CN114159557B

CN114159557B - Combined medicine composition for treating tumor diseases and application thereof

Info

Publication number: CN114159557B
Application number: CN202111318017.8A
Authority: CN
Inventors: 白义
Original assignee: Beijing Dongfang Baitai Biotechnology Co ltd
Current assignee: Beijing Dongfang Baitai Biotechnology Co ltd
Priority date: 2021-11-09
Filing date: 2021-11-09
Publication date: 2024-07-19
Anticipated expiration: 2041-11-09
Also published as: CN114159557A

Abstract

The invention relates to the field of biological medicine, and in particular provides a combined medicine composition for treating tumor diseases, which comprises an anti-PD-1 monoclonal antibody taking PD-1 as a target point and an anti-EGFR monoclonal antibody taking EGFR as a target point. The combined application of the PD-1 monoclonal antibody and the EGFR monoclonal antibody medicine has higher medication safety, effectively improves the treatment response duration, prolongs the progression-free survival time and the total survival time of cancer patients, improves the health condition and the survival quality of the patients, and can be used for treating non-small cell lung cancer, metastatic non-small cell lung cancer, glioma, colorectal cancer, liver cancer, hepatocellular carcinoma, metastatic hepatocellular carcinoma, HER2 negative metastatic breast cancer, metastatic gastric adenocarcinoma, metastatic colorectal cancer, metastatic melanoma, metastatic renal cell carcinoma, advanced esophageal squamous carcinoma, advanced squamous non-small cell lung cancer or advanced head and neck squamous carcinoma.

Description

Combined medicine composition for treating tumor diseases and application thereof

Technical Field

The invention relates to the technical field of biological medicines, in particular to a combined medicine composition for treating tumor diseases and application thereof.

Background

Immunotherapy is a popular field of tumor treatment, and PD-1 targets are popular targets for current immunotherapy. PD-1 is a key immunocheckpoint receptor expressed on the surface of activated T cells, inhibits activation of T cells after encountering PD-L1 and PD-L2 ligand molecules on the surface of tumor cells, and invades tumor cells to escape from monitoring the immune system, while the use of anti-PD-1 monoclonal antibody inhibitor releases inhibition of activation of T cells by tumor cells, so that the T cells of the autoimmune system of a patient are activated in large quantity, thereby killing the tumor cells. At present, various monoclonal antibodies which take PD-1 as targets are marketed at home and abroad, and meanwhile, a plurality of anti-PD-1 monoclonal antibodies are in a preclinical research or clinical test stage, for example, a monoclonal antibody which is anti-PD-1 is disclosed in patent application No. CN201510312910.8 and an obtaining method thereof, the anti-PD-1 monoclonal antibody disclosed in the patent is declared to be clinically researched at present, and the safety and the effectiveness in vivo of the monoclonal antibody are verified through preclinical data. However, in order to further improve the cancer treatment effect, the anti-PD-1 monoclonal antibody combined drug research is a new research hotspot.

There are many drugs that can be combined with anti-PD-1 mab, but there is no approved market situation for the combination of anti-PD-1 mab and anti-EGFR mab. EGFR (Epidermal Growth Factor Receptor) are receptors for Epithelial Growth Factor (EGF) cell proliferation and signaling. EGFR is overexpressed in various solid tumors, such as head and neck cancer, lung cancer and colorectal cancer, and EGFR overexpression phenomenon exists. Overexpression of EGFR is highly correlated with high invasiveness, high metastatic properties, and poor prognosis of tumors. Currently, many anti-EGFR drugs that target EGFR have been marketed, such as Cetuximab (Cetuximab), nituzumab (Nimotuzumab), panitumumab (Panitumumab), and the like, but how to further improve the anti-EGFR drug cancer treatment effect on the basis of anti-EGFR mab has become the research direction of many drug enterprises.

Therefore, in the large environment with higher development heat of the antibody combination drug, in order to improve the treatment effect of the anti-PD-1 monoclonal antibody and the anti-EGFR monoclonal antibody on cancers, the invention needs to develop a composition for treating tumor diseases by combining the anti-PD-1 monoclonal antibody and the anti-EGFR monoclonal antibody.

Disclosure of Invention

In order to solve the problem that the prior art does not aim at the situation that the combined drug of the anti-PD-1 monoclonal antibody and the anti-EGFR monoclonal antibody is marketed in batches, and to improve the development and development progress of antibody drugs as soon as possible and improve the cancer treatment effect of the anti-PD-1 monoclonal antibody, development of the combined drug of the monoclonal antibody with clinical value is needed, and therefore, the invention discloses a combined drug composition for treating tumor diseases and application thereof.

The specific technical scheme of the invention is as follows:

The invention provides a combined medicine composition for treating tumor diseases, which comprises an anti-PD-1 monoclonal antibody taking PD-1 as a target spot and an anti-EGFR monoclonal antibody taking EGFR as a target spot.

Further, the anti-PD-1 monoclonal antibodies include DFPD1-9, DFPD1-10, DFPD1-11, DFPD1-12, DFPD1-13, nivolumab, pembrolizumab, terep Li Shan, xindi Li Shan, tirelimumab, carelimumab, pa An Puli mab, or Sipalmumab.

Further, the anti-EGFR monoclonal antibodies include cetuximab, nituzumab, and panitumumab.

Further, the mode of administration of the anti-PD-1 monoclonal antibody and the anti-EGFR monoclonal antibody includes simultaneous, concurrent, sequential, alternating, or separate administration;

preferably, the administration is sequential.

Further, the anti-PD-1 monoclonal antibody and the anti-EGFR monoclonal antibody are both intravenously infused.

Further, the anti-PD-1 monoclonal antibody is DFPD1-10, wherein the DFPD1-10 comprises a light chain variable region shown as SEQ ID No. 3 and a heavy chain variable region shown as SEQ ID No. 1;

the dosage of the DFPD1-10 is 100-300 mg;

preferably, the DFPD1-10 is used in an amount of 200mg.

Further, the anti-EGFR monoclonal antibody is nituzumab; the dosage of the nituzumab is 100 mg-500 mg;

Preferably, the use amount of the nituzumab is 200mg;

Preferably, the use amount of the nituzumab is 400mg.

Further, the dosing cycle of DFPD1-10 is once every two weeks, and the dosing cycle of nituzumab is once a week.

The invention also provides application of the combined medicine composition in preparing medicines for treating tumor diseases;

Preferably, the neoplastic disease comprises non-small cell lung cancer, glioma, colorectal cancer, liver cancer, HER2 negative metastatic breast cancer, metastatic gastric adenocarcinoma, metastatic melanoma, metastatic renal cell carcinoma, advanced esophageal squamous carcinoma, advanced head and neck squamous carcinoma;

Preferably, the neoplastic disease includes advanced esophageal squamous carcinoma, non-small cell lung cancer, advanced head and neck squamous carcinoma.

The beneficial effects of the invention are as follows: the combined application of the PD-1 monoclonal antibody and the EGFR monoclonal antibody drug has higher medication safety, effectively improves the treatment response duration, prolongs the progression-free survival time and the total survival time of cancer patients, improves the health condition and the survival quality of the patients, and can be used for treating various cancers such as non-small cell lung cancer, glioma, colorectal cancer, liver cancer, HER2 negative metastatic breast cancer, metastatic gastric adenocarcinoma, metastatic melanoma, metastatic renal cell carcinoma, advanced esophageal squamous cell carcinoma, advanced head and neck squamous cell carcinoma and the like.

Drawings

FIG. 1 is a graph showing tumor volume increase of the combination composition of Experimental example 1 of the present invention versus FaDu head and neck squamous cell carcinoma model;

FIG. 2 is a graph showing the inhibition of tumor weight in FaDu head and neck squamous carcinoma model by the combination of the compositions of Experimental example 1 of the present invention.

Detailed Description

The invention will be described in further detail with reference to the following examples.

Example 1

The embodiment 1 of the invention provides a combined medicine composition for treating tumor diseases, which comprises an anti-PD-1 monoclonal antibody taking PD-1 as a target spot and an anti-EGFR monoclonal antibody taking EGFR as a target spot.

Anti-PD-1 monoclonal antibodies include DFPD1-9, DFPD1-10, DFPD1-11, DFPD1-12, DFPD1-13, nivolumab, pembrolizumab, terlipressin Li Shan, xindi Li Shan, tirelizumab, carilizumab, pa An Puli or Sipalizumab.

Wherein Nivolumab, pembrolizumab, terlipressin Li Shan, xindi Li Shan, tirelimumab, karilimumab, pe An Puli mab or sapalimumab are all marketed products.

Wherein, DFPD1-9, DFPD1-10, DFPD1-11, DFPD1-12, DFPD1-13 are monoclonal antibodies against PD-1 provided by application number CN201510312910.8, DFPD1-9 comprises a light chain variable region as shown in SEQ ID No. 2 and a heavy chain variable region as shown in SEQ ID No. 1; DFPD1-10 includes a light chain variable region as shown in SEQ ID No. 3 and a heavy chain variable region as shown in SEQ ID No. 1; DFPD1-11 comprises a light chain variable region as shown in SEQ ID No. 2 and a heavy chain variable region as shown in SEQ ID No. 4; DFPD1-12 comprises a light chain variable region as shown in SEQ ID No. 2 and a heavy chain variable region as shown in SEQ ID No. 5; DFPD1-13 includes a light chain variable region as shown in SEQ ID No. 3 and a heavy chain variable region as shown in SEQ ID No. 4, and the specific sequences are as follows:

SEQ ID No. 1, sequence as follows:

QVQLVESGGGVVQPGRSLRLDCKASGITFSNYGMHWVRQAPGKGLEWV AVIWYDSSRKYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNN DYWGQGTLVTVSS;

SEQ ID No. 2, sequence as follows:

DIQMTQSPSSLSASVGDRVTITCRASQSIHNYLDWYQQKPGKAPKLLIYNASTRATGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQELHLPLTFGQGTKVE IK;

SEQ ID No. 3, sequence as follows:

DIQMTQSPSSLSASVGDRVTITCRASQSVSNYLDWYQQKPGKAPKLLIYD ASTRATGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQNMQLPLTFGQGTKV EIK;

SEQ ID No. 4, sequence as follows:

QVQLVESGGGVVQPGRSLRLDCKASGITFSNNGMHWVRQAPGKGLEWV AVIWYDSSRKYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNN DYWGQGTLVTVSS;

SEQ ID No. 5, sequence as follows:

QVQLVESGGGVVQPGRSLRLDCKASGITFSNYGMHWVRQAPGKGLEWV AVIWYDGSKKYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNN DYWGQGTLVTVSS.

In addition, the anti-PD-1 monoclonal antibody further comprises a heavy chain constant region and a light chain constant region, wherein the heavy chain constant region and the light chain constant region have the same sequence as the heavy chain constant region and the light chain constant region of the anti-PD-1 monoclonal antibody provided in patent application No. CN 201510312910.8.

Anti-EGFR monoclonal antibodies include cetuximab, nituzumab, and panitumumab.

Administration of the anti-PD-1 monoclonal antibody and the anti-EGFR monoclonal antibody includes simultaneous, concurrent, sequential, alternating, or separate administration.

The anti-PD-1 monoclonal antibody and the anti-EGFR monoclonal antibody are both intravenous infusions.

Example 2

The embodiment 2 of the invention provides a combined medicine composition for treating tumor diseases based on the embodiment 1, which comprises an anti-PD-1 monoclonal antibody taking PD-1 as a target point and an anti-EGFR monoclonal antibody taking EGFR as a target point.

The anti-PD-1 monoclonal antibody is DFPD1-10, and the DFPD1-10 comprises a light chain variable region shown as SEQ ID No. 3 and a heavy chain variable region shown as SEQ ID No.1.

The anti-EGFR monoclonal antibody is nituzumab.

The mode of administration of DFPD1-10 and nitobuzumab was sequential.

DFPD1-10 and nituzumab were both infused intravenously.

The dosage of DFPD1-10 is 100mg;

the dosage of the nimuzumab is 100mg;

the dosing period for DFPD1-10 was once every two weeks, and the dosing period for nitobuzumab was once per week.

Example 3

Inventive example 3 further defined the amount of DFPD1-10 modified to 200mg based on example 2; nituzumab was used in an amount of 200mg, with the remainder of the procedure being as in example 2.

Example 4

Inventive example 4 further defined on the basis of example 2 that the amount of DFPD1-10 was modified to 200mg; nituzumab was used in an amount of 400mg, with the remainder of the procedure being as in example 2.

Example 5

Inventive example 5 further defined the amount of DFPD1-10 modified to 300mg based on example 2; nituzumab was used in an amount of 500mg, with the remainder of the procedure being as in example 2.

Example 6

The embodiment 6 of the invention further provides application of the combination drug composition in preparing drugs for treating tumor diseases on the basis of the above embodiments 1 to 5;

tumor diseases include non-small cell lung cancer, glioma, colorectal cancer, liver cancer, HER2 negative metastatic breast cancer, metastatic gastric adenocarcinoma, metastatic melanoma, metastatic renal cell carcinoma, advanced esophageal squamous carcinoma, advanced head and neck squamous carcinoma.

Non-small cell lung cancer includes, but is not limited to, metastatic non-small cell lung cancer, advanced squamous non-small cell lung cancer; colorectal cancer includes, but is not limited to, metastatic colorectal cancer; liver cancer includes, but is not limited to, hepatocellular carcinoma and metastatic stem cell carcinoma.

Example 7

Example 8 of the present invention further defines neoplastic diseases including advanced esophageal squamous carcinoma, non-small cell lung carcinoma, advanced head and neck squamous carcinoma on the basis of example 7 above. Non-small cell lung cancer includes, but is not limited to, metastatic non-small cell lung cancer, advanced squamous non-small cell lung cancer.

Experimental example 1: in vivo efficacy experiment of combined pharmaceutical composition on FaDu head and neck squamous cell carcinoma model

1. Test agent

Test drug 1: the anti-PD-1 monoclonal antibody is DFPD1-10 (with the number JY 034), and the DFPD1-10 comprises a light chain variable region shown as SEQ ID No. 3 and a heavy chain variable region shown as SEQ ID No. 1.

Production unit: beijing Oriental Biotech Co., ltd;

quantity: 10ml,10mg/ml,3 bottles, 300mg total;

Traits: a liquid;

test drug 2: opdivo (Nivolumab);

The purchasing manufacturer: bai Shi Mei Gui Bao;

Quantity: 100mg/10ml,1 branch;

Traits: a liquid;

Test drug 3: toxinsheng (nimuzumab injection);

Providing units: beijing Oriental Biotech Co., ltd;

Quantity: about 10ml (49 mg), 4.9mg/ml,1 bottle;

Traits: a liquid;

2. experimental animals:

Species strain: musMusculus, NCG;

Gender: a male;

Weight of: 18-22g;

Quantity: 105;

experimental animal provider: jiangsu Jiyaokang biotechnology Co., ltd;

production license number: SCXK (su) 2018-0008;

Quality certification number: 202013758.

3. The experimental method comprises the following steps:

3.1 cell culture

FaDu tumor cells (YK-CL-222) were cultured in an RPMI-1640 medium containing inactivated 10% fetal bovine serum, 100U/ml penicillin and 100. Mu.g/ml streptomycin and 2mM glutamine in an incubator at 37℃with 5% CO2, and the cells were bottled for passage every 3 to 4 days after they were grown up and used for in vivo tumor inoculation in the logarithmic phase.

3.2 Tumor cell seeding and grouping

PBMC (human peripheral blood mononuclear cells) derived from normal human peripheral blood (Donor #: SLB 200234) were inoculated into tumor-bearing mice eight days prior to FaDu tumor cell inoculation, 2X 10 ⁶/mouse; PBS-resuspended FaDu tumor cells at a concentration of 5X 10 ⁷/ml were inoculated subcutaneously in the right flank of the experimental animals, 100. Mu.l/dose were grouped when tumors grew to around 53mm ³ (day of the notation PG-D0), for a total of 8 groups of 10, each, with specific dosing regimens as shown in Table 1.

3.3 Measurement and Experimental indicators of mouse body weight

The tumor volume was measured 2 times per week using a vernier caliper, the body weight of the mice was weighed with an electronic balance, and the long and short diameters of the tumor were measured, and the volume calculation formula was: volume = 0.5 x long diameter x short diameter ². T/C values were calculated from tumor volumes, where T is the average of the Relative Tumor Volumes (RTV) for each subject treatment group, C is the average of the Relative Tumor Volumes (RTV) for the control group, and RTV is the ratio of tumor volumes after and before administration. Tumor growth inhibition rate TGITV (%) = (1-T/C) ×100%.

At the end of the experiment, animals were euthanized, the exfoliated tumors weighed, placed in order, photographed, and the tumor weight inhibition TGITW (%) was calculated as (1-T/C) ×100%, where T/c=the average of TW in the treatment group/the average of TW in the control group.

In principle, the evaluation criteria were: T/C (%) >40% is not effective; T/C (%) is less than or equal to 40%, and P <0.05 is effective after statistical treatment.

3.4 Dosing regimen

Table 1 dosing regimen table

Note that: * The dosage of each group can be adjusted according to the weight of animals according to the dosage of 10 mu l/g when the weight is reduced by 15-20%;

i.p.: injecting into the abdominal cavity; tiw x 3ks: three times per week for 9 times; biw x 3wks: twice weekly, 6 times in total.

3.5 Statistical analysis

The tumor volumes and tumor weights were statistically analyzed between groups using IBM SPSS STATISTICS 22.0.0 statistical software, using One-Way ANOVA test, with p <0.05 considered significant differences.

3.6 Tumor growth inhibition results are shown in table 2 below:

TABLE 2 tumor inhibiting effect of the test substances on PBMC humanized head and neck squamous carcinoma FaDu model animals

Note that: one-Way ANOVA single-factor analysis of variance was performed for each group and Bonferroni was used for post hoc multiple comparisons; ^a. Mean ± standard error; ^b. Compare to Isotype groups; ^c. Comparison to the Opdivo group; ^d. Comparing with the Thixin group; ^e. Compared with JY034 low dose+Thioxin group. No significant statistical difference (p > 0.05) was seen for the comparisons between treatment groups.

3.7 Tumor weight results are shown in table 3 below:

TABLE 3 tumor inhibiting effect of the test substances on tumor weights of PBMC humanized head and neck squamous carcinoma FaDu model animals

In summary, as shown in the above data and fig. 1 and 2, in the PBMC humanized head and neck squamous carcinoma FaDu model, the combined treatment of the test drug JY034 and tiaxin produced a clear anti-tumor effect, the efficacy was superior to that of the single drug JY034 or tiaxin treatment, and the tumor inhibition effect was optimal in each treatment group.

Experimental example 2: safety test of anti-PD-1 monoclonal antibody

1. Test drug:

The anti-PD-1 monoclonal antibody is DFPD1-10 (with the number JY 034), and the DFPD1-10 comprises a light chain variable region shown as SEQ ID No. 3 and a heavy chain variable region shown as SEQ ID No. 1.

Production and supply units: beijing Oriental Baitai Biotech Co.Ltd

Specification of: 12 mL/tube, 40 mL/tube;

Protein concentration/content: 14.2mg/mL.

2. Experimental animal

Species & strain: cynomolgus monkey (Macacafascicularis)

Animal grade: common grade

Number of animals: 40 animals (20/sex).

Age at start of administration (D1): the regimen requires animals with ages 2.5-5 years at the beginning of dosing, animals with ages 3.5-4.8 years male at the beginning of dosing, and females with ages 2.8-3.6 years.

Initial dosing body weight (D-1): 2-5 kg of scheme; actual male body weight: 2.76-4.55 kg, female body weight: 2.12-3.24kg.

Experimental animal sources: guangxi androstane primate laboratory animal culture development Co.Ltd

Animal production license: SCXK (cinnamon) 2016-0003

Experimental animal pass number: 0002908, 0002916, 0002917

Issuing unit: guangxi Zhuang autonomous area science and technology hall

3. Route of administration: intravenous injection.

4. Frequency and period of dosing: the administration was 1 time per week, 13 weeks continuously, and 14 times in total. Calculated from the last dose, recovery period was 6 weeks.

5. The experimental method comprises the following steps: the test uses 40 cynomolgus monkeys, which are randomly divided into 1-4 groups according to sex zones, namely an auxiliary material control group and a low, medium and high dose group of the test product, and 5 cynomolgus monkeys/sex/group. Administering JY034 preparation buffer to animals in an auxiliary material control group; the test low, medium and high dose animals were given JY034 at 5, 15 and 50mg/kg respectively, at concentrations of 0.5, 1.5 and 5mg/mL respectively, and at volumes of 10mL/kg. The administration route is intravenous infusion administration, the administration speed is about 0.5mL/kg/min, the administration is carried out 1 time per week, the continuous administration is carried out for 13 weeks, and the total administration is carried out 14 times; and blood was collected at various time points before and after the first, 5 th and 13 th administrations for toxicodendron analysis. Animals were euthanized the next day after last dose (D93) and at the end of 6 weeks of recovery (D134), all animals were subjected to gross anatomic observations and bone marrow smear readings, weighed for major viscera, calculated for visceral and brain ratios and subjected to multiple histopathological examinations.

6. Experimental results: under the conditions of this experiment, JY034 was repeatedly administered to cynomolgus monkeys by intravenous infusion at doses of 5, 15, 50mg/kg, 1 time per week for 13 weeks and 14 times in total, and the animals in each dose group had elevated cytokines IL-6, MCP-1 and IP-10 associated with immune function, and the other animals did not have systemic toxicity.

Experimental example 3: clinical trials of anti-PD-1 monoclonal antibody Single drug and Nituzumab-combined treatment of advanced solid tumors

1. Test drug name:

(1) anti-PD-1 monoclonal antibody, the anti-PD-1 monoclonal antibody is DFPD1-10, and the DFPD1-10 comprises a light chain variable region shown as SEQ ID No. 3 and a heavy chain variable region shown as SEQ ID No. 1.

Dosage form: injection preparation

Specification of: 100 mg/bottle (10 mL)

The manufacturing factory: beijing Oriental Biotech Co., ltd;

(2) Nituzumab injections, trade name: taixin Sheng

Dosage form: injection preparation

Specification of: 50 mg/bottle (10 mL)

The manufacturing factory: baitai biopharmaceutical company limited.

2. Target population: advanced esophageal squamous carcinoma, squamous non-small cell lung cancer and head and neck squamous carcinoma;

Sample size: phase1a: 9 to 18 patients are simulated to be in a group in the single dose increasing stage; the single medicament dose expansion stage is fit for 45-90 patients; phase1b: anti-PD-1 monoclonal antibodies in combination with Nituzumab on-dose escalation stage were enrolled in 6-20 patients; anti-PD-1 monoclonal antibodies were dosed in combination with Nituzumab on 45-90 subjects in the extended phase.

Co-planning into groups: 105 to 218 patients.

3. The subject must meet all of the following conditions to qualify for the study:

(1) With a full understanding of the purpose of the test, the investigator judged to be able to follow the test protocol, voluntarily signing a written informed consent.

(2) The age of the people who sign the informed consent is more than or equal to 18 years old, and the people can both men and women.

(3) The subject can provide tumor tissue and blood samples for tumor mutation burden (Tumor Mutational Burden, TMB), PD-L1 expression level determination. And (3) injection: it is suggested to provide a formalin-fixed paraffin-embedded tumor tissue sample (ormalin fixed paraffn embed-ded, FFPE) or an undyed freshly cut serial tissue section (slide) taken at a non-radiotherapy site within 6 months prior to the first study administration, and must also be able to provide relevant pathology reporting of the above specimens. Freshly collected specimens, resections, hollow core biopsies, resections, incisions, punches or jaw biopsies are all within acceptable limits (with newly obtained tissue being preferred). No needle samples (i.e., samples lacking intact tissue structures that only provide a cell suspension and/or a cell smear), brush samples, cell pellet samples from pleural or peritoneal effusions.

(4) Advanced solid tumors diagnosed histologically or cytologically:

phase1a part a: advanced esophageal squamous carcinoma, squamous non-small cell lung carcinoma or head and neck squamous carcinoma that has been histologically or cytologically confirmed to be non-standard, refused to receive standard, ineffective or intolerable of standard treatment;

Phase1a part B：

① Esophageal squamous carcinoma cohort: patients with advanced stage, recurrent esophageal squamous carcinoma, histologically or cytologically confirmed inoperable or unsuitable for local treatment, failed at least by first-line chemotherapy;

② Squamous non-small cell lung cancer cohort: patients with histologically or cytologically proven, non-surgically or non-topically treatable locally advanced, recurrent metastatic squamous non-small cell lung cancer, failed at least by first line chemotherapy;

③ Head and neck squamous carcinoma queues: patients with advanced stage of local stage, recurrent metastatic head and neck squamous carcinoma, which is not operable or suitable for local treatment as confirmed by histology or cytology, and failure of squamous non-small cell lung carcinoma by at least one-line chemotherapy;

Phase1b: patients with histologically or cytologically proven, non-surgically or non-suitable local treatment of advanced stage, recurrent metastatic esophageal squamous carcinoma, squamous non-small cell lung cancer or head and neck squamous carcinoma, failed at least one-line chemotherapy or intolerant of first-line platinum-containing chemotherapy;

Note that: patients who have developed tumors within 6 months after radical or adjuvant or neoadjuvant chemotherapy treatment may also be considered as first line chemotherapy failure.

(5) At least one measurable tumor lesion (recistv 1.1) is present;

(6) ECOG score 0 or 1 within 7 days prior to initiation of treatment;

(7) The expected survival time is more than or equal to 3 months;

(8) The major organs and bone marrow function were essentially normal, requiring no transfusion, no use of hematopoietic stimulators (including G-CSF, GM-CSF, EPO, TPO, etc.), and no infusion of human albumin preparation within 14 days prior to screening. Laboratory test results are within 7 days of initiation of treatment:

① The coagulation function INR is less than or equal to 1.5 XULN, aPTT is less than or equal to 1.5 XULN (if the subject is undergoing anticoagulant therapy, PT or aPTT is within the expected treatment range of anticoagulant drugs);

② The liver and kidney functions of the subject meet the following conditions: total bilirubin is less than or equal to 1.5 XULN, AST and ALT are less than or equal to 2.5 XULN (liver metastasis is less than or equal to 5 XULN); creatinine is less than or equal to 1.5 XULN;

③ Blood routine satisfies: the neutrophil count is more than or equal to 1.5X10 ⁹/L, the platelet is more than or equal to 100X 10 ⁹/L, and the hemoglobin is more than or equal to 9g/dL.

(9) Adverse events caused by previous treatments were restored to ∈ctcae grade 1 (except hair loss).

(10) The screening period was negative for HIV detection.

(11) Effective contraceptive measures were taken throughout the study period until 12 weeks after the last administration.

4. And (3) test design:

The test is designed as a multicenter, open, dose escalation and dose extension phase I clinical trial comprising two parts, phase1a and phase1b, with the main objective of determining the safety, efficacy, PK/PD profile of anti-PD-1 monoclonal antibody single and combined nituzumab in patients with advanced solid tumors.

Phase1a partA: the anti-PD-1 monoclonal antibody Single dose escalation stage selects 3 doses of 1mg/kg, 3mg/kg, 10mg/kg of anti-PD-1 monoclonal antibody, and DLT is evaluated within 28 days after the first dose, using a conventional "3+3" dose escalation test, followed by Q2W dosing until intolerable toxicity or disease progression. Incremental testing from low to high dose is performed, and if a dose group is less than or equal to 1 subject with DLT, then the next dose group is incrementally tested, and if less than or equal to 2 subjects with DLT, then the previous dose group is considered to be the MTD.

Phase1a partB: the single dose expansion stage of the anti-PD-1 monoclonal antibody is respectively added into 15-30 cases of advanced esophageal squamous carcinoma, advanced squamous non-small cell lung carcinoma and late date of maturity cervical squamous carcinoma, if no DLT appears in partA, 200mg of the anti-PD-1 monoclonal antibody is administered in this part, and the administration is continued until intolerable toxicity or disease progress.

After the phase1a test is completed, the phase1b test can be started timely to evaluate the safety and preliminary effectiveness of the anti-PD-1 monoclonal antibody combined with Nituzumab.

Phase1b partA: the anti-PD-1 monoclonal antibody is combined with Nituzumab in the dose-increasing stage, 2 doses of Nituzumab are to be selected, 200mg of Nituzumab and 400mg of QW are tentatively administered in combination with 200mgQ W of anti-PD-1 monoclonal antibody respectively, and each dose is organized into 3-10 patients. The first 3 patients in each dose group were observed for evaluation of DLT within 14 days after the first administration, followed by 200mg of nituzumab and 400mg of QW in combination with 200mg of anti-PD-1 monoclonal antibody and 2 w; the next 7 patients were dosed directly with 200mg of nituzumab and 400mg of QW in combination with 200mg of anti-PD-1 monoclonal antibody q2w until intolerable toxicity or disease progression.

Phase1b partB: the anti-PD-1 monoclonal antibody is combined with the nimuzumab in a dose expansion stage, one administration dose RP2D QW of the nimuzumab is selected from a phase1b partA part, 200mg of the anti-PD-1 monoclonal antibody is combined with the anti-PD-1 monoclonal antibody to be administered in Q2W, and 15 to 30 cases of late esophageal squamous cell lung cancer, late squamous non-small cell lung cancer and late date of maturity cervical squamous cell carcinoma are respectively added, and the administration is continued until intolerable toxicity or disease progress.

5. Dosing regimen:

The part of Phase1a is administered by anti-PD-1 monoclonal antibody single drug, the dose is 4, the experimental design part, DLT is observed within 28 days after the first administration of Phase1a partA (C1D 1), and then a administration period is formed every 2 weeks, and D1 is administered every period; phase1a partB was scheduled to be administered as 200mg q2w of anti-PD-1 monoclonal antibody. The time of the first intravenous infusion of the anti-PD-1 monoclonal antibody is 60+/-15 min. If the first infusion is well tolerated, the time for the second infusion can be shortened to 30.+ -. 10min. If the patient is well tolerated for 30 minutes of infusion, all subsequent infusions can be completed at 30 minutes. Dose adjustments were not made to the anti-PD-1 monoclonal antibody mab during the trial.

Phase1b fraction was dosed with anti-PD-1 monoclonal antibody in combination with nimuzumab, the dose was seen in 4, experimental design section, DLT was observed within 14 days after first dose of Phase1b partA (C1D 1), followed by one dosing cycle every 2 weeks, dosing of nimuzumab D1 and D8 every cycle, anti-PD-1 monoclonal antibody at D1; phase1b partB schedule dose was 200mg q2w of anti-PD-1 monoclonal antibody, nimuzumab was administered according to the recommended RP2D dose QW of partA section. The anti-PD-1 monoclonal antibody injection is firstly given to the patient each time, and the Nituzumab injection is then given to the patient at intervals of more than 10 minutes. The time of the first intravenous infusion of the anti-PD-1 monoclonal antibody and the nitobuzumab is 60+/-15 min. If the first infusion is well tolerated, the time for the second infusion can be shortened to 30.+ -. 10min. If the patient is well tolerated for 30 minutes of infusion, all subsequent infusions can be completed at 30 minutes.

The present application is not limited to the above-described preferred embodiments, and any person who has come to various other forms of products in the light of the present application can, however, make any changes in shape or structure, and all that is within the scope of protection same as or similar to the present application.

Sequence listing

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<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 1

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

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

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Trp Tyr Asp Ser Ser Arg 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 Phe

65 70 75 80

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

85 90 95

Ala Thr Asn Asn Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

100 105 110

Ser

<210> 2

<211> 107

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 2

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 Ser Ile His Asn Tyr

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Glu Leu His Leu Pro Leu

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 3

<211> 107

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 3

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 Ser Val Ser Asn Tyr

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 4

<211> 113

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 4

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

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

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Trp Tyr Asp Ser Ser Arg 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 Phe

65 70 75 80

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

85 90 95

Ala Thr Asn Asn Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

100 105 110

Ser

<210> 5

<211> 113

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 5

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

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

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Trp Tyr Asp Gly Ser Lys 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 Phe

65 70 75 80

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

85 90 95

Ala Thr Asn Asn Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

100 105 110

Ser

Claims

1. The application of the combined medicine composition in preparing a medicine for treating cervical squamous cell carcinoma of late date of maturity is characterized in that the combined medicine composition comprises an anti-PD-1 monoclonal antibody taking PD-1 as a target point and an anti-EGFR monoclonal antibody taking EGFR as a target point; wherein the anti-EGFR monoclonal antibody is Nituzumab, the anti-PD-1 monoclonal antibody is DFPD1-10, and the DFPD1-10 comprises a light chain variable region shown as SEQ ID No. 3 and a heavy chain variable region shown as SEQ ID No. 1.

2. The use of claim 1, wherein the administration of the anti-PD-1 monoclonal antibody and the anti-EGFR monoclonal antibody comprises simultaneous, concurrent, sequential, alternating, or separate administration.

3. The use of claim 1, wherein the anti-PD-1 monoclonal antibody and the anti-EGFR monoclonal antibody are both administered intravenously.

4. The use of claim 1, wherein the DFPD1-10 is administered once every two weeks and the nituzumab is administered once a week.