CN111944050A - anti-B7-H3 antibody and application thereof - Google Patents

Abstract

The invention provides an anti-B7-H3 antibody, wherein a heavy chain complementarity determining region of the anti-B7-H3 antibody comprises HCDR1, HCDR2 or HCDR 3; the anti-B7-H3 antibody light chain complementarity determining region comprises LCDR1, LCDR2 or LCDR 3; the HCDR1 amino acid sequence is SEQ ID NO: 3, the amino acid sequence of the HCDR2 is SEQ ID NO: 4, the amino acid sequence of the HCDR3 is SEQ ID NO: 5; the LCDR1 amino acid sequence is SEQ ID NO: 6, the LCDR2 amino acid sequence is SEQ ID NO: 7, the LCDR3 amino acid sequence is SEQ ID NO: 8. the antibody can exert an antitumor effect by an antibody-mediated ADCC effect after binding to a B7-H3 protein molecule with high affinity.

Description

anti-B7-H3 antibody and application thereof

Technical Field

The invention belongs to the field of biological medicines, and particularly relates to an anti-B7-H3 antibody and application thereof in cancer treatment.

Background

The B7-H3 protein (B7 homolog 3 protein), also known as CD276, is an important immune checkpoint molecule of the B7-CD28 family. Charova et al were first found in human dendritic cells and identified as members of the B7 family (Andrei I chapova, et al, "B7-H3: A diagnostic module for T cell activation and IFN- γ production," Nature Immunology 2.3(2001): 269-274.). The B7-H3 molecules have abnormal high expression in various tumors, including gastric cancer, lung cancer, prostate cancer, renal cancer, pancreatic cancer, ovarian cancer, breast cancer, endometrial cancer, liver cancer, colorectal cancer, oral cancer, bladder cancer, osteosarcoma and hematological malignancies, and the B7-H3 molecules are associated with tumor growth, metastasis, poor prognosis and the like. At present, the action mechanism of B7-H3 molecules is not clear, and the B7-H3 molecules are generally considered to be immune co-suppression molecules, play a role in immune negative regulation in tumor immunity, and can suppress the function of T cells and reduce the killing effect of NK cells.

There are currently a number of studies on drug development against B7-H3 molecules, researchers in patents CN2018000004340, CN2012000030835, CN2017000048428 and US20100143245a1 developed monoclonal antibodies against B7-H3 molecules and validated the binding or killing ability of antibodies against cancer cells at the in vitro or in vivo level. The antibody against B7-H3 reported in WO2011109400 exerts antitumor activity through antibody-dependent cell-mediated cytotoxicity (ADCC), is currently undergoing a clinical test (clinical test number NCT01391143) and shows a good antitumor effect, and the curative effect is more remarkable in patients with high expression of B7-H3.

Antibody drugs against B7-H3 molecules are still in the research stage at present, no officially approved drugs are available on the market, and due to the fact that B7-H3 is highly expressed in various cancer patients, the development of novel anti-B7-H3 antibody drugs has important practical significance for treating cancers.

Disclosure of Invention

The invention provides a novel monoclonal antibody capable of binding human B7-H3, compared with the prior art, the affinity of the monoclonal antibody is greatly improved, and the novel antibody can effectively play an anti-tumor role.

Interpretation of terms:

in the present invention, unless defined otherwise, scientific and technical terms used in connection with the present invention shall have the meanings that are commonly understood by those of ordinary skill in the art. Furthermore, unless the context requires otherwise, singular terms shall include the plural and plural terms shall include the singular.

Exemplary techniques for use in conjunction with recombinant DNA, oligonucleotide synthesis, tissue culture and transformation (e.g., electroporation, lipofection), enzymatic reactions, and purification techniques are known in the art. Many such techniques and procedures are described, for example, in Sambrook et al, Molecular Cloning: a Laboratory Manual (2 nd edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)), and many others. In addition, exemplary techniques for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and patient treatment are also known in the art.

In this application, the use of "or" means "and/or" unless stated otherwise. In the case of multiple dependent claims, the use of "or" in the alternative merely refers to more than one of the foregoing independent or dependent claims.

As described herein, any concentration range, percentage range, ratio range, or integer range is to be understood as including the value of any integer within the range, and where appropriate including fractions thereof (such as tenths and hundredths of integers), unless otherwise indicated.

Units, prefixes, and symbols are denoted in their international system of units (SI) recognized form. Numerical ranges include the numbers defining the range. The headings provided herein are not limitations of the various aspects of the disclosure which can be had by reference to the specification as a whole.

Unless otherwise indicated, as used in accordance with this disclosure, the following terms are to be understood to have the following meanings:

an "antibody" refers to a molecule comprising at least the complementarity determining regions CDR1, CDR2 and CDR3 of the heavy chain and at least the CDR1, CDR2 and CDR3 of the light chain, wherein the molecule is capable of binding to an antigen. The term antibody includes, but is not limited to, fragments capable of binding antigen, such as Fv, single chain Fv (scFv), Fab ', and (Fab') 2. The term antibody also includes, but is not limited to, chimeric antibodies, humanized antibodies, and antibodies of various species, such as mouse, human, cynomolgus monkey, alpaca, and the like.

In some embodiments, the antibody comprises a heavy chain variable region and a light chain variable region. In some embodiments, the antibody comprises: at least one heavy chain comprising a heavy chain variable region and at least a portion of a heavy chain constant region, and at least one light chain comprising a light chain variable region and at least a portion of a light chain constant region. In some embodiments, the antibody comprises two heavy chains and two light chains, wherein each heavy chain comprises a heavy chain variable region and at least a portion of a heavy chain constant region, and wherein each light chain comprises a light chain variable region and at least a portion of a light chain constant region. Any other antibody comprising a single polypeptide chain comprising, for example, all six CDRs (three heavy chain CDRs and three light chain CDRs) is considered to have one heavy chain and one light chain. In some such embodiments, the heavy chain is an antibody region comprising three heavy chain CDRs and the light chain is an antibody region comprising three light chain CDRs.

"complementarity determining regions" or "CDRs" are regions in an antibody variable domain that are mutated in sequence and form structurally defined loops ("hypervariable loops") and/or regions containing antigen-contacting residues ("antigen-contacting points"). The CDRs are primarily responsible for binding to an epitope of the antigen. The CDRs of the heavy and light chains are commonly referred to as CDR1, CDR2 and CDR3, numbered sequentially from the N-terminus. The CDRs located within the antibody heavy chain variable domain are referred to as HCDR1, HCDR2 and HCDR3, while the CDRs located within the antibody light chain variable domain are referred to as LCDR1, LCDR2 and LCDR 3. In a given light chain variable region or heavy chain variable region amino acid sequence, the precise amino acid sequence boundaries of each CDR can be determined using any one or combination of a number of well-known antibody CDR assignment systems, including, for example: chothia (Chothia et Al (1989) Nature 342:877-883, Al-Lazikani et Al, "Standard constraints for the structural of the canonical structures of immunology", Journal of Molecular Biology,273,927-948(1997)), Kabat (Kabat et Al, Sequences of Proteins of Immunological Interest, 4 th edition, U.S. Depatory of Health and Human Services, National Institutes of Health (1987)), AbM (correlation of Bath), Unit of University (Collection of London), International Munongensis database (IMGT), and the clustering of the CDRs using the bulk crystal structure (normal propagation). The CDR partitioning according to the present invention is performed according to the Kabat et al protocol.

"Fc" or "Fc fragment" refers to the "fragment crystallizable" region of an immunoglobulin heavy chain. In general, an Fc domain is capable of interacting with another Fc domain to form a dimeric complex. The Fc domain may bind to a cell surface receptor (Fc receptor) and/or a protein of the complement system, or may be modified to impair or enhance such binding activity. The Fc domain may be derived from IgG, IgA, IgD, IgM, or IgE antibodies and produce immune functions such as opsonization, cell lysis, mast cell degranulation, among other Fc receptor dependent processes. The original immunoglobulin source of the native Fc is preferably an immunoglobulin of human origin, preferably IgG1, IgG2 and IgG 4. In the present invention, the Fc domain is preferably composed of CH2 and CH3 of human IgG 1.

In certain embodiments, an "Fc mutant" refers to a molecule or sequence that is modified by native Fc and can still bind an Fc receptor. "Fc mutants" include molecules or sequences humanized from non-human native Fc "Fc domains" include molecules or sequences of native Fc and Fc variants as described above, which comprise molecules in monomeric or multimeric form, which can be obtained by whole antibody dissociation, or by gene recombinant expression or by other means.

"ADCC" is antibody-dependent cell-mediated cytotoxicity (ADCC), and refers to binding of Fab fragment of antibody to epitope of virus-infected cell or tumor cell, binding of Fc fragment to FcR on surface of killer cell (NK cell, macrophage, etc.) and direct killing of target cell by killer cell. NK cells, mononuclear macrophages and neutrophils can kill tumor cells in an ADCC mode under the mediation of specific IgG. The NK cells have broad-spectrum anti-swelling and pain functions, and can kill and dissolve tumor cells coated by specific IgG through an ADCC mode, and can directly kill the tumor cells through close contact with the tumor cells. ADCC effects can be enhanced or attenuated by engineering of the native Fc region (Wang X, Mathieu M, Brezski R J, et al. IgG Fc engineering to modulated antibody effects [ J ]. proteins & cells, 2018,9(1): 63-73.).

"bispecific antibody" refers to an antibody molecule that specifically binds two antigens or two epitopes simultaneously, and can exert the synergistic effect of the combination of the two monoclonal antibodies. Bispecific antibodies do not exist in nature and need to be prepared artificially by recombinant DNA or cell fusion techniques. The typical bispecific antibody concept is to redirect cytotoxic activity of effector T cells to specifically eliminate tumor cells. In this way, the T cell establishes a physical link with the tumor cell via a bispecific antibody consisting of a T cell binding domain and a tumor binding domain. These bispecific antibodies (T cell-engaging bsAb, bsTCE) that T cells participate activate T cells primarily by binding to CD3 in the TCR complex. It is also common practice to construct bispecific antibodies using antibodies directed against immune checkpoints, including PD-1/PD-L1, CTLA4, CD137, CD47, LAG3, TIM3, and the like. Bispecific antibody construction can occur in Fc-containing or non-Fc-containing formats, e.g., current bispecific antibody formats include BiTE, KiH, CrossMab, DART, DVD-Ig, FIT-Ig, YBody, Duobody, κ λ -body, Bi-nanobody, and the like.

An antibody-drug conjugate (ADC) consists of an antibody, cytotoxin (cytotoxin payload) and a linker, and has high specificity of the antibody and high toxicity of the cytotoxin to tumors. Wherein the antibody specifically recognizes and directs the drug to the lesion, and the conjugate is generally cleaved at the site of the lesion at pH, thereby releasing the therapeutic cytotoxin. Cytotoxins mainly target the DNA and tubulin of cancer cells and block the proliferation function of tumor cells, so that the cytotoxins commonly used in the development of ADC drugs at present for inducing apoptosis comprise anti-DNA: spinosyns (calicheamicins) and duocarmycins, anti-tubulins: orlistatin (MMAE; MMAF) and maytansine (maytansines, DM1, DM 4).

"host cell" refers to a cell that may be or has been the recipient of a vector or isolated polynucleotide. The host cell may be a prokaryotic cell or a eukaryotic cell. Exemplary eukaryotic cells include mammalian cells, such as primate or non-primate cells; fungal cells, such as yeast; a plant cell; and insect cells. Non-limiting exemplary mammalian cells include, but are not limited to, NSO cells, and 293 and CHO cells and derivatives thereof, such as 293-6E and DG44 cells, respectively.

"cancer" is customarily used to refer broadly to all malignant tumors. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. More specific non-limiting examples of such cancers include squamous cell cancer, small-cell lung cancer, pituitary cancer, esophageal cancer, astrocytoma, soft tissue sarcoma, non-small cell lung cancer (including squamous cell non-small cell lung cancer), adenocarcinoma of the lung, squamous cell carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer (liver cancer), bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine cancer, salivary gland carcinoma, kidney cancer, renal cell carcinoma, liver cancer, prostate cancer, vulval cancer, thyroid cancer, liver cancer (hepatoma), brain cancer, endometrial cancer, testicular cancer, cholangiocarcinoma, gallbladder cancer, gastric cancer, melanoma, and various types of head and neck cancer (including head and neck squamous cell carcinoma).

"treatment" refers to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) the targeted pathological condition or disorder. In certain embodiments, the term "treatment" encompasses any administration or use of a disease therapeutic agent in a mammal (including a human), and includes inhibiting or slowing disease or disease progression; partial or complete response to disease, e.g., by causing regression, or recovery or repair of lost, lost or defective function; a stimulus-ineffective process; or to smooth the disease to a reduced severity. The term "treating" also includes reducing the severity of any phenotypic feature and/or reducing the incidence, extent or likelihood of that feature. The subject in need of treatment includes those already having the disorder as well as those predisposed to having the disorder, or those in which the disorder is to be prevented.

In one aspect, the invention provides an anti-B7-H3 antibody.

The B7-H3 is B7-H3 protein, and the sequence of the protein is shown as SEQ ID NO: shown at 13.

The heavy chain complementarity determining region of the anti-B7-H3 antibody comprises one or more of HCDR1, HCDR2 or HCDR 3; preferably, the heavy chain complementarity determining regions of the anti-B7-H3 antibody include HCDR1, HCDR2 and HCDR 3.

The anti-B7-H3 antibody light chain complementarity determining region comprises one or more of LCDR1, LCDR2 or LCDR 3; preferably, the anti-B7-H3 antibody light chain complementarity determining regions include LCDR1, LCDR2 and LCDR 3.

The HCDR1 amino acid sequence is SEQ ID NO: 3, the amino acid sequence of the HCDR2 is SEQ ID NO: 4, the amino acid sequence of the HCDR3 is SEQ ID NO: 5.

the LCDR1 amino acid sequence is SEQ ID NO: 6, the LCDR2 amino acid sequence is SEQ ID NO: 7, the LCDR3 amino acid sequence is SEQ ID NO: 8.

further, the heavy chain of the anti-B7-H3 antibody comprises a heavy chain variable region SEQ ID NO: 1.

further, the light chain of the anti-B7-H3 antibody comprises the light chain variable region SEQ ID NO: 2.

preferably, the heavy chain sequence of the anti-B7-H3 antibody is SEQ ID NO: 9, the light chain sequence of the anti-B7-H3 antibody is SEQ ID NO: 10.

the Fc sequence of the anti-B7-H3 antibody is selected from human IgG1, human IgG2 or human IgG 4; preferably selected from human IgG1 or human IgG1 mutants.

In another aspect, the invention provides biomaterials associated with the aforementioned anti-B7-H3 antibodies.

The biological material includes but is not limited to nucleic acid molecules, biological expression vectors, and host cells.

The nucleic acid molecule encodes the anti-B7-H3 antibody sequence described above.

The biological expression vector comprises the nucleic acid molecule described above.

The host cell comprises the aforementioned biological expression vector.

In still another aspect, the invention provides the use of the aforementioned anti-B7-H3 antibody, nucleic acid molecule, biological expression vector or host cell in the preparation of a medicament for the treatment of cancer.

The cancer is kidney cancer, liver cancer, lung cancer, stomach cancer, esophagus cancer, colorectal cancer, bladder cancer, urothelial cell cancer, lymph node cancer, head and neck cancer, prostatic cancer, pancreatic cancer, breast cancer, melanoma, lymphoma, glioma, neuroblastoma, retinoblastoma, rhabdomyosarcoma, wilms' tumor or osteosarcoma.

The drug includes but is not limited to an antibody conjugate drug or a bispecific antibody drug.

The antibody coupling drug comprises an anti-B7-H3 antibody, a linker and a small molecule cytotoxic drug.

The linker in the antibody conjugated drug includes but is not limited to hydrazone bond, disulfide bond and peptide bond.

The small molecular cytotoxic drug in the antibody coupling drug is a chemotherapy drug which is generally used clinically.

The antibody conjugate drug is combined with 2-4 toxic compounds on each antibody.

The bispecific antibody comprises an anti-B7-H3 antibody and any other antibody, such as: anti-CD 3 antibody, anti-CD 19 antibody, anti-EGFR antibody, and the like.

The preparation method of the bispecific antibody includes but is not limited to a two-hybridoma cell method, a chemical coupling method, recombinant gene preparation and the like.

Drawings

FIG. 1 shows the detection result of anti-B7-H3 antibody SEC-HPLC.

FIG. 2 shows the SEC-HPLC detection result of the control antibody.

FIG. 3 is a graph showing the tumor growth in mice after the administration of the tumor mice was initiated.

Detailed Description

The present invention will be further illustrated in detail with reference to the following specific examples, which are not intended to limit the present invention but are merely illustrative thereof. The experimental methods used in the following examples are not specifically described, and the materials, reagents and the like used in the following examples are generally commercially available under the usual conditions without specific descriptions.

Example 1 Gene construction and expression of anti-B7-H3 antibody

Combinations according to the following table:

combination of heavy and light chain sequences for expression of antibodies

Sample (I)	Heavy chain sequence	Light chain sequence
			anti-B7-H3 antibody	SEQ ID NO:9	SEQ ID NO:10
Control antibody	SEQ ID NO:11	SEQ ID NO:12

By adopting a conventional molecular biology construction scheme, adding a signal peptide suitable for eukaryotic cell expression, respectively constructing synthesized heavy chain and light chain fragments of an antibody gene into a pTT5 vector, co-transfecting the heavy chain and light chain vectors of the antibody gene by adopting an HEK293 cell line, culturing the cells, separating and purifying cell culture supernatant by using protein A filler after culturing for 5 days, and analyzing the protein purity of the purified sample by using a SEC-HPLC (Secondary-evaporative light chromatography) technical method, wherein the results are shown in figure 1 and figure 2.

FIG. 1 shows the detection result of anti-B7-H3 antibody SEC-HPLC, and FIG. 2 shows the detection result of control antibody SEC-HPLC.

Example 2 binding of anti-B7-H3 antibodies to different species of B7-H3 protein

The anti-B7-H3 antibody obtained in example 1 was subjected to affinity detection with human B7-H3 Protein (available from Novoprotein, cat # CK62) and monkey B7-H3 Protein (available from Novoprotein, cat # CA61) using a GatorTM non-labeled bioanalyzer, and a Protein A biosensor was selected to capture an antibody sample, and then the captured antibody sample was subjected to kinetic detection of binding and dissociation with B7-H3 Protein. Kinetics were performed using 1:1 the fitting analysis is performed in conjunction with the model. The brief steps are shown in the following table:

step (ii) of	Time of day
		Protein loading	200s
Bonding of	180s
		Dissociation	300s
Regeneration	30s

Using Gator^TMThe affinity data measured by the instrument are shown in the following table:

the detection result shows that the anti-B7-H3 antibody can be combined with human B7-H3 protein or monkey B7-H3 protein with high affinity.

According to the reported information (ASCO 2016, Abstract 165034), the control antibody binds to human B7-H3 protein with an affinity of about 7E-9M, and the disclosed anti-B7-H3 antibody binds to human B7-H3 protein with an affinity about 15.9 times that of the control antibody. This shows that the antibody disclosed in the patent can better bind with B7-H3 on the surface of tumor, and the high affinity is more favorable for the antibody molecule to exert the anti-tumor efficacy in tumor treatment.

Example 3 detection of the thermostability of anti-B7-H3 antibody

Using Applied Biosystems^TM7500 real-time fluorescent quantitative PCR System Using

The thermal stability assay was performed after staining the protein with dye from Orange (5,000X concentratin DMSO) (available from Thermofisiher, cat # S6650).

The brief method is as follows: will be provided with

After the Orange reagent and the anti-B7-H3 antibody (1mg/mL dissolved in PBS buffer) are fully mixed according to the volume of 1:1, 100uL of sample is placed in a sample plate, the temperature variation range is set to be 20-100 ℃, and the temperature is increased at the speed of 0.5 ℃/min. After the experiment is finished, data are collected, fitting treatment is carried out by using a Melt Curve functional module with software of an instrument, and the Tm value of the anti-B7-H3 antibody obtained after treatment is 67.3 ℃, which indicates that the antibody molecule disclosed by the patent has good thermal stability.

Example 4 verification of antitumor Effect of anti-B7-H3 antibody

(1) Cell culture

A498 cells were cultured in MEM +0.01mM NEAA medium containing 10% fetal bovine serum. A498 cells were harvested in exponential growth phase and resuspended in PBS to appropriate concentration for subcutaneous tumor inoculation in BALB/c nude mice (Shanghai Jie laboratory animals Co., Ltd.).

(2) Animal model

Experimental mice were inoculated subcutaneously on the right back with 5X 10⁶A498 cells, resuspended in 1:1 (0.1 mL/piece) in PBS and matrigel, and observing the growth of tumor until the average volume of tumor grows to 80-120mm³(about 100 mm)³) The administration was randomized and divided into groups according to the tumor size and the mouse body weight. The day of tumor cell inoculation was defined as day 0. Before the start of dosing, all animals were weighed and tumor volume was measured with a vernier caliper. Since tumor volume affects the effectiveness of the treatment, mice were grouped according to their tumor volume using a random grouping design to ensure similar tumor volumes between groups.

The number of animals per group and the detailed route, dosage and schedule of administration are given in the table below.

Administration route, dosage and scheme in subcutaneous xenograft model of human renal carcinoma A498 cell line

Group of

Number of

Administration set

Dosage form

Mode of administration

Administration cycle

1

8

Vehicle control

--

Tail vein injection

Once a week for 5 times

2

8

Control antibody

20mg/kg

Tail vein injection

Once a week for 5 times

3

8

anti-B7-H3 antibody

20mg/kg

Tail vein injection

Once a week for 5 times

(3) Experimental observations and data Collection

After tumor cell inoculation, the influence of tumor growth and treatment on the normal behavior of animals is monitored conventionally, and the specific contents comprise: mobility, feeding and drinking conditions, weight gain or loss, eyes, hair and other abnormal conditions in the test animal. After the start of the administration, the body weight and tumor size of the mice were measured twice a week. Tumor volume calculation formula: tumor volume (mm)³)＝1/2×(a×b²) (wherein a represents a long diameter and b represents a short diameter).

After the experiment, the collected data were processed and analyzed as shown in fig. 3. The results show that the anti-B7-H3 antibody and the control antibody can obviously inhibit the growth of tumors in established tumor models, and the anti-B7-H3 antibody shows the same anti-tumor effect as the control antibody.

Finally, it should be noted that the above-mentioned contents are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, and that the simple modifications or equivalent substitutions of the technical solutions of the present invention by those of ordinary skill in the art can be made without departing from the spirit and scope of the technical solutions of the present invention.

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50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Asp Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Gly Arg Gly Arg Glu Asn Ile Tyr Tyr Gly Ser Arg Leu Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Val

225 230 235 240

Gly Gly Pro Ser Val Phe Leu Leu Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Pro Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Leu Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Leu Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly Lys

450

<210> 12

<211> 214

<212> PRT

<213> Artificial sequence (artificial sequence)

<400> 12

Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Asp Thr Asn

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile

35 40 45

Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Asn Tyr Pro Phe

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 13

<211> 506

<212> PRT

<213> Artificial sequence (artificial sequence)

<400> 13

Leu Glu Val Gln Val Pro Glu Asp Pro Val Val Ala Leu Val Gly Thr

1 5 10 15

Asp Ala Thr Leu Cys Cys Ser Phe Ser Pro Glu Pro Gly Phe Ser Leu

20 25 30

Ala Gln Leu Asn Leu Ile Trp Gln Leu Thr Asp Thr Lys Gln Leu Val

35 40 45

His Ser Phe Ala Glu Gly Gln Asp Gln Gly Ser Ala Tyr Ala Asn Arg

50 55 60

Thr Ala Leu Phe Pro Asp Leu Leu Ala Gln Gly Asn Ala Ser Leu Arg

65 70 75 80

Leu Gln Arg Val Arg Val Ala Asp Glu Gly Ser Phe Thr Cys Phe Val

85 90 95

Ser Ile Arg Asp Phe Gly Ser Ala Ala Val Ser Leu Gln Val Ala Ala

100 105 110

Pro Tyr Ser Lys Pro Ser Met Thr Leu Glu Pro Asn Lys Asp Leu Arg

115 120 125

Pro Gly Asp Thr Val Thr Ile Thr Cys Ser Ser Tyr Gln Gly Tyr Pro

130 135 140

Glu Ala Glu Val Phe Trp Gln Asp Gly Gln Gly Val Pro Leu Thr Gly

145 150 155 160

Asn Val Thr Thr Ser Gln Met Ala Asn Glu Gln Gly Leu Phe Asp Val

165 170 175

His Ser Ile Leu Arg Val Val Leu Gly Ala Asn Gly Thr Tyr Ser Cys

180 185 190

Leu Val Arg Asn Pro Val Leu Gln Gln Asp Ala His Ser Ser Val Thr

195 200 205

Ile Thr Pro Gln Arg Ser Pro Thr Gly Ala Val Glu Val Gln Val Pro

210 215 220

Glu Asp Pro Val Val Ala Leu Val Gly Thr Asp Ala Thr Leu Arg Cys

225 230 235 240

Ser Phe Ser Pro Glu Pro Gly Phe Ser Leu Ala Gln Leu Asn Leu Ile

245 250 255

Trp Gln Leu Thr Asp Thr Lys Gln Leu Val His Ser Phe Thr Glu Gly

260 265 270

Arg Asp Gln Gly Ser Ala Tyr Ala Asn Arg Thr Ala Leu Phe Pro Asp

275 280 285

Leu Leu Ala Gln Gly Asn Ala Ser Leu Arg Leu Gln Arg Val Arg Val

290 295 300

Ala Asp Glu Gly Ser Phe Thr Cys Phe Val Ser Ile Arg Asp Phe Gly

305 310 315 320

Ser Ala Ala Val Ser Leu Gln Val Ala Ala Pro Tyr Ser Lys Pro Ser

325 330 335

Met Thr Leu Glu Pro Asn Lys Asp Leu Arg Pro Gly Asp Thr Val Thr

340 345 350

Ile Thr Cys Ser Ser Tyr Arg Gly Tyr Pro Glu Ala Glu Val Phe Trp

355 360 365

Gln Asp Gly Gln Gly Val Pro Leu Thr Gly Asn Val Thr Thr Ser Gln

370 375 380

Met Ala Asn Glu Gln Gly Leu Phe Asp Val His Ser Val Leu Arg Val

385 390 395 400

Val Leu Gly Ala Asn Gly Thr Tyr Ser Cys Leu Val Arg Asn Pro Val

405 410 415

Leu Gln Gln Asp Ala His Gly Ser Val Thr Ile Thr Gly Gln Pro Met

420 425 430

Thr Phe Pro Pro Glu Ala Leu Trp Val Thr Val Gly Leu Ser Val Cys

435 440 445

Leu Ile Ala Leu Leu Val Ala Leu Ala Phe Val Cys Trp Arg Lys Ile

450 455 460

Lys Gln Ser Cys Glu Glu Glu Asn Ala Gly Ala Glu Asp Gln Asp Gly

465 470 475 480

Glu Gly Glu Gly Ser Lys Thr Ala Leu Gln Pro Leu Lys His Ser Asp

485 490 495

Ser Lys Glu Asp Asp Gly Gln Glu Ile Ala

500 505

Claims (10)

1. An anti-B7-H3 antibody, wherein the heavy chain complementarity determining region of the anti-B7-H3 antibody comprises one or more of HCDR1, HCDR2 or HCDR 3; the anti-B7-H3 antibody light chain complementarity determining region comprises one or more of LCDR1, LCDR2 or LCDR 3; the HCDR1 amino acid sequence is SEQ ID NO: 3, the amino acid sequence of the HCDR2 is SEQ ID NO: 4, the amino acid sequence of the HCDR3 is SEQ ID NO: 5; the LCDR1 amino acid sequence is SEQ ID NO: 6, the LCDR2 amino acid sequence is SEQ ID NO: 7, the LCDR3 amino acid sequence is SEQ ID NO: 8.

2. the anti-B7-H3 antibody of claim 1, wherein said anti-B7-H3 heavy chain comprises a heavy chain variable region of SEQ ID NO: 1; the anti-B7-H3 antibody light chain comprises a light chain variable region SEQ ID NO: 2.

3. the anti-B7-H3 antibody of claim 1, wherein the Fc sequence of the anti-B7-H3 antibody is selected from the group consisting of human IgG1, human IgG2, and human IgG 4; preferably selected from human IgG1 or human IgG1 mutants.

4. The anti-B7-H3 antibody of claim 1, wherein the heavy chain sequence of said anti-B7-H3 antibody is SEQ ID NO: 9, the light chain sequence of the anti-B7-H3 antibody is SEQ ID NO: 10.

5. a nucleic acid molecule encoding the anti-B7-H3 antibody sequence of any one of claims 1-3.

6. A biological expression vector comprising the nucleic acid molecule of claim 6.

7. A host cell comprising the biological expression vector of claim 7.

8. Use of the anti-B7-H3 antibody of any one of claims 1-3, the nucleic acid molecule of claim 6, the biological expression vector of claim 7, or the host cell of claim 8 in the manufacture of a medicament for treating cancer.

9. The use of claim 8, wherein the drug is an antibody-conjugated drug or a bispecific antibody drug.

10. The use of claim 9, wherein the cancer is renal cancer, liver cancer, lung cancer, stomach cancer, esophageal cancer, colorectal cancer, bladder cancer, urothelial cell cancer, lymph node cancer, head and neck cancer, prostate cancer, pancreatic cancer, breast cancer, melanoma, lymphoma, glioma, neuroblastoma, retinoblastoma, rhabdomyosarcoma, wilms' tumor, or osteosarcoma.

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