Use of anti-pd-1 antibodies in combination

Abstract

The present application discloses the application of anti-PD-1 antibody in combination, the treatment method includes the administration of effective amount of anti-PD-1 antibody and therapeutic agent to the patient in need.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biological medicine, and particularly relates to application of anti-PD-1 antibody in combination therapy. BACKGROUND

[0002] The incidence of tumors is increasing year by year, and the use of anti-tumor drugs is also gradually increasing. In recent years, great progress has been made in the treatment of tumors, especially some antibody drugs have shown good efficacy in the treatment of malignant tumors. However, only a small number of patients can achieve long-term survival. In order to further improve the clinical efficacy of anti-tumor drugs, combination therapy can be used when using anti-tumor drugs.

[0003] Programmed death receptor-1 (PD-1) is an immunosuppressive receptor expressed on activated T cells, B cells and myeloid cells, and is a member of the CD28 immunoglobulin superfamily. PD-1 is a 55kDa type I transmembrane glycoprotein, which contains an Ig variable domain that binds to a ligand and a cytoplasmic tail responsible for binding signal transduction molecules. The cytoplasmic tail of PD-1 contains two tyrosine-based signal transduction motifs ITIM (immunoreceptor tyrosine-based inhibitory motif) and ITSM (immunoreceptor tyrosine-based switch motif).

[0004] A large number of studies have shown that the interaction between PD-1 and PD-L1 (programmed death ligand 1) leads to a decrease in lymphocytes infiltrating tumors, a decrease in T cell receptor-mediated proliferation, and immune evasion of cancer cells. Blocking the interaction between PD-1 and PD-L1 can increase T cell proliferation and cytokine production, and improve tumor-specific CD8 + Immunity of T cells.

[0005] Therefore, anti-PD-1 antibody plays an important role in the combination therapy of anti-tumor. SUMMARY

[0006] The present application discloses a method or use of anti-PD-1 antibody for combination therapy of tumors or cancers. In some embodiments, the anti-PD-1 antibody and a therapeutic agent are combined for treating tumors or cancers. In some embodiments, the therapeutic agent is an antibody or an antibody drug conjugate.

[0007] In some embodiments, the anti-PD-1 antibody comprises at least one of HCDR1 shown in SEQ ID NO: 1, HCDR2 shown in SEQ ID NO: 2, HCDR3 shown in SEQ ID NO: 3, LCDR1 shown in SEQ ID NO: 4, LCDR2 shown in SEQ ID NO: 5, and LCDR3 shown in SEQ ID NO: 6.

[0008] In some embodiments, the anti-PD-1 antibody comprises a HCDR1 as set forth in SEQ ID NO: 1, a HCDR2 as set forth in SEQ ID NO: 2, a HCDR3 as set forth in SEQ ID NO: 3, a LCDR1 as set forth in SEQ ID NO: 4, a LCDR2 as set forth in SEQ ID NO: 5, and a LCDR3 as set forth in SEQ ID NO: 6.

[0009] In some embodiments, the heavy chain variable region of the anti-PD-1 antibody comprises a sequence as set forth in SEQ ID NO: 7, a sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 7, or an amino acid sequence that has one or more conservative amino acid substitutions compared to the sequence set forth in SEQ ID NO: 7; and / or

[0010] the light chain variable region of the anti-PD-1 antibody comprises a sequence as set forth in SEQ ID NO: 8, a sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 8, or an amino acid sequence that has one or more conservative amino acid substitutions compared to the sequence set forth in SEQ ID NO: 8.

[0011] In some embodiments, the heavy chain variable region of the anti-PD-1 antibody comprises a sequence as set forth in SEQ ID NO: 7, and the light chain variable region of the anti-PD-1 antibody comprises a sequence as set forth in SEQ ID NO: 8.

[0012] In some embodiments, the heavy chain of the anti-PD-1 antibody comprises a sequence as set forth in SEQ ID NO: 9, a sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 9, or an amino acid sequence that has one or more conservative amino acid substitutions compared to the sequence set forth in SEQ ID NO: 9; and / or

[0013] the light chain of the anti-PD-1 antibody comprises a sequence as set forth in SEQ ID NO: 10, a sequence that is at least 80% identical to the sequence set forth in SEQ ID NO: 10, or an amino acid sequence that has one or more conservative amino acid substitutions compared to the sequence set forth in SEQ ID NO: 10.

[0014] In some embodiments, the anti-PD-1 antibody is Antibody A, the heavy chain of Antibody A comprises a sequence as set forth in SEQ ID NO: 9, the light chain of Antibody A comprises a sequence as set forth in SEQ ID NO: 10; Antibody A contains two identical sequences of heavy chain and two identical sequences of light chain.

[0015] In some embodiments, the therapeutic agent is selected from the group consisting of an antibody or an antibody drug conjugate (ADC) against the following targets: EGFR (epidermal growth factor receptor), VEGF (vascular endothelial growth factor), VEGFR2 (vascular endothelial growth factor receptor 2), CTLA4 (cytotoxic T-lymphocyte-associated protein 4), PD-L1, HER2 (human epidermal growth factor receptor 2), CD20 (cluster of differentiation 20), Trop2 (human trophoblast cell surface antigen 2), Lag3 (lymphocyte activation gene-3 molecule), TIGIT (T cell Ig and ITIM domain), CD27 (cluster of differentiation 27), OX40 (tumor necrosis factor receptor superfamily member 4), ICOS (inducible costimulator), BTLA (B and T lymphocyte attenuator), TIM3 (T cell immunoglobulin mucin 3), BCMA (B cell maturation antigen), c-MET (mesenchymal epithelial transition factor), and TAA-1 / 2 / 3 (tumor associated antigen). In some embodiments, the antibody is an inhibitory antibody or an agonistic antibody.

[0016] In some embodiments, the therapeutic agent is selected from the group consisting of an anti-EGFR antibody, an anti-VEGF antibody, an anti-VEGFR2 antibody, an anti-CTLA4 antibody, an anti-PD-L1 antibody, an anti-HER2 antibody, an anti-CD20 antibody, an anti-Trop2 antibody, an anti-TIGIT antibody, an anti-OX40 antibody, or an anti-ICOS antibody.

[0017] In some embodiments, the therapeutic agent is an anti-CTLA4 antibody. In some embodiments, the anti-CTLA4 antibody is antibody C, the heavy chain of antibody C comprises a sequence as set forth in SEQ ID NO: 15, the light chain of antibody C comprises a sequence as set forth in SEQ ID NO: 16; antibody C contains two identical sequences of heavy chains and two identical sequences of light chains. In some embodiments, antibody C is expressed by CHO cells with knock-out of a-(1, 6)-fucosyltransferase gene, such as the CHO-BAT-KF cell line disclosed in PCT / CN2018 / 100008.

[0018] In some embodiments, the total amount of high mannose glycoforms of the Fc region in the antibody C is < 5% and / or the total amount of sialylated glycoforms is < 3%. In some embodiments, the total amount of high mannose glycoforms of the Fc region in the antibody C is about 0.1%, about 0.3%, about 0.9%, about 1.18%, about 1.7%, about 2.6%, about 3.3%, about 4.1%, about 4.9%, about 4.99%, or a range between any two of these values, inclusive of the endpoints, or any value therein. In some embodiments, the total amount of sialylated glycoforms of the Fc region in the antibody C is about 0.1%, 0.2%, about 0.36%, about 0.8%, about 1.5%, about 2.2%, about 2.7%, about 2.9%, 2.99%, or a range between any two of these values, inclusive of the endpoints, or any value therein.

[0019] In some embodiments, the total amount of high mannose glycoforms of the Fc region in the antibody C is < 2% and / or the total amount of sialylated glycoforms is < 1%.

[0020] In some embodiments, the level of fucosylation of the Fc region in the antibody C is 0-10%. In some embodiments, the level of fucosylation of the Fc region in the antibody C is 0-5%. In some embodiments, the level of fucosylation of the Fc region in the antibody C is about 0, about 0.1%, about 0.3%, about 0.4%, about 0.6%, about 1.3%, about 1.9%, about 2.2%, about 2.8%, about 3.3%, about 3.7%, about 4.1%, about 4.5%, about 5%, or a range between any two of these values, inclusive of the endpoints, or any value therein.

[0021] In some embodiments, the therapeutic agent is an anti-HER2 antibody drug conjugate or an anti-Trop2 antibody drug conjugate. In some embodiments, the therapeutic agent is an anti-HER2-antibody drug conjugate (HER2-ADC). In some embodiments, the therapeutic agent is an anti-Trop2-antibody drug conjugate (Trop2-ADC).

[0022] In some embodiments, the Trop2-ADC is a conjugate of BAT0807 or BAT0808 in the patent application CN109078181A and a drug.

[0023] In some embodiments, the therapeutic agent is an antibody drug conjugate (ADC) as shown in Formula I or a pharmaceutically acceptable salt thereof:

[0024]

[0025] wherein Abu is an anti-HER2 antibody or an anti-Trop2 antibody, and p is selected from 1-10;

[0026] X is -H or a halogen group;

[0027] Y is selected from -H, C1-C6alkyl, C3-C6cycloalkyl, or -C(=O)R 5 ;

[0028] R 1 is selected from -H, -OH, -OC(=O)R 5 or -OR 5 groups;

[0029] R 2 is -H or C1-C6alkyl;

[0030] R 3 is methyl, -CH2OH, or -CH2OC(=O)R 6 ;

[0031] R 4 is -OH or -SH;

[0032] R 5 is C1-C6alkyl or benzyl;

[0033] R 6 is C1-C6alkyl, phenyl, or benzyl;

[0034] R 7 is -H, C1-C6alkyl, or an amino acid side chain;

[0035] R 8 is -H or C1-C6alkyl;

[0036] Z is independently -H or C1-C4hydrocarbyl, or two Zs, together with the carbon atom to which they are attached, form a carbonyl group;

[0037] L is selected from optionally substituted C1-C20hydrocarbylene or C3-C8cyclohydrocarbylene, wherein one or more -CH2- groups are independently optionally replaced by a group selected from C3-C8cyclohydrocarbylene, -O-, -S-, -NR 8 -, -C(=O)-, -C(=O)NR 8 -, -NR 8 C(=O)-, -SO2NR 8 -, or -NR 8 SO2-.

[0038] In some embodiments, L is -(CH2) mwherein m is an integer, m is 1-20. In some embodiments, m is 1-10. In some embodiments, m is 5-10. In some embodiments, m is 1, 2, 3, 4, 5, 7, 9, 11, 12, 13, 15, 16, 18, 19, or 20, or a range between any two of these values, inclusive or any value therein.

[0039] In some embodiments, the C1-C20 alkylene in L is substituted with one to four -SO3H, -P(=O)(OH)2, or R 23 substituted with one or two groups independently selected from -SH, -S-C 23 each independently is C1-C6 alkyl optionally substituted with one or two groups independently selected from -SH, -S-C 1-4 alkyl, -CONR 11 R 11 and -NR 11 R 11 In some embodiments, R 11 is selected from -H, alkyl, aryl, cycloalkyl, cycloalkenyl, heteroaryl, and heterocycle, or two R 11 and the nitrogen form a heterocycle, wherein the carbons in the heterocycle can optionally be substituted with one or two oxygens.

[0040] In some embodiments, the therapeutic agent is an antibody drug conjugate (ADC) as shown in Formula II, or a pharmaceutically acceptable salt thereof:

[0041]

[0042] wherein Abu is an anti-HER2 antibody or an anti-Trop2 antibody, and p is selected from 1-10.

[0043] p is the number of small molecule drugs to which an antibody binds in an ADC, i.e., the drug binding number of an antibody, or the drug to antibody coupling ratio (DAR). In some embodiments, p is about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10. In some embodiments, p is 2-3. In some embodiments, p is about 2.

[0044] In some embodiments, p is the average number of small molecule drug molecules bound, i.e., the average drug-to-antibody ratio (DAR). In some embodiments, p is 1-10, and p can be a non-integer. In some embodiments, p is about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, or a range (including endpoints) of any two of these values, or any value therein. In some embodiments, p is selected from 2-8. In some embodiments, p is selected from 3-5. In some embodiments, p is selected from 3.3-3.7. In some embodiments, p is about 3.5. In some embodiments, p is about 2.1.

[0045] In some embodiments, the heavy chain of the anti-HER2 antibody comprises the sequence shown in SEQ ID NO:11, a sequence having at least 80% identity with the sequence shown in SEQ ID NO:11, or an amino acid sequence having one or more conserved amino acid substitutions compared to the sequence shown in SEQ ID NO:11; and / or

[0046] The light chain of the anti-HER2 antibody comprises the sequence shown in SEQ ID NO:12, a sequence having at least 80% identity with the sequence shown in SEQ ID NO:12, or an amino acid sequence having one or more conserved amino acid substitutions compared to the sequence shown in SEQ ID NO:12.

[0047] In some embodiments, the anti-HER2 antibody is antibody B, the heavy chain of antibody B comprising the sequence shown in SEQ ID NO:11, and the light chain of antibody B comprising the sequence shown in SEQ ID NO:12.

[0048] In some embodiments, the anti-Trop2 antibody is BAT0807 or BAT0808 as described in patent application CN109078181A. In some embodiments, p is about 2. In some embodiments, p is about 2.1.

[0049] In some implementations, the HER2-ADC is a HER2-ADC B HER2-ADC B The compound represented by Formula II, HER2-ADC B The antibody is antibody B, which contains two identical heavy chains and two identical light chains; the p-value is 3.3-3.7, or about 3.5.

[0050] The antibody protein can be expressed in CHO cells by genetic engineering and obtained by purification; the purification can be performed by conventional methods, for example, centrifuging the cell suspension and collecting the supernatant, and further removing impurities by centrifugation again. Methods such as Protein A affinity column and ion exchange column can be used for purifying the antibody protein.

[0051] In some embodiments, the method or use comprises: administering to a patient in need thereof an effective amount of an anti-PD-1 antibody and a therapeutic agent. In some embodiments, the anti-PD-1 antibody is antibody A. In some embodiments, the therapeutic agent is HER2-ADC B In some embodiments, the effective dose of the anti-PD-1 antibody administration is about 50 mg to 600 mg per dose. In some embodiments, the effective dose of the HER2-ADC administration is about 70 mg to 400 mg per dose. In some embodiments, the therapeutic agent is an anti-CTLA4 antibody. In some embodiments, the therapeutic agent is antibody C. In some embodiments, the effective dose of the anti-CTLA4 administration is about 6 mg to 600 mg per dose.

[0052] In some embodiments, the anti-PD-1 antibody and the therapeutic agent are independent administration units, combined for administration. In some embodiments, the anti-PD-1 antibody can be administered before the administration of the therapeutic agent, after the administration of the therapeutic agent, or simultaneously with the administration of the therapeutic agent.

[0053] In some embodiments, the anti-PD-1 antibody and the therapeutic agent form a combined administration unit, combined for administration.

[0054] In some embodiments, the patient has a tumor or cancer. In some embodiments, the tumor or cancer includes, but is not limited to, head and neck tumor, Hodgkin's lymphoma, non-Hodgkin's lymphoma, pre-lymphoblastic lymphoma, small unclipped cell lymphoma, Burkitt's lymphoma, non-Burkitt's lymphoma, diffuse large B-cell lymphoma, anaplastic large cell lymphoma, kidney tumor, nephroblastoma, Wilms' tumor, renal clear cell carcinoma, renal rhabdoid tumor, renal clear cell sarcoma, renal primitive neuroectodermal tumor, neuroblastoma, nodal cell neuroblastoma, nodal cell neuroma, extracranial germ cell tumor, mature teratoma, immature teratoma, endodermal sinus tumor, yolk sac tumor, seminoma, dysgerminoma, choriocarcinoma, embryonal carcinoma, osteosarcoma, chondrosarcoma, rhabdomyosarcoma, soft tissue sarcoma, fibrosarcoma, malignant fibrous histiocytoma, liposarcoma, leiomyosarcoma, angiosarcoma, lymphangiosarcoma, malignant schwannoma, alveolar soft part sarcoma, epithelioid sarcoma, clear cell sarcoma, malignant melanoma, synovial sarcoma, desmoplastic small round cell tumor, Ewing's sarcoma, primitive neuroectodermal tumor, liver tumor, hepatoblastoma, retinoblastoma, posterior fossa medulloblastoma, thymoma, pulmonary blastoma, pancreatic blastoma, islet cell tumor, ileocecal carcinoid, mesothelioma, melanoma, mesenchymoma, myeloma, brain astrocytoma, nasopharyngeal carcinoma, thyroid papillary carcinoma, intestinal carcinoma, breast carcinoma, stomach carcinoma, liver carcinoma, prostate carcinoma, breast carcinoma, lung carcinoma, cervical carcinoma, ovarian carcinoma, renal carcinoma, lymphoma, leukemia, skin carcinoma, and esophageal squamous carcinoma, etc.

[0055] In some embodiments, the present disclosure provides a method for treating a tumor or cancer in a patient in need thereof, comprising administering an effective amount of an anti-PD-1 antibody and a therapeutic agent, wherein the effective amount of the anti-PD-1 administration is about 50 mg to 600 mg per treatment cycle. In some embodiments, one treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values, inclusive of the endpoints, or any value therein. In some embodiments, the anti-PD-1 antibody is Antibody A.

[0056] In some embodiments, the therapeutic agent is a monoclonal antibody that specifically binds to the extracellular dimerization domain of human epidermal growth factor receptor 2 (HER2) (subdomain II) (anti-HER2 antibody), such as pertuzumab, which has a molecular weight of about 148 kDa. Pertuzumab can be expressed in cells (e.g., CHO) by genetic engineering and obtained by purification; the purification can be performed by conventional methods, for example, centrifuging the cell suspension and collecting the supernatant, and centrifuging again to further remove impurities. Methods such as Protein A affinity column and ion exchange column can be used for purifying the antibody.

[0057] In some embodiments, the therapeutic agent is pertuzumab, pertuzumab includes or a biosimilar thereof or an ADCC effector-enhancing mAb or afucosylated mAb. In some embodiments, the effective amount of pertuzumab administration is about 40 mg to 900 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (inclusive of the endpoints) or any value therein. In some embodiments, the effective amount of pertuzumab administration is about 840 mg initially, followed by 420 mg every 3 weeks.

[0058] In some embodiments, the therapeutic agent is a recombinant humanized immunoglobulin G1 (IgG1) monoclonal antibody that can bind to VEGF-A, inhibiting its binding to VEGF receptor-2 (VEGFR-2) (anti-VEGF antibody), such as bevacizumab. Bevacizumab can be expressed by genetic engineering in cells (such as CHO) and obtained by purification; purification can be carried out by conventional methods, for example, centrifuging the cell suspension and collecting the supernatant, and centrifuging again to further remove impurities. Methods such as Protein A affinity column and ion exchange column can be used for purification of antibodies.

[0059] In some embodiments, the therapeutic agent is bevacizumab, bevacizumab includes or a biosimilar thereof, such as or BAT1706, or an ADCC effector-enhancing mAb or afucosylated mAb.

[0060] In some embodiments, the effective amount of bevacizumab administration is about 50 mg to 400 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (inclusive of the endpoints) or any value therein. In some embodiments, the effective amount of bevacizumab administration is about 5 mg / kg to 15 mg / kg every 2 weeks or every 3 weeks. In some embodiments, the effective amount of bevacizumab administration is about 5 mg / kg, 7.5 mg / kg, 10 mg / kg, or 15 mg / kg every 2 weeks or every 3 weeks. In some embodiments, the effective amount of bevacizumab administration is about 5 mg / kg every 2 weeks, 10 mg / kg every 2 weeks, 7.5 mg / kg every 3 weeks, 15 mg / kg every 3 weeks.

[0061] In some embodiments, the therapeutic agent is a targeted CD20 antibody (anti-CD20 antibody), such as ofatumumab, which is a fully human IgGl kappa monoclonal antibody, or obinutuzumab. Ofatumumab and obinutuzumab can be expressed by genetic engineering in cells (such as CHO) and obtained by purification; the purification can be carried out by conventional methods, for example, centrifuging the cell suspension and collecting the supernatant, and centrifuging again to further remove impurities. Methods such as Protein A affinity column and ion exchange column can be used for purification of antibodies.

[0062] In some embodiments, the therapeutic agent is ofatumumab, which includes or or its biosimilar or ADCC effect-enhanced monoclonal antibody or defucosylated monoclonal antibody, such as BAT4406F disclosed in CN109096399A. In some embodiments, the effective amount of ofatumumab administration is about 10 mg to 2000 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including the endpoints) or any value therein. In some embodiments, the effective amount of ofatumumab administration is about 20 mg once a week or once a month. In some embodiments, the effective amount of ofatumumab administration is about initial 300 mg, 1000 mg after 1 week, and then 1000 mg once every 4 weeks or every 8 weeks. In some embodiments, the effective amount of ofatumumab administration is about initial 300 mg, 2000 mg after 1 week, and then 2000 mg once every 1 week or every 4 weeks.

[0063] In some embodiments, the therapeutic agent is obinutuzumab, which includes or its biosimilar, or ADCC effect-enhanced monoclonal antibody or defucosylated monoclonal antibody, such as BAT4306F described in CN109096399A. In some embodiments, the effective amount of obinutuzumab administration is about 10 mg to 2000 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including the endpoints) or any value therein. In some embodiments, the effective amount of obinutuzumab administration is 100 mg on day 1, 900 mg on day 2, 1,000 mg on day 8, day 15, and then 1,000 mg per course of treatment. In some embodiments, the effective amount of obinutuzumab administration is 1,000 mg on day 1, day 8, day 15, and then 1,000 mg per course of treatment. Each course of treatment can be 1 month or 2 months.

[0064] In some embodiments, the therapeutic agent is an antibody targeting cytotoxic T-lymphocyte-associated antigen 4 (CTLA4) (anti-CTLA4 antibody), such as ipilimumab, which is an IgG1 kappa immunoglobulin with a molecular weight of about 148 kDa. Ipilimumab can be expressed in cells (such as CHO) by genetic engineering and obtained by purification; the purification can be performed using conventional methods, for example, centrifuging the cell suspension and collecting the supernatant, and centrifuging again to further remove impurities. Methods such as Protein A affinity column and ion exchange column can be used for purifying the antibody.

[0065] In some embodiments, the therapeutic agent is ipilimumab, which includes Yervoy TM or a biosimilar or ADCC effect-enhancing mAb or afutuzumab, as described in WO2014089113. In some embodiments, the effective amount of ipilimumab administration is about 30 mg to 300 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including the endpoints) or any value therein. In some embodiments, the effective amount of ipilimumab administration is 1 mg / kg, 3 mg / kg or 10 mg / kg once every 3 weeks, 6 weeks, or 12 weeks. In some embodiments, the effective amount of ipilimumab administration is 1 mg / kg once every 3 weeks or 6 weeks. In some embodiments, the effective amount of ipilimumab administration is 3 mg / kg once every 3 weeks or 6 weeks. In some embodiments, the effective amount of ipilimumab administration is 10 mg / kg once every 3 weeks or every 12 weeks.

[0066] In some embodiments, the therapeutic agent is a Trop2-ADC described herein. In some embodiments, the effective amount of Trop2-ADC administration is about 60 mg to 400 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values (including the endpoints) or any value therein. In some embodiments, the effective amount of Trop2-ADC administration is 0.2, 0.5, 1, 2, 4, 6, 8 or 10 mg / kg once every week to every 3 weeks. In some embodiments, the effective amount of Trop2-ADC administration is 0.2, 0.5, 1, 2, 4, 6, 8 or 10 mg / kg once every week or every 3 weeks.

[0067] In some embodiments, the therapeutic agent is a HER2-ADC. In some embodiments, the effective amount of HER2-ADC administration is about 70 mg to 400 mg per treatment cycle. In some embodiments, a treatment cycle is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, or a range between any two of these values, inclusive of the endpoints, or any value therein. In some embodiments, the HER2-ADC is HERCEPTIN®-ADC B .

[0068] In some embodiments, the anti-PD-1 antibody and the therapeutic agent (or a combination of the PD-1 antibody and the therapeutic agent) can be formulated separately into pharmaceutical compositions and administered to a patient in a variety of forms adapted to the chosen route of administration, e.g., parenterally, intravenously (iv), intramuscularly, topically, or subcutaneously. In some embodiments, the anti-PD-1 antibody and the therapeutic agent (or a combination of the PD-1 antibody and the therapeutic agent) can be infused intravenously. The amount of the anti-PD-1 antibody and the therapeutic agent will depend on the nature of the drug, the extent of internalization of the cell surface trigger of the drug, the extent of transport and release, the disease being treated, the condition of the patient (e.g., age, sex, weight, etc.).

[0069] In some embodiments, the anti-PD-1 antibody is administered at about 1 mg / kg to 10 mg / kg per administration or a formulation containing such a dose of the anti-PD-1 antibody. In some embodiments, the anti-PD-1 antibody is administered at about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, or a range between any two of these values, inclusive of the endpoints, or any value therein, per administration or a formulation containing such a dose of the anti-PD-1 antibody.

[0070] In some embodiments, a therapeutically effective amount of pertuzumab and an anti-PD-1 antibody are applied to a subject patient separately or simultaneously. The dosing cycles of pertuzumab and the anti-PD-1 antibody can be the same or different.

[0071] In some embodiments, the paclitaxel is administered at about 1 mg / m2to 100 mg / m2or a formulation containing such an amount of paclitaxel per administration. In some embodiments, the paclitaxel is administered at about 1 mg / m2, about 1.2 mg / m2, about 2 mg / m2, about 2.4 mg / m2, about 3 mg / m2, about 3.6 mg / m2, about 4 mg / m2, about 4.8 mg / m2, about 5 mg / m2, about 5.5 mg / m2, about 6 mg / m2, about 6.9 mg / m2, about 7 mg / m2, about 8.4 mg / m2, about 9 mg / m2, about 11 mg / m2, about 12 mg / m2, about 15 mg / m2, about 18 mg / m2, about 20 mg / m2, about 22 mg / m2, about 24 mg / m2, about 26 mg / m2, about 28 mg / m2, about 30 mg / m2, about 32 mg / m2, about 36 mg / m2, about 40 mg / m2, about 44 mg / m2, about 48 mg / m2, about 50 mg / m2, about 52 mg / m2, about 54 mg / m2, about 56 mg / m2, about 60 mg / m2, about 64 mg / m2, about 68 mg / m2, about 70 mg / m2, about 72 mg / m2, about 74 mg / m2, about 76 mg / m2, about 80 mg / m2, about 84 mg / m2, about 88 mg / m2, about 90 mg / m2, about 92 mg / m2, about 94 mg / m2, about 96 mg / m2, about 98 mg / m2, or a range between any two of these values (including endpoints) or any value therein, or a formulation containing such an amount of paclitaxel per administration.

[0072] In some embodiments, a therapeutically effective amount of bevacizumab and an anti-PD-1 antibody are administered separately or concurrently to a subject patient. The dosing cycles of bevacizumab and the anti-PD-1 antibody can be the same or different.

[0073] In some embodiments, the bevacizumab is administered at about 1 mg / kg to 9 mg / kg or a formulation containing such an amount of bevacizumab per administration. In some embodiments, the bevacizumab is administered at about 1 mg / kg, about 1.2 mg / kg, about 2 mg / kg, about 2.4 mg / kg, about 3 mg / kg, about 3.6 mg / kg, about 4 mg / kg, about 4.8 mg / kg, about 5 mg / kg, about 5.5 mg / kg, about 6 mg / kg, about 6.9 mg / kg, about 7 mg / kg, about 8.4 mg / kg, about 9 mg / kg, or a range between any two of these values (including endpoints) or any value therein, or a formulation containing such an amount of bevacizumab per administration.

[0074] In some embodiments, a therapeutically effective amount of ofatumumab and an anti-PD-1 antibody are administered separately or concurrently to a subject patient. The dosing cycles of ofatumumab and the anti-PD-1 antibody can be the same or different.

[0075] In some embodiments, the ofatumumab administered per dose is about 0.5 mg / kg to 18 mg / kg or a formulation containing such a dose of ofatumumab. In some embodiments, the ofatumumab administered per dose is about 0.5 mg / kg, about 1 mg / kg, about 1.2 mg / kg, about 2 mg / kg, about 2.4 mg / kg, about 3 mg / kg, about 3.6 mg / kg, about 4 mg / kg, about 4.8 mg / kg, about 5 mg / kg, about 5.5 mg / kg, about 6 mg / kg, about 6.9 mg / kg, about 7 mg / kg, about 8.4 mg / kg, about 9 mg / kg, about 11 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, about 17 mg / kg, about 18 mg / kg, or a range between any two of these values, inclusive of the endpoints, or any value therein, or a formulation containing such a dose of ofatumumab.

[0076] In some embodiments, a therapeutically effective amount of obinutuzumab and an anti-PD-1 antibody are administered separately or concurrently to a subject patient. The dosing schedules of obinutuzumab and the anti-PD-1 antibody can be the same or different.

[0077] In some embodiments, the obinutuzumab administered per dose is about 0.5 mg / kg to 15 mg / kg or a formulation containing such a dose of ofatumumab. In some embodiments, the ofatumumab administered per dose is about 0.5 mg / kg, about 1 mg / kg, about 1.2 mg / kg, about 2 mg / kg, about 2.4 mg / kg, about 3 mg / kg, about 3.6 mg / kg, about 4 mg / kg, about 4.8 mg / kg, about 5 mg / kg, about 5.5 mg / kg, about 6 mg / kg, about 6.9 mg / kg, about 7 mg / kg, about 8.4 mg / kg, about 9 mg / kg, about 11 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, or a range between any two of these values, inclusive of the endpoints, or any value therein, or a formulation containing such a dose of ofatumumab.

[0078] In some embodiments, a therapeutically effective amount of an anti-CTLA4 antibody (such as ipilimumab or afutumumab) and an anti-PD-1 antibody are administered separately or concurrently to a subject patient. The dosing schedules of the anti-CTLA4 antibody (such as ipilimumab or afutumumab) and the anti-PD-1 antibody can be the same or different.

[0079] In some embodiments, the anti-CTLA4 antibody (such as ipilimumab or afutumumab) administered per dose is about 0.1 mg / kg to 10 mg / kg, or about 0.5 mg / kg to 10 mg / kg or a formulation containing such a dose of anti-CTLA4 antibody (such as ipilimumab or afutumumab). In some embodiments, the ipilimumab administered per dose is about 0.5 mg / kg, about 1 mg / kg, about 1.2 mg / kg, about 2 mg / kg, about 2.4 mg / kg, about 3 mg / kg, about 3.6 mg / kg, about 4 mg / kg, about 4.8 mg / kg, about 5 mg / kg, about 5.5 mg / kg, about 6 mg / kg, about 6.9 mg / kg, about 7 mg / kg, about 8.4 mg / kg, about 9 mg / kg, about 10 mg / kg, or a range between any two of these values (including endpoints) or any value therein, or a formulation containing such a dose of ipilimumab.

[0080] In some embodiments, a therapeutically effective amount of the Trop2-ADC and the anti-PD-1 antibody are applied to the subject patient separately or simultaneously. The dosing cycles of the Trop2-ADC and the anti-PD-1 antibody can be the same or different.

[0081] In some embodiments, the Trop2-ADC administered per dose is about 0.5 mg / kg to 7 mg / kg or a formulation containing such a dose of Trop2-ADC. In some embodiments, the Trop2-ADC administered per dose is about 1 mg / kg, about 1.2 mg / kg, about 2 mg / kg, about 2.4 mg / kg, about 3 mg / kg, about 3.6 mg / kg, about 4 mg / kg, about 4.8 mg / kg, about 5 mg / kg, about 5.5 mg / kg, about 7 mg / kg, or a range between any two of these values (including endpoints) or any value therein, or a formulation containing such a dose of Trop2-ADC.

[0082] In some embodiments, a therapeutically effective amount of the HER2-ADC and the anti-PD-1 antibody are applied to the subject patient separately or simultaneously. The dosing cycles of the HER2-ADC and the anti-PD-1 antibody can be the same or different.

[0083] In some embodiments, the HER2-ADC is about 1 mg / kg to 6 mg / kg per administration or a formulation containing such a dose of HER2-ADC. In some embodiments, the HER2-ADC is about 1 mg / kg, about 1.2 mg / kg, about 2 mg / kg, about 2.4 mg / kg, about 3 mg / kg, about 3.6 mg / kg, about 4 mg / kg, about 4.8 mg / kg, about 5 mg / kg, about 5.5 mg / kg, about 6 mg / kg, or a range between any two of these values (including the endpoints) or any value therein, per administration or a formulation containing such a dose of HER2-ADC.

[0084] In some embodiments, the present disclosure provides a method of treating a tumor or cancer, comprising administering to a patient in need thereof an effective amount of an anti-PD-1 antibody (or a formulation thereof) and a therapeutic agent (or a formulation thereof); wherein the effective amount of the anti-PD-1 antibody is about 50 mg to 600 mg (or a formulation containing such a dose of anti-PD-1 antibody) per administration. In some embodiments, the therapeutic agent is a HER2-ADC. In some embodiments, the effective amount of the HER2-ADC is about 70 mg to 400 mg (or a formulation containing such a dose of HER2-ADC) per administration. In some embodiments, the therapeutic agent is an anti-CTLA4 antibody. In some embodiments, the anti-CTLA4 antibody is Antibody C. In some embodiments, the effective amount of the anti-CTLA4 antibody is about 6 mg to 600 mg (or a formulation containing such a dose of anti-CTLA4 antibody) per administration. The dosage schedule and mode of administration depend on the risk-benefit assessment of the anti-PD-1 antibody (or a formulation thereof), the HER2-ADC (or a formulation thereof), or the anti-CTLA4 antibody (or a formulation thereof) in certain patient populations and general clinical practice guidelines.

[0085] In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 50 mg to 600 mg of the anti-PD-1 antibody (or a formulation containing such a dose of anti-PD-1 antibody), and the effective amount of the HER2-ADC administered to the patient per treatment cycle is about 70 mg to 400 mg (or a formulation containing such a dose of HER2-ADC).

[0086] In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 50 mg to 600 mg of the anti-PD-1 antibody (or a formulation containing such a dose of anti-PD-1 antibody), and the effective amount of the anti-CTLA4 administered to the patient per treatment cycle is about 6 mg to 600 mg (or a formulation containing such a dose of anti-CTLA4 antibody).

[0087] In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 50 mg, about 60 mg, about 80 mg, about 120 mg, about 200 mg, about 250 mg, about 290 mg, about 300 mg, about 330 mg, about 380 mg, about 400 mg, about 434 mg, about 480 mg, about 500 mg, about 567 mg, about 580 mg, about 600 mg, or a range between any two of these values, inclusive of the endpoints, or any of these values, or a formulation containing such a dose of the anti-PD-1 antibody. In some embodiments, one treatment cycle is a single administration of 1 week to 7 weeks. In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is 100 mg to 200 mg, or a formulation containing such a dose of the anti-PD-1 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, or a range between any two of these values, inclusive of the endpoints, or any of these values. In some embodiments, one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 200 mg to about 300 mg, or a formulation containing such a dose of the anti-PD-1 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, or a range between any two of these values, inclusive of the endpoints, or any of these values, or a formulation containing such a dose of the anti-PD-1 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks.

[0088] In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 100 mg, or a formulation containing such a dose of the anti-PD-1 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 90 mg to 110 mg, or a formulation containing such a dose of the anti-PD-1 antibody; such as about 100 mg administered once.

[0089] In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 120 mg, or a formulation containing this amount of the anti-PD-1 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 100 mg to 140 mg, or a formulation containing this amount of the anti-PD-1 antibody; such as about 120 mg administered once.

[0090] In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 160 mg, or a formulation containing this amount of the anti-PD-1 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 150 mg to 190 mg, or a formulation containing this amount of the anti-PD-1 antibody; such as about 160 mg administered once.

[0091] In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 200 mg, or a formulation containing this amount of the anti-PD-1 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the effective amount of the anti-PD-1 antibody administered to the patient per treatment cycle is about 190 mg to 230 mg, or a formulation containing this amount of the anti-PD-1 antibody; such as about 200 mg administered once.

[0092] In some embodiments, the effective amount of the anti-PD-1 antibody is about 100 mg to 600 mg once every 3 weeks. In some embodiments, the effective amount of the anti-PD-1 antibody is about 100 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, or about 600 mg once every 3 weeks. In some embodiments, the effective amount of the anti-PD-1 antibody is about 100 mg, about 300 mg, or about 600 mg once every 3 weeks.

[0093] In some embodiments, the effective amount of the HER2-ADC administered to the patient per treatment cycle is about 70 mg, about 90 mg, about 120 mg, about 160 mg, about 180 mg, about 200 mg, about 230 mg, about 250 mg, about 280 mg, about 300 mg, about 310 mg, about 334 mg, about 350 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, or a range between any two of these values, inclusive as to endpoints, or any value therein, or a formulation containing such a dose of the HER2-ADC. In some embodiments, one treatment cycle is a 1 -week to 4-week administration once. In some embodiments, the effective amount of the HER2-ADC administered to the patient per treatment cycle is 100 mg to 200 mg, or a formulation containing such a dose of the HER2-ADC; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, or a range between any two of these values, inclusive as to endpoints, or any value therein. In some embodiments, the effective amount of the HER2-ADC administered to the patient per treatment cycle is about 200 mg to about 300 mg, or a formulation containing such a dose of the HER2-ADC; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the effective amount of the HER2-ADC administered to the patient per treatment cycle is about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, or a range between any two of these values, inclusive as to endpoints, or any value therein, or a formulation containing such a dose of the HER2-ADC; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks.

[0094] In some embodiments, the effective amount of the HER2-ADC administered to the patient per treatment cycle is about 100 mg, or a formulation containing such a dose of the HER2-ADC; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the effective amount of the HER2-ADC administered to the patient per treatment cycle is about 90 mg to 110 mg, or a formulation containing such a dose of the HER2-ADC; such as about 100 mg administered once.

[0095] In some embodiments, the effective amount of the HER2-ADC administered to the patient per treatment cycle is about 130 mg, or a formulation containing such a dose of the HER2-ADC; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the effective amount of the HER2-ADC administered to the patient per treatment cycle is about 100 mg to 140 mg, or a formulation containing such a dose of the HER2-ADC; such as about 130 mg administered once.

[0096] In some embodiments, the patient is administered an effective amount of the HER2-ADC is about 170 mg, or a formulation containing such a dose of the HER2-ADC, per treatment cycle; wherein a treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the patient is administered an effective amount of the HER2-ADC is about 150 mg to 190 mg, or a formulation containing such a dose of the HER2-ADC, per treatment cycle; such as about 170 mg administered once.

[0097] In some embodiments, the patient is administered an effective amount of the HER2-ADC is about 200 mg, or a formulation containing such a dose of the HER2-ADC, per treatment cycle; wherein a treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the patient is administered an effective amount of the HER2-ADC is about 190 mg to 230 mg, or a formulation containing such a dose of the HER2-ADC, per treatment cycle; such as about 200 mg administered once.

[0098] In some embodiments, the effective amount of the anti-CTLA4 antibody administered to the patient per treatment cycle is about 6 mg, about 8 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 50 mg, about 60 mg, about 80 mg, about 120 mg, about 200 mg, about 250 mg, about 290 mg, about 300 mg, about 330 mg, about 380 mg, about 400 mg, about 434 mg, about 480 mg, about 500 mg, about 567 mg, about 580 mg, about 600 mg, or a range between any two of these values, inclusive of the endpoints, or any of these values, or a formulation containing such a dose of the anti-CTLA4 antibody. In some embodiments, one treatment cycle is one administration per week for 1 to 7 weeks. In some embodiments, the effective amount of the anti-CTLA4 antibody administered to the patient per treatment cycle is 6 mg to 80 mg, or a formulation containing such a dose of the anti-CTLA4 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, or a range between any two of these values, inclusive of the endpoints, or any of these values. In some embodiments, the effective amount of the anti-CTLA4 antibody administered to the patient per treatment cycle is 100 mg to 200 mg, or a formulation containing such a dose of the anti-CTLA4 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, or a range between any two of these values, inclusive of the endpoints, or any of these values. In some embodiments, one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the effective amount of the anti-CTLA4 antibody administered to the patient per treatment cycle is about 200 mg to about 300 mg, or a formulation containing such a dose of the anti-CTLA4 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the effective amount of the anti-CTLA4 antibody administered to the patient per treatment cycle is about about 6 mg, about 8 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 50 mg, about 60 mg, 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, or a range between any two of these values, inclusive of the endpoints, or any of these values, or a formulation containing such a dose of the anti-CTLA4 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks.

[0099] In some embodiments, the effective amount of the anti-CTLA4 antibody administered to the patient per treatment cycle is about 6 mg, or a formulation containing this amount of the anti-CTLA4 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the about 6 mg is administered once per treatment cycle.

[0100] In some embodiments, the effective amount of the anti-CTLA4 antibody administered to the patient per treatment cycle is about 15 mg, or a formulation containing this amount of the anti-CTLA4 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the about 15 mg is administered once per treatment cycle.

[0101] In some embodiments, the effective amount of the anti-CTLA4 antibody administered to the patient per treatment cycle is about 23 mg, or a formulation containing this amount of the anti-CTLA4 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the about 23 mg is administered once per treatment cycle.

[0102] In some embodiments, the effective amount of the anti-CTLA4 antibody administered to the patient per treatment cycle is about 60 mg, or a formulation containing this amount of the anti-CTLA4 antibody; wherein one treatment cycle is about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some embodiments, the about 60 mg is administered once per treatment cycle.

[0103] In some embodiments, the effective amount of the anti-CTLA4 antibody is about 6 mg to 600 mg once every 3 weeks. In some embodiments, the effective amount of the anti-CTLA4 antibody is about 6 mg, about 20 mg, about 30 mg, about 100 mg, about 200 mg, about 230 mg, about 300 mg, about 400 mg, about 500 mg, or about 600 mg once every 3 weeks. In some embodiments, the effective amount of the anti-CTLA4 antibody is about 6 mg, about 30 mg, or about 60 mg once every 3 weeks.

[0104] In some embodiments, the anti-PD-1 antibody and the therapeutic agent (or the combination of the anti-PD-1 antibody and the therapeutic agent) are each administered once per treatment cycle. In some embodiments, the anti-PD-1 antibody and the therapeutic agent (or the combination of the anti-PD-1 antibody and the therapeutic agent) are each administered multiple times per treatment cycle, e.g., 2 times, 3 times, 4 times, or 5 times.

[0105] In some implementations, patients receive only one or four doses per treatment cycle. In some implementations, patients receive one treatment cycle. In some implementations, patients receive multiple treatment cycles (e.g., two, three, or four). In some implementations, patients receive treatment until their symptoms are relieved and treatment is no longer necessary. In some implementations, patients receive multiple cycles of anti-PD-1 antibody treatment and four cycles of anti-CTLA4 antibody treatment.

[0106] In some embodiments, a method for treating tumors or cancer includes: administering to a patient in need an anti-PD-1 antibody, such as about 50 mg to 200 mg, about 200 mg to 300 mg, about 300 mg to 400 mg, or about 400 mg to 600 mg, for example about 100 mg, about 120 mg, about 200 mg, or about 400 mg, a formulation containing this dose of the anti-PD-1 antibody; and further administering to a patient in need a HER2-ADC, such as about 70 mg to 100 mg, about 100 mg to 200 mg, about 200 mg to 300 mg, or about 300 mg to 400 mg, for example about 100 mg, about 120 mg, about 150 mg, or about 200 mg, a formulation containing this dose of the HER2-ADC. In some embodiments, the patient receives a single dose of the anti-PD-1 antibody and a single dose of the HER2-ADC. In some embodiments, the patient receives a single dose of a combination of the anti-PD-1 antibody and the HER2-ADC.

[0107] In some implementations, 200 mg of anti-PD-1 antibody and 2.4 mg / kg of HER2-ADC are administered every 3 weeks. In some implementations, 200 mg of anti-PD-1 antibody and 3.6 mg / kg of HER2-ADC are administered every 3 weeks.

[0108] In some embodiments, a method for treating tumors or cancer includes: administering to a patient in need an anti-PD-1 antibody at a dose of approximately 50 mg to 200 mg, approximately 200 mg to 300 mg, approximately 300 mg to 400 mg, or approximately 400 mg to 600 mg, such as approximately 100 mg, approximately 120 mg, approximately 200 mg, or approximately 400 mg, a formulation containing this dose of the anti-PD-1 antibody; and further administering to a patient in need an anti-CTLA4 antibody at a dose of approximately 6 mg to 80 mg, approximately 80 mg to 200 mg, approximately 200 mg to 300 mg, or approximately 300 mg to 600 mg, such as approximately 6 mg, approximately 12 mg, approximately 25 mg, approximately 35 mg, approximately 50 mg, approximately 75 mg, approximately 125 mg, or approximately 200 mg, a formulation containing this dose of the anti-CTLA4 antibody. In some embodiments, the patient receives a single dose of the anti-PD-1 antibody and a single dose of the anti-CTLA4 antibody. In some implementations, patients receive a single dose of a combination of anti-PD-1 antibody and anti-CTLA4 antibody.

[0109] In some embodiments, 200 mg of anti-PD-1 antibody and 0.1 mg / kg of anti-CTLA4 antibody are administered every 3 weeks. In some embodiments, 200 mg of anti-PD-1 antibody and 1 mg / kg of anti-CTLA4 antibody are administered every 3 weeks. In some embodiments, the patient's symptoms are relieved after a single dose. In some embodiments, if the patient's symptoms do not improve as expected after a single dose, the patient is subsequently administered approximately 50 mg to 600 mg of anti-PD-1 antibody and approximately 70 mg to 400 mg of HER2-ADC, respectively. In some embodiments, if the patient's symptoms do not improve as expected after a single dose, the patient is subsequently administered a combination of approximately 50 mg to 600 mg of anti-PD-1 antibody and approximately 70 mg to 400 mg of HER2-ADC, respectively. In some embodiments, if the patient's symptoms do not improve as expected after a single dose, the patient is subsequently administered approximately 50 mg to 600 mg of anti-PD-1 antibody and approximately 6 mg to 600 mg of anti-CTLA4 antibody, respectively. In some implementations, if the patient's symptoms do not improve as expected after a single dose, the patient is then given a combination of approximately 50 mg to 600 mg of anti-PD-1 antibody and approximately 6 mg to 600 mg of anti-CTLA4 antibody.

[0110] In some embodiments, the anti-PD-1 antibody (or formulation), HER2-ADC (or formulation), and anti-CTLA4 antibody (or formulation) are administered via subcutaneous (sc), intraperitoneal (ip), parenteral, intra-arterial, or intravenous (iv) injection. In some embodiments, the anti-PD-1 antibody (or formulation) and HER2-ADC (or formulation) are administered via intravenous infusion. In some embodiments, the anti-PD-1 antibody (or formulation) and HER2-ADC (or formulation) are administered via bolus injection. In some embodiments, the anti-PD-1 antibody (or formulation) and anti-CTLA4 antibody (or formulation) are administered via intravenous infusion. In some embodiments, the anti-PD-1 antibody (or formulation) and anti-CTLA4 antibody (or formulation) are administered via bolus injection.

[0111] In some embodiments, the anti-PD-1 antibody (or formulation), HER2-ADC (or formulation), and anti-CTLA4 antibody (or formulation) are administered via intravenous (iv) infusion. In some embodiments, the duration of intravenous infusion is approximately 50 minutes, approximately 55 minutes, approximately 60 minutes, approximately 65 minutes, approximately 70 minutes, approximately 75 minutes, approximately 81 minutes, approximately 87 minutes, approximately 90 minutes, approximately 95 minutes, or a range (including endpoints) of any two of these values, or any value thereof.

[0112] In some implementations, anti-PD-1 antibodies (or formulations), therapeutic agents (or formulations) are used in combination with other treatments to treat tumors or cancers, such as chemotherapy, radiotherapy, and surgery.

[0113] On the other hand, this invention discloses the use of anti-PD-1 antibodies and therapeutic agents in the preparation of medicaments for treating tumors or cancer. In some embodiments, the therapeutic agent is selected from the group consisting of: anti-EGRR antibodies, anti-VEGF antibodies, anti-VEGFR2 antibodies, anti-CTLA4 antibodies, anti-PD-L1 antibodies, anti-HER2 antibodies, anti-CD20 antibodies, anti-Trop2 antibodies, anti-TIGIT antibodies, anti-OX40 antibodies, and anti-ICOS antibodies, anti-HER2 antibody-drug conjugates, and anti-Trop2 antibody-drug conjugates. In some embodiments, the medicament for treating tumors or cancer comprises an anti-PD-1 antibody and a therapeutic agent. In some embodiments, the anti-PD-1 antibody is antibody A. In some embodiments, the therapeutic agent is a HER2-ADC. In some embodiments, the HER2-ADC is a HER2-ADC. B In some embodiments, the therapeutic agent is an anti-CTLA4 antibody. In some embodiments, the anti-CTLA4 antibody is antibody C.

[0114] On the other hand, the present invention also discloses a kit comprising an anti-PD-1 antibody (or formulation), a therapeutic agent (or formulation), and instructions for administering the anti-PD-1 antibody (or formulation) and the therapeutic agent (or formulation) to patients in need. In some embodiments, the present invention also discloses a kit comprising a composition (or formulation) of an anti-PD-1 antibody and a therapeutic agent, and instructions for administering the composition (or formulation) of the anti-PD-1 antibody and the therapeutic agent to patients in need.

[0115] On the other hand, the present invention also discloses injectable pharmaceutical compositions comprising an anti-PD-1 antibody and a therapeutic agent, such as bolus-type or infusion (drip)-type pharmaceutical compositions. In some embodiments, the pharmaceutical composition comprises at least 0.1% of an anti-PD-1 antibody and 0.1% of a therapeutic agent. The percentages of antibody and therapeutic agent can vary and are between about 2% and about 90% by weight of a given dosage form. The amount of anti-PD-1 antibody and therapeutic agent in such therapeutically useful pharmaceutical compositions can be an effective amount for administration.

[0116] On the other hand, the present invention also discloses a method for preparing the above-described pharmaceutical composition: mixing the anti-PD-1 antibody and therapeutic agent (or a composition of anti-PD-1 antibody and therapeutic agent) described herein with a pharmaceutically acceptable carrier suitable for injection (e.g., water for injection, physiological saline, etc.). The method for mixing the above-described anti-PD-1 antibody and therapeutic agent with a pharmaceutically acceptable carrier is generally known in the art.

[0117] This invention utilizes anti-PD-1 antibodies (or formulations) and therapeutic agents (or formulations) in the treatment of tumors or cancer, which can alleviate symptoms. Attached Figure Description

[0118] Figure 1 The combined administration of anti-PD-1 antibody and HER2-ADC showed that it inhibited the proliferation of tumor cells.

[0119] Figure 2 The changes in tumor volume after combined administration of antibody A and antibody C are shown; mean body weight is expressed as mean ± SEM.

[0120] Figure 3 This chart shows tumor weight on day 27 after combined administration of antibody A and antibody C; data are expressed as mean ± SEM. For group G5, the independent samples t-test was used for all groups except G5. For group G5, due to the non-normal distribution of data, the Mann-Whitney test was used for two independent samples. * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001.

[0121] Figure 4This shows the changes in mouse body weight after combined administration of antibody A and antibody C.

[0122] Terminology

[0123] Unless otherwise stated, each of the following terms shall have the meaning described below.

[0124] definition

[0125] It should be noted that the term “a” entity refers to one or more of the same entity. For example, “an antibody” should be understood as one or more antibodies. Therefore, the terms “a” (or “an”), “one or more” and “at least one” can be used interchangeably in this document.

[0126] As used herein, the terms “comprising” or “including” mean that compositions and methods include the listed elements, such as components or steps, but do not exclude others. “Substantially composed of” means that compositions and methods exclude other elements that have a fundamental effect on the characterization of the composition, but do not exclude elements that do not substantially affect the composition or method. “Composed of” means excluding elements not specifically listed.

[0127] The term "polypeptide" is intended to encompass both the singular and plural forms of "polypeptide" and refers to a molecule formed by amino acid monomers linearly linked by amide bonds (also known as peptide bonds). The term "polypeptide" refers to any single or multiple chains of two or more amino acids and does not imply a specific length of the product. Therefore, the definition of "polypeptide" includes peptide, dipeptide, tripeptide, oligopeptide, "protein," "amino acid chain," or any other term used to refer to two or more amino acid chains, and the term "polypeptide" can be used in place of any of the foregoing terms or interchangeably with any of the foregoing terms. The term "polypeptide" is also intended to refer to products modified after polypeptide expression, including but not limited to glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting / blocking groups, proteolytic cleavage, or non-naturally occurring amino acid modifications. Polypeptides can be derived from natural biological sources or produced through recombinant technologies, but they do not necessarily have to be translated from a specified nucleic acid sequence; they can be produced in any manner, including chemical synthesis.

[0128] An amino acid is an organic compound containing both an amino group and a carboxyl group, such as an α-amino acid, which can be encoded by nucleic acids directly or in its precursor form. A single amino acid is encoded by a nucleic acid consisting of three nucleotides (so-called codons or base triplets). Each amino acid is encoded by at least one codon. The fact that the same amino acid is encoded by different codons is called "degeneracy of the genetic code." Amino acids include both natural and non-natural amino acids. Natural amino acids include alanine (three-letter code: ala, one-letter code: A), arginine (arg, R), asparagine (asn, N), aspartic acid (asp, D), cysteine ​​(cys, C), glutamine (gln, Q), glutamic acid (glu, E), glycine (gly, G), histidine (his, H), isoleucine (ile, I), leucine (leu, L), lysine (lys, K), methionine (met, M), phenylalanine (phe, F), proline (pro, P), serine (ser, S), threonine (thr, T), tryptophan (trp, W), tyrosine (tyr, Y), and valine (val, V).

[0129] "Conservative amino acid substitution" refers to the replacement of one amino acid residue with another amino acid residue containing a side chain (R group) with similar chemical properties (such as charge or hydrophobicity). Generally, conservative amino acid substitution does not substantially change the functional properties of a protein. Examples of amino acid classes containing chemically similar side chains include: 1) Aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; 2) Aliphatic hydroxyl side chains: serine and threonine; 3) Amide-containing side chains: asparagine and glutamine; 4) Aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) Basic side chains: lysine, arginine, and histidine; 6) Acidic side chains: aspartic acid and glutamic acid.

[0130] The number of amino acids for “conserved amino acid substitutions in VL and VH” can be about 1, about 2, about 3, about 4, about 5, about 6, about 8, about 9, about 10, about 11, about 13, about 14, or about 15 conserved amino acid substitutions, or a range between any two of these values ​​(including endpoints) or any of these values. The number of amino acids for “conserved amino acid substitutions in the heavy or light chain” can be about 1, about 2, about 3, about 4, about 5, about 6, about 8, about 9, about 10, about 11, about 13, about 14, about 15, about 18, about 19, about 22, about 24, about 25, about 29, about 31, about 35, about 38, about 41, or about 45 conserved amino acid substitutions, or a range between any two of these values ​​(including endpoints) or any of these values.

[0131] When the term “encoding” is applied to polynucleotides, it refers to a polynucleotide called the “encoding” polypeptide that, in its natural state or when manipulated by methods known to those skilled in the art, can be transcribed and / or translated to produce the polypeptide and / or fragments thereof.

[0132] The term “recombination” refers to polypeptides or polynucleotides, meaning forms of polypeptides or polynucleotides that do not exist naturally. Unrestricted embodiments can be created by combining polynucleotides or polypeptides that do not normally exist.

[0133] "Homology," "identity," or "similarity" refers to the sequence similarity between two peptides or two nucleic acid molecules. Homology can be determined by comparing comparable positions in each sequence. Molecules are homologous at those positions when the positions in the compared sequences are occupied by the same bases or amino acids. The degree of homology between sequences is a function of the number of shared matching or homologous positions.

[0134] "At least 80% identity" means approximately 80% identity, approximately 81% identity, approximately 82% identity, approximately 83% identity, approximately 85% identity, approximately 86% identity, approximately 87% identity, approximately 88% identity, approximately 90% identity, approximately 91% identity, approximately 92% identity, approximately 94% identity, approximately 95% identity, approximately 98% identity, approximately 99% identity, or a range (including endpoints) between any two of these values ​​or any value therein.

[0135] "Identity" or "sequence identity" of a polynucleotide or polynucleotide sequence (or polypeptide or antibody sequence) with another sequence at a certain percentage (e.g., 90%, 95%, 98%, or 99%) means that, when sequence alignment is performed, that percentage of bases (or amino acids) are identical in the two compared sequences. This alignment identity percentage or sequence identity can be determined visually or using software programs known in the art, such as those described in Ausubel et al. eds. (2007) in Current Protocols in Molecular Biology. Alignment is preferably performed using default parameters. One alignment procedure is BLAST using default parameters, such as BLASTN and BLASTP, which use the following default parameters: Geneticcode=standard; filter=none; strand=both; cutoff=60; expect=10; Matrix=BLOSUM62; Descriptions=50 sequences; sortby=HIGHSCORE; Databases=non-redundant; GenBank+EMBL+DDBJ+PDB+GenBankCDStranslations+SwissProtein+SPupdate+PIR. Biologically equivalent polynucleotides are polynucleotides that have the above-specified percentages of identity and encode polypeptides with the same or similar biological activities.

[0136] "Antibody" or "antigen-binding fragment" refers to a polypeptide or polypeptide complex that specifically recognizes and binds to an antigen. An antibody can be a complete antibody, any antigen-binding fragment, or a single chain thereof. Therefore, the term "antibody" includes any protein or peptide containing at least a portion of an immunoglobulin molecule that has biological activity of binding to an antigen. Antibody and antigen-binding fragments include, but are not limited to, the complementarity-determining region (CDR), heavy chain variable region (VH), light chain variable region (VL), heavy chain constant region (CH), light chain constant region (CL), framework region (FR), or any portion thereof of the heavy chain or light chain or its ligand-binding moiety, or at least a portion of the binding protein. The CDR region includes the CDR regions of the light chain (LCDR1-3) and the CDR regions of the heavy chain (HCDR1-3).

[0137] The term "antibody" encompasses a wide range of polypeptides that can be distinguished biochemically. Those skilled in the art will understand that heavy chains are categorized as gamma, mu, alpha, delta, or epsilon (γ, μ, α, δ, ε), with further subclasses (e.g., γ1-γ4). The properties of this chain determine the "type" of the antibody, such as IgG, IgM, IgA, IgG, or IgE. Immunoglobulin subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgG5, etc., have been well characterized and their assigned functional specificities are known. All immunoglobulin types are within the scope of this invention. In some embodiments, the immunoglobulin molecule is of the IgG type. The four chains are linked by disulfide bonds in a "Y" configuration, wherein the light chain begins at the "Y" junction and continues through a variable region surrounding the heavy chain.

[0138] The antibodies, antigen-binding fragments or derivatives disclosed in this invention include, but are not limited to, polyclonal, monoclonal, multispecific, fully human, humanized, primate-like, chimeric antibodies, single-chain antibodies, epitope-binding fragments (e.g., Fab-like, Fab'-like and F(ab')2-like), and single-chain Fvs (scFv).

[0139] Light chains can be divided into kappa (κ) or lambda (λ). Each heavy chain can bind to either a κ or λ light chain. Generally, when immunoglobulins are produced by hybridomas, B cells, or genetically engineered host cells, their light and heavy chains are covalently bonded, and the "tail" portions of the two heavy chains are linked by covalent disulfide bonds or non-covalent bonds. In the heavy chain, the amino acid sequence extends from the N-terminus of the Y-configuration forked end to the C-terminus at the bottom of each chain. The variable region of the immunoglobulin κ light chain is Vκ; the variable region of the immunoglobulin λ light chain is V... λ .

[0140] The variable regions of the light chain (VL) and heavy chain (VH) determine antigen recognition and specificity. The constant regions (CL) of the light chain and (CH) of the heavy chain impart important biological properties, such as secretion, transplacental migration, Fc receptor binding, and complement binding. Conventionally, the constant regions are numbered as they move further away from the antibody's antigen-binding site or N-terminus. The N-terminal region is the variable region, and the C-terminal region is the constant region; the CH3 and CL domains actually contain the carboxyl terms of the heavy and light chains, respectively.

[0141] Where a term used and / or accepted in this field has two or more definitions, the definition of the term used herein includes all of those meanings unless explicitly stated otherwise. A concrete example is the use of the term “complementarity-determining region” (“CDR”) to describe a discontinuous antigen-binding site found within the variable region of heavy and light chain polypeptides. This specific region is described in Kabat et al., USDept. of Health and Human Services, Sequences of Proteins of Immunological Interest (1983) and Chothia et al. in J. Mol. Biol. 196:901-917 (1987), which are incorporated herein by reference in their entirety.

[0142] According to the definitions of Kabat and Chothia, a CDR includes overlaps or subsets of amino acid residues when compared with each other. Nevertheless, the application of either definition to refer to a CDR of an antibody or a variant thereof is within the scope of this invention. The exact residue numbering containing a particular CDR will vary depending on the sequence and size of the CDR. Those skilled in the art can typically determine which specific residues a CDR contains based on the amino acid sequence of the variable region of the antibody.

[0143] Kabat et al. also defined a numbering system applicable to the variable region sequence of any antibody. Those skilled in the art can apply this "Kabat numbering" system to any variable region sequence without relying on experimental data other than the sequence itself. "Kabat numbering" refers to the numbering system proposed by Kabat et al., USDept. of Health and Human Services in "Sequence of Proteins of Immunological Interest" (1983). Antibodies can also be numbered using the EU or Chothia numbering systems.

[0144] In this invention, the term "antibody-drug conjugate" or "ADC" refers to an antibody or antigen-binding fragment thereof chemically linked to one or more chemical drugs (which may optionally be therapeutic agents or cytotoxic agents). In some embodiments, an ADC includes an antibody, a cytotoxic or therapeutic drug, and a connector that enables the drug to be linked or conjugated to the antibody. ADCs typically have 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 drugs conjugated to the antibody. Drugs that may be included in an ADC include, but are not limited to: mitotic inhibitors, antitumor antibiotics, immunomodulators, gene therapy vectors, alkylating agents, antiangiogenic agents, antimetabolites, boron-containing reagents, chemotherapeutic agents, hormones, antihormonal agents, corticosteroids, photoactive therapeutic agents, oligonucleotides, radionuclide reagents, topoisomerase inhibitors, tyrosine kinase inhibitors, and radiosensitizers. In some embodiments, the drug included in an ADC maytansine-like drugs. In some embodiments, the drug included in an ADC may be a compound of Formula I as described in this application or a pharmaceutically acceptable salt thereof. In some implementations, in an ADC, the antibody conjugates to the drug via a disulfide bond formed by its own cysteine ​​or a sulfhydrylated amino acid such as sulfhydrylated lysine.

[0145] As used herein, the terms "alkyl" and "alkylene" refer to saturated aliphatic hydrocarbon groups, including branched and straight-chain groups with a specific number of carbon atoms. For example, the definition of "C1-C6" in "C1-C6 alkyl" includes groups with 1, 2, 3, 4, 5, or 6 carbon atoms arranged in a straight or branched chain. Specifically, "C1-C6" includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl (including 8 isomers), and hexyl (including 23 isomers). The term "cycloalkyl" refers to a monocyclic saturated aliphatic hydrocarbon group with a specific number of carbon atoms. For example, "cycloalkyl" includes cyclopropyl, methyl-cyclopropyl, 2,2-dimethyl-cyclobutyl, 2-ethyl-cyclopentyl, cyclohexyl, etc.

[0146] The term "halogen" as used in this article includes fluorine, chlorine, bromine, and iodine.

[0147] As used herein, the term "amino acid side chain" refers to a substituent that replaces a group (such as a hydrogen atom) in an amino acid; for example, a glycine side chain is a substituent that replaces a hydrogen atom on the methylene group of glycine. Examples of amino acid side chains include, but are not limited to, natural amino acid side chains.

[0148] Antibody-drug conjugates can form a wide variety of pharmaceutically acceptable salts, including but not limited to: acid addition salts formed with organic acids, such as aliphatic monocarboxylic acids and dicarboxylic acids, phenyl-substituted alkyl acids, hydroxyalkyl acids, alkyl diacids, aromatic acids, aliphatic and aromatic sulfonic acids, amino acids, etc., such as acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.; acid addition salts formed by reactions with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid, hydrofluoric acid, phosphorous acid, etc.; and salts formed with metal ions (e.g., alkali metal ions (e.g., sodium or potassium), alkaline earth metal ions (e.g., calcium or magnesium) or aluminum ions) or with organic bases such as diethanolamine, triethanolamine, N-methylglucosamine, etc. The antibody-drug conjugates described in this article include their pharmaceutically acceptable salts.

[0149] "Treatment" refers to therapeutic treatments and preventative or preventative measures aimed at preventing, mitigating, improving, and stopping adverse physiological changes or disorders, such as disease progression, including but not limited to the following, whether detectable or undetectable: symptom relief, reduction in disease severity, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or mitigation of the disease state, reduction or disappearance (whether partial or complete), and prolongation of expected survival without treatment. Patients requiring treatment include those already suffering from the condition or disorder, those susceptible to the condition or disorder, or those needing prevention of the condition or disorder, as well as those who can or are expected to benefit from the application of the antibodies or compositions disclosed in this invention for detection, diagnostic procedures, and / or treatment.

[0150] "Patient" refers to any mammal that requires diagnosis, prognosis, or treatment, including humans, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, etc.

[0151] "Approximately" refers to a typical error range for the corresponding value that is readily known to those skilled in the art. In some embodiments, "approximately" as used herein refers to the described value and its range of ±10%, ±5%, or ±1%.

[0152] "Effective amount" refers to the amount of an active compound or agent that elicits a biological or medical response in a tissue, system, animal, individual, or human; effective amount is sought by researchers, veterinarians, physicians, or other clinicians.

[0153] As used herein, the phrase “in need” means that a patient has been identified as requiring a particular method or treatment. In some embodiments, this identification can be made through any diagnostic approach. In any of the methods and treatments described herein, the patient may require [the specific method or treatment].

[0154] DNA encoding the antibody can be designed and synthesized according to the antibody amino acid sequence described herein using conventional methods. This DNA can then be placed into an expression vector, transfected into host cells, and cultured in a culture medium to produce monoclonal antibodies. In some embodiments, the antibody expression vector includes at least one promoter element, an antibody-coding sequence, a transcription termination signal, and a polyA tail. Other elements include an enhancer, a Kozak sequence, and donor and acceptor sites for RNA splicing flanking the insert sequence. Efficient transcription can be achieved using early and late promoters of SV40, early promoters from long terminal repeat sequences of retroviruses such as RSV, HTLV1, HIV, and cytomegalovirus, or other cellular promoters such as the actin promoter. Suitable expression vectors may include pIRES1neo, pRetro-Off, pRetro-On, PLXSN, or Plncx, pcDNA3.1(+ / -), pcDNA / Zeo(+ / -), pcDNA3.1 / Hygro(+ / -), PSVL, PMSG, pRSVcat, pSV2dhfr, pBC12MI, and pCS2, etc. Commonly used mammalian cell lines include 293 cells, Cos1 cells, Cos7 cells, CV1 cells, mouse L cells, and CHO cells, etc.

[0155] EC 50 "The half-maximal effect concentration (EC50) is the concentration that can cause 50% of the maximum effect." Detailed Implementation

[0156] The following specific embodiments further illustrate the technical solution of the present invention. These specific embodiments do not represent a limitation on the scope of protection of the present invention. Non-essential modifications and adjustments made by others based on the concept of the present invention still fall within the scope of protection of the present invention.

[0157] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0158] Method of preparing the antibody of Example 1

[0159] 1) Construct DNA sequences for the light and light chains based on the amino acid sequences of the antibody heavy and light chains. Modify the 5' and 3' ends of the DNA sequences using PCR primers designed to add appropriate leader sequences to each chain. Then clone the DNA into an existing recombinant antibody expression vector and verify the correct vector construction through sequencing analysis. Express the expression vector in CHO cells and obtain the antibody through purification. The preparation method of antibody A can be found in the invention patent application with application number PCT / CN2020 / 083954. The amino acid sequences of antibodies A, B, and C are shown in Table 1, and the nucleic acid sequences of antibodies A and C are shown in Table 2.

[0160] Table 1 Amino acid sequences

[0161]

[0162]

[0163]

[0164] Table 2 Nucleic acid sequences of antibody A and antibody C

[0165]

[0166]

[0167] 2) Antibody C was prepared using CHO-BAT-KF cells. After purification, the glycoform content of antibody C was determined, as shown in Table 3.

[0168] Table 3 shows the percentage content of some sugar types.

[0169]

[0170]

[0171] Example 2 HER2-ADC B Preparation method

[0172] HER2-ADC B The preparation method can be referred to Examples 1-3 and Example 9 of the invention patent with publication number WO2014094527A.

[0173] Example 3 Antibody A and HER2-ADC B In vitro combined administration inhibited the proliferation of NCI-N87 cells (human gastric cancer cells).

[0174] The concentration of antibody A was fixed (25 μg / mL), and the HER2-ADC concentration was changed. BThe concentration (starting at 10 nM, serially diluted 3-fold) was added to PBMC cells (Red Biotech Co., Ltd.), and SEB (staphylococcal enterotoxin B, Academy of Military Medical Sciences SL008) was added simultaneously to activate PBMC cells. After culturing for 72 h, the inhibitory effect of the combined drug on the proliferation of HER2 and PD-L1 double-positive cells was measured in vitro using the CCK method (CCK8 kit from Dojindo).

[0175] 1) Cell seeding in 96-well plates, with NCI-N87 cells adjusted to a density of 2.0 × 10⁻⁶. 5 Cells / ml, 10,000 cells / well, 50 μL per well;

[0176] 2) After culturing cells for 3-5 hours, add PMBC cells and adjust the PBMC density to 8.0 × 10⁶ cells / mL using medium containing 100 ng / mL SEB. 5 cells / ml (PBMC cells to NCI-N87 cells ratio of 4:1), volume 50μL;

[0177] 3) Gradual dilution of HER2-ADC B Starting at 10 nM, use a 3-fold serial dilution for a total of 9 gradients (10 nM, 3.3 nM, 1.1 nM, 0.37 nM, 0.12 nM, 0.04 nM, 0.01 nM, 0.004 nM, 0.001 nM). The 10th well contains 0 concentration, 50 μl per well. Add 25 μg / mL antibody A to the top three wells, 50 μl per well. Add culture medium to the bottom three wells as a control, 50 μl per well. Plate arrangement is shown in the table below. Antibody A and HER2-ADC B Diluted with RPMI 1640 (Gibco) + 10% FBS (Excell Bio).

[0178] 4) Incubate at 37℃ with 5% CO2 for about 72 hours, remove the supernatant, and add culture medium containing 10% CCK8.

[0179] 5) Incubate at 37℃ for 4 hours; read the plate using an ELISA reader at an absorbance of 450nm.

[0180] The results are as follows Figure 1 As shown, in the presence of PBMCs, antibody A (anti-PD-1 antibody) can enhance HER2-ADC to a certain extent. B Inhibitory effect on the proliferation of NCI-N87 tumor cells; by calculation, antibody A and HER2-ADC B EC with combination therapy 50 HER2-ADC has a capacity of 0.032 nM. B EC alone 50 It is 0.045 nM.

[0181] Example 4: Combined administration of antibody A and antibody C inhibits the proliferation of cancer cells.

[0182] This embodiment evaluates the efficacy of antibody A and antibody C (antibody C is expressed by CHO-BAT-KF cells) in a humanized mouse model of C57BL / 6-hPD1 / hCTLA4 subcutaneously injected with MC38 colon cancer tumors.

[0183] 1) Tumor cell inoculation

[0184] Mouse colon cancer cells MC38 (Jiangsu Jicui Yaokang Biotechnology Co., Ltd.) were resuscitated. MC38 cells in the logarithmic growth phase were collected, the culture medium was removed, and the cells were washed twice with PBS before inoculation (cell viability before and after tumor bearing was 99.6% and 97.8%, respectively). The inoculation density was 1×10⁶ cells / year. 6 / 100μL / animal, administered via axillary or intraperitoneal injection.

[0185] 2) Grouped administration

[0186] On day 6 post-inoculation, the average tumor volume reached 89.19 mm. 3 At that time, 60 female mice were randomly divided into 6 groups of 10 mice each according to tumor volume. The day of grouping was defined as day D0, and the drug was administered on day D0. The grouping and administration regimen is shown in Table 4. The administration dates were: D0, D4, D7, D11, D14, and D18.

[0187] 3) Experimental observation and data collection

[0188] Following cell inoculation, the impact of the tumor on the animals' normal behavior was routinely monitored weekly. Specific indicators included mouse activity, food and water intake, weight gain or loss, and any abnormalities in the eyes, coat, or other areas.

[0189] After drug administration, tumor size was observed and mouse weight was measured on days 0, 3, 6, 10, 13, 17, 20, 24, and 27. Tumor volume was calculated as: tumor volume (mm). 3 = 0.5 × (tumor long diameter × tumor short diameter) 2 ).

[0190] 4) Statistics

[0191] Tumor volume, body weight, and tumor mass in each group of mice are expressed as mean ± standard error (mean ± SEM). Independent samples t-tests were used to compare the differences between the different treatment groups and the control group. Data were analyzed using SPSS. P < 0.05 was considered statistically significant.

[0192] The formula for calculating TGItv (inhibition rate relative to tumor volume) is as follows:

[0193] RTV n =V nt / V n0 V nt : Tumor volume of mouse number n on day t, V n0 Tumor volume (RTV) of mouse number n on day 0 n The relative tumor volume of mouse number n on day t

[0194] TGItv = (1 - (mean RTV group) / (mean RTV) 对照组 ))×100%; mean RTV: mean RTV of the treatment group 对照组 : The average RTV of the control group;

[0195] Formula for calculating TGItw (tumor weight inhibition rate):

[0196] TGItw=(1-(mean TW treatment group) / (mean TW) 对照组 ))×100%; Mean TW treatment group: the average tumor weight of mice in the treatment group at the endpoint treatment, Mean TW 对照组 : The average tumor weight of control mice at the endpoint treatment.

[0197] Table 4 Dosage Regimen

[0198]

[0199] Note: N is the number of animals; BIW x 3W, administered twice a week for 3 weeks; intraperitoneal administration 6 times; G5 and G6 groups are combination therapy groups, antibody A is administered first, followed by antibody C about 2 hours later, the concentration of the administered antibody is not doubled and is consistent with the concentration of the single drug group.

[0200] like Figure 2 , Figure 3 As shown in Tables 5 and 6, single-drug antibody A showed significant antitumor effects, and the high-dose group of antibody C showed significant antitumor efficacy, with antibody C exhibiting a certain dose-dependent effect. The combined high-dose administration of antibody A and antibody C showed significant antitumor efficacy, and the antitumor effect was superior to that of single-drug antibody A and single-drug antibody C alone (no complete tumor regression was observed in mice in groups G1 and G2, complete tumor regression was observed in 2 mice in each of groups G3 and G6, complete tumor regression was observed in 1 mouse in group G4, and complete tumor regression was observed in 6 mice in group G5). Figure 4As shown, no significant difference was found in the body weight of mice among the groups, and no mouse weighed less than 10% of its body weight on the day of grouping during the experiment, indicating that the mice had good tolerance to the drugs in the current system.

[0201] Table 5. Tumor volume inhibition rate (TGItv) in different groups

[0202]

[0203] Table 6. Statistical analysis of tumor weight inhibition rate (TGItw) and p-value (vs G1) in different groups on day 27.

[0204] Group Tumor weight (g) TGItw P value G1 1.4662±0.2701 - - G2 0.7601±0.1302 48.16％ 0.030* G3 0.3144±0.1050 78.56％ 0.002** G4 1.0236±0.2542 30.19％ 0.248 G5 0.0796±0.0500 94.57％ <0.001*** G6 0.9008±0.2827 38.57％ 0.165 sequence list <110> Bio-Thera Solutions, Ltd. <120> Application of anti-PD-1 antibodies in combination therapy <150> PCT / CN2020 / 120262 <151> 2020-10-11 <160> 18 <170> SIPOSequenceListing 1.0 <210> 1 <211> 6 <212> PRT <213> Artificial Sequence <400> 1 Asn Tyr Tyr Met Tyr Trp 1 5 <210> 2 <211> 15 <212> PRT <213> Artificial Sequence <400> 2 Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe 1 5 10 15 <210> 3 <211> 12 <212> PRT <213> Artificial Sequence <400> 3 Ala Arg Asp Tyr Arg Leu Asp Met Gly Phe Glu Phe 1 5 10 <210> 4 <211> 14 <212> PRT <213> Artificial Sequence <400> 4 Ala Ser Lys Gly Val Ser Thr Ser Gly Tyr Ser Tyr Leu His 1 5 10 <210> 5 <211> 6 <212> PRT <213> Artificial Sequence <400> 5 Leu Ala Ser Tyr Leu Glu 1 5 <210> 6 <211> 11 <212> PRT <213> Artificial Sequence <400> 6 Tyr Cys Gln His Ala Tyr Asp Leu Pro Leu Thr 1 5 10 <210> 7 <211> 120 <212> PRT <213> Artificial Sequence <400> 7 Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe 50 55 60 Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr 65 70 75 80 Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Ala Arg Asp Tyr Arg Leu Asp Met Gly Phe Glu Phe Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 8 <211> 111 <212> PRT <213> Artificial Sequence <400> 8 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Lys Gly Val Ser Thr Ser 20 25 30 Gly Tyr Ser Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 35 40 45 Arg Leu Leu Ile Tyr Leu Ala Ser Tyr Leu Glu Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 65 70 75 80 Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Ala Tyr 85 90 95 Asp Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 9 <211> 447 <212> PRT <213> Artificial Sequence <400> 9 Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe 50 55 60 Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr 65 70 75 80 Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Ala Arg Asp Tyr Arg Leu Asp Met Gly Phe Glu Phe Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro 210 215 220 Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val 225 230 235 240 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255 Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu 260 265 270 Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285 Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser 290 295 300 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 305 310 315 320 Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile 325 330 335 Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345 350 Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 385 390 395 400 Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg 405 410 415 Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys 435 440 445 <210> 10 <211> 218 <212> PRT <213> Artificial Sequence <400> 10 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Lys Gly Val Ser Thr Ser 20 25 30 Gly Tyr Ser Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 35 40 45 Arg Leu Leu Ile Tyr Leu Ala Ser Tyr Leu Glu Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 65 70 75 80 Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Ala Tyr 85 90 95 Asp Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 100 105 110 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 115 120 125 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 145 150 155 160 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170 175 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 180 185 190 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 195 200 205 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 11 <211> 450 <212> PRT <213> Artificial Sequence <400> 11 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr 20 25 30 Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <210> 12 <211> 214 <212> PRT <213> Artificial Sequence <400> 12 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Arg 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 His Tyr Thr Thr Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val 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> 1341 <212> DNA <213> Artificial Sequence <400> 13 caggtgcagc tggtgcagtc cggcgtggag gtgaagaagc ctggcgccag cgtgaaggtg 60 tcctgtaagg ccagcggcta caccttcacc aattactata tgtattgggt gcggcaggcc 120 cccggccagg gactggagtg gatgggaggc atcaatccca gcaacggcgg caccaacttc 180 aatgagaagt ttaagaaccg ggtgaccctg accaccgata gcagcaccac caccgcttac 240 atggagctga agagcctgca gtttgacgat accgctgtgt actattgcgc tgcccgggat 300 tacaggctgg acatgggctt cgagttctgg ggccagggca ccaccgtgac cgtgtccagc 360 gctagcacca agggcccttc cgtgttcccc ctggccccct gtagccggtc cacctctgag 420 agcaccgctg ctctgggctg tctggtgaag gattactttc ccgaaccggt gaccgtgtca 480 tggaactccg gggctctgac atccggtgtc cacacttttc ctgcagtgct gcagtcatcc 540 ggcctgtaca gcctgagctc tgtggtcaca gtcccaagtt catccctggg aaccaagaca 600 tatacttgca acgtggatca taaacccagc aatactaagg tcgacaaacg agtggagtct 660 aagtacggac caccttgccc accatgtcca gcacctgagt tcctgggagg accaagcgtg 720 ttcctgtttc ctccaaagcc taaagatacc ctgatgatca gtcggactcc cgaggtcacc 780 tgcgtggtcg tggacgtgtc ccaggaggac cctgaagtcc agttcaactg gtacgtggac 840 ggcgtcgaag tgcacaatgc taagacaaaa cctcgagagg aacagtttaa ctccacatac 900 cgtgtcgtga gcgtcctgac tgtgctgcat caggattggc tgaacggcaa ggagtataag 960 tgcaaagtga gcaataaggg actgccaagc tctatcgaga aaactatttc taaggctaaa 1020 ggacagccta gggaaccaca ggtgtacacc ctgcccccta gtcaggagga aatgactaag 1080 aaccaggtct cactgacctg tctggtgaaa gggttctatc cttcagatat tgcagtggag 1140 tgggaatcca atggtcagcc agagaacaat tacaagacaa ctccacccgt gctggacagc 1200 gatgggtctt tctttctgta ttctagactg accgtggaca aaagtcgctg gcaggagggt 1260 aatgtctttt cttgtagtgt gatgcacgaa gccctgcaca accactacac tcagaaaagc 1320 ctgtcactgt ccctgggtaa a 1341 <210> 14 <211> 654 <212> DNA <213> Artificial Sequence <400> 14 gagatcgtgc tgacccagtc ccccgctacc ctgagcctgt cccccggaga gcgggctacc 60 ctgtcttgtc gggcctccaa gggcgtgagc accagcggat actcctatct gcactggtac 120 cagcagaagc ccggccaggc tcccaggctg ctgatctacc tggcttccta cctggagagc 180 ggcgtgcccg ctaggtttag cggcagcggc agcggaaccg atttcaccct gaccatcagc 240 tccctggagc ccgaggattt tgccgtgtac tactgccagc acgcttacga cctgcccctg 300 acctttggcg gcggcaccaa ggtggagatc aagcgtacgg tggctgcacc atctgtcttc 360 atcttcccgc catctgatga gcagttgaaa tctggaactg cctctgttgt gtgcctgctg 420 aataacttct atcccagaga ggccaaagta cagtggaagg tggataacgc cctccaatcg 480 ggtaactccc aggagagtgt cacagagcag gacagcaagg acagcaccta cagcctcagc 540 agcaccctga cgctgagcaa agcagactac gagaaacaca aagtctacgc ctgcgaagtc 600 acccatcagg gcctgagctc gcccgtcaca aagagcttca acaggggaga gtgt 654 <210> 15 <211> 448 <212> PRT <213> Artificial Sequence <400> 15 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 16 <211> 215 <212> PRT <213> Artificial Sequence <400> 16 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Phe Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95 Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205 Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 17 <211> 1344 <212> DNA <213> Artificial Sequence <400> 17 caggtccagc tggtcgaatc tggcggcggc gtcgtccagc ctggaagatc tctgagactg 60 tcttgcgctg cttctggctt taccttctct tcttacacaa tgcattgggt cagacaggct 120 180 gccgattctg tcaagggcag attcaccatc tccagagaca actccaagaa caccctgtac 240 ctgcagatga actctctgag agctgaggat accgctatct actattgcgc tagaacaggc 300 tggctggac cttttgatta ctggggcag ggaacactgg tcacagtctc ttctgcttct 360 accaaaggac cttctgtctt tcctctggct ccttcttcta agtctacctc tggcggaaca 420 gctgctctgg gttgtctggt caaggattac ttccctgaac cggtgacagt gtcttggaat 480 tctggagctc tgacctcagg agtccataca tttcctgctg tcctgcagtc ttctggcctg 540 tattctctgt cctctgtggt gacagtccct tcttcttctc tgggaacaca gacctacatc 600 tgcaacgtca accacaagcc ttccaacacc aaggtcgaca agagagtgga gcctaagtct 660 tgcgacaaga cacatacatg tccaccgtgc ccagcacctg aactcctggg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 1080 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 1140 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1200 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1260 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1320 agcctctccc tgtctccggg taaa 1344 <210> 18 <211> 645 <212> DNA <213> Artificial Sequence <400> 18 gaaattgtcc tgacacagtc tcctggaaca ctgtctctgt ctcctggaga gagagctaca 60 ctgtcttgta gggcttctca gtctgtggga tcttcttacc tggcttggta tcagcagaaa 120 cctggacagg ctcctagact gctgatctac ggcgcttttt ctagagctac cggaatccct 180 gacagattct ctggatctgg ctctggaacc gatttcaccc tgaccatctc tagactggaa 240 cctgaggact ttgctgtcta ctactgtcag cagtacggct cttctccttg gacatttgga 300 cagggaacca aggtcgagat caagcgtacg gtggctgcac catctgtctt catcttcccg 360 ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 420 tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 480 caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 540 acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 600 ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgt 645

Claims

1. Use of an anti-PD-1 antibody combination therapeutic agent in the manufacture of a medicament for treating cancer, characterized in that, the anti-PD-1 antibody comprises a HCDR1 as set forth in SEQ ID NO: 1, a HCDR2 as set forth in SEQ ID NO: 2, a HCDR3 as set forth in SEQ ID NO: 3, a LCDR1 as set forth in SEQ ID NO: 4, a LCDR2 as set forth in SEQ ID NO: 5, and a LCDR3 as set forth in SEQ ID NO: 6, the therapeutic agent is an anti-CTLA4 antibody, a sequence of a heavy chain of the anti-CTLA4 antibody is a sequence as set forth in SEQ ID NO: 15, and a sequence of a light chain of the anti-CTLA4 antibody is a sequence as set forth in SEQ ID NO: 16, the cancer is colon cancer; or, the therapeutic agent is an anti-HER2 antibody drug conjugate, the anti-HER2 antibody drug conjugate is an antibody drug conjugate as set forth in Formula II or a pharmaceutically acceptable salt thereof: wherein, Abu is an anti-HER2 antibody, p is selected from 1-10; a sequence of a heavy chain of the anti-HER2 antibody is a sequence as set forth in SEQ ID NO: 11, and a sequence of a light chain of the anti-HER2 antibody is a sequence as set forth in SEQ ID NO: 12, the cancer is gastric cancer.

2. Use according to claim 1, characterized in that, a sequence of a heavy chain variable region of the anti-PD-1 antibody is a sequence as set forth in SEQ ID NO: 7, a sequence having about 90% identity, about 91% identity, about 92% identity, about 94% identity, about 95% identity, about 98% identity, about 99% identity to the sequence as set forth in SEQ ID NO: 7, or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence as set forth in SEQ ID NO:

7.

3. Use according to claim 1, characterized in that, a sequence of a light chain variable region of the anti-PD-1 antibody is a sequence as set forth in SEQ ID NO:

8.

4. The use according to claim 1, characterized in that, a sequence of a heavy chain variable region of the anti-PD-1 antibody is a sequence as set forth in SEQ ID NO: 7, a sequence having about 90% identity, about 91% identity, about 92% identity, about 94% identity, about 95% identity, about 98% identity, about 99% identity to the sequence as set forth in SEQ ID NO: 7, and a sequence of a light chain variable region of the anti-PD-1 antibody is a sequence as set forth in SEQ ID NO: 8, a sequence having about 90% identity, about 91% identity, about 92% identity, about 94% identity, about 95% identity, about 98% identity, about 99% identity to the sequence as set forth in SEQ ID NO:

8.

5. The use according to claim 1, characterized in that, a sequence of a heavy chain variable region of the anti-PD-1 antibody is a sequence as set forth in SEQ ID NO: 7, and a sequence of a light chain variable region of the anti-PD-1 antibody is a sequence as set forth in SEQ ID NO:

8.

6. The use according to claim 1, characterized in that, the sequence of the heavy chain of the anti-PD-1 antibody is the sequence set forth in SEQ ID NO: 9, a sequence having at least 80% identity to the sequence set forth in SEQ ID NO: 9, or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence set forth in SEQ ID NO: 9; and / or the sequence of the light chain of the anti-PD-1 antibody is the sequence set forth in SEQ ID NO: 10, a sequence having at least 80% identity to the sequence set forth in SEQ ID NO: 10, or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence set forth in SEQ ID NO:

10.

7. Use according to claim 6, characterized in that, the sequence of the heavy chain of the anti-PD-1 antibody is the sequence set forth in SEQ ID NO: 9, and the sequence of the light chain of the anti-PD-1 antibody is the sequence set forth in SEQ ID NO:

10.

8. Use according to any one of claims 1 to 7, characterized in that, the anti-HER2 antibody drug conjugate is an antibody drug conjugate as shown in Formula II or a pharmaceutically acceptable salt thereof: wherein, p is 3.3-3.

7.

9. Use according to claim 8, characterized in that, the effective amount of the anti-HER2 antibody drug conjugate administered in each treatment cycle is 70 mg to 400 mg.

10. The use according to claim 8, characterized in that, the amount of the anti-HER2 antibody drug conjugate administered in each administration is 1-6 mg / kg.

11. Use according to any one of claims 1 to 7, characterized in that, the anti-CTLA4 antibody is expressed by a CHO cell line in which the alpha-(1, 6)-fucosyltransferase gene is knocked out.

12. Use according to any one of claims 1 to 7, characterized in that, the effective amount of the anti-CTLA4 antibody administered in each treatment cycle is 6 mg to 600 mg.

13. Use according to any one of claims 1 to 7, characterized in that, the amount of the anti-CTLA4 antibody administered in each administration is 0.1-10 mg / kg.

14. Use according to any one of claims 1 to 7, characterized in that, the effective amount of the anti-PD-1 antibody administered in each treatment cycle is 50 mg to 600 mg.

15. Use according to any one of claims 1 to 7, characterized in that, the amount of the anti-PD-1 antibody administered in each administration is 1-10 mg / kg.

16. Use according to any one of claims 1 to 7, characterized in that, one treatment cycle of administration to the patient is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 5 weeks, 6 weeks, or 7 weeks.

17. Use according to any one of claims 1 to 7, characterized in that the patient receives multiple treatment cycles until the condition is in remission and no longer requires treatment.

18. The use of any one of claims 1-7, wherein the patient is administered by intravenous infusion.

19. A kit comprising, an anti-PD-1 antibody, a therapeutic agent, and instructions for administering the anti-PD-1 antibody and the therapeutic agent to a patient in need thereof; the anti-PD-1 antibody comprises a HCDR1 set forth in SEQ ID NO: 1, a HCDR2 set forth in SEQ ID NO: 2, a HCDR3 set forth in SEQ ID NO: 3, a LCDR1 set forth in SEQ ID NO: 4, a LCDR2 set forth in SEQ ID NO: 5, and a LCDR3 set forth in SEQ ID NO: 6, the therapeutic agent is an anti-CTLA4 antibody, the sequence of the heavy chain of the anti-CTLA4 antibody is the sequence set forth in SEQ ID NO: 15, and the sequence of the light chain of the anti-CTLA4 antibody is the sequence set forth in SEQ ID NO: 16, the patient has colon cancer; or, the therapeutic agent is an anti-HER2 antibody drug conjugate, the anti-HER2 antibody drug conjugate is an antibody drug conjugate as shown in Formula II or a pharmaceutically acceptable salt thereof: wherein Abu is an anti-HER2 antibody, p is selected from 1-10; the sequence of the heavy chain of the anti-HER2 antibody is the sequence set forth in SEQ ID NO: 11, the sequence of the light chain of the anti-HER2 antibody is the sequence set forth in SEQ ID NO: 12, the patient has gastric cancer.

20. The kit of claim 19, wherein the anti-HER2 antibody drug conjugate is an antibody drug conjugate of Formula II or a pharmaceutically acceptable salt thereof: wherein p is 3.3-3.

7.

21. The kit of claim 19, wherein the anti-CTLA4 antibody is expressed by a CHO cell line knocked out of alpha-(1,6)-fucosyltransferase gene.

22. A pharmaceutical composition comprising an anti-PD-1 antibody and a therapeutic agent, characterized in that, the anti-PD-1 antibody comprises HCDR1 set forth in SEQ ID NO: 1, HCDR2 set forth in SEQ ID NO: 2, HCDR3 set forth in SEQ ID NO: 3, LCDR1 set forth in SEQ ID NO: 4, LCDR2 set forth in SEQ ID NO: 5, and LCDR3 set forth in SEQ ID NO: 6, the therapeutic agent is an anti-CTLA4 antibody, the sequence of the heavy chain of the anti-CTLA4 antibody is the sequence set forth in SEQ ID NO: 15, the sequence of the light chain of the anti-CTLA4 antibody is the sequence set forth in SEQ ID NO: 16, and the pharmaceutical composition is used for treating colon cancer; or, the therapeutic agent is an anti-HER2 antibody drug conjugate, the anti-HER2 antibody drug conjugate is an antibody drug conjugate of Formula II or a pharmaceutically acceptable salt thereof: wherein Abu is an anti-HER2 antibody, p is selected from 1-10; the sequence of the heavy chain of the anti-HER2 antibody is the sequence set forth in SEQ ID NO: 11, the sequence of the light chain of the anti-HER2 antibody is the sequence set forth in SEQ ID NO: 12, the pharmaceutical composition is used for treating gastric cancer.

23. The pharmaceutical composition of claim 22, wherein the anti-HER2 antibody drug conjugate is an antibody drug conjugate of Formula II or a pharmaceutically acceptable salt thereof: wherein p is 3.3-3.

7.

24. The pharmaceutical composition of claim 22, wherein the anti-CTLA4 antibody is expressed by a CHO cell line knocked out of alpha-(1,6)-fucosyltransferase gene.

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