WO2022063193A1 - MOLÉCULE BIFONCTIONNELLE CIBLANT SIMULTANÉMENT PD-L1 ET TGFβ ET SON UTILISATION MÉDICALE - Google Patents

MOLÉCULE BIFONCTIONNELLE CIBLANT SIMULTANÉMENT PD-L1 ET TGFβ ET SON UTILISATION MÉDICALE Download PDF

Info

Publication number
WO2022063193A1
WO2022063193A1 PCT/CN2021/120006 CN2021120006W WO2022063193A1 WO 2022063193 A1 WO2022063193 A1 WO 2022063193A1 CN 2021120006 W CN2021120006 W CN 2021120006W WO 2022063193 A1 WO2022063193 A1 WO 2022063193A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
cancer
sequence
bifunctional molecule
antibody
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2021/120006
Other languages
English (en)
Chinese (zh)
Inventor
陈奇
蒋家骅
周传初
顾津明
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Qilu Pharmaceutical Research and Development Centre Ltd
Original Assignee
Shanghai Qilu Pharmaceutical Research and Development Centre Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shanghai Qilu Pharmaceutical Research and Development Centre Ltd filed Critical Shanghai Qilu Pharmaceutical Research and Development Centre Ltd
Priority to CN202180064731.0A priority Critical patent/CN116323657B/zh
Publication of WO2022063193A1 publication Critical patent/WO2022063193A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/495—Transforming growth factor [TGF]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/71—Receptors; Cell surface antigens; Cell surface determinants for growth factors; for growth regulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes

Definitions

  • the invention belongs to the field of biomedicine, in particular to a bifunctional molecule that simultaneously targets PD-L1 and TGF ⁇ and its medical application.
  • PD-1 and PD-L1 are the core proteins of the immune check signaling pathway, which can protect their own tissues against normal conditions. Attacked by T cells, but also used by tumor cells to evade clearance by the immune system.
  • Antibody drugs targeting PD-1 or PD-L1 can significantly inhibit tumor growth, and patients with multiple cancer types who respond to PD-1/PD-L1 therapy have significant survival benefits.
  • the response rate of tumor patients to PD-1/PD-L1 therapy is not high, one of the reasons is that the complex microenvironment of the tumor has multiple immunosuppressive signaling pathways. Therefore, bispecific antibodies targeting PD-1/PD-L1 and another immune checkpoint at the same time have more therapeutic potential.
  • TGF ⁇ transforming growth factor beta
  • TGF ⁇ acts as an immunoregulatory factor to protect its own tissues from the attack of the immune system, but in the tumor microenvironment, TGF ⁇ acts on the immune system to help tumor cells achieve immune escape and accelerate tumor progression.
  • TGF ⁇ has three subtypes, namely TGF ⁇ 1, TGF ⁇ 2 and TGF ⁇ 3, which are highly expressed in various tumor tissues, and their high expression in serum is also associated with poor prognosis.
  • TGF ⁇ 1 can directly prevent the differentiation of T cells and inhibit the function of T cells and NK cells to kill cancer cells.
  • drugs that block both the PD-1/PD-L1 signaling pathway and the TGF ⁇ signaling pathway are expected to further improve the response rate of immunotherapy compared with drugs targeting a single signaling pathway.
  • bifunctional molecules composed of PD-L1 antibody and TGF ⁇ RII fusion protein have been disclosed, such as WO2015118175A2, WO2018205985A1 and the like.
  • WO2015118175A2 WO2018205985A1
  • the present invention provides a technical solution with better stability in vivo and in vitro.
  • the present invention provides a bifunctional molecule targeting PD-L1 and TGF ⁇ at the same time, comprising a PD-L1-targeting part and a TGF- ⁇ receptor part, and the PD-L1-targeting part is a PD-L1 antibody,
  • the TGF- ⁇ receptor part is the N-terminal truncated form of the extracellular region of TGF ⁇ RII (TGF-beta receptor type-2), and the C-terminus of each heavy chain of the PD-L1 antibody is connected to a TGF ⁇ RII extracellular region
  • TGF ⁇ RII TGF-beta receptor type-2
  • the N-terminal truncated form of the PD-L1 antibody, the light chain and heavy chain variable region CDR sequences of the PD-L1 antibody are as follows:
  • LCDR1 The sequence of LCDR1 is shown in SEQ ID NO: 9;
  • LCDR2 is shown in SEQ ID NO: 10;
  • LCDR3 The sequence of LCDR3 is shown in SEQ ID NO: 11;
  • HCDR1 The sequence of HCDR1 is shown in SEQ ID NO: 12;
  • HCDR2 The sequence of HCDR2 is shown in SEQ ID NO: 13;
  • HCDR3 The sequence of HCDR3 is shown in SEQ ID NO:14.
  • the full-length sequence of the extracellular region of TGF ⁇ RII is shown in SEQ ID NO: 1, and the N-terminal truncated form is a truncation of 17-27 amino acids.
  • the sequence of the TGF- ⁇ receptor part is shown in SEQ ID NO: 2.
  • the light chain variable region sequence of the PD-L1 antibody is shown in SEQ ID NO: 15; the heavy chain variable region sequence is shown in SEQ ID NO: 16.
  • the light chain sequence of the PD-L1 antibody is shown in SEQ ID NO: 5; the heavy chain sequence is shown in SEQ ID NO: 6, and the K at the last position of the C-terminal of the heavy chain is mutated into A.
  • the C-terminal of the heavy chain of the PD-L1 antibody is connected to the N-terminal truncated form of the extracellular region of TGF ⁇ RII through a linking peptide.
  • the connecting peptide is (G 4 S) X G, and the x is 3-6, preferably 4-5.
  • the light chain sequence of the PD-L1 antibody is shown in SEQ ID NO: 7, and the overall sequence of the heavy chain and the N-terminal truncated form of the extracellular region of TGF ⁇ RII is shown in SEQ ID NO: 8.
  • the present invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising the above-mentioned bifunctional molecule and a pharmaceutically acceptable carrier.
  • the present invention also provides a nucleic acid molecule encoding the above-mentioned bifunctional molecule.
  • the present invention also provides an expression vector, which contains the above-mentioned nucleic acid molecule.
  • the present invention also provides a host cell comprising the above-mentioned expression vector, the host cell is selected from bacteria, yeast and mammalian cells; preferably mammalian cells; more preferably HEK293E cells, expi293 cells or CHO cells.
  • the present invention also provides the use of the above-mentioned bifunctional molecule in the preparation of a medicament for treating cancer.
  • the cancer is a PD-L1 positive tumor.
  • the cancer is selected from the group consisting of lung cancer, gastric cancer, melanoma, kidney cancer, breast cancer, bowel cancer, liver cancer, ovarian cancer, cervical cancer, bladder cancer, esophageal cancer, pancreatic cancer and head and neck cancer.
  • the present invention also provides a method for treating and preventing tumors, comprising administering to a patient in need a therapeutically effective amount of the above-mentioned bifunctional molecule or the above-mentioned pharmaceutical composition.
  • Figure 1 Schematic diagram of the bifunctional molecular structure.
  • Figure 2 Results of in vitro binding of bifunctional molecules to human PD-L1.
  • Figure 3 Results of in vitro binding of bifunctional molecules to human TGF ⁇ 1.
  • Figure 4 Detection of bifunctional molecule 6 binding to human PD-L1 and human TGF ⁇ 1 using SPR technology.
  • Figure 5 The ability of bifunctional molecules to block the PD-1/PD-L1 signaling pathway was tested at the cellular level.
  • Figure 7 Effect of bifunctional molecules on tumor weight in the MDA-MB-231 model.
  • Figure 8 Effect of bifunctional molecules on tumor volume and tumor weight in a humanized MC-38 model.
  • the term “about” is meant to include ⁇ 20% of the specified value, or in some cases ⁇ 10%, or in some cases ⁇ 5%, or within ⁇ 1% in some cases, or ⁇ 0.1% in some cases.
  • antibody typically refers to a Y-type tetramer comprising two heavy (H) polypeptide chains and two light (L) polypeptide chains held together by covalent disulfide bonds and non-covalent interactions protein.
  • Natural IgG antibodies have such a structure. Each light chain consists of a variable domain (VL) and a constant domain (CL). Each heavy chain contains a variable domain (VH) and constant region.
  • IgA immunoglobulin A
  • IgD immunoglobulin D
  • IgE immunoglobulin G
  • IgG immunoglobulin M
  • the corresponding heavy chain constant domains are called ⁇ , ⁇ , ⁇ , ⁇ and ⁇ , respectively
  • IgG and IgA can be further divided into different
  • the subclasses, such as IgG can be divided into IgG1, IgG2, IgG3, IgG4, and IgA can be divided into IgA1 and IgA2.
  • the light chains of antibodies from any vertebrate species can be assigned to one of two distinct types, called kappa and lambda, based on the amino acid sequence of their constant domains.
  • the constant region comprises three domains called CH1, CH2 and CH3 (IgM and IgE have a fourth domain CH4).
  • the CH1 and CH2 domains are separated by a flexible hinge region, which is a variable length proline and cysteine rich segment.
  • Each class of antibodies further comprises interchain and intrachain disulfide bonds formed by paired cysteine residues.
  • variable region exhibit significant changes in amino acid composition from one antibody to another, and are primarily responsible for antigen recognition and binding.
  • the variable regions of each light/heavy chain pair form the antibody binding site such that an intact IgG antibody has two binding sites (ie it is bivalent).
  • the variable region (VH) of the heavy chain and the variable region (VL) domain of the light chain each contain three regions of extreme variability known as hypervariable regions (HVRs), or more commonly, known as Complementarity determining regions (CDRs), VH and VL each have four framework regions FR, which are represented by FR1, FR2, FR3, and FR4, respectively.
  • CDR and FR sequences typically occur in the following sequences of the heavy chain variable domain (or light chain variable domain): FR1-HCDR1(LCDR1)-FR2-HCDR2(LCDR2)-FR3-HCDR3(LCDR3)- FR4.
  • Fc is used to define the C-terminal region of an immunoglobulin heavy chain, which region comprises at least a portion of the constant region.
  • the term includes native sequence Fc regions and variant Fc regions.
  • antibodies in a broad sense may include, for example, polyclonal antibodies, monoclonal antibodies, chimeric antibodies, humanized antibodies, and primatized antibodies, CDR-grafted antibodies (CDR- grafted antibodies), human antibodies (including recombinantly produced human antibodies), recombinantly produced antibodies, intrabodies, multispecific antibodies, bispecific antibodies, monovalent antibodies, multivalent antibodies, anti-idiotypic antibodies, synthetic antibodies (including mutant proteins and their variants) and so on.
  • monoclonal antibody refers to a substantially homogeneous antibody produced by a single cell clone directed against only a particular epitope.
  • Monoclonal antibodies can be prepared using a variety of techniques known in the art, including hybridoma techniques, recombinant techniques, phage display techniques, transgenic animals, synthetic techniques, or a combination of the foregoing, and the like.
  • antibody fragment encompasses at least a portion of an intact antibody.
  • a “fragment” of an antibody molecule includes an "antigen-binding fragment” of an antibody, and the term “antigen-binding fragment” refers to an immunoglobulin or antibody that specifically binds a selected antigen or an immunogenic determinant thereof Or reacted polypeptide fragments, or fusion protein products further derived from such fragments, such as single-chain antibodies, extracellular binding domains in chimeric antigen receptors, and the like.
  • Exemplary antibody fragments or antigen-binding fragments thereof include, but are not limited to, variable light chain fragments, variable heavy chain fragments, Fab fragments, F(ab') 2 fragments, Fd fragments, Fv fragments, single domain antibodies, linear Antibodies, single chain antibodies (scFv) and bispecific or multispecific antibodies formed from antibody fragments, etc.
  • an antigen refers to a substance that is recognized and specifically bound by an antibody or antibody-binding fragment.
  • an antigen can include any immunogenic fragment or determinant of a selected target, including mono-epitopes, poly-epitopes, mono-structures domain, multi-domain, intact extracellular domain (ECD) or protein.
  • ECD extracellular domain
  • Peptides, proteins, glycoproteins, polysaccharides, and lipids, parts thereof, and combinations thereof can constitute antigens.
  • Non-limiting exemplary antigens include tumor antigens or pathogen antigens, and the like.
  • Antigen can also refer to a molecule that elicits an immune response.
  • the antigen can be an isolated full-length protein, a cell surface protein (eg, immunized with a cell expressing at least a portion of the antigen on its surface), or a soluble protein (eg, immunized with only the ECD portion of the protein), or a protein Constructs (eg, Fc antigens).
  • the antigen can be produced in genetically modified cells. Any of the foregoing antigens may be used alone or in combination with one or more immunogenicity enhancing adjuvants known in the art.
  • the DNA encoding the antigen can be genomic or non-genomic (eg, cDNA) and can encode at least a portion of the ECD sufficient to elicit an immunogenic response.
  • Cells in which the antigen is expressed can be transformed using any vector including, but not limited to, adenoviral vectors, lentiviral vectors, plasmids, and non-viral vectors such as cationic lipids.
  • epitope refers to the site on an antigen to which an immunoglobulin or antibody specifically binds.
  • Epitopes can be formed by adjacent amino acids, or non-adjacent amino acids juxtaposed by tertiary folding of the protein. Epitopes formed by adjacent amino acids are typically retained upon exposure to denaturing solvents, whereas epitopes formed by tertiary folding are typically lost upon treatment with denaturing solvents. Epitopes typically exist in unique spatial conformations and include at least 3-15 amino acids.
  • Methods for determining the epitope to which a given antibody binds are well known in the art and include immunoblotting and immunoprecipitation assays, among others. Methods for determining the spatial conformation of epitopes include techniques in the art, such as X-ray crystallography and two-dimensional nuclear magnetic resonance, among others.
  • affinity refers to the strength of the sum of all non-covalent interactions between a single binding site of a molecule (eg, an antibody) and its binding partner (eg, an antigen).
  • KD refers to the dissociation constant for a particular antibody-antigen interaction. Binding affinity can be determined using various techniques known in the art, such as surface plasmon resonance, biolayer interferometry, dual polarization interferometry, static light scattering, dynamic light scattering, isothermal titration calorimetry, ELISA, analytical ultrafast Centrifugation and flow cytometry, etc.
  • biological activity refers to the ability of an antibody to bind an antigen and cause a measurable biological response, which can be measured in vitro or in vivo.
  • pharmaceutical formulation or “formulation” or “formulation formulation” means a product that exists in a form that allows the biological activity of the active ingredient to be effective and that does not contain other components that are toxic to the subject to which the formulation is to be administered .
  • solution formulation means a formulation that is liquid at a temperature of at least about 2°C to about 8°C at atmospheric pressure.
  • deamidation means that one or more asparagine residues in an antibody have been derivatized, eg, aspartic acid or iso-aspartic acid.
  • aggregated antibody is an antibody that has been found to aggregate with other antibody molecules, especially after freezing and/or agitation.
  • stable formulation is one in which the protein substantially retains its physical and/or chemical stability and/or biological activity after storage.
  • the formulation substantially retains its physical and chemical stability, as well as its biological activity, after storage.
  • the shelf life is generally selected based on the shelf life of the formulation.
  • Various analytical techniques for measuring protein stability are available in the art. Stability can be measured at a selected temperature for a selected time.
  • Stability can be assessed qualitatively and/or quantitatively in many different ways, including assessing aggregate formation (eg, by size exclusion chromatography, by measuring turbidity, and/or by visual observation); by use of cation exchange chromatography or capillary partition electrophoresis to assess charge heterogeneity; amino-terminal or carboxy-terminal sequence analysis; mass spectrometry analysis; SDS-PAGE analysis to compare reduced and intact antibodies; peptide mapping analysis; assessment of biological activity or antigen-binding function of antibodies; etc.
  • Instability may include any one or more of the following: aggregation, deamidation (eg, Asn deamidation), oxidation (eg, Met oxidation), isomerization (eg, Asp isomerization), shearing Cleavage/hydrolysis/fragmentation (eg hinge region fragmentation), succinimide formation, unpaired cysteines, N-terminal extension, C-terminal processing, differential glycosylation, etc.
  • deamidation eg, Asn deamidation
  • oxidation eg, Met oxidation
  • isomerization eg, Asp isomerization
  • shearing Cleavage/hydrolysis/fragmentation eg hinge region fragmentation
  • succinimide formation unpaired cysteines, N-terminal extension, C-terminal processing, differential glycosylation, etc.
  • buffer refers to a pharmaceutically acceptable excipient that stabilizes the pH of a pharmaceutical formulation.
  • Suitable buffers are well known in the art and can be found in the literature.
  • Preferred pharmaceutically acceptable buffers include, but are not limited to: histidine buffer, citrate buffer, succinate buffer, acetate buffer, arginine buffer, phosphate buffer, or the like mixture, etc.
  • the pH of the buffer is adjusted with acids or bases known in the art, the pH can be adjusted to a value in the range of 4.5-6.0, in particular to a value in the range of 4.5-5.5, most particularly to pH 5.5. 2.
  • stabilizer means a pharmaceutically acceptable excipient which protects the active pharmaceutical ingredient and/or formulation from chemical and/or physical degradation during manufacture, storage and application.
  • Stabilizers include, but are not limited to, sugars, amino acids, polyols, cyclodextrins, and the like.
  • surfactant refers to a pharmaceutically acceptable excipient used to protect a protein formulation against physical stress such as stirring and shearing.
  • Pharmaceutically acceptable surfactants include: polyoxyethylene sorbitan fatty acid esters (Tween), polyoxyethylene alkyl ethers (such as those sold under the trademark Brij TM ) and polyoxyethylene-polyoxypropylene Copolymer (Poloxamer, Pluronic).
  • Polyoxyethylene sorbitan-fatty acid esters include polysorbate 20 (sold under the trademark Tween 20 TM ) and polysorbate 80 (sold under the trademark Tween 80 TM ).
  • combination drug refers to a combination comprising two or more pharmaceutical formulations, each having an active ingredient, which is required to be used in combination when administered to a subject.
  • the active ingredients can be mixed together to form a single administration unit, or they can be separately formed into administration units and used separately.
  • an effective amount refers to the dose of a pharmaceutical formulation of an antibody or fragment of the invention which, after administration to the patient in single or multiple doses, produces the desired effect in the treated patient.
  • An effective amount can be readily determined by the attending physician, who is skilled in the art, by taking into account a variety of factors such as ethnic differences; weight, age, and health; the specific disease involved; the severity of the disease; the individual patient's response; The particular antibody administered; the mode of administration; the bioavailability characteristics of the administered formulation; the chosen dosing regimen; and the use of any concomitant therapy.
  • kit includes an effective amount of one or more pharmaceutical formulations or combinations of the invention in unit dosage form.
  • the kit may contain a sterile container of the therapeutic or prophylactic composition; such container may be a box, ampule, bottle, vial, tube, bag, blister pack, or other suitable known in the art container form.
  • Such containers may be made of plastic, glass, laminated paper, metal foil, or other materials suitable for holding drugs.
  • the kit also includes instructions for administering the pharmaceutical formulation or combination of the present invention to an individual. A method of treating or preventing a disease using the pharmaceutical formulation or combination of the present invention is generally included in the specification.
  • the engineered antibodies or antigen-binding fragments thereof of the present invention can be prepared and purified using conventional methods.
  • cDNA sequences encoding heavy and light chains can be cloned and recombined into expression vectors.
  • the recombinant immunoglobulin expression vector can be stably transfected into CHO cells.
  • mammalian-like expression systems lead to glycosylation of antibodies, especially at the highly conserved N-terminus of the Fc region.
  • Stable clones are obtained by expressing antibodies that specifically bind to human antigens. Positive clones were expanded in serum-free medium in bioreactors for antibody production.
  • the antibody-secreted culture medium can be purified and collected by conventional techniques.
  • Antibodies can be filtered and concentrated by conventional methods. Soluble mixtures and polymers can also be removed by conventional methods, such as molecular sieves, ion exchange.
  • Subjects of the present invention refers to any animal, such as a mammal or a marsupial.
  • Subjects of the present invention include, but are not limited to, humans, non-human primates (eg, cynomolgus or rhesus monkeys or other types of rhesus monkeys), mice, pigs, horses, donkeys, cattle, sheep, rats, and any species of poultry.
  • tumor refers to a disease characterized by the pathological proliferation of cells or tissues, and its subsequent migration or invasion of other tissues or organs. Tumor growth is usually uncontrolled and progressive, and does not induce or inhibit normal cell proliferation. Tumors can affect a variety of cells, tissues or organs, including but not limited to those selected from the group consisting of bladder, bone, brain, breast, cartilage, glial cells, esophagus, fallopian tubes, gallbladder, heart, intestine, kidney, liver, lung, lymph nodes, Nervous tissue, ovary, pancreas, prostate, skeletal muscle, skin, spinal cord, spleen, stomach, testis, thymus, thyroid, trachea, ureter, urethra, uterus, vaginal organs, or tissue or corresponding cells.
  • bladder bone, brain, breast, cartilage, glial cells, esophagus, fallopian tubes, gallbladder, heart, intestine, kidney, liver, lung, lymph nodes, Nerv
  • Tumors include cancers such as sarcomas, carcinomas, or plasmacytomas (malignant tumors of plasma cells).
  • the tumors described in the present invention may include, but are not limited to, leukemia (such as acute leukemia, acute lymphocytic leukemia, acute myeloid leukemia, acute myeloid leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, Acute monocytic leukemia, chronic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, polycythemia vera), lymphoma (Hodgkin's disease, non-Hodgkin's disease), primary macroglobulinemia, severe Chain disease, solid tumors such as sarcomas and cancers (eg, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, endothelial sar
  • the "tumor” includes but is not limited to: pancreatic cancer, liver cancer, lung cancer, gastric cancer, esophageal cancer, head and neck squamous cell carcinoma, prostate cancer, colon cancer, breast cancer, lymphoma, gallbladder cancer, Kidney cancer, leukemia, multiple myeloma, ovarian cancer, cervical cancer and glioma.
  • disease or “condition” or “disorder” and the like refer to any alteration or disorder that impairs or interferes with the normal function of a cell, tissue or organ.
  • diseases include, but are not limited to, tumors, pathogen infections, autoimmune diseases, T cell dysfunctional diseases, or defective immune tolerance (e.g., transplant rejection).
  • treatment refers to clinical intervention in an attempt to alter an individual or to manipulate a cell-induced disease process, either prophylactically or in a clinical pathological process.
  • Therapeutic effects include, but are not limited to, preventing the occurrence or recurrence of the disease, alleviating symptoms, reducing the direct or indirect pathological consequences of any disease, preventing metastasis, slowing the rate of disease progression, improving or relieving the condition, relieving or improving the prognosis, etc.
  • pharmaceutical composition refers to a mixture comprising one or more of the compounds described herein, or a physiological/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, such as physiological/pharmaceutically acceptable salts or prodrugs.
  • Pharmaceutically acceptable carriers and excipients are included in the pharmaceutical composition. The purpose of the pharmaceutical composition. The purpose of the pharmaceutical composition is to facilitate the administration to the organism, facilitate the absorption of the active ingredient and then exert the biological activity.
  • the structure of the bifunctional molecule is to connect the extracellular domain of TGF ⁇ RII protein at the C-terminus of the heavy chain of the PD-L1 antibody, as shown in Figure 1.
  • the full length of the extracellular domain of TGF ⁇ RII protein is composed of 136 amino acids. It has been found that truncation of the N-terminal less than 27 amino acids will not affect the binding ability of TGF ⁇ cytokines, nor its function, and can improve the dual function.
  • the stability of the molecule is preferably 17-27 amino acid truncation, more preferably the N-terminal truncated 22 amino acid form, wherein the full-length sequence ECD (1-136) of the extracellular region of the TGF ⁇ RII protein is as SEQ ID NO: 1 As shown, the N-terminal truncated 22 amino acid sequence ECD (23-136) of the extracellular region of the TGF ⁇ RII protein is shown in SEQ ID NO: 2.
  • the CDR sequences of the light chain and heavy chain variable regions of the PD-L1 antibody are as follows:
  • LCDR1 The sequence of LCDR1 is shown in SEQ ID NO: 9;
  • LCDR2 is shown in SEQ ID NO: 10;
  • LCDR3 The sequence of LCDR3 is shown in SEQ ID NO: 11;
  • HCDR1 The sequence of HCDR1 is shown in SEQ ID NO: 12;
  • HCDR2 The sequence of HCDR2 is shown in SEQ ID NO: 13;
  • HCDR3 The sequence of HCDR3 is shown in SEQ ID NO:14.
  • the light chain variable region sequence of the PD-L1 antibody is shown in SEQ ID NO: 15; the heavy chain variable region sequence is shown in SEQ ID NO: 16.
  • the light chain sequence of the PD-L1 antibody is shown in SEQ ID NO: 5; the heavy chain sequence is shown in SEQ ID NO: 6.
  • TGF ⁇ RII protein The extracellular domain of different truncated forms of TGF ⁇ RII protein is linked to the C-terminus of the heavy chain of the PD-L1 antibody through (G 4 S) x G, and the K at the last position of the C-terminus of the heavy chain of the original antibody is mutated to A, and the light Chain together, expressed by HEK293E or expi293 cell system, to obtain bifunctional molecules as shown in Table 1 below:
  • bifunctional molecules sequence description Number of N-terminal truncated amino acids bifunctional molecule 1 anti-PDL1-(G 4 S) 4 G-ECD(18-136,N19S) 17 bifunctional molecule 2 anti-PDL1-(G 4 S) 4 G-ECD(19-136,N19S) 18 bifunctional molecule 3 anti-PDL1-(G 4 S) 4 G-ECD(20-136) 19 bifunctional molecule 4 anti-PDL1-(G 4 S) 4 G-ECD(21-136) 20 bifunctional molecule 5 anti-PDL1-(G 4 S) 5 G-ECD(21-136) 20 bifunctional molecule 6 anti-PDL1-(G 4 S) 5 G-ECD(23-136) twenty two bifunctional molecule 7 anti-PDL1-(G 4 S) 5 G-ECD(26-136) 25 bifunctional molecule 8 anti-PDL1-(G 4 S) 5 G-ECD(28-136) 27
  • ECD(n-136) in the sequence is the truncated form of the extracellular domain of TGF ⁇ RII protein, n is the sequence number of the truncated starting amino acid; N19S means that the 19th amino acid is mutated to S.
  • the light chain sequence of the above-mentioned bifunctional molecule 6 is shown in SEQ ID NO: 7, and the overall sequence of the heavy chain and the N-terminal truncated form of the extracellular region of TGF ⁇ RII is shown in SEQ ID NO: 8.
  • the bifunctional molecules expressed in cells were purified by Example 2, and the resulting proteins were used in the experiments described below.
  • the cell culture medium was centrifuged at 4500 g for 30 min to collect the supernatant, and filtered with a 0.22 ⁇ m filter.
  • the supernatant was purified using MabSelect SuRe Protein A column (GE Healthcare); the equilibration buffer was 1 ⁇ PBS, equilibrated for 10 column volumes, and the cell supernatant was loaded with Protein A combined. and 0.1% Triton X114 in 1 ⁇ PBS for 10 column volumes, then with 1 ⁇ PBS for 10 column volumes, then with 100mM sodium acetate (pH3.5) elution buffer, according to A280 UV
  • the eluted sample was collected from the absorption peak, and the collected eluted sample was neutralized with 1 M Tris-HCl (pH 9.0).
  • the neutralized eluted samples were filtered with a 0.22 ⁇ m filter membrane, concentrated by ultrafiltration, and then subjected to molecular sieve chromatography with HiLoadTM26/600SuperdexTM200pg (GE Healthcare), the buffer was 1 ⁇ PBS, and the target protein peaks were merged according to A280 ultraviolet absorption.
  • the collected protein samples were identified by SEC-HPLC with a purity greater than 95%; the collected protein samples were detected by LAL (Endosafe nexgen-PTS) method for endotoxin, and the result was less than 1EU/mg.
  • the light chain sequence of the PD-L1 monoclonal antibody in the following experiments is shown in SEQ ID NO: 5; the heavy chain sequence is shown in SEQ ID NO: 6.
  • the positive control molecule is SHR-1701, which is a bifunctional molecule targeting PD-L1 and TGF ⁇ of Hengrui Medicine.
  • the sequence is shown in Patent WO2018205985A1.
  • its light chain sequence is as shown in SEQ ID NO:3.
  • the overall sequence of the heavy chain and the N-terminal truncated form of the extracellular region of TGF ⁇ RII is shown in SEQ ID NO: 4. Expression purification was also performed as described above.
  • Examples 3-5 are binding activity evaluation experiments
  • Examples 6-7 are cell function evaluation experiments
  • Example 8 is a pharmacokinetic evaluation experiment
  • Examples 9-10 are in vivo pharmacodynamic evaluation experiments.
  • Example 3 ELISA detection of PD-L1/TGF ⁇ bifunctional molecule combined with human PD-L1 experiment
  • the M5 plate reader of MD Company detects the absorbance value at the wavelength of 450 nm, and uses the softmax software to process the data.
  • the experimental results of ELISA binding are shown in Figure 2.
  • the bifunctional molecule 6 retains the binding activity to human PD-L1, and the binding ability is comparable to that of the positive control molecule.
  • Example 4 ELISA detection of PD-L1/TGF ⁇ bifunctional molecule combined with human TGF ⁇ 1 experiment
  • the M5 plate reader of MD Company detects the absorbance value at the wavelength of 450 nm, and uses the softmax software to process the data.
  • Biacore detects the affinity and kinetic properties of PD-L1/TGF ⁇ bifunctional molecule and antigen
  • the affinity and kinetic properties of the bifunctional molecule to human PD-L1 and human TGF ⁇ 1 were analyzed using a Biacore 8K instrument.
  • the CM5 chip was first activated with EDC and NHS, then anti-human Fc mouse mAb was immobilized, and then blocked with ethanolamine.
  • the bifunctional molecule was diluted to 5 ⁇ g/mL with HBS-EP+ (10 mM HEPES, pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.05% P20) buffer at 10 ⁇ L/mL The flow rate of min was captured for 30s.
  • Human PD-L1 was diluted to serial concentrations (5nM-0.078nM) two-fold stepwise, and was bound for 180s at a flow rate of 30 ⁇ L/min, and dissociated for 300s.
  • the bifunctional molecule was diluted to 1 ⁇ g/mL with HBS-EP+ buffer and captured at a flow rate of 10 ⁇ L/min for 60 s.
  • Human TGF ⁇ 1 was two-fold serially diluted to serial concentrations (2nM-0.00156nM), bound for 120s at a flow rate of 30 ⁇ L/min, and dissociated for 1200s.
  • Ligand (ligand) Analyte ka(1/Ms) kd(1/s) KD(M) bifunctional molecule 6 Human PD-L1 1.41E+06 7.44E-04 5.26E-10 bifunctional molecule 6 Human TGF ⁇ 1 7.44E+08 3.65E-04 4.90E-13
  • Example 6 Experiment to detect bifunctional molecules blocking PD-1/PD-L1 signaling pathway
  • the biological activity of the molecule was detected by reporter gene method, and CHO cells transfected with PD-L1 and anti-CD3-single-chain antibody fragment (scFv) were used as target cells to transfect PD-1 and NFAT element-regulated CHO cells.
  • Jurkat cells with the luciferase gene were used as effector cells.
  • the anti-CD3-scFv on the CHO cell membrane binds to CD3 on the surface of Jurkat cells, it will present an activation signal to Jurkat cells, thereby expressing luciferase; PD-L1 on the surface of CHO cells binds to PD-1 on the surface of Jurkat cells.
  • the inhibitory signal is delivered to Jurkat cells to inhibit the expression of luciferase; while the bifunctional molecule can block the binding of PD-1 and PD-L1, thereby releasing the delivery of the inhibitory signal, restoring the expression of luciferase, and generating a fluorescent signal.
  • CHO-PDL1-CD3L cells were harvested, washed and resuspended to 4E5 cells/mL;
  • Example 7 Experiment to detect bifunctional molecules blocking TGF ⁇ /SMAD signaling pathway
  • the SBE Reporter-HEK293 cell line purchased from BPS Bioscience was used to monitor the activity of TGF ⁇ /SMAD signaling pathway.
  • TGF ⁇ protein binds to cell surface receptors and initiates a signaling cascade leading to phosphorylation and activation of SMAD2 and SMAD3, which then form a complex with SMAD4.
  • the SMAD complex then translocates to the nucleus and binds to the SMAD-binding element (SBE) in the nucleus, resulting in the transcription and expression of TGF ⁇ /SMAD-responsive genes, including the luciferase gene transfected into the cell, resulting in a fluorescent signal.
  • SBE SMAD-binding element
  • the bifunctional molecule contains TGF ⁇ RII fusion protein, which can prevent the binding of TGF ⁇ protein to cell surface receptors, thereby inhibiting the expression of fluorescent signals.
  • TGF ⁇ protein was added after 4 hours, the final concentration was 10ng/mL;
  • the dosage of bifunctional molecule and positive control was 10mg/kg, and the intravenous infusion was completed in 30min.
  • Blood collection time points were Pre-dose, 30min, 1hr, 6hr, 24hr, 2d, 4d, 7d, 10d, 14d, 21d, 28d, 35d, 42d. After the whole blood was collected, it was left to stand at room temperature for half an hour, and the supernatant was collected by centrifugation (6000 rpm, 8 minutes, 4°C) to collect serum.
  • the concentration of drug molecules in serum was detected by ELISA, and the detection process was described as follows:
  • the M5 plate reader of MD Company detects the absorbance value at the wavelength of 450 nm, and uses the softmax software to process the data.
  • the measured monkey serum concentration was calculated by Phoenix Winnolin software to obtain pharmacokinetic parameters, as shown in Table 3 below.
  • the units of the concentrations in the table are all ⁇ g/mL, and BLQ is below the detection limit.
  • the units of concentrations in the above table are all ⁇ g/mL, and BLQ is below the detection limit.
  • bifunctional molecules have target-mediated drug clearance in vivo, and there are certain immunogens. Therefore, the bifunctional molecule has good pharmacokinetic properties, stable properties in cynomolgus monkeys, and no obvious off-target binding.
  • bifunctional molecule 6 is of the IgG1 subtype and the positive control is of the IgG4 subtype, and antibodies of the IgG1 subtype are generally considered to be more stable in vivo than those of the IgG4 subtype. Therefore, bifunctional molecule 6 is expected to achieve a longer half-life than the positive control in subsequent clinical experiments, further reducing the frequency of administration and the cost of medication.
  • Example 9 Evaluation of the tumor inhibition rate of bifunctional molecules in the subcutaneous xenograft model of human breast cancer MDA-MB-231 mice
  • MDA-MB-231 cells were cultured in L-15 medium containing 10% fetal bovine serum (FBS). MDA-MB-231 cells in logarithmic growth phase were collected and resuspended in HBSS to an appropriate concentration for subcutaneous tumor inoculation of NCG mice. Normal human peripheral blood was taken, and human PBMCs were separated by density gradient centrifugation. The separated PBMCs were added to MDA-MB-231 cells treated with Mitomycin C, and PBMCs and MDA-MB-231 cells were co-cultured for 6 days. IL-2 and 10% FBS in RPMI 1640 medium.
  • FBS fetal bovine serum
  • the cultured PBMCs were collected, and the PBMCs were mixed with freshly digested MDA-MB-231 cells and inoculated into the right subcutaneous of 64 NCG mice (Jiangsu JiCui Yaokang Biotechnology Co., Ltd.), with 8 mice in each group. 8 groups. After inoculation, the mice were randomly administered into groups according to their body weight. The detailed administration method, dose and route of administration were shown in Table 5. The day of group administration was Day 0.
  • Tumor growth inhibition rate TGI (%) [1-(T i -T 0 )/( V i -V 0 )] ⁇ 100, where Ti is the average tumor volume after the compound group started to be administered, and T 0 is the compound The mean tumor volume of the group at the first administration, V 0 is the mean tumor volume of the vehicle control group at the first administration, and Vi is the mean tumor volume of the vehicle control group after the start of administration. Body weights of all tumor-bearing mice were measured twice a week.
  • the tumor mass was weighed and photographed. The results are shown in Table 6 below:
  • the results of tumor weighing are shown in Figure 7. It can be seen that the bifunctional molecule 6 has obvious tumor inhibitory effect. At the doses of 5mg/kg, 10mg/kg and 20mg/kg, the tumor inhibition rates were 59.92%, 62.78% and 71.48%, respectively. The tumor inhibition rates increased with the dose. increase with the increase. At the level of equimolar dose, the efficacy of bifunctional molecule 6 is better than that of PD-L1 monoclonal antibody and the combination of PD-L1 monoclonal antibody and TGFbRII fusion protein. At the same time, the mice in the bifunctional molecule 6 administration group did not die, and the weight of the mice increased steadily, indicating that the bifunctional molecule 6 is safe in mice.
  • Example 10 Evaluation of the tumor inhibition rate of bifunctional molecules in the human colon cancer MC-38 mouse subcutaneously transplanted tumor model
  • MC38 (humanized PDL1) cells were subcutaneously inoculated into the right axilla of PD-L1 humanized mice at 1 ⁇ 10 6 cells/0.1 mL, totaling 60 mice.
  • the first group was IgG1 isotype antibody (20 mg/kg)
  • the second group is bifunctional molecule 6 (5mg/kg)
  • the third group is bifunctional molecule 6 (10mg/kg)
  • the fourth group is bifunctional molecule 6 (20mg/kg)
  • the fifth group is positive control SHR -1701 (20mg/kg).
  • the animals were grouped on the day of administration, and the administration volume was 10 mL/kg, and the administration method was intraperitoneal injection (ip). 2 times a week for a total of 6 doses.
  • the body weight and tumor growth status of the experimental animals were continuously observed, the tumor was measured and weighed twice a week, and the tumor volume and tumor growth inhibition rate were calculated.
  • tumor tissue was collected and photographed, the tumor tissue was weighed and the tumor weight inhibition rate was calculated (see Figure 8).
  • the tumor growth curve showed that the bifunctional molecule 6 could significantly inhibit the growth of the MC38 tumor model at doses of 5, 10 and 20 mg/kg (P ⁇ 0.01), and showed a significant dose-dependent manner. Tumor growth inhibition rates at endpoint were 41.0%, 52.9%, and 60.8%, respectively.
  • bifunctional molecule 6 and positive control antibody SHR-1701 had comparable tumor inhibitory effects (P>0.05), and their TGIs were 60.8% and 63.2%, respectively.
  • the tumor tissue weight at the end of the experiment showed that the bifunctional molecule 6 could significantly increase the tumor tissue growth at doses of 5, 10 and 20 mg/kg (P ⁇ 0.01), and showed a significant dose-dependence, and its tumor weight inhibition rates were respectively were 43.2%, 54.4% and 64.8%.
  • the bifunctional molecule 6 and the positive control antibody SHR-1701 showed similar anti-tumor activity (P>0.05) at the same dose (20 mg/kg), and their tumor weight inhibition rates (IR%) were 64.8% and 61.0, respectively. %.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Zoology (AREA)
  • Biophysics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Immunology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Toxicology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne une molécule bifonctionnelle ciblant simultanément PD-L1 et TGFβ et une composition pharmaceutique comprenant la molécule bifonctionnelle. L'invention concerne en outre l'utilisation de la molécule bifonctionnelle ciblant simultanément PD-L1 et TGFβ dans le traitement et la prévention de cancers.
PCT/CN2021/120006 2020-09-24 2021-09-23 MOLÉCULE BIFONCTIONNELLE CIBLANT SIMULTANÉMENT PD-L1 ET TGFβ ET SON UTILISATION MÉDICALE Ceased WO2022063193A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202180064731.0A CN116323657B (zh) 2020-09-24 2021-09-23 同时靶向PD-L1和TGFβ的双功能分子及其医药用途

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202011012844 2020-09-24
CN202011012844.X 2020-09-24

Publications (1)

Publication Number Publication Date
WO2022063193A1 true WO2022063193A1 (fr) 2022-03-31

Family

ID=80844952

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/120006 Ceased WO2022063193A1 (fr) 2020-09-24 2021-09-23 MOLÉCULE BIFONCTIONNELLE CIBLANT SIMULTANÉMENT PD-L1 ET TGFβ ET SON UTILISATION MÉDICALE

Country Status (2)

Country Link
CN (1) CN116323657B (fr)
WO (1) WO2022063193A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020215020A1 (fr) * 2019-04-18 2020-10-22 Qlsf Biotherapeutics Inc. Anticorps anti-pd-l1 humanisés
CN116688115B (zh) * 2022-03-18 2024-02-06 上海齐鲁制药研究中心有限公司 一种PD-L1/TGF-β双功能融合蛋白制剂及其用途

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106103488A (zh) * 2014-02-10 2016-11-09 默克专利有限公司 靶向TGFβ抑制
WO2018205985A1 (fr) * 2017-05-12 2018-11-15 江苏恒瑞医药股份有限公司 PROTÉINE DE FUSION CONTENANT UN RÉCEPTEUR DE TGF-β ET UTILISATIONS MÉDICALES ASSOCIÉES
CN109641963A (zh) * 2016-08-12 2019-04-16 默克专利有限公司 癌症的联合治疗
CN110198738A (zh) * 2017-01-07 2019-09-03 默克专利有限公司 用于靶向的TGF-β抑制的给药方案和剂型
WO2019206281A1 (fr) * 2018-04-28 2019-10-31 齐鲁制药有限公司 Formulation d'anticorps monoclonal contre la pd-1 humaine, médicament d'association, et utilisation de ces derniers

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3770171A1 (fr) * 2015-04-03 2021-01-27 XOMA Technology Ltd. Traitement du cancer à l'aide d'inhibiteurs de tgf-bêta et pd-1
BR112021012027A2 (pt) * 2018-12-21 2021-11-03 Ose Immunotherapeutics Molécula bifuncional direcionada contra pd-1 humano

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106103488A (zh) * 2014-02-10 2016-11-09 默克专利有限公司 靶向TGFβ抑制
CN109641963A (zh) * 2016-08-12 2019-04-16 默克专利有限公司 癌症的联合治疗
CN110198738A (zh) * 2017-01-07 2019-09-03 默克专利有限公司 用于靶向的TGF-β抑制的给药方案和剂型
WO2018205985A1 (fr) * 2017-05-12 2018-11-15 江苏恒瑞医药股份有限公司 PROTÉINE DE FUSION CONTENANT UN RÉCEPTEUR DE TGF-β ET UTILISATIONS MÉDICALES ASSOCIÉES
WO2019206281A1 (fr) * 2018-04-28 2019-10-31 齐鲁制药有限公司 Formulation d'anticorps monoclonal contre la pd-1 humaine, médicament d'association, et utilisation de ces derniers

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
JULIUS STRAUSS, CHRISTOPHER R. HEERY, JEFFREY SCHLOM, RAVI A. MADAN, LIANG CAO, ZHIGANG KANG, ELIZABETH LAMPING, JENNIFER L. MART&: "Phase I Trial of M7824 (MSB0011359C), a Bifunctional Fusion Protein Targeting PD-L1 and TGFβ, in Advanced Solid Tumors", CLINICAL CANCER RESEARCH, ASSOCIATION FOR CANCER RESEARCH, US, vol. 24, no. 6, 15 March 2018 (2018-03-15), US, pages 1287 - 1295, XP055488645, ISSN: 1078-0432, DOI: 10.1158/1078-0432.CCR-17-2653 *
JUSTIN M. DAVID, CHARLI DOMINGUEZ, KRISTEN K. MCCAMPBELL, JAMES L. GULLEY, JEFFREY SCHLOM, CLAUDIA PALENA: "A novel bifunctional anti-PD-L1/TGF-β Trap fusion protein (M7824) efficiently reverts mesenchymalization of human lung cancer cells", ONCOIMMUNOLOGY, vol. 6, no. 10, 3 October 2017 (2017-10-03), pages e1349589, XP055665439, DOI: 10.1080/2162402X.2017.1349589 *
KARIN M. KNUDSON, KRISTIN C. HICKS, XIAOLING LUO, JIN-QIU CHEN, JEFFREY SCHLOM, SOFIA R. GAMEIRO: "M7824, a novel bifunctional anti-PD-L1/TGFβ Trap fusion protein, promotes anti-tumor efficacy as monotherapy and in combination with vaccine", ONCOIMMUNOLOGY, vol. 7, no. 5, 4 May 2018 (2018-05-04), pages 1 - 14, XP055665440, ISSN: 2162-402X, DOI: 10.1080/2162402X.2018.1426519 *

Also Published As

Publication number Publication date
CN116323657B (zh) 2023-12-08
CN116323657A (zh) 2023-06-23

Similar Documents

Publication Publication Date Title
CN111971062B (zh) 一种抗人pd-1的单克隆抗体制剂、联合用药物及其用途
CN109963591B (zh) B7h3抗体-药物偶联物及其医药用途
EP3119432B1 (fr) Anticorps contre des glycopeptides immunogènes, composition les comprenant et son utilisation
CN105026428B (zh) PD‑l抗体、其抗原结合片段及其医药用途
JP2022514693A (ja) Ｍｕｃ１８に特異的な抗体
EP3725802A2 (fr) Anticorps anti-synucléine et leurs utilisations
JP7711208B2 (ja) Ｄｌｌ３に対する結合分子及びその使用
US20220031842A1 (en) Lag-3 antibody pharmaceutical composition and use thereof
WO2019117684A1 (fr) Anticorps bispécifique contre a-syn/igf1r et utilisation associée
EP4477670A1 (fr) Molécule bispécifique de liaison à l'antigène et son utilisation
JP2022514786A (ja) Ｍｕｃ１８に特異的な抗体
CN112243443B (zh) 抗trop-2抗体、其抗原结合片段及其医药用途
CN116323657B (zh) 同时靶向PD-L1和TGFβ的双功能分子及其医药用途
EP3990488A1 (fr) Anticorps semaphorine 3a et leurs utilisations
US12460004B2 (en) Antibodies that bind CD277 and uses thereof
US20250179208A1 (en) Anti-cd26 antibodies and use thereof
CN121311504A (zh) 用针对cd3和cd20的双特异性抗体治疗淋巴瘤的方法
WO2022068894A1 (fr) Molécule bifonctionnelle ciblant simultanément pd-l1 et vegf et son utilisation médicale
WO2022237819A1 (fr) Anticorps anti-siglec-15 et son utilisation
WO2022121846A1 (fr) Anticorps se liant au pd-l1 et son application
CN113842456B (zh) 一种抗人4-1bb的单克隆抗体制剂及其用途
WO2024235239A1 (fr) Molécule de liaison à un antigène bispécifique et son utilisation
CN116688115A (zh) 一种PD-L1/TGF-β双功能融合蛋白制剂及其用途
WO2023017149A1 (fr) Anticorps de thiorédoxine 1
WO2024125352A1 (fr) Anticorps monoclonal ciblant cd47 et son utilisation

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21871562

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21871562

Country of ref document: EP

Kind code of ref document: A1