 °C and 40 °C and subjected to stability tests and repeated freeze-thaw stability tests. Petition 870250091928, dated 08 / 10 / 2025, pages 152 / 175 145 / 155 Table 29. Results of screening for anti-FAP / CD40 bispecific antibody auxiliary material. Test item Condition F19 F2 0 F21 F22 F23 F24 F25 Appearance to LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF 40 °C-2 W LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF 40 °C-4 W LY,SO,AFP LY,SO,PF LY,SO,AFP LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF 25 °C-4 WN / A LY,SO,PF N / A LY,SO,PF N / AN / A LY,SO,PF 2~8 °C-4 WN / A LY,SO,PF N / A LY,SO,PF N / AN / A LY,SO,PF Freeze-thaw - 3 rounds LY,SO,PF LY,SO,PF LY,SO,AFP LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF Freeze-thaw - 5 rounds LY,SO,PF LY,SO,PF LY,O, AFP LY,SO,PF LY,SO,PF LY,SO,PF LY,SO,PF SEC to % Aggregate 2.23 2.12 2.10 2.23 2.14 2.07 2.22 % Monomer 97.77 97.88 97.90 97.77 97.86 97.93 97.78 % Fragment 0.00 0.00 0.00 0.00 0.00 0.00 0.00 40 °C-2 W % Aggregate 3.71 3.56 3.62 2.26 3.66 2.25 2.02 % Monomer 96.29 96.44 96.38 97.74 96.34 97.75 97.98 % Fragment 0.00 0.00 0.00 0.00 0.00 0.00 0.00 40 °C-4 W % Aggregate 2.98 3.15 2.93 1.58 3.06 1.32 1.25 % Monomer 96.98 96.85 97.04 98.30 96.91 98.68 98.75% 0.04 0.00 0.03 0.12 0.03 0.00 0.00 Petition 870250091928, dated 08 / 10 / 2025, pp. 153 / 175 146 / 155 Fragment 25 °C -4 W % Aggregate N / A 1.53 N / A 1.54 N / AN / A 1.71 % Monomer N / A 98.47 N / A 98.46 N / AN / A 98.29 % Fragment N / A 0.00 N / A 0.00 N / AN / A 0.00 2~8 °C -4 W % Aggregate N / A 1.49 N / A 1.54 N / AN / A 1.40 % Monomer N / A 98.51 N / A 98.46 N / AN / A 98.60 % Fragment N / A 0.00 N / A 0.00 N / AN / A 0.00 5 freeze-thaw cycles % Aggregate 2.18 2.09 3.72 2.20 N / AN / A 2.18% Monomer 97.82 97.91 96.28 97.80 N / AN / A 97.82% Fragment 0.00 0.00 0.00 0.00 N / AN / A 0.00 CEX to % Acid peak 35.92 36.37 36.44 35.36 36.62 35.77 3 6.56% Main peak 61.04 59.09 59.40 61.67 59.21 61.29 59.15% Basic peak 3.04 4.54 4.16 2.97 4.17 2.94 4.29 40 °C-2 W% Acid peak 50.89 48.24 49.00 46.16 47.48 47.70 47.04 % Main peak 47.15 50.00 48.99 52.31 50.40 50.45 51.14 % Basic peak 1.96 1.75 2.01 1.53 2.13 1.84 1.82 40 °C-4 W % Acid peak 63.82 60.35 60.25 57.22 59.14 57.71 61.73 % Peak 33.83 37.62 37.61 41.33 38.80 40.77 35.90 Petition 870250091928, dated 08 / 10 / 2025, pages 154 / 175 147 / 155 main % Basic peak 2.36 2.03 2.14 1.45 2.06 1.52 2.36 25 °C-4 W % N / A acid peak 40.16 N / A 40.05 N / AN / A 28.01 % N / 56 Major peak, N8 / A N / 2 59.64 % N / A basic peak 3.01 N / A 2.69 N / AN / A 2.35 2~8 °C -4 W % N / A acid peak 37.50 N / A 36.56 N / AN / A 36.41 % N / A main peak, 184 N / A 59 61.19 % N / A basic peak 2.65 N / A 2.26 N / AN / A 2.38 5 freeze-thaw rounds % Acid peak 38.58 38.59 38.18 38.61 N / AN / A 58.89.63 % main peak 59.11 N / AN / A 59.11 % Basic Peak 2.30 2.22 2.92 2.28 N / AN / A 2.25 NR-EC % T0 95.19 95.32 ° 95.14 94.96 95.62 95.54 94 94.69 94.79 93.72 94.37 95.00 94.89 94.66 40 °C-4 W 93.04 93.85 93.95 94.15 94.16 93.93 95.57 / 94 WN 25 °C N / AN / A 95.52 2~8 °C-4 WN / A 95.68 N / A 95.67 N / AN / A 95.67 Freezedefrost - 5 rounds 95.39 95.18 95.36 95.35 N / AN / A 95.24 Note: LY: visible; AFP = light yellow; SO = slightly opalescent; O = opalescent; PF = particle-free, some particles. N / A: not detected. Petition 870250091928, dated 08 / 10 / 2025, pages 155 / 175 148 / 155 Experimental results

[00236] Appearance results show that: after freezing and thawing, visible particles were observed in formulation F21, the degree of opalescence gradually increased with the increase in the number of freeze-thaw cycles, and no visible particles were observed in the other formulations. After storage at 40 °C for 4 weeks, some visible particles were observed in formulations F19 and F21, and no visible particles were observed in the other formulations. After storage at 2-8 °C and 25 °C for 4 weeks, all formulations showed no visible particles.

[00237] SEC results show that: after storage at 40 °C for 4 weeks, all formulations showed no significant change in monomer content. After 5 freeze-thaw cycles, the SEC monomer content showed no significant change.

[00238] CEX results show that: after storage at 40 °C for 4 weeks, the main peaks of formulations F22-F24 decreased slightly, and the main peaks of the other formulations decreased more significantly. After 5 freeze-thaw cycles, the levels of the main, acid and basic CEX peaks did not show significant changes. Petition 870250091928, dated 08 / 10 / 2025, pages 156 / 175 149 / 155

[00239] NR-CE results show that: after storage at 40 °C for 4 weeks, formulation F19 showed the greatest decrease in purity, while the other formulations showed slight decreases in purity, but the decreases were comparable. After 5 freeze-thaw cycles, all formulations showed no significant changes in purity, and the difference between the formulations was not large.

[00240] In summary, as can be observed in the appearance results, after 5 rounds of freeze-thaw cycles, some particles were observed and the degree of opalescence was increased in formulation F21 compared to the other formulations; after incubation at 40 °C for 4 weeks, some particles were observed in formulations F19 and F21, and no significant changes were observed in the other formulations. The purity of the SEC monomer in each formulation did not show significant changes. As can be observed in the CEX results, after the samples were incubated at 40 °C for 4 weeks, the main peak contents of formulations F20-F24 decreased less than those of the other formulations. As can be observed in the NR-CE results, after incubation at 40 °C for 4 weeks, formulation F19 showed the greatest decrease in purity, and the other formulations showed slight decreases in purity. Therefore, formulation F20 (10 mM of) was selected. Petition 870250091928, dated 08 / 10 / 2025, pages 157 / 175 150 / 155 histidine buffer, 8% sucrose, 0.04% PS80, pH 5.5, with a protein concentration of 25 mg / mL. Example 8. Screening of Protein Concentration in Anti-FAP / CD40 Bispecific Antibody Formulations

[00241] Acetic acid-histidine buffer systems 10 mM pH 5.0 and pH 5.5 were selected, and bispecific anti-FAP / CD40 antibody formulations containing 8% sucrose and 0.8 mg / mL polysorbate 80 with different antibody concentrations were prepared: 26) 10 mM acetic acid-histidine pH 5.0, 50 mg / mL of bispecific anti-FAP / CD40 antibody; 27) 10 mM acetic acid-histidine pH 5.0, 80 mg / mL of bispecific anti-FAP / CD40 antibody; 28) 10 mM acetic acid-histidine pH 5.0, 100 mg / mL of bispecific anti-FAP / CD40 antibody; 29) 10 mM acetic acid-histidine pH 5.5, 50 mg / mL of bispecific anti-FAP / CD40 antibody.

[00242] The stability of each formulation at 5 °C and 25 °C was investigated. Table 30. Results of screening for the concentration of bispecific anti-FAP / CD40 antibodies. Test Item Condition F2 6 F27 F2 8 F2 9 Appearance Light yellow, almost colorless, clear, free from Light yellow, almost colorless, clear, free from Light yellow, almost colorless, clear, free from Light yellow, almost colorless, clear, free from Petition 870250091928, dated 08 / 10 / 2025, pages 158 / 175 151 / 155 Particles Particles Particles Particles 5 °C-3 M Light yellow, almost colorless, clear, free of particles Light yellow, almost colorless, clear, free of particles Light yellow, almost colorless, clear, free of particles Light yellow, almost colorless, clear, free of particles 25 °C-3 M Light yellow, almost colorless, clear, free of particles Light yellow, almost colorless, clear, free of particles Light yellow, almost colorless, clear, free of particles Light yellow, almost colorless, clear, free of particles SEC to % Aggregate 0.66 0.72 0.76 0.72 % Monomer 99.29 99.23 99.2 99.23 % Fragment 0.05 0.05 0.04 0.05 5 °C-3 M % Aggregate 0.85 1.02 1.12 0.97 % Monomer 99.04 98.89 98.7 98.9 % Fragment 0.11 0.09 0.18 0.13 25 °C- 3 M % Aggregate 1.48 3.57 5.15 1.5 % Monomer 98.27 96.21 94.73 98.34 % Fragment 0.25 0.22 0.12 0.16 CEX to % Acid peak 34.01 33.43 33.82 34.18 % Main peak 63.87 64.31 64.03 63.66 % Basic peak 2.12 2.26 2.15 2.15 5 °C-3 M % Acid peak 37.59 37.52 37,34 37.69 % Main peak 59.93 59.93 60.57 60.33 % Basic peak 2.47 2.55 2.09 1.98 25 °C- 3 M % Acidic peak 44.21 44.65 44.83 44.81 % Main peak 54.24 53.39 51.79 54.38 % Basic peak 1.56 1.96 3.38 0.8 % Main peak NR-CE to 97.98 98.01 98.11 98.05 5 °C-3 M 98.98 98.98 98.35 98.95 25 °C-3 M 97.92 97.72 97.64 93.61 Petition 870250091928, dated 08 / 10 / 2025, pages 159 / 175 152 / 155

[00243] The results of the appearance analysis show that: after being left to stand at 5 °C and 25 °C for 3 months, the formulations with different antibody concentrations did not show significant changes and demonstrated relatively good stability.

[00244] SEC results show that: the high-concentration formulations F27 and F28 showed a decreasing trend in monomer content after being left to stand at 25 °C for 3 months, and the higher the concentration, the more significantly the monomer content decreased; formulation F28 (100 mg / mL) showed a decrease in monomer purity to approximately 95% after being left to stand at 25 °C for 3 months. After being left to stand at 5 °C for 3 months, the formulations did not show significant changes.

[00245] CEX results show that: all formulations showed slight decreases in the levels of the main peaks after being left to stand at 5 °C for 3 months or at 25 °C for 3 months, and the difference between the formulations was not large.

[00246] NR-CE results show that: after remaining at rest at 25 °C for 3 months, formulation F29 showed a decrease in the main peak of about 5%, and the other formulations did not show a significant decreasing trend. Petition 870250091928, dated 08 / 10 / 2025, pages 160 / 175 153 / 155

[00247] In summary, stability was relatively good when the protein concentration was up to 100 mg / mL and the pH was 5.0. Example 9. Other Alternative Formulation Formulas

[00248] In addition, the present description also provides pharmaceutical formulations of bispecific anti-FAP / CD40 antibodies of other formulation formulas, wherein the bispecific anti-FAP / CD40 antibody is Ab10-A297V3-4 of the present application, and its heavy and light chain sequences are SEQ ID Nos: 22 and 20, respectively. The formulas include, but are not limited to: (1) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.6 mg / mL of polysorbate 80 and 10 mM of histidine-histidine hydrochloride buffer pH 5.5; (2) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.6 mg / mL of polysorbate 80 and 10 mM of histidine-histidine hydrochloride buffer pH 5.3; (3) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.6 mg / mL of polysorbate 80 and 10 mM of histidine-histidine hydrochloride buffer pH 5.2; (4) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.6 mg / mL of polysorbate 80 and 10 mM of histidine-histidine hydrochloride buffer pH 5.0; (5) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.4 mg / mL of polysorbate 80 and 10 mM of Petition 870250091928, dated 08 / 10 / 2025, pages 161 / 175 154 / 155 histidine buffer - histidine hydrochloride pH 5.5; (6) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.4 mg / mL of polysorbate 80 and 10 mM of histidine-histidine hydrochloride buffer pH 5.3; (7) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.4 mg / mL of polysorbate 80 and 10 mM of histidine-histidine hydrochloride buffer pH 5.2; (8) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.4 mg / mL of polysorbate 80 and 10 mM of histidine-histidine hydrochloride buffer pH 5.0; (9) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.4 mg / mL of polysorbate 80 and 10 mM of histidine-acetic acid buffer pH 5.0; (10) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.4 mg / mL of polysorbate 80 and 10 mM of histidine-acetic acid buffer pH 5.5; (11) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.4 mg / mL of polysorbate 80 and 10 mM of histidine-acetic acid buffer pH 5.3; (12) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.4 mg / mL of polysorbate 80 and 10 mM of histidine-acetic acid buffer pH 5.2; (13) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.6 mg / mL of polysorbate 80 and 10 mM of histidine-acetic acid buffer pH 5.0; Petition 870250091928, dated 08 / 10 / 2025, pp. 162 / 175 155 / 155 (14) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.6 mg / mL of polysorbate 80 and 10 mM of histidine-acetic acid buffer pH 5.5; (15) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.6 mg / mL of polysorbate 80 and 10 mM of histidine-acetic acid buffer pH 5.3; (16) 25 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.6 mg / mL of polysorbate 80 and 10 mM of histidine-acetic acid buffer pH 5.2; (17) 10 mg / mL of FAP / CD40 bispecific antibody, 80 mg / mL of sucrose, 0.4 mg / mL of polysorbate 80 and 10 mM of histidine-histidine hydrochloride buffer pH 5.5.

[00249] Experimental results demonstrate that the FAP / CD40 bispecific antibody formulations described above exhibit good stability; the formulations can be applied to the preparation of FAP / CD40 bispecific antibody drugs. Petition 870250091928, dated 08 / 10 / 2025, pages 163 / 175

Claims

1 / 23 CLAIMS 1. Pharmaceutical composition, characterized in that it comprises a FAP / CD40 binding molecule and a buffer, wherein the FAP / CD40 binding molecule comprises a first antigen-binding domain that specifically binds to FAP and a second antigen-binding domain that specifically binds to CD40; the first antigen-binding domain that specifically binds to FAP comprises a variable region of the heavy chain and a variable region of the light chain; the variable region of the heavy chain comprises an HCDR1, an HCDR2 and an HCDR3, and the variable region of the light chain comprises an LCDR1, an LCDR2 and an LCDR3, wherein: 1) HCDR1, HCDR2 and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 3, 4 and 5, respectively; LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences established in SEQ ID NOS: 6, 7, and 8, respectively; or 2) HCDR1,HCDR2 and HCDR3 comprise the amino acid sequences established in SEQ ID NOs: 3, 4, and 5, respectively; LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences established in SEQ ID NOs: 6, 32, and 8, respectively; the buffer is selected from the group consisting of an acetate buffer, a succinate buffer, a histidine salt buffer, and a phosphate buffer; preferably, the buffer is a histidine salt buffer; more preferably, the buffer is a histidine hydrochloride buffer or a histidine acetate buffer; In a much more preferred manner, the buffer is a histidine-histidine hydrochloride buffer or a histidine-acetic acid buffer.

2. Pharmaceutical composition according to claim 1,characterized in that the second antigen-binding domain that specifically binds to CD40 in the FAP / CD40-binding molecule comprises at least one unique immunoglobulin variable domain, and the unique immunoglobulin variable domain comprises three complementarity-determining regions CDR1, CDR2, and CDR3, wherein CDR1, CDR2, and CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 12, 13, and 14, respectively.

3. Pharmaceutical composition, according to claim 1 or 2, characterized in that, in the first antigen-binding domain that specifically binds to FAP in the FAP / CD40 binding molecule, the variable region of the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 1 or an amino acid sequence with at least 90% identity thereto, and the variable region of the light chain comprises the amino acid sequence set forth in SEQ ID NO: 2 or Petition 870250073483, dated 20 / 08 / 2025,Page 19 / 42 3 / 23 an amino acid sequence with at least 90% identity to it.

4. Pharmaceutical composition, according to claim 2 or 3, characterized in that the single variable immunoglobulin domain in the second antigen-binding domain that specifically binds to CD40 in the FAP / CD40 binding molecule comprises any of the amino acid sequences set forth in SEQ ID NOs: 11 and 16 to 19 or an amino acid sequence with at least 90% sequence identity to any of the SEQ ID NOs: 11 and 16 to 19.

5. Pharmaceutical composition, according to any of claims 2 to 4, characterized in that the second antigen-binding domain that specifically binds to CD40 in the FAP / CD40 binding molecule comprises 2, 3, 4, 5 or 6 single variable immunoglobulin domains.

6. Pharmaceutical composition, according to any one of claims 2 to 5,characterized by the fact that the first antigen-binding domain that specifically binds to FAP in the FAP / CD40 binding molecule comprises a variable heavy chain region and a variable light chain region, wherein: the single immunoglobulin variable domain of the second antigen-binding domain that specifically binds Petition 870250073483, dated 08 / 20 / 2025,pg. 20 / 42 4 / 23 to CD40 is located at the N-terminus of the variable region of the heavy chain of the first antigen-binding domain that specifically binds to FAP; the single immunoglobulin variable domain of the second antigen-binding domain that specifically binds to CD40 is located at the C-terminus of the variable region of the heavy chain of the first antigen-binding domain that specifically binds to FAP; the single immunoglobulin variable domain of the second antigen-binding domain that specifically binds to CD40 is located at the N-terminus of the variable region of the light chain of the first antigen-binding domain that specifically binds to FAP; and / or the single immunoglobulin variable domain of the second antigen-binding domain that specifically binds to CD40 is located at the C-terminus of the variable region of the light chain of the first antigen-binding domain that specifically binds to FAP.

7. Pharmaceutical composition,according to any one of claims 2 to 6, characterized in that the single variable immunoglobulin domain of the second antigen-binding domain that specifically binds to CD40 in the FAP / CD40 binding molecule is linked, directly or by a linker, to the first antigen-binding domain that specifically binds to FAP; Petition 870250073483, dated 20 / 08 / 2025, p. 21 / 42 5 / 23 preferably, the linker is an amino acid sequence represented by (G4S)x, where x is independently selected from the group consisting of integers from 1 to 20; more preferably, the linker is an amino acid sequence represented by (G4S)2, (G4S)3, or (G4S)4.

8. Pharmaceutical composition, according to any one of claims 1 to 7, characterized in that the FAP / CD40 binding molecule further comprises a human immunoglobulin Fc region, wherein: preferably,The Fc region is an Fc region of human IgG1 or IgG4; more preferably, the human IgG1 comprises a mutation that removes or reduces the effector function of Fc; much more preferably, the human IgG1 comprises a mutation selected from the group consisting of N297A, D265A / N297A, L234A / L235A, L234A / L235A / P329G, L234E, L234F, L234E / L235F and L234E / L235F / P329G.

9. Pharmaceutical composition, according to any one of claims 1 to 8, characterized in that the first antigen-binding domain that specifically binds to FAP in the FAP / CD40 binding molecule comprises a heavy chain and a light chain, wherein: preferably, the heavy chain is of the IgG1 or IgG4 isotype, and the light chain is of the Kappa isotype; Petition 870250073483, dated 20 / 08 / 2025, p. 22 / 42 6 / 23 more preferably, the heavy chain is the amino acid sequence established in SEQ ID NO: 24 or an amino acid sequence with at least 90% identity to it,and the light chain is the amino acid sequence established in SEQ ID NO: 25 or an amino acid sequence with at least 90% identity to it.

10. Pharmaceutical composition, according to any one of claims 1 to 9, characterized in that the FAP / CD40 binding molecule comprises a first polypeptide chain and a second polypeptide chain, wherein: the first polypeptide chain comprises the amino acid sequence established in any of the SEQ ID NOs: 21-23 or an amino acid sequence with at least 90% identity to it, and the second polypeptide chain comprises the amino acid sequence established in SEQ ID NO: 20 or an amino acid sequence with at least 90% identity to it; preferably, the FAP / CD40 binding molecule comprises two identical first polypeptide chains and two identical second polypeptide chains.

11. Pharmaceutical composition,according to any one of claims 1 to 10, characterized in that the FAP / CD40 binding molecule is a bispecific anti-FAP / CD40 antibody.

12. Pharmaceutical composition, according to any one of claims 1 to 11, characterized in that it further comprises a surfactant, wherein, preferably, the surfactant is one or more surfactants selected from the group consisting of a polysorbate and a poloxamer; more preferably, the surfactant is a polysorbate; In a much more preferred manner, the surfactant is polysorbate 80.

13. Pharmaceutical composition, according to any one of claims 1 to 12, characterized in that it further comprises a sugar, wherein, preferably, the sugar is one or more sugars selected from the group consisting of sucrose, glucose, trehalose and maltose; more preferably, the sugar is sucrose.

14. Pharmaceutical composition,according to any one of claims 1 to 13, characterized in that it further comprises one or more auxiliary materials selected from the group consisting of a polyol and a metal chelating agent, wherein, preferably, the polyol is one or more polyols selected from the group consisting of glycerol, mannitol and sorbitol; the metal chelating agent is selected from the group consisting of ethylenediaminetetraacetic acid and a pharmaceutically acceptable salt thereof; more preferably, the polyol is mannitol; the metal chelating agent is ethylenediaminetetraacetic acid. Petition 870250073483, dated 20 / 08 / 2025, p. 24 / 42 8 / 23 15. Pharmaceutical composition, according to any one of claims 1 to 14, characterized in that the pharmaceutical composition has a pH of 3.5 to 7, preferably 4 to 6.5, more preferably 4.2 to 6.2 and much more preferably 4.5 to 6.

16. Pharmaceutical composition,According to any one of claims 1 to 15, characterized in that the FAP / CD40 binding molecule is at a concentration of 0.01 mg / mL to 500 mg / mL, preferably 0.1 mg / mL to 400 mg / mL, more preferably 0.5 mg / mL to 200 mg / mL, and much more preferably 1 mg / mL to 150 mg / mL.

17. Pharmaceutical composition, according to any one of claims 1 to 16, characterized in that the buffer is at a concentration of 0.1 mM to 50 mM, preferably 0.5 mM to 40 mM, more preferably 1 mM to 30 mM, and much more preferably 5 mM to 20 mM.

18. Pharmaceutical composition, according to any one of claims 12 to 17, characterized in that the surfactant is in a concentration of 0.01 mg / mL to 10 mg / mL, preferably 0.05 mg / mL to 5 mg / mL, more preferably 0.1 mg / mL to 3 mg / mL and much more preferably 0.2 mg / mL to 2 mg / mL.

19. Pharmaceutical composition,according to any of claims 13 to 18, characterized in that the sugar is in a concentration of 1 mg / mL to 200 mg / mL, as per Petition 870250073483, dated 08 / 20 / 2025, page 25 / 42 9 / 23, in a preferred manner 10 mg / mL to 150 mg / mL, in a more preferred manner 30 mg / mL to 120 mg / mL and in a much more preferred manner 50 mg / mL to 10 mg / mL.

20. Pharmaceutical composition, according to any one of claims 14 to 1, characterized in that the auxiliary material is in a concentration of 0.001% w / v 20% w / v, preferably 0.005% w / v 15% w / v, more preferably 0.008% w / v 10% w / v, much more preferably 0.01% w / v 8% w / v.

21. Pharmaceutical composition, characterized in that it comprises: the FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; a histidine salt buffer, preferably a histidine hydrochloride buffer or a histidine acetate buffer,and more preferably a histidine-histidine hydrochloride buffer or a histidine-acetic acid buffer; a polysorbate; sucrose; wherein, optionally, the composition further comprises mannitol or ethylenediaminetetraacetic acid.

22. Pharmaceutical composition according to claim 21, characterized in that it comprises any of the following groups 1) to 4): Petition 870250073483, dated 20 / 08 / 2025, page 26 / 42 10 / 23 1) 0.01 mg / mL to 500 mg / mL of said FAP / CD40 binding molecule, as defined in any of claims 1 to 11; 0.1 mM to 50 mM of histidine salt buffer, preferably a histidine hydrochloride buffer or a histidine acetate buffer, and more preferably a histidine-histidine hydrochloride buffer or a histidine-acetic acid buffer; 0.01 mg / mL to 10 mg / mL of polysorbate; 1 mg / mL to 200 mg / mL of sucrose; wherein, optionally, the pharmaceutical composition further comprises 0.1% w / v 15% w / v of mannitol or 0.001% w / v 5% w / v of ethylenediaminetetraacetic acid; and the pharmaceutical composition has a pH of 3.5 to 7; 2) 0.1 mg / mL to 400 mg / mL of said FAP / CD40 binding molecule, as defined in any of claims 1 to 11; 0.5 mM to 40 mM of histidine salt buffer, preferably a histidine hydrochloride buffer or a histidine acetate buffer, and more preferably a histidine-histidine hydrochloride buffer or a histidine-acetic acid buffer; 0.05 mg / mL to 5 mg / mL of polysorbate; 10 mg / mL to 150 mg / mL of sucrose; wherein, optionally, the pharmaceutical composition Petition 870250073483, dated 20 / 08 / 2025, p. 27 / 42 11 / 23 additionally comprises 0.5% w / v 12% w / v of mannitol or 0.001% w / v 1% w / v of ethylenediaminetetraacetic acid; and the pharmaceutical composition has a pH of 4 to 6.5; 3) 0.5 mg / mL to 200 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; 1 mM to 30 mM of histidine salt buffer, preferably a histidine hydrochloride buffer or a histidine acetate buffer, and more preferably a histidine-histidine hydrochloride buffer or a histidine-acetic acid buffer; 0.1 mg / mL to 3 mg / mL of polysorbate; 30 mg / mL to 120 mg / mL of sucrose; wherein, optionally, the pharmaceutical composition further comprises 1% w / v to 10% w / v of mannitol or 0.005% w / v to 0.1% w / v of ethylenediaminetetraacetic acid; and the pharmaceutical composition has a pH of 4.2 to 6.2; 4) 1 mg / mL to 150 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; 5 mM to 20 mM of histidine salt buffer, preferably a histidine hydrochloride buffer or a histidine acetate buffer, and more preferably a histidine-histidine hydrochloride buffer or a histidine-acetic acid buffer; Petition 870250073483, dated 20 / 08 / 2025, p. 28 / 42 12 / 23 0.2 mg / mL to 2 mg / mL of polysorbate; 50 mg / mL to 100 mg / mL of sucrose; wherein, optionally, the pharmaceutical composition additionally comprises 1% w / w 10% w / w of mannitol or 0.005% w / w 0.1% w / w of ethylenediaminetetraacetic acid; and the pharmaceutical composition has a pH of 4.5 to 6.

23. Pharmaceutical composition according to claim 21, characterized in that it comprises any one of the following groups 1) to 21): 1) 1 to 150 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11;approximately 10 mM of histidine hydrochloride buffer or histidine acetate buffer, preferably a histidine-histidine hydrochloride buffer or a histidine-acetic acid buffer; 0.2 mg / mL to 1 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of 4.5 to 6; 2) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL, preferably approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL, of said FAP / CD40 binding molecule, as defined in any of claims 1 to 11; Petition 870250073483, dated 20 / 08 / 2025, p. 29 / 42 13 / 23 approximately 10 mM of histidine hydrochloride buffer, preferably a histidine-histidine hydrochloride buffer; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of approximately 5.5;3) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL, preferably approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL, of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; approximately 10 mM of histidine hydrochloride buffer, preferably a histidine-histidine hydrochloride buffer; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH around 5.3; 4) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL, preferably approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL, of said FAP / CD40 binding molecule, as defined in any of claims 1 to 11;approximately 10 mM of histidine hydrochloride buffer, preferably a histidine-histidine hydrochloride buffer; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of approximately 5.2; 5) about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, about 25 mg / mL, about 50 mg / mL, about 80 mg / mL or about 100 mg / mL, preferably about 5 mg / mL, about 10 mg / mL, about 20 mg / mL or about 25 mg / mL, of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; about 10 mM of histidine hydrochloride buffer, preferably a histidine-histidine hydrochloride buffer; about 0.4 mg / mL of polysorbate 80; about 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of about 5; 6) about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, about 25 mg / mL, about 50 mg / mL, about 80 mg / mL or about 100 mg / mL, preferably about 5 mg / mL, about 10 mg / mL, about 20 mg / mL or about 25 mg / mL, of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; about 10 mM of histidine hydrochloride buffer, preferably a histidine-histidine hydrochloride buffer; about 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of around 5.5; 7) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL, preferably approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL, of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; approximately 10 mM of histidine hydrochloride buffer, preferably a histidine-histidine hydrochloride buffer; approximately 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of approximately 5.3; 8) about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, about 25 mg / mL, about 50 mg / mL, about 80 mg / mL or about 100 mg / mL, preferably about 5 mg / mL, about 10 mg / mL, about 20 mg / mL or about 25 mg / mL, of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; about 10 mM of histidine hydrochloride buffer, preferably a histidine-histidine hydrochloride buffer; about 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of approximately 5.2; 9) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL, preferably approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL or approximately 25 mg / mL, of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; approximately 10 mM of histidine hydrochloride buffer, preferably a histidine-histidine hydrochloride buffer; approximately 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of approximately 5; 10) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; approximately 10 mM of histidine-acetic acid buffer; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of approximately 5; 11) about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, about 25 mg / mL, about 50 mg / mL, about 80 mg / mL or about 100 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; about 10 mM of histidine-acetic acid buffer; about 0.4 mg / mL of polysorbate 80; about 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of about 5.2; 12) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; approximately 10 mM of histidine-acetic acid buffer; approximately 0.4 mg / mL of polysorbate 80; Petition 870250073483, dated 20 / 08 / 2025, p. 34 / 42 18 / 23 approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH around 5.3; 13) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; approximately 10 mM of histidine-acetic acid buffer; approximately 0.4 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH around 5.5; 14) about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, about 25 mg / mL, about 50 mg / mL, about 80 mg / mL or about 100 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; about 10 mM of histidine-acetic acid buffer; about 0.6 mg / mL of polysorbate 80; about 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of about 5; 15) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; approximately 10 mM of histidine-acetic acid buffer; approximately 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of approximately 5.2; 16) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; approximately 10 mM of histidine-acetic acid buffer; approximately 0.6 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH around 5.3; 17) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; approximately 10 mM of histidine-acetic acid buffer; approximately 0.6 mg / mL of polysorbate 80; Petition 870250073483, dated 20 / 08 / 2025, p. 36 / 42 20 / 23 approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH around 5.5; 18) about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, about 25 mg / mL, about 50 mg / mL, about 80 mg / mL or about 100 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; about 10 mM of histidine-acetic acid buffer; about 0.8 mg / mL of polysorbate 80; about 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of about 5; 19) about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, about 25 mg / mL, about 50 mg / mL, about 80 mg / mL or about 100 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; about 10 mM of histidine-acetic acid buffer; about 0.8 mg / mL of polysorbate 80; about 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of about 5.2; 20) approximately 5 mg / mL, approximately 10 mg / mL, approximately 20 mg / mL, approximately 25 mg / mL, approximately 50 mg / mL, approximately 80 mg / mL or approximately 100 mg / mL of said FAP / CD40 binding molecule, as defined in any of claims 1 to 11; approximately 10 mM of histidine-acetic acid buffer; approximately 0.8 mg / mL of polysorbate 80; approximately 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of approximately 5.3; 21) about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, about 25 mg / mL, about 50 mg / mL, about 80 mg / mL or about 100 mg / mL of said FAP / CD40 binding molecule, as defined in any one of claims 1 to 11; about 10 mM of histidine-acetic acid buffer; about 0.8 mg / mL of polysorbate 80; about 80 mg / mL of sucrose; wherein the pharmaceutical composition has a pH of about 5.

5.

24. Lyophilized formulation, characterized in that the lyophilized formulation is capable of forming the pharmaceutical composition as defined in any one of claims 1 to 23 after reconstitution, or the lyophilized formulation is obtained by lyophilizing the pharmaceutical composition as defined in any one of claims 1 to 23. Petition 870250073483, dated 08 / 20 / 2025, pp. 38 / 42 22 / 23 25.Reconstituted solution, characterized in that the reconstituted solution is prepared by reconstituting the lyophilized formulation as defined in claim 24.

26. Article of manufacture, characterized in that it comprises a container, wherein the container contains the pharmaceutical composition as defined in any one of claims 1 to 23, the lyophilized formulation as defined in claim 24, or the reconstituted solution as defined in claim 25. 27.Use of the pharmaceutical composition as defined in any of claims 1 to 23, of the lyophilized formulation as defined in claim 24, or of the reconstituted solution as defined in claim 25, characterized in that it is used in the manufacture of a medicament to treat or improve a disease or disorder, wherein, preferably, the disease or disorder is a tumor or cancer; more preferably, the disease or disorder is lung cancer, prostate cancer, breast cancer, head and neck cancer, esophageal cancer, gastric cancer, colorectal cancer, bladder cancer, cervical cancer, uterine cancer, ovarian cancer, liver cancer, melanoma, renal cancer, squamous cell carcinoma or hematological cancer. Petition 870250073483, dated 20 / 08 / 2025, pp. 39 / 42 23 / 23A method for treating or improving a tumor or cancer, characterized in that it comprises: administering to an individual in need a therapeutically effective amount of the pharmaceutical composition as defined in any one of claims 1 to 23, of the lyophilized formulation as defined in claim 24, or of the reconstituted solution as defined in claim 25, wherein, preferably, the tumor or cancer is lung cancer, prostate cancer, breast cancer, head and neck cancer, esophageal cancer, gastric cancer, colorectal cancer, bladder cancer, cervical cancer, uterine cancer, ovarian cancer, liver cancer, melanoma, renal cancer, squamous cell carcinoma, or hematologic cancer. Petition 870250073483, dated 20 / 08 / 2025, pp. 40 / 42.

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