Drug combination of quinoline derivatives and anti-PD-1 monoclonal antibody

Abstract

Provided is a pharmaceutical combination of a quinoline derivative and a PD-1 monoclonal antibody, which comprises a tyrosine kinase inhibitor and an immune checkpoint inhibitor, wherein the tyrosine kinase inhibitor is a compound of formula I or a pharmaceutically acceptable salt thereof. The pharmaceutical combination has good anti-gastrointestinal tumor, urinary system tumor or neuroendocrine tumor activity.

Description

Technical Field

[0001] This application belongs to the field of medical technology and relates to combination therapy for tumors. Specifically, this application relates to a combination of quinoline derivatives and PD-1 monoclonal antibodies and their use in treating digestive tract tumors, urinary system tumors, and neuroendocrine tumors. Background Art

[0002] Tyrosine kinases are a group of enzymes that catalyze the phosphorylation of protein tyrosine residues. They play a crucial role in intracellular signal transduction, participating in the regulation, signaling, and development of normal cells. They are also closely associated with the proliferation, differentiation, migration, and apoptosis of tumor cells. Many receptor tyrosine kinases are associated with tumor formation. Based on the structure of their extracellular domains, they can be divided into epidermal growth factor receptor (EGFR), platelet-derived growth factor receptor (PDGFR), vascular endothelial growth factor receptor (VEGFR), and fibroblast growth factor receptor (FGFR).

[0003] Anlotinib is a quinoline derivative tyrosine kinase inhibitor that acts as a multi-target tyrosine kinase inhibitor (TKI) in influencing tumor angiogenesis and proliferation signal transduction. Its main targets include receptor tyrosine kinases vascular endothelial growth factor receptors (VEGFR) 1 to 3, epidermal growth factor receptor (EGFR), fibroblast growth factor receptors (FGFR) 1 to 4, platelet-derived growth factor receptors (PDGFR) α and β, and stem cell factor receptors (SCFR) 7, 8, and 9. A phase 2 trial showed that anlotinib can improve progression-free survival and has a potential benefit in overall survival (Han B, et al. Br J Cancer. 2018; 118(5): 654-661). A multicenter, double-blind, phase 3 randomized clinical trial showed that anlotinib resulted in prolonged overall survival and progression-free survival in Chinese patients. This finding suggests that anlotinib is well tolerated and is a potential third-line or further treatment for patients with advanced NSCLC (Han B, et al. JAMA Oncol. 2018 Nov; 4(11): 1569-1575).

[0004] Document WO2008112407 discloses in Example 24 a quinoline derivative tyrosine kinase inhibitor, 1-[[[4-(4-fluoro-2-methyl-1H-indol-5-yl)oxy-6-methoxyquinolin-7-yl]oxy]methyl]cyclopropylamine, and a preparation method thereof. Its structural formula is shown in Formula I:

[0005]

[0006] PD-1 (programmed death-1) is a key immune checkpoint receptor expressed by activated T and B lymphocytes and mediates immunosuppression. Its ligands include at least PD-L1 and PD-L2. PD-L1 (Programmed death-ligand 1), also known as CD274 or B7-H1, is a 40 kDa type I transmembrane protein encoded by the CD274 gene and is a ligand of PD-1. Both PD-L1 and PD-1 belong to the immunoglobulin superfamily and consist of two extracellular Ig domains, namely the N-terminal V domain and the C-terminal constant domain. The binding interfaces of PD-L1 with programmed death receptor-1 (PD-1) and B7-1 (CD80) are on the IgV-like domain (Lin et al. (2008) PNAS 105: 3011-3016). PD-L1 contains a conserved short intracellular tail region (about 30 amino acids), and PD-1 contains two cytoplasmic tyrosine-based signaling motifs, namely immunoreceptor tyrosine-based inhibitory motif (ITIM) and immunoreceptor tyrosine-based switch motif (ITSM). After T cell stimulation, PD-1 recruits the tyrosine phosphatase SHP-2 to the ITSM motif within its cytoplasmic tail, resulting in dephosphorylation of effector molecules involved in the CD3+ T cell signaling cascade, such as CD3ζ, PKCθ, and ZAP70 (Freeman et al. (2000) J Exp Med 192: 1027-34; Latchman et al. (2001) Nat Immunol 2: 261-8; Carter et al. (2002) Eur J Immunol 32: 634-43). PD-L1 is widely distributed not only on leukocytes and non-hematopoietic cells in lymphoid and non-lymphoid tissues but also on various cancer cells, highly expressed on the surface of a variety of tumor cells, and the malignancy and poor prognosis of tumors are closely related to the expression level of PD-L1. There is clinical data indicating that high tumor expression of PD-L1 is associated with increased tumor invasiveness and poor prognosis.The formation of the PD-1 / PD-L1 complex transmits inhibitory signals and negatively regulates T cell immune responses; it inhibits TCR-mediated T cell activation, cytokine production, and T cell proliferation (Fife et al. (2011) Nature Immunology 10: 1185-1193); induces exhaustion or anergy among antigen-specific T cells (Hofmeyer et al. (2011) Journal of Biomedicine and Biotechnology 2011: 1-9); promotes the differentiation of Th1 cells into Foxp3+ regulatory T cells (Armanath et al. (2011) Science TransMed 3: 1-13; Francisco et al. (2009) J. Exp. Med. 206: 3015-3029); and induces apoptosis of effector T cells. Disruption of the PD-L1 gene results in upregulated T cell responses and the generation of autoreactive T cells (Latchman et al. (2004) PNAS 101: 10691-10696). Blockade of PD-1 or PD-L1 antibodies leads to increased anti-tumor immunity (Iwai et al. (2002) PNAS 99: 12293-12297).

[0007] Chinese Patent Document CN106977602A discloses a PD-1 monoclonal antibody 14C12H1L1, which can effectively block the binding of PD1 to PDL1 and shows good anti-tumor activity.

[0008] The biggest challenge encountered by predecessors in the process of tumor immunotherapy is the poor efficacy caused by tumor immune tolerance and escape. Therefore, it has important theoretical significance and application value to break the immune tolerance established by the body against tumor cells through the combined use of small molecule anti-tumor compounds and anti-PD-1 / PD-L1 antibodies. Summary of the Invention

[0009] The object of the present invention is to provide at least a drug combination, which comprises a tyrosine kinase inhibitor and a human PD-1 antibody, the human PD-1 antibody comprising a light chain and a heavy chain, wherein the light chain comprises light chain complementarity determining regions LCDR1, LCDR2 and LCDR3, the light chain complementarity determining regions are respectively composed of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, and wherein the heavy chain comprises heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3, the heavy chain complementarity determining regions are respectively composed of the amino acid sequences shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6.

[0010] In some embodiments, the tyrosine kinase inhibitor is a compound of formula I or a pharmaceutically acceptable salt thereof. In some specific embodiments, the tyrosine kinase inhibitor is the hydrochloride salt of the compound of formula I, namely anlotinib hydrochloride.

[0011] In some embodiments, the human PD-1 antibody comprises a light chain variable region having the amino acid sequence as shown in SEQ ID NO:7 and a heavy chain variable region as shown in SEQ ID NO:8.

[0012] In some embodiments, the human PD-1 antibody is 14C12H1L1.

[0013] In some embodiments, the compound of formula I may exist in the form of a pharmaceutically acceptable salt or a pharmaceutically acceptable formulation thereof, preferably in the form of its hydrochloride salt.

[0014] In some specific embodiments, the compound is the hydrochloride salt of 1-[[[4-(4-fluoro-2-methyl-1H-indol-5-yl)oxy-6-methoxyquinolin-7-yl]oxy]methyl]cyclopropanamine, namely anlotinib hydrochloride.

[0015] In some embodiments, the pharmaceutical combination comprises: a compound of formula I or its hydrochloride salt (such as the dihydrochloride); and the 14C12H1L1 monoclonal antibody or an antigen-binding fragment thereof.

[0016] The object of the present invention further lies in providing at least a use of a pharmaceutical combination for treating tumors, the pharmaceutical combination comprising a tyrosine kinase inhibitor and a human PD-1 antibody, the human PD-1 antibody comprising a light chain and a heavy chain, wherein the light chain comprises light chain complementarity determining regions LCDR1, LCDR2 and LCDR3, the light chain complementarity determining regions are respectively composed of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, and wherein the heavy chain comprises heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3, the heavy chain complementarity determining regions are respectively composed of the amino acid sequences shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6.

[0017] In some embodiments, the tyrosine kinase inhibitor is a compound of formula I or its hydrochloride salt.

[0018] In some embodiments, the human PD-1 antibody comprises a light chain variable region having the amino acid sequence as shown in SEQ ID NO:7 and a heavy chain variable region as shown in SEQ ID NO:8. In some embodiments, the human PD-1 antibody is 14C12H1L1.

[0019] The present invention also provides a method for treating a subject having a gastrointestinal tumor, or a urinary system tumor, or a neuroendocrine tumor, which comprises administering to the subject a therapeutically effective amount of a tyrosine kinase inhibitor and a therapeutically effective amount of a human PD-1 antibody, wherein the human PD-1 antibody comprises a light chain and a heavy chain, wherein the light chain comprises light chain complementarity determining regions LCDR1, LCDR2 and LCDR3, and the light chain complementarity determining regions are respectively composed of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, and wherein the heavy chain comprises heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3, and the heavy chain complementarity determining regions are respectively composed of the amino acid sequences shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6. In some embodiments, the tyrosine kinase inhibitor is a compound of formula I or its hydrochloride salt. In some embodiments, the human PD-1 antibody comprises a light chain variable region having the amino acid sequence as shown in SEQ ID NO:7 and a heavy chain variable region as shown in SEQ ID NO:8. In some embodiments, the human PD-1 antibody is 14C12H1L1.

[0020] The present invention also provides a combination therapy for treating a subject having a gastrointestinal tumor, or a urinary system tumor, or a neuroendocrine tumor, the method comprising separately administering to the subject a therapeutically effective amount of a tyrosine kinase inhibitor and a therapeutically effective amount of a human PD-1 antibody, wherein the human PD-1 antibody comprises a light chain and a heavy chain, wherein the light chain comprises light chain complementarity determining regions LCDR1, LCDR2 and LCDR3, and the light chain complementarity determining regions are respectively composed of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, and wherein the heavy chain comprises heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3, and the heavy chain complementarity determining regions are respectively composed of the amino acid sequences shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6.

[0021] The present invention also provides a method for treating a subject having a cancer or tumor, which cancer or tumor is a gastrointestinal tumor, or a urinary system tumor, or a neuroendocrine tumor, the method comprising: (i) measuring the levels of PD-1 and / or PD-L1 in a sample of the subject, wherein the subject is PD-1 and / or PD-L1 positive, and (ii) administering to the subject a therapeutically effective amount of an anti-PD-1 and / or PD-L1 antibody or an antigen-binding portion thereof.

[0022] The present invention also provides a method for treating a subject suffering from cancer or a tumor. In certain embodiments, the subject is a patient diagnosed with a gastrointestinal tumor, or a urinary system tumor, or a neuroendocrine tumor, such as a patient diagnosed with biliary tract cancer, or intrahepatic cholangiocarcinoma, or extrahepatic cholangiocarcinoma, or gallbladder cancer, or colorectal cancer, or gastric or gastroesophageal junction (GEJ) adenocarcinoma, or urothelial carcinoma.

[0023] In some embodiments of the present invention, the subject has previously received surgery, chemotherapy, and / or radiotherapy. In some specific embodiments, the subject experiences disease progression again after achieving a complete remission after surgery, chemotherapy, and / or radiotherapy. In some specific embodiments, the subject fails to achieve a complete remission or a partial remission after surgery, chemotherapy, and / or radiotherapy.

[0024] In some embodiments of the present invention, the subject has not previously received systemic chemotherapy. In some embodiments, the subject has previously received surgical treatment, radiotherapy, induction chemotherapy, and / or adjuvant chemotherapy, or the subject receives concurrent chemotherapy. In some specific embodiments, the subject has not previously received systemic chemotherapy, but has received surgical treatment, radiotherapy, induction chemotherapy, and / or adjuvant chemotherapy or will receive concurrent chemotherapy. In some specific embodiments, the subject experiences disease progression again after achieving a complete remission after surgical treatment, radiotherapy, induction chemotherapy, concurrent chemotherapy, and / or adjuvant chemotherapy. In some specific embodiments, the subject fails to achieve a complete remission or a partial remission after surgical treatment, radiotherapy, induction chemotherapy, concurrent chemotherapy, and / or adjuvant chemotherapy. In some specific embodiments, the cancer metastasizes after surgical treatment, radiotherapy, induction chemotherapy, concurrent chemotherapy, and / or adjuvant chemotherapy.

[0025] In some embodiments, the use for treating a tumor is for treating a gastrointestinal tumor. In some embodiments, the gastrointestinal tumor has previously failed first-line or more than first-line chemotherapy. In some embodiments, the gastrointestinal tumor has a deficiency in mismatch repair genes (dMMR) or high microsatellite instability (MSI-H). In some embodiments, the gastrointestinal tumor is not suitable for surgical resection. In some embodiments, it is a recurrent or metastatic gastrointestinal tumor. In some embodiments, the gastrointestinal tumor is a gastrointestinal tumor that has not previously received treatment for recurrence or metastasis.

[0026] In some embodiments, the gastrointestinal tumor is biliary tract cancer. In some embodiments, the biliary tract cancer is inoperable biliary tract cancer or metastatic biliary tract cancer. In some embodiments, the biliary tract cancer has previously failed first-line or more than first-line chemotherapy. In some embodiments, the biliary tract cancer is cholangiocarcinoma. In some embodiments, the biliary tract cancer is intrahepatic cholangiocarcinoma (IHCC), or extrahepatic cholangiocarcinoma (EHCC), or gallbladder cancer (GBC). In some embodiments, the biliary cancer is intrahepatic cholangiocarcinoma (IHCC), or extrahepatic cholangiocarcinoma (EHCC), or gallbladder cancer (GBC) that has previously failed first-line or more than first-line chemotherapy. In some embodiments, the extrahepatic cholangiocarcinoma includes hilar cholangiocarcinoma (also known as Klatskin tumor) and distal cholangiocarcinoma. In some embodiments, the biliary tract cancer includes adenocarcinoma-type gallbladder cancer, adenocarcinoma-type intrahepatic cholangiocarcinoma, or adenocarcinoma-type extrahepatic cholangiocarcinoma. In some embodiments, the biliary tract cancer is diffuse infiltrating adenocarcinoma of the bile duct.

[0027] In some embodiments, the gastrointestinal tumor is colorectal cancer. In some embodiments, the colorectal cancer has a deficiency in mismatch repair genes (dMMR) or high microsatellite instability (MSI-H). In some embodiments, the colorectal cancer is not suitable for surgical resection. In some embodiments, the colorectal cancer is recurrent or metastatic colorectal cancer. In some embodiments, the colorectal cancer is colorectal cancer that has not previously received treatment for recurrence or metastasis. In some embodiments, the gastrointestinal tumor is intestinal cancer. In some embodiments, the gastrointestinal tumor is metastatic intestinal cancer.

[0028] In some embodiments, the gastrointestinal tumor is gastric or gastroesophageal junction (GEJ) adenocarcinoma. In some embodiments, the gastrointestinal tumor is metastatic or recurrent gastric or gastroesophageal junction adenocarcinoma. In some embodiments, the gastrointestinal tumor is gastric or gastroesophageal junction adenocarcinoma that has not previously received treatment for recurrence or metastasis.

[0029] In some embodiments, the use for treating a tumor is for treating urinary system tumors. In some embodiments, the urinary system tumor is not suitable for surgical resection. In some embodiments, the urinary system tumor is locally advanced or metastatic urinary system tumor. In some embodiments, the urinary system tumor is a urinary system tumor that has previously failed at least one systemic standard chemotherapy regimen. In some embodiments, the urinary system tumor is urothelial carcinoma. In some embodiments, the urinary system tumor is urothelial carcinoma originating from the bladder, or ureter, or renal pelvis, or urethra. In some embodiments, the urinary system tumor is locally advanced or metastatic urothelial carcinoma. In some embodiments, the urinary system tumor is urothelial carcinoma that has previously failed at least one systemic standard chemotherapy regimen.

[0030] In some embodiments, the use for treating tumors is for treating neuroendocrine tumors. In some embodiments, the neuroendocrine tumors are advanced neuroendocrine tumors of low grade (G1) or intermediate grade (G2). In some embodiments, the neuroendocrine tumors are not amenable to surgical resection. In some embodiments, the neuroendocrine tumors are locally advanced or distant metastatic neuroendocrine tumors. In some embodiments, the neuroendocrine tumors are neuroendocrine tumors that have previously received at least one systemic anti-tumor drug treatment for advanced neuroendocrine tumors. Preferably, the systemic anti-tumor drug treatment includes treatment with mTOR inhibitors, chemotherapy, long-acting somatostatin analogs, interferons, PRRT (peptide receptor radionuclide), etc.

[0031] In some embodiments, the neuroendocrine tumors are gastroenteropancreatic neuroendocrine tumors. In some embodiments, the gastroenteropancreatic neuroendocrine tumors are advanced gastroenteropancreatic neuroendocrine tumors of low grade (G1) or intermediate grade (G2). In some embodiments, the gastroenteropancreatic neuroendocrine tumors are not amenable to surgical resection. In some embodiments, the gastroenteropancreatic neuroendocrine tumors are locally advanced or distant metastatic gastroenteropancreatic neuroendocrine tumors. In some embodiments, the gastroenteropancreatic neuroendocrine tumors are gastroenteropancreatic neuroendocrine tumors that have previously received at least one systemic anti-tumor drug treatment for advanced gastroenteropancreatic neuroendocrine tumors. Preferably, the systemic anti-tumor drug treatment includes treatment with mTOR inhibitors, chemotherapy, long-acting somatostatin analogs, interferons, PRRT (peptide receptor radionuclide), etc.

[0032] In some embodiments of the present application, the drug combination is a fixed combination. In some embodiments, the fixed combination is in the form of a solid pharmaceutical composition or a liquid pharmaceutical composition.

[0033] In some embodiments of the present application, the drug combination is a non-fixed combination. In some embodiments, the human PD-1 antibody and the compound of formula I in the non-fixed combination are each in the form of a pharmaceutical composition.

[0034] Another object of the present application is to provide at least a drug pack, which separately contains single-packaged pharmaceutical compositions in separate containers, wherein one container contains a pharmaceutical composition containing a compound of formula I or a pharmaceutically acceptable salt thereof, and the second container contains a pharmaceutical composition containing a human PD-1 antibody.

[0035] In some embodiments of the present application, the pharmaceutical composition comprises a Compound of Formula I in an amount of 6 to 168 mg. In some embodiments, the pharmaceutical composition comprises a Compound of Formula I in an amount selected from 6 mg, 8 mg, 10 mg, 12 mg, 15 mg, 20 mg, 30 mg, 50 mg, 56 mg, 70 mg, 84 mg, 112 mg, 140 mg, 168 mg, or a range formed by any of the above values. In some embodiments, the pharmaceutical composition comprises a Compound of Formula I in an amount of 10 mg - 12 mg. In some embodiments, the pharmaceutical composition comprises a Compound of Formula I in an amount of 10 mg. In some embodiments, the pharmaceutical composition comprises a Compound of Formula I in an amount of 12 mg.

[0036] In some embodiments, the human PD-1 antibody is administered in one or more unitary doses effective to treat the cancer. In some specific embodiments, the unitary dose is in the range of about 10 mg to about 1000 mg of the human PD-1 antibody. In some specific embodiments, the unitary dose is selected from about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, or about 1000 mg of the human PD-1 antibody. In some specific embodiments, the unitary dose is selected from about 200 mg of the human PD-1 antibody.

[0037] In some embodiments, the administration treatment of the human PD-1 antibody is in a cycle of 2 weeks (14 days) or 3 weeks (21 days), and preferably the human PD-1 antibody is intravenously administered on the first day (D1) of each cycle. That is, the anti-PD-1 antibody is administered at a frequency of once every two weeks (q2w) or once every three weeks (q3w).

[0038] The present application also aims to provide a unit dosage form, wherein the unit dosage form comprises: a compound component, 6 - 12 mg of a Compound of Formula I or its hydrochloride; and an antibody component, 50 - 350 mg of a human PD-1 antibody or its antigen-binding fragment; wherein the compound component and the antibody component are separately packaged.

[0039] In some embodiments, the unit dosage form comprises: a compound component, 8 mg, 10 mg, or 12 mg of a Compound of Formula I or its hydrochloride; and an antibody component, 100 mg or 200 mg of a human PD-1 antibody or its antigen-binding fragment; wherein the compound component and the antibody component are separately packaged.

[0040] The object of the present application is also to provide a method for preventing or treating cancer or tumor, wherein one or more of the above-mentioned unit preparations are administered to a subject in need. Preferably, the compound component and the antibody component in the unit preparation are administered separately. Preferably, the cancer or tumor is a digestive tract tumor, or a urinary system tumor, or a neuroendocrine tumor.

[0041] In some embodiments of the present application, anlotinib is administered in combination with 14C12H1L1, and each treatment cycle is 21 days. The drug is administered as follows: anlotinib is administered at 10 mg or 12 mg per time from D1 to D14, and 14C12H1L1 is administered at 200 mg on D1.

[0042] In some embodiments of the present application, anlotinib is administered in combination with 14C12H1L1. The administration method of anlotinib hydrochloride capsules is once a day, 12 mg or 10 mg each time. Oral administration is continued for 2 weeks and then stopped for 1 week, that is, 3 weeks (21 days) is a treatment cycle. The administration method of 14C12H1L1 injection is once every 3 weeks, 200 mg each time, and is administered by intravenous infusion. The infusion time is 60 ± 10 min.

[0043] The drug combination of the present invention shows better efficacy than existing therapies in patients with urothelial carcinoma who have relapsed or metastasized after surgery and / or chemotherapy. The chemotherapy is, for example, doxorubicin and / or gemcitabine combined with cisplatin treatment. An example of the ureteral cancer is ureteral urothelial carcinoma, bladder cancer, etc. The metastasis is, for example, bone metastasis.

[0044] The drug combination of the present invention shows better efficacy than existing therapies in patients with rectal cancer who have relapsed or metastasized after surgery and / or chemotherapy. The chemotherapy is, for example, tegafur, temozolomide and / or octreotide acetate treatment. The metastasis can be multiple metastases, for example, multiple liver metastases.

[0045] The drug combination of the present invention shows better efficacy than existing therapies in patients with colon cancer who have relapsed or metastasized after surgery and / or chemotherapy. The chemotherapy is, for example, tegafur, temozolomide and / or octreotide acetate treatment. The metastasis can be multiple metastases, for example, multiple liver metastases.

[0046] The drug combination of the present invention shows better efficacy than existing therapies in patients with gastric cancer who have relapsed or metastasized after surgery and / or chemotherapy. The metastasis can be multiple metastases, for example, duodenal metastasis.

[0047] The pharmaceutical combination of the present invention shows superior efficacy compared to existing therapies in patients with cholangiocarcinoma or gallbladder cancer who have relapsed or metastasized after surgery and / or chemotherapy. The chemotherapy may be, for example, capecitabine, oxaliplatin combined with capecitabine, paclitaxel combined with tegafur, or paclitaxel combined with capecitabine. The metastasis may be multiple metastases, such as liver, peritoneal lymph nodes, peritoneal lymph nodes, mesenteric lymph nodes, kidney, or bone metastases.

[0048] Anlotinib

[0049] As used in the present application, the chemical name of the anlotinib (i.e., the compound of formula I) is 1-[[[4-(4-fluoro-2-methyl-1H-indol-5-yl)oxy-6-methoxyquinolin-7-yl]oxy]methyl]cyclopropanamine, and it has the following structural formula:

[0050]

[0051] As used in the present application, the anlotinib includes its non-salt form (e.g., free acid or free base), and also includes its pharmaceutically acceptable salts. Both the non-salt form and the salts are within the scope of protection of the present application. For example, the pharmaceutically acceptable salt of the anlotinib may be hydrochloride or dihydrochloride. The doses of anlotinib or its salts involved in the present application are calculated based on anlotinib free base unless otherwise specified.

[0052] 14C12H1L1

[0053] As used in the present application, 14C12H1L1 or 14C12H1L1 monoclonal antibody is an anti-PD-1 monoclonal antibody, and its sequence and structure can be referred to in the literature (CN106977602A). In the 14C12H1L1 monoclonal antibody, LCDR1 contains the sequence QDINTY (SEQ ID NO:1), LCDR2 contains the sequence RAN (SEQ ID NO:2), LCDR3 contains the sequence LQYDEFPLT (SEQ ID NO:3), HCDR1 contains the sequence GFAFSSYD (SEQ ID NO:4), HCDR2 contains the sequence ISGGGRYT (SEQ ID NO:5), and HCDR3 contains the sequence ANRYGEAWFAY (SEQ ID NO:6).

[0054] The amino acid sequence of its light chain variable region is:

[0055] DIQMTQSPSSMSASVGDRVTFTCRASQDINTYLSWFQQKPGKSPKTLIYRANRLVSGVPSRFSGSGSGQDYTLTISSLQPEDMATYYCLQYDEFPLTFGAGTKLELK (SEQ ID NO:7).

[0056] The amino acid sequence of its heavy chain variable region is:

[0057] EVQLVESGGGLVQPGGSLRLSCAASGFAFSSYDMSWVRQAPGKGLDWVATISGGGRYTYYPDSVKGRFTISRDNSKNNLYLQMNSLRAEDTALYYCANRYGEAWFAYWGQGTLVTVSS (SEQ ID NO:8).

[0058] Definitions and descriptions

[0059] Unless otherwise indicated, the following terms used in this application have the following meanings. A particular term should not be considered indefinite or unclear without a specific definition, but should be understood according to its ordinary meaning in the art. When a trade name appears in this application, it is intended to refer to the corresponding product, composition, or its active ingredient.

[0060] As used herein, the term "antibody" refers to an antigen-binding protein having at least one antigen-binding domain. The antibodies and fragments thereof of this application can be whole antibodies or any fragment thereof. Thus, the antibodies and fragments thereof of this application include monoclonal antibodies or fragments thereof and antibody variants or fragments thereof, as well as immunoconjugates. Examples of antibody fragments include Fab fragments, Fab' fragments, F(ab)' fragments, Fv fragments, isolated CDR regions, single-chain Fv molecules (scFv), and other antibody fragments known in the art. Antibodies and fragments thereof can also include recombinant polypeptides, fusion proteins, and bispecific antibodies. The anti-PD-L1 antibodies and fragments thereof disclosed herein can be of the IgG1, IgG2, IgG3, or IgG4 isotype.

[0061] The term "isotype" refers to the class of antibodies encoded by the heavy chain constant region genes. In one embodiment, the anti-PD-1 / PD-L1 antibodies and fragments thereof disclosed herein are of the IgG1 or IgG4 isotype. The anti-PD-1 / PD-L1 antibodies and fragments thereof of this application can be derived from any species, including but not limited to mice, rats, rabbits, primates, llamas, and humans. The PD-1 / PD-L1 antibodies and fragments thereof can be chimeric antibodies, humanized antibodies, or fully human antibodies.

[0062] The term "humanized antibody" refers to an antibody in which the antigen-binding site is derived from a non-human species and the variable region framework is derived from human immunoglobulin sequences. A humanized antibody may contain substitutions in the framework region such that the framework may not be an exact copy of the expressed human immunoglobulin or germline gene sequence.

[0063] "Isolated antibody" means an antibody that is substantially free of other antibodies having different antigen specificities (e.g., an isolated antibody that specifically binds PD-1 / PD-L1 is substantially free of antibodies that specifically bind antigens other than PD-1 / PD-L1). However, an isolated antibody that specifically binds PD-1 / PD-L1 may have cross-reactivity with other antigens, such as PD-1 / PD-L1 molecules from different species. In addition, an isolated antibody may be substantially free of other cellular materials and / or chemicals.

[0064] The "antigen-binding portion" (also referred to as "antigen-binding fragment") of an antibody means one or more fragments of the antibody that retain the ability to specifically bind the antigen to which the intact antibody binds.

[0065] As used herein, the term "derived," when used in reference to a molecule or polypeptide relative to a reference antibody or other binding protein, means a molecule or polypeptide that is capable of specifically binding the same epitope as the reference antibody or other binding protein.

[0066] As used herein, the term "EC50" refers to the effective concentration, the 50% maximal response of an antibody. As used herein, the term "IC50" refers to the inhibitory concentration, the 50% maximal response of an antibody. Both EC50 and IC50 can be measured by ELISA or FACS analysis or any other method known in the art.

[0067] The term "treatment" generally refers to an operation that obtains a desired pharmacological and / or physiological effect. The effect can be prophylactic, in that it completely or partially prevents a disease or its symptoms; and / or therapeutic, in that it partially or completely stabilizes or cures a disease and / or the side effects resulting from the disease. "Treatment" as used herein encompasses any treatment of a disease in a patient, including: (a) preventing a disease or symptom from occurring in a patient who is predisposed to the disease or symptom but has not yet been diagnosed as having the disease; (b) inhibiting the symptoms of a disease, i.e., preventing its progression; or (c) alleviating the symptoms of a disease, i.e., causing regression of the disease or symptom.

[0068] As used herein, the term "systemic treatment" refers to a treatment in which a pharmaceutical substance is delivered via the bloodstream and reaches and affects cells throughout the body.

[0069] As used herein, the term "systemic chemotherapy" refers to systemic chemotherapy that does not include chemotherapy for locally advanced disease as part of a multimodal treatment, where chemotherapy for locally advanced disease includes induction chemotherapy, chemotherapy concurrent with radiotherapy, and adjuvant chemotherapy.

[0070] As used herein, the term "subject" refers to a mammal, such as a rodent, feline, canine, and primate. Preferably, the subject according to the present application is a human.

[0071] "Administering" means physically introducing a composition comprising a therapeutic agent to a subject using any of a variety of methods and delivery systems known to those of skill in the art. Routes of administration of immune checkpoint inhibitors (e.g., anti-PD-1 antibodies or anti-PD-L1 antibodies) include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes of administration, such as by injection or infusion. The phrase "parenteral administration" as used herein refers to a mode of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, and in vivo electroporation. In certain embodiments, the immune checkpoint inhibitor (e.g., anti-PD-1 antibody or anti-PD-L1 antibody) is administered by a non-parenteral route, and in certain embodiments, orally. Other non-parenteral routes include topical, epidermal, or mucosal routes of administration, such as intranasally, vaginally, rectally, sublingually, or topically. Administration can also be carried out, for example, once, multiple times, and / or over one or more extended time periods.

[0072] "Adverse event" (AE) as used herein is any adverse and usually unintended or undesired sign (including abnormal laboratory findings), symptom, or disease associated with the application of a medical treatment. For example, an adverse event can be associated with activation of the immune system or expansion of immune system cells (e.g., T cells) in response to treatment. A medical treatment can have one or more associated AEs, and each AE can have the same or different severity levels. A reference to a method that can "alter an adverse event" refers to a treatment regimen that reduces the incidence and / or severity of one or more AEs associated with the application of different treatment regimens.

[0073] "Dosing interval" as used herein refers to the amount of time elapsed between multiple doses of a formulation disclosed herein administered to a subject. Thus, the dosing interval can be indicated as a range.

[0074] As used herein, the term "administration frequency" refers to the frequency of administration of the dosage of the formulations disclosed herein over a given period of time. The administration frequency can be indicated as the number of administrations per given period of time, e.g., once a week or once every two weeks.

[0075] The application of the term "flat dose" refers to a dose administered to a patient without regard to the patient's weight or body surface area (BSA). Thus, a flat dose is defined as the absolute amount of a medicament (e.g., an anti-PD-1 antibody), rather than as a mg / kg dose. For example, a 60 kg person and a 100 kg person will receive the same dose of antibody (e.g., 240 mg anti-PD-1 antibody).

[0076] The application of the term "fixed dose" with respect to the compositions of the present application refers to two or more different antibodies in a single composition being present in the composition in a specific (fixed) ratio to each other. In certain embodiments, the fixed dose is based on the weight of the antibody (e.g., mg). In certain embodiments, the fixed dose is based on the concentration of the antibody (e.g., mg / ml). In certain embodiments, the ratio of mg of the first antibody:mg of the second antibody is at least about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, about 1:100, about 1:120, about 1:140, about 1:160, about 1:180, about 1:200, about 200:1, about 180:1, about 160:1, about 140:1, about 120:1, about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1 or about 2:1. For example, a 3:1 ratio of the first antibody to the second antibody can mean that the vial can contain about 240 mg of the first antibody and 80 mg of the second antibody, or about 3 mg / ml of the first antibody and 1 mg / ml of the second antibody.

[0077] The term "weight-based dose" as referred to herein means a dose administered to a patient calculated based on the patient's weight. For example, when a patient with a body weight of 60 kg requires 3 mg / kg of anti-PD-1 antibody and 1 mg / kg of anti-CTLA-4 antibody, one can withdraw an appropriate amount of anti-PD-1 antibody (i.e., 180 mg) and anti-CTLA-4 antibody (i.e., 60 mg) at one time from a fixed-dose formulation of the anti-PD-1 antibody and anti-CTLA-4 antibody in a 3:1 ratio.

[0078] The term "immunotherapy" refers to a method of treating a subject suffering from a disease or at risk of infection or disease recurrence, the method comprising inducing, enhancing, suppressing or otherwise altering an immune response. "Treatment" or "therapy" of a subject refers to any type of intervention or process performed on the subject, or the administration of an active agent to the subject, with the aim of reversing, alleviating, improving, suppressing, slowing or preventing the onset, progression, development, severity or recurrence of symptoms, complications or disorders, or biochemical markers associated with the disease.

[0079] As used herein, "PD1 / PD-L1 positive" may be used interchangeably with "at least about 1% PD-1 / PD-L1 expression". In one embodiment, PD-1 / PD-L1 expression may be used by any method known in the art. In another embodiment, PD-1 / PD-L1 expression is measured by automated IHC. In certain embodiments, "PD-1 / PD-L1 positive" refers to the presence of at least 100 cells expressing PD-1 / PD-L1 on the cell surface.

[0080] "Programmed death receptor-1 (PD-1)" refers to an immunosuppressive receptor belonging to the CD28 family. PD-1 is mainly expressed on previously activated T cells in vivo and binds two ligands, PD-L1 and PD-L2. The term "PD-1" as used herein includes human PD-1 (hPD-1), variants of hPD-1, allotypes and species homologues, and analogues having at least one common epitope with hPD-1.

[0081] "Programmed death ligand-1 (PD-L1)" is one of two cell surface glycoprotein ligands for PD-1 (the other being PD-L2), which downregulates T cell activation and cytokine secretion upon binding to PD-1.

[0082] "Subject" includes any human or non-human animal. The term "non-human animal" includes, but is not limited to, vertebrates such as non-human primates, sheep, dogs, and rodents such as mice, rats, and guinea pigs. In certain embodiments, the subject is a human. The terms "subject", "subject" and "patient" may be used interchangeably in certain contexts herein.

[0083] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is any amount of the drug that, when used alone or in combination with another therapeutic agent, protects a subject from the onset of a disease or promotes the regression of a disease, as evidenced by a decrease in the severity of disease symptoms, an increase in the frequency and duration of disease-free periods, or the prevention of damage or disability caused by affliction with the disease. The ability of a therapeutic agent to promote disease regression can be evaluated using a variety of methods known to skilled practitioners, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by measuring the activity of the agent in in vitro assays.

[0084] As used herein, a "sub-therapeutic dose" refers to a dose of a therapeutic compound (e.g., an antibody) that is below the usual or typical dose of the therapeutic compound when administered alone for the treatment of a proliferative disorder (e.g., cancer).

[0085] As an example, an "anticancer drug" promotes cancer regression in a subject or inhibits further tumor growth. In certain embodiments, a therapeutically effective amount of a drug promotes cancer regression to the point of cancer elimination. "Promoting cancer regression" means administering an effective amount of a drug, either alone or in combination with an anti-tumor agent, that results in a decrease in tumor growth or size, tumor necrosis, a decrease in the severity of at least one disease symptom, an increase in the frequency and duration of disease-free periods, or the prevention of damage or disability caused by affliction with the disease. In addition, the terms "effective" and "efficacy" with respect to treatment include pharmacological efficacy and physiological safety. Pharmacological efficacy represents the ability of a drug to promote cancer regression in a patient. Physiological safety represents the level of toxicity or other adverse physiological effects (adverse reactions) at the cellular, organ, and / or organism level caused by drug administration.

[0086] As an example for the treatment of tumors, a therapeutically effective amount of an anticancer drug can inhibit cell growth or tumor growth by at least about 10%, at least about 20%, at least about 40%, at least about 60%, or at least about 80% relative to an untreated subject, or, in certain embodiments, relative to a patient treated with standard of care therapy. In other embodiments of the present application, tumor regression can be observed and sustained for a period of at least about 20 days, at least about 40 days, or at least about 60 days. Despite these ultimate measures of treatment efficacy, the evaluation of immunotherapeutic drugs must also consider "immune-related" response patterns.

[0087] An "immune-related" response pattern refers to a clinical response pattern frequently observed in cancer patients treated with immunotherapeutic agents that exert antitumor effects by inducing cancer-specific immune responses or by altering innate immune processes. This response pattern is characterized by a beneficial therapeutic effect following an initial increase in tumor burden or the appearance of new lesions, which would be classified as disease progression in the evaluation of conventional chemotherapeutic agents and would be synonymous with drug failure. Thus, appropriate evaluation of immunotherapeutic agents may require long-term monitoring of the effects of these agents on the target disease.

[0088] A therapeutically effective amount of a drug includes a "preventive effective amount", which is any amount of a drug that, when administered alone or in combination with an antitumor agent to a subject at risk of developing cancer (e.g., a subject with a pre-malignant condition) or a subject at risk of cancer recurrence, inhibits the occurrence or recurrence of cancer. In certain embodiments, the preventive effective amount completely prevents the occurrence or recurrence of cancer. "Inhibiting" the occurrence or recurrence of cancer means reducing the likelihood of the occurrence or recurrence of cancer, or completely preventing the occurrence or recurrence of cancer.

[0089] "Recurrent" cancer is cancer that regenerates at the original site or at a distant site after responding to initial treatment (e.g., surgery). "Locally recurrent" cancer is cancer that appears at the same location as the previously treated cancer after treatment.

[0090] "Unresectable" cancer cannot be removed by surgery.

[0091] "Metastatic" cancer refers to cancer that has spread from one part of the body (e.g., the lungs) to another part of the body.

[0092] The use of alternatives (e.g., "or") should be understood to mean any one, two, or any combination of the alternatives. The indefinite article "a" or "an" as used herein should be understood to mean "one or more / one or more kinds" of any listed or enumerated component.

[0093] "Failure of platinum-containing chemotherapy regimen" means disease progression or intolerable toxic side effects during or after first-line chemotherapy or chemoradiotherapy with a platinum-containing regimen.

[0094] The definition of "failure of a systemic standard chemotherapy" is: disease progression during the treatment or after the last treatment, or intolerable toxic side effects during the treatment.

[0095] The definition of "failure of first-line or higher-line chemotherapy" is: disease progression during the treatment or after the last treatment; or intolerable toxic side effects during the treatment.

[0096] The definition of low grade (G1) is a mitotic count <2 / 10 high power fields [HPF] and / or a Ki-67 proliferation index <3%; the definition of intermediate grade (G2) is a mitotic count of 2-20 / 10 high power fields [HPF] and / or a Ki-67 proliferation index of 3-20%. If the mitotic count and Ki-67 index of the same tumor tissue correspond to different grades, the higher grade shall prevail.

[0097] The terms "about", "approximately" or "substantially comprise" mean a value or a composition within an acceptable error range of a specific value or a composition determined by a person of ordinary skill in the art, which will depend in part on how the value or the composition is measured or determined, i.e., the limitations of the measuring system. For example, "about", "approximately" or "substantially comprise" may mean within 1 or more standard deviations in accordance with the practice in the art. Alternatively, "about" or "substantially comprise" may mean a range that differs from the parameter or value being modified by at most 10% or 20% (i.e., ±10% or ±20%). For example, about 3 mg may include any number between 2.7 mg and 3.3 mg (for 10%) or between 2.4 mg and 3.6 mg (for 20%). In addition, particularly with respect to biological systems or processes, the term may mean up to one order of magnitude or up to 5 times the value. When a specific value or composition is provided in the present application and the claims, unless otherwise stated, the meaning of "about" or "substantially comprise" should be assumed to be within the acceptable error range of the specific value or composition.

[0098] As used herein, the terms "about once a week", "about once every two weeks" or any other similar dosing interval terms mean approximate values. "About once a week" may include every 7 days ±1 day, i.e., every 6 days to every 8 days. "About once every two weeks" may include every 14 days ±3 days, i.e., every 11 days to every 17 days. Similar approximations apply, for example, to about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, and about once every 12 weeks. In certain embodiments, a dosing interval of about once every 6 weeks or about once every 12 weeks means that the first dose can be administered on any day of the first week, and then the second dose can be administered on any day of the sixth or twelfth week, respectively. In other embodiments, a dosing interval of about once every 6 weeks or about once every 12 weeks means that the first dose is administered on a specific day of the first week (e.g., Monday), and then the second dose is administered on the same day (i.e., Monday) of the sixth or twelfth week, respectively. Similar principles apply to phrases including, but not limited to, "about once every 2 weeks", "about once a month", etc.

[0099] As described herein, any concentration range, percentage range, ratio range or integer range should be understood to include any integer value within the recited range, and when appropriate, include its fractions (such as tenths and hundredths of an integer), unless otherwise indicated.

[0100] Unless otherwise specified, "about" or "approximately" in this application means fluctuating within the range of ±5% of the given specific numerical range, preferably within the range of ±2%, and more preferably within the range of ±1%. For example, a pH value of about 5.5 means a pH of 5.5 ± 5%, preferably a pH of 5.5 ± 2%, and more preferably a pH of 5.5 ± 1%.

[0101] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions, and / or dosage forms that are within the scope of sound medical judgment, suitable for contact with human and animal tissues, and without excessive toxicity, irritation, allergic reaction, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0102] The term "pharmaceutically acceptable salt" includes salts formed by basic radicals and free acids or acid radicals and free bases. For example, it includes hydrochloride, hydrobromide, nitrate, sulfate, phosphate, formate, acetate, trifluoroacetate, fumarate, oxalate, maleate, citrate, succinate, methanesulfonate, benzenesulfonate, or p-toluenesulfonate. Preferably, it includes hydrochloride, hydrobromide, sulfate, formate, acetate, trifluoroacetate, fumarate, maleate, methanesulfonate, p-toluenesulfonate, sodium salt, potassium salt, ammonium salt, amino acid salt, etc. In this application, when forming a pharmaceutically acceptable salt, the molar ratio of the free acid to the basic radical is about 1:0.5 to 1:5, preferably 1:0.5, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, or 1:8. In this application, when forming a pharmaceutically acceptable salt, the molar ratio of the free base to the acid radical is about 1:0.5 to 1:5, preferably 1:0.5, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, or 1:8.

[0103] The term "fixed combination" refers to the active components (such as anti-PD-1 antibody or compound of formula I) being administered to a subject simultaneously in a fixed total dose or dose ratio, or in the form of a single entity, pharmaceutical composition, or preparation.

[0104] The term "non-fixed combination" refers to two or more active components being administered to a subject simultaneously, in parallel, or sequentially without a specific time limit as independent entities (such as pharmaceutical compositions, preparations), where the active ingredients administered to the subject reach a therapeutically effective amount level. Examples of non-fixed combinations that can be cited are cocktail therapies, such as administering 3 or more active components. In a non-fixed combination, the individual active components can be packaged, sold, or administered as completely independent pharmaceutical compositions. The "non-fixed combination" also includes the combined use between "fixed combinations", or between a "fixed combination" and an independent entity of any one or more active components.

[0105] As used herein, "administered in combination" or "used in combination" means that two or more active substances can be administered to a subject together in a mixture, simultaneously as a single formulation, or sequentially as a single formulation in any order.

[0106] The term "pharmaceutical composition" refers to a mixture composed of one or more active ingredients (e.g., anti-PD-1 antibody or compound of formula I) of the present application or their pharmaceutical combinations and pharmaceutically acceptable excipients. The purpose of the pharmaceutical composition is to facilitate the administration of the compounds or their pharmaceutical combinations of the present application to a subject.

[0107] The term "synergistic effect" refers to the effect (e.g., inhibiting the growth of colon cancer or alleviating the symptoms of colon cancer) produced by two or more components (e.g., anti-PD-1 antibody or compound of formula I) being greater than the simple addition of the effects of the components when administered alone.

[0108] Route of administration

[0109] The following does not limit the administration modes of the pharmaceutical combinations of the present application.

[0110] The components in the pharmaceutical combinations of the present application can be formulated separately, or some or all of them can be formulated together. In one embodiment, the pharmaceutical combinations of the present application can be formulated into pharmaceutical compositions suitable for single or multiple administrations.

[0111] The components in the pharmaceutical combinations of the present application can be administered separately, or some or all of them can be administered together. The components in the pharmaceutical combinations of the present application can be administered substantially non-simultaneously, or some or all of them can be administered substantially simultaneously.

[0112] The components in the pharmaceutical combinations of the present application can be administered independently or some or all of them can be administered together via various suitable routes, including but not limited to oral or parenteral (by intravenous, intramuscular, topical or subcutaneous routes). In some embodiments, the components of the pharmaceutical combinations of the present application can be administered independently or some or all of them can be administered together orally or by injection, such as intravenous injection or intraperitoneal injection.

[0113] The components in the pharmaceutical combinations of the present application can be independently or some or all of them can be together suitable dosage forms, including but not limited to tablets, lozenges, pills, capsules (e.g., hard capsules, soft capsules, enteric-coated capsules, microcapsules), elixirs, granules, syrups, injections (intramuscular, intravenous, intraperitoneal), granules, emulsions, suspensions, solutions, dispersants and dosage forms of sustained-release preparations for oral or non-oral administration.

[0114] The components in the pharmaceutical combination of the present application can each independently, or some or all of them together, contain a pharmaceutically acceptable carrier and / or excipient.

[0115] The pharmaceutical combination of the present application may further contain additional therapeutic agents. In one embodiment, the additional therapeutic agent may be a cancer therapeutic agent known in the art.

[0116] The present application also includes the following schemes:

[0117] 1. A pharmaceutical combination comprising:

[0118] a) A human PD-1 antibody, the human PD-1 antibody comprising a light chain and a heavy chain, wherein the light chain comprises light chain complementary determining regions LCDR1, LCDR2, and LCDR3, the light chain complementary determining regions are respectively composed of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, and wherein the heavy chain comprises heavy chain complementary determining regions HCDR1, HCDR2, and HCDR3, the heavy chain complementary determining regions are respectively composed of the amino acid sequences shown in SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, and

[0119] b) A tyrosine kinase inhibitor, wherein the tyrosine kinase inhibitor is a compound of formula I or a pharmaceutically acceptable salt thereof,

[0120]

[0121] 2. The pharmaceutical combination according to item 1, wherein the pharmaceutically acceptable salt of the compound of formula I is the hydrochloride salt of 1-[[[4-(4-fluoro-2-methyl-1H-indol-5-yl)oxy-6-methoxyquinolin-7-yl]oxy]methyl]cyclopropanamine, preferably the dihydrochloride salt.

[0122] 3. The pharmaceutical combination according to any one of the foregoing, wherein the human PD-1 antibody comprises a light chain variable region having the amino acid sequence shown in SEQ ID NO:7 and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO:8.

[0123] 4. The pharmaceutical combination according to any one of the foregoing, wherein the human PD-1 antibody is 14C12H1L1.

[0124] 5. The pharmaceutical combination according to any one of the foregoing, the pharmaceutical combination being a non-fixed combination.

[0125] 6. The pharmaceutical combination according to any one of the foregoing, wherein the human PD-1 antibody and the compound of formula I or a pharmaceutically acceptable salt thereof in the non-fixed combination are each in the form of a pharmaceutical composition.

[0126] 7. Use of the pharmaceutical combination according to any one of the preceding items in the treatment or prevention of digestive tract tumors, or urinary system tumors, or neuroendocrine tumors.

[0127] 8. The use according to any one of the preceding items, wherein the pharmaceutical combination is used for the treatment of digestive tract tumors, or urinary system tumors, or neuroendocrine tumors that are not suitable for surgical resection, or recurrent or metastatic, or have failed first-line or more than first-line chemotherapy, or are locally advanced.

[0128] 9. The use according to any one of the preceding items, wherein the pharmaceutical combination is used for the treatment of cholangiocarcinoma, or diffusely infiltrating adenocarcinoma of the bile duct, or cholangiocellular carcinoma, or intrahepatic cholangiocellular carcinoma, or extrahepatic cholangiocellular carcinoma, or gallbladder cancer, or intestinal cancer, or colorectal cancer, or adenocarcinoma of the stomach or gastroesophageal junction, or urothelial carcinoma.

[0129] 10. The use according to any one of the preceding items, wherein the pharmaceutical combination is used for the treatment of cholangiocarcinoma that cannot be surgically resected or is metastatic, or has failed first-line or more than first-line chemotherapy.

[0130] 11. The use according to any one of the preceding items, wherein the pharmaceutical combination is used for the treatment of colorectal cancer with deficient mismatch repair genes or high microsatellite instability, or recurrent or metastatic colorectal cancer, or colorectal cancer that has not previously received treatment for recurrence or metastasis.

[0131] 12. The use according to any one of the preceding items, wherein the pharmaceutical combination is used for the treatment of recurrent or metastatic adenocarcinoma of the stomach or gastroesophageal junction, or adenocarcinoma of the stomach or gastroesophageal junction that has not previously received treatment for recurrence or metastasis.

[0132] 13. The use according to any one of claims 7-9, wherein the pharmaceutical combination is used for the treatment of locally advanced or metastatic urothelial carcinoma, or urothelial carcinoma that has failed at least one systemic standardized chemotherapy regimen, or urothelial carcinoma originating from the bladder, ureter, renal pelvis, or urethra.

[0133] 14. The use according to any one of the preceding items, wherein the pharmaceutical combination is used for the treatment of low-grade, or intermediate-grade, or advanced, or inoperable, or locally advanced, or distant metastatic, or gastroenteropancreatic neuroendocrine tumors that have previously received at least one systemic anti-tumor drug treatment for advanced gastroenteropancreatic neuroendocrine tumors.

[0134] 15. The use according to any one of the preceding items, wherein the unified administration dose of the hydrochloride salt of the compound of formula I in the pharmaceutical combination is about 10 mg / time or 12 mg / time, and the unified administration dose of 14C12H1L1 is about 200 mg / time.

[0135] 16. The use according to any one of the foregoing, wherein the hydrochloride of the compound of formula I in the pharmaceutical combination is administered once a day, 12 mg or 10 mg each time, orally for 2 consecutive weeks followed by a 1-week break, and the 14C12H1L1 injection is administered once every 3 weeks, 200 mg each time.

[0136] 17. A product comprising a container containing a fixed dose of the hydrochloride of the compound of formula I and the 14C12H1L1 antibody, wherein the fixed dose of the hydrochloride of the compound of formula I is selected from about 8 mg, about 10 mg and about 12 mg, and the fixed dose of the 14C12H1L1 antibody is selected from about 100 mg and about 200 mg. Detailed implementation manners

[0137] The present application will be further described below in conjunction with specific embodiments. However, these embodiments in the present application are only used for illustration and do not limit the scope of the present application. Similarly, the present application is not limited to any specific preferred implementation manner described herein. Those skilled in the art should understand that equivalent replacements or corresponding improvements made to the technical features of the present application still fall within the protection scope of the present application. Unless otherwise specified, the reagents used in the following examples are all commercially available products, and the preparation of solutions can adopt conventional techniques in the art.

[0138] Table 1 Abbreviation table

[0139]

[0140]

[0141] "14C12H1L1 injection" refers to a medical preparation for injection containing the 14C12H1L1 monoclonal antibody, which is usually administered to patients via the intravenous infusion route. In a specific embodiment, the expression "14C12H1L1 injection, 200 mg each time" can be understood in the usual way in the art as a liquid medical preparation for injection containing 200 mg of the 14C12H1L1 monoclonal antibody administered to patients each time.

[0142] Example 1 Clinical research protocol - Research criteria and endpoints

[0143] 1.1 Inclusion and exclusion criteria

[0144] Inclusion criteria: Those who meet the following inclusion items can be included in this trial

[0145] 1) Meet all the conditions of any one of the following cohorts:

[0146] Cohort 1:

[0147] a) Subjects with pathologically confirmed unresectable or metastatic biliary tract cancer, including intrahepatic cholangiocarcinoma (IHCC), extrahepatic cholangiocarcinoma (EHCC), and gallbladder cancer (GBC);

[0148] b) Having failed first-line or more than first-line chemotherapy. The definition of failure of first-line or more than first-line chemotherapy is: disease progression during treatment or after the last treatment; or intolerance to toxic and side effects during treatment.

[0149] Cohort 2:

[0150] Pathologically confirmed recurrent or metastatic colorectal cancer that is not suitable for surgical resection and has MSI-H or dMMR. Has not received prior systemic treatment for recurrent or metastatic colorectal cancer.

[0151] Cohort 3:

[0152] Pathologically confirmed metastatic or recurrent gastric or gastroesophageal junction (GEJ) adenocarcinoma. Has not received prior systemic treatment for metastatic or recurrent gastric or gastroesophageal junction (GEJ) adenocarcinoma, with PD-L1 expression ≥ 1% (CPS).

[0153] Cohort 4:

[0154] Pathologically confirmed locally advanced or metastatic urothelial cancer (including those originating from the bladder, ureter, renal pelvis, and urethra) that cannot be completely resected surgically. Has failed at least 1 prior systemic standard chemotherapy regimen. The definition of failure of 1 systemic standard chemotherapy is: disease progression during treatment or after the last treatment, or intolerance to toxic and side effects during treatment.

[0155] Cohort 5:

[0156] a) Subjects with pathologically confirmed low- or intermediate-grade (G1 or G2) advanced (locally advanced unresectable or distant metastasis) gastroenteropancreatic neuroendocrine tumors (NET).

[0157] The definition of low grade (G1) is a mitotic count < 2 / 10 high-power fields [HPF] and / or a Ki-67 proliferation index < 3%; the definition of intermediate grade (G2) is a mitotic count of 2 - 20 / 10 high-power fields [HPF] and / or a Ki-67 proliferation index of 3 - 20%. If the mitotic count and Ki-67 index of the same tumor tissue correspond to different grades, follow the higher grade.

[0158] b) Having received ≥ 1 prior systemic anti-tumor drug treatment for advanced NET: including mTOR inhibitors, chemotherapy, long-acting somatostatin analogs, interferon, PRRT (peptide receptor radionuclide), etc. Advanced treatment-naive patients who cannot accept or refuse the above treatment can also be enrolled;

[0159] c) The patient had evidence of radiologically confirmed disease progression within 12 months before the first administration.

[0160] 2) At least 18 years old; ECOG performance status: 0 - 1; Prognosis of survival more than 3 months;

[0161] 3) There are measurable lesions defined by RECIST 1.1 criteria. If the previously irradiated lesions show definite progression after radiotherapy and this previously irradiated lesion is not the only lesion, then this lesion can be considered as a measurable lesion.

[0162] 4) Major organ functions are normal.

[0163] 5) The patient voluntarily participates in this study, signs the informed consent form and has good compliance.

[0164] Exclusion criteria: Subjects with any of the following conditions will not be enrolled in this study

[0165] 1) If the patient has previously received local radiotherapy, they can be enrolled if the following conditions are met: More than 4 weeks (more than 2 weeks for brain radiotherapy) have passed since the end of radiotherapy until the start of the study treatment; and the target lesions selected for this study are not within the radiotherapy area; or the target lesions are within the radiotherapy area but have been confirmed to have progressed.

[0166] 2) Brain metastases with symptoms or symptom control time less than 2 months.

[0167] 3) The patient had or had other malignancies in the past 5 years. Except for the following two situations: Other malignancies treated by single surgery, achieving continuous 5 - year disease - free survival (DFS); Cured cervical carcinoma in situ, non - melanoma skin cancer, and superficial bladder tumors [Ta (non - invasive tumor), Tis (in situ carcinoma), and T1 (tumor invading the basement membrane)].

[0168] 4) Those with multiple factors affecting oral medications (such as inability to swallow, after gastrointestinal resection, chronic diarrhea, and intestinal obstruction, etc.);

[0169] 5) Imaging (CT or MRI) shows that the tumor invades large blood vessels or the boundary with blood vessels is unclear;

[0170] 6) Received major surgical treatment, incisional biopsy, or obvious traumatic injury within 28 days before the first administration;

[0171] 7) Had an arterial / venous thrombosis event within 6 months before the first administration, such as cerebrovascular accident (including transient ischemic attack), deep vein thrombosis, and pulmonary embolism;

[0172] 8) Those with a history of abuse of psychotropic drugs and unable to quit or with mental disorders;

[0173] 9) Participated in other clinical trials within four weeks;

[0174] 10) Patients with concomitant diseases that, according to the investigator's judgment, seriously endanger patient safety or affect the patient's completion of the study.

[0175] 1.2 Exit Criteria

[0176] 1) Disease progression occurs, and the investigator determines that the subject will not benefit from continued treatment;

[0177] 2) Adverse events occur and cannot be tolerated or relieved;

[0178] 3) Subjects who have experienced serious adverse events and are not suitable to continue participating in the study;

[0179] 4) Those who seriously deviate from or violate the protocol and affect the safety or efficacy evaluation of the drug;

[0180] 5) The subject withdraws informed consent;

[0181] 6) Those who are unable to complete the follow-up on time due to various reasons.

[0182] 1.3 Study Endpoints

[0183] Phase 1: Safe introduction period

[0184] Primary endpoint:

[0185] Safety and tolerability of the first cycle

[0186] Secondary endpoints:

[0187] Objective response rate (ORR), disease control rate (DCR), duration of response (DOR), progression-free survival (PFS), overall survival (OS), etc.

[0188] Phase II: Formal Trial

[0189] Primary End Point

[0190] Objective response rate (ORR)

[0191] Secondary End Points

[0192] Disease control rate (DCR), duration of response (DOR), progression-free survival (PFS), overall survival (OS), etc.

[0193] The incidence and severity of adverse events (AEs) and serious adverse events (SAEs), as well as abnormal laboratory test indicators.

[0194] Example 2. Clinical trial design

[0195] A single-arm, open-label, multi-cohort, multi-center phase II clinical trial was adopted.

[0196] 2.1 Sample size

[0197] There are a total of 5 cohorts in this study, with 20 - 30 cases included in each cohort (adjusted according to specific trial results).

[0198] 2.2 Imaging evaluation design

[0199] The primary efficacy endpoint of this study is ORR, and the results evaluated by the researchers at each research center are adopted. In this study, an independent imaging group is additionally set up to conduct a review of the imaging efficacy evaluation.

[0200] 2.3 Administration plan design

[0201] The subjects of administration are all patients recruited in Cohort 1 to Cohort 5 in Example 1.

[0202] The study is divided into two phases:

[0203] The first phase is a single-arm study, which is a safety lead-in period.

[0204] Patients will receive anlotinib combined with 14C12H1L1. Every 21 days is a treatment cycle. After collecting safety information to determine the RP2D of anlotinib combined with 14C12H1L1 in patients with digestive tract tumors, urinary system tumors, and neuroendocrine tumors, a formal trial will be carried out. The lead-in period includes the following 2 dose groups:

[0205] Dose group A: Anlotinib 10mg D1 - D14 combined with 14C12H1L1 200mg once every 3 weeks

[0206] Dose group B: Anlotinib 12mg D1 - D14 combined with 14C12H1L1 200mg once every 3 weeks

[0207] It is planned to enroll 3 - 6 patients in each dose group to observe the safety of the combined medication. Based on the safety data of the large sample clinical study of anlotinib capsules in the past, the dose exploration will start from the dosing plan of dose group A in the lead-in period (if the 3 - 6 patients enrolled in dose group A have good tolerance in the first cycle, 3 - 6 patients in dose group B will be explored). After the lead-in trial ends, it will be discussed by the principal investigator and the sponsor to determine the RP2D of anlotinib combined with 14C12H1L1 in patients with digestive tract tumors, urinary system tumors, and neuroendocrine tumors.

[0208] The second phase is a formal trial, and patients will receive anlotinib combined with 14C12H1L1 treatment:

[0209] Anlotinib hydrochloride capsules:

[0210] Once daily (it is recommended to take on an empty stomach within ±60 minutes before or after the start of the infusion of 14C12H1L1 injection), 12 mg each time (the final dose is determined during the safety lead-in period, tentatively 12 mg). Take orally continuously for 2 weeks and then stop for 1 week, that is, 3 weeks (21 days) is one treatment cycle. If a dose is missed during the medication period, and it is confirmed that the time to the next dose is less than 12 hours, do not take a supplementary dose.

[0211] 14C12H1L1 injection:

[0212] Administer once every 3 weeks, 200 mg each time, by intravenous infusion. The infusion time is 60 ± 10 minutes.

[0213] Medication cycle:

[0214] Each 21 days is one treatment cycle. Other anti-tumor treatments cannot be carried out during the medication period; for patients with disease control (CR + PR + SD) and tolerable adverse reactions, continue the medication until disease progression or intolerance.

[0215] Evaluate the efficacy once every 2 cycles, and once every 4 cycles after 16 cycles. For patients with disease control (CR + PR + SD) and tolerable adverse reactions, the medication can be continued until the loss of clinical benefit, intolerable toxicity, the efficacy evaluation is PD, or the researcher deems it inappropriate to continue the medication, then the study ends.

[0216] Dose adjustment of Anlotinib Hydrochloride Capsules:

[0217] During the study, the dose of Anlotinib Hydrochloride can be down-regulated due to drug-related adverse events in patients (down-regulated in sequence of 12 mg - 10 mg - 8 mg, cross-dose adjustment is not allowed). For patients who cannot tolerate at the 8 mg dose level, the study needs to be terminated. For patients with Anlotinib Hydrochloride Capsules whose doses have been down-regulated to 10 mg or 8 mg, after taking the medication for some time, if the researcher determines that there may be disease progression and the patient's safety is stable, the dose can be up-regulated once. Each patient can only have one dose up-regulation at most, and cross-dose group adjustment is not allowed.

[0218] 14C12H1L1 injection has a fixed dose and no dose down-regulation plan.

[0219] 2.4 Method of combined administration

[0220] 14C12H1L1 injection:

[0221] Dilute 200 mg with normal saline to 100 mL, and the infusion time is 60 ± 10 min. Administer the drug once every 21 days. The infusion time starts from the start of the infusion of 14C12H1L1 and ends when the infusion of 14C12H1L1 is completed and the normal saline flushing of the tube is completed (it is recommended to flush the tube with 20 mL of normal saline). For subjects who cannot tolerate a 60-min infusion, the infusion time can be extended up to 120 minutes (±15 minutes).

[0222] Anlotinib Hydrochloride Capsules:

[0223] Take orally once a day (on an empty stomach before breakfast), 1 capsule (12 mg) each time (the final dose is determined by the safety lead-in period, tentatively 12 mg), and it is recommended to take it on an empty stomach within ±60 min of the start of the infusion of 14C12H1L1 injection. Take orally continuously for 2 weeks and then stop for 1 week, that is, 3 weeks (21 days) is a treatment cycle. Under normal circumstances, it is recommended to take the medicine at a fixed time every day. On the day of blood collection or safety inspection, take the medicine after blood collection. If a dose is missed during the medication period, and it is confirmed that the time until the next dose is less than 12 hours, do not take a supplementary dose.

[0224] Medication cycle:

[0225] Each 21 days is a treatment cycle, and other anti-tumor treatments cannot be carried out during the medication period; for patients with disease control (CR + PR + SD) and tolerable adverse reactions, continue the medication until the disease progresses or cannot be tolerated.

[0226] The subjects to be administered the drug are all patients recruited from cohort 1 to cohort 5 of Example 1.

[0227] 2.5 Delayed administration and dose adjustment

[0228] If the adverse events caused by the 14C12H1L1 injection result in delayed medication and the medication cannot be resumed after more than 12 weeks, the treatment with the 14C12H1L1 injection should be permanently terminated. For the treatment suggestions for the adverse events caused by the 14C12H1L1 injection, refer to "Treatment Suggestions for Immune-Related Adverse Events Caused by Immune Checkpoint Inhibitors". The 14C12H1L1 injection allows delayed administration without dose adjustment.

[0229] During each cycle of Anlotinib Hydrochloride Capsules, if an adverse reaction related to anlotinib occurs during the medication period (days 1 - 14) and dose delay is required, the maximum delay time shall not exceed 5 days. If the patient is still unable to continue taking the medication after 5 days, the remaining anlotinib for this cycle shall no longer be used. When starting the next cycle of medication, if dose delay is still possible due to adverse reactions, the maximum delay time shall not exceed 2 weeks. For patients who are still unable to take anlotinib (including dose reduction) after 2 weeks, anlotinib shall be permanently discontinued (except for events not caused by safety reasons). During this study, if necessary, Anlotinib Hydrochloride Capsules can be delayed or the dose can be adjusted.

[0230] During the study, when an adverse event related to anlotinib occurs, dose reduction is allowed: 12 mg → 10 mg and 10 mg → 8 mg. If the patient is still unable to tolerate the drug at the 8 - mg level, the medication shall be terminated. After dose reduction for some time, when considering the possible progression of the disease and controllable safety, dose increase to the original level is allowed starting from the next cycle.

[0231] 2.6 Disease Progression and Efficacy Evaluation

[0232] During the clinical use of drugs similar to 14C12H1L1 injection, pseudo - progression occurred in the subjects. The efficacy evaluation criteria for this study are based on RECIST 1.1 (Response Evaluation Criteria in Solid Tumors). At the same time, the iRECIST standard (Evaluation Criteria Related to Cancer Immunotherapy) is used to confirm the efficacy. That is, for subjects determined to have disease progression (PD) according to the RECIST 1.1 standard, further confirmation is carried out according to the iRECIST standard to decide whether to continue drug observation.

[0233] Starting from the first day of the first cycle, the efficacy is evaluated every 2 cycles. After 16 cycles, the efficacy is evaluated every 4 cycles until the subject develops disease progression confirmed by tumor imaging. This evaluation will not change the evaluation frequency due to the subject's treatment delay or interruption. If the subject terminates the study treatment for reasons other than disease progression, the subject still needs to continue to receive tumor imaging evaluations at the above - mentioned frequency until the subject starts a new anti - tumor treatment, or there is disease progression evidenced by imaging, or the subject withdraws voluntarily, or the subject dies, whichever occurs first.

[0234] The imaging evaluation methods for tumors can use CT or MRI, but the evaluation methods, machines, and technical parameters should be consistent throughout the study period; if there are no contraindications, contrast agents should be used. If tumor evaluation has been performed within 14 days before the first dose and the same method is used in the same hospital, it can be used as the baseline tumor evaluation. The baseline tumor evaluation should include CT or MRI of the chest, abdomen, and pelvis. A plain scan + enhanced / enhanced MRI of the brain should be performed during the screening period. Imaging examinations should be performed on all suspicious lesion sites. For patients with bone metastases, bone scans should be used to follow up the lesions. For patients with bone metastases, if there is no aggravation of clinical symptoms, there is no need to recheck during each tumor assessment. If there is aggravation of clinical symptoms, reexamination should be performed in a timely manner. For cases suspected of disease progression before the start of the planned next evaluation, an unplanned tumor assessment should be performed. During the trial, imaging examinations should only be performed on the lesion sites. If there are suspicious sites during the period, corresponding site imaging can be added for examination.

[0235] Example 3. Collection of biological samples

[0236] 3.1 Detection of serum anti-14C12H1L1 antibody (ADA)

[0237] The time points for immunogenicity monitoring are based on the administration time of 14C12H1L1 injection; when 14C12H1L1 is administered with a delay, the immunogenicity blood sampling is correspondingly delayed. If the ADA of the subject is detected to be positive, neutralizing antibodies are additionally detected.

[0238] Samples are collected before dosing (-60 min) at the 1st, 2nd, 4th, 8th cycles and every 6 cycles thereafter. At the same time, samples are collected 30 min (±5 min) after the infusion ends at the 1st cycle and the 8th cycle, and 30 days (±7 days) and 90 days (±7 days) after the last dose. Each time, 5 mL of venous blood is required and placed in a blood collection tube containing clot-promoting separating gel. After standing at room temperature for 30 min and natural coagulation, centrifuge at 3000 g for 10 min, and evenly divide and transfer into 4 cryotubes (3 test tubes and 1 backup tube, each test tube not less than 0.5 mL). Take the serum, label it, and store it in a refrigerator at -40 to -80 °C for detecting immunogenicity and the blood drug concentration of 14C12H1L1.

[0239] During the trial, if an unexpected immune-related adverse event occurs, an additional blood sampling is required in a timely manner after the adverse event is confirmed to detect immunogenicity and the blood drug concentration of 14C12H1L1. However, if the time since the most recent blood sampling is less than 24 h, sampling can be omitted.

[0240] Note: According to the ADA results, neutralizing antibodies may be additionally detected if necessary in the follow-up.

[0241] 3.2 Biomarker determination

[0242] For this study, tumor tissue specimens are required for biomarker research, including detection of PD-L1 expression, mismatch repair / microsatellite instability (MMR / MSI), etc.

[0243] For samples for biomarker detection, fresh biopsy tissue samples within 1 month before enrollment are preferred. When taking fresh tissue samples, percutaneous puncture is performed with 1 or more needles. If fresh biopsy tissue samples cannot be obtained, archived tissue samples can be collected and used. Take 10 unstained pathological tissue sections (anti - detachment) with a thickness of 4 - 6 μm, which are freshly cut and prepared white slides. If they cannot be sent out in time (within one month), wax sealing treatment is required.

[0244] Example 4. Efficacy and safety evaluation

[0245] 4.1. Analysis of primary efficacy indicators

[0246] 4.1.1. Objective response rate (ORR):

[0247] Calculate the ratio of the number of objective response cases (PR + CR) to the total number of cases in each group and its 95% CI. The 95% CI of ORR is calculated based on the exact binomial method of the F distribution.

[0248] 4.1.2. Analysis of secondary efficacy indicators

[0249] 4.1.2.1. Progression - free survival (PFS)

[0250] Estimate the median PFS using the Kaplan - Meier method and draw a survival curve.

[0251] 4.1.2.2. Overall survival (OS)

[0252] Estimate the median OS using the Kaplan - Meier method and draw a survival curve.

[0253] 4.1.2.3. Duration of response (DOR)

[0254] Estimate the median PFS and its 95% CI using the Kaplan - Meier method and draw a survival curve.

[0255] 4.1.2.4. Disease control rate (DCR):

[0256] Calculate the ratio of the number of disease control cases (CR + PR + SD) to the total number of cases and its 95% CI. The 95% CI of DCR is calculated based on the exact binomial method of the F distribution.

[0257] 4.2. Safety evaluation

[0258] 4.2.1. Drug Exposure and Compliance

[0259] The drug exposure was described by mean, standard deviation, maximum value, minimum value, and median.

[0260] Summarize the situation of subjects exposed to the study drug treatment, the number of cycles completed by patients, the dose adjustment during treatment, the cumulative number of dose adjustments during treatment, etc.

[0261] Statistically describe the treatment time of the study drug during the treatment period, the total dose of the study drug taken, the daily average dose, and the dose compliance of the study drug.

[0262] The dose compliance of the study drug will be calculated based on the total actual dose of the study drug taken every day recorded in the eCRF and the total dose of the study drug specified in the protocol.

[0263] One-way ANOVA was used to compare the treatment time of each study drug, the total dose of the study drug taken, the daily average dose, and the dose compliance of the study drug. Chi-square test or Fisher's exact probability method was used to compare the compliance classification.

[0264] 4.2.2. Adverse Events

[0265] Summarize the number of cases, number of patients, and incidence rate of adverse events, adverse events before the first dose, adverse events during treatment, unexpected adverse events during treatment, serious adverse events during treatment, adverse events of special concern during treatment, grade 3 and above adverse events during treatment, serious adverse events during treatment, adverse events related to the study drug during treatment, SAEs related to the study drug during treatment, adverse events that led to dose adjustment, permanent treatment discontinuation, trial termination, and patient death during treatment, and summarize them according to SOC and PT.

[0266] Summarize the adverse events and drug-related adverse events with an incidence rate ≥ 5% during the treatment period according to PT classification.

[0267] Summarize the drug-related adverse events with CTC AE grade 3 or 4 during the treatment period according to PT classification.

[0268] Summarize the adverse events and drug-related adverse events with an incidence rate ≥ 10% during the treatment period according to PT classification.

[0269] Median time to first occurrence of adverse events of special concern.

[0270] 4.2.3. Vital Signs

[0271] The measured values and change values before and after treatment were described by mean ± standard deviation, maximum value, minimum value, and median.

[0272] 4.2.4. Laboratory test indicators

[0273] Table 2 List of examination items

[0274]

[0275] For blood routine, blood biochemistry, thyroid function, coagulation function, amylase, and lipase, the measured values and change values before and after treatment were described using mean ± standard deviation, maximum value, minimum value, and median. Paired t-tests were used for within-group comparisons. The changes in normal and abnormal conditions before and after treatment were described using cross-classification tables.

[0276] Urine routine: The changes in normal and abnormal conditions before and after treatment were described using cross-classification tables.

[0277] Stool routine: The changes in normal and abnormal conditions before and after treatment were described using cross-classification tables.

[0278] Describe the proportion of "abnormal and clinically significant" among the subjects with abnormal changes, where the clinical significance of the abnormality was determined by the researcher.

[0279] 4.2.5. Electrocardiogram

[0280] Electrocardiogram: Based on the normality and abnormality determined by the researcher, the changes in normal and abnormal conditions before and after treatment were described.

[0281] For heart rate, PR interval, QRS interval, QT interval, and QTc, the measured values and change values before and after medication were described using mean ± standard deviation, maximum value, minimum value, and median. The overall evaluation results of the electrocardiogram were described using cross-classification tables for the changes in normal and abnormal conditions before and after medication. Describe the proportion of "clinically significant abnormalities" among the subjects with abnormal changes, where the clinical significance of the abnormality was determined by the researcher. List the abnormal list after administration.

[0282] 4.2.6. Physical examination

[0283] Describe the changes in normal and abnormal conditions before and after treatment.

[0284] Example 5. Clinical application

[0285] 1. Patient: c01012 - urothelial carcinoma

[0286] Past medical history: A 63-year-old female. On May 14, 2019, the patient underwent a lower abdominal CT, showing dilatation of the left ureter. Further CTU of the urinary system was performed, revealing a soft tissue density mass (about 16*20 mm) in the lower segment of the left ureter, suggesting possible ureteral cancer; left hydronephrosis. On May 29, 2019, laparoscopic radical resection of left ureteral cancer + ureteroscopy + renal adhesion lysis were performed. Postoperative pathology: (left kidney + left ureter + part of the bladder) high-grade urothelial carcinoma, with local squamous differentiation, and mucoid stroma in some areas, infiltrating the extra-muscular fibrosa, with the deepest infiltration of 0.75 cm. From May 31 to July 18, 2019, epirubicin bladder perfusion was performed seven times (specific volume unknown), and there were no chemotherapy contraindications. From August 14 to November 29, 2019, adjuvant chemotherapy with gemcitabine + cisplatin regimen was performed for 6 cycles after surgery. Specifically: gemcitabine 1.0 g d1, 8 Ivgtt, cisplatin 30 mg d2-4 Ivgtt, with a 21-day cycle. On August 15, 2020, chest and abdomen CT plain scan: The chest was symmetric, and a soft tissue-like shadow with a diameter of about 45 cm was seen at the left thoracic inlet; the transparency of both lung fields was slightly enhanced, the bronchial wall was thickened, and there were cord-like shadows, micronodules and small nodules in both lung fields. The largest one was located in the middle lobe of the right lung, about 10*7.5 mm, which was a mixed ground-glass nodule with a little solid shadow inside; multiple cord-like shadows were seen in both lungs. CT plain scan of the whole abdomen: The liver was normal in size and shape, and several slightly low-density shadows with a diameter of less than 17 mm were seen. Several soft tissue-like nodular shadows with a diameter of 20 mm were seen in the retroperitoneum, with rough edges. The left acetabulum was shallow, the shape of the left femoral head and neck was irregular, and the left femur was displaced upward; the soft tissues around the left hip joint were swollen. Metastasis was considered. On August 24, 2020, pathological examination of the puncture of the left neck mass showed urothelial carcinoma metastasis.

[0287] On September 10, 2020, treatment with oral anlotinib hydrochloride capsules at a dose of 12 mg once a day was started, taking for 2 weeks and then stopping for 1 week. 14C12H1L1: Administered once every 3 weeks, 200 mg per dose, by intravenous infusion. Each 3-week period was one treatment cycle until disease progression or intolerance. The patient had good tolerance during the medication period and could continue the medication.

[0288] Pathology report: On September 2, 2020, left neck puncture pathology: High-grade urothelial carcinoma.

[0289] Dosage: Anlotinib hydrochloride: 12 mg, 14C12H1L1: 200 mg

[0290] Efficacy evaluation:

[0291] Screening period: Target lesions: 68 mm; Non-target lesions: None.

[0292] After the second cycle of drug administration: Target lesions: 27 mm; Non-target lesions: None.

[0293] 2. Patient: c01012 - Rectal cancer

[0294] Past medical history: A 44-year-old male. On February 10, 2018, colonoscopy showed rectal cancer; on February 13, 2018, pathological diagnosis was highly suspected adenocarcinoma, poorly differentiated (rectum). On February 24, 2018, imaging diagnosis by pelvic rectal MRI showed cancer in the middle and lower segments of the rectum, infiltrating the outer membrane surface and invading the prostate. On February 26, 2018, endoscopic diagnosis by colonoscopy: rectal cancer (about 4 cm from the anal verge), involving the anal canal. Imaging diagnosis by enhanced CT of chest, abdomen and pelvis: cancer in the middle and lower segments of the rectum, penetrating the outer membrane surface, invading the prostate, multiple low-density lesions in the liver, considered metastatic tumors. Nodule in the lower lobe of the left lung, highly vigilant for lung metastasis. On February 28, 2018, biopsy pathology showed neuroendocrine tumor with necrosis, low Ki-67 proliferation index, consistent with NET G2, Ki-67 (15%), SSTR2 (3+), PD-L (+, 5%). On March 5, 2018, imaging diagnosis by abdominal MRI showed multiple metastatic tumors in the liver. From March 11, 2018 to the end of August 2018, 6 cycles of chemotherapy were performed. The specific regimen was: Tegafur 60 mg Bid d1 - 14; Temozolomide 250 mg po d10 - 14; repeated every 21 days. From April 26, 2018 to July 25, 2018, Octreotide acetate microsphere 20 mg was given by intramuscular injection once a month; CT reexamination in September 2018 showed effective treatment. From November 2018 to September 2019, single-agent S1 maintenance treatment was performed for 6 cycles. The specific regimen was: Tegafur 60 mg po bid d1 - 14; repeated every 21 days. During this period, traditional Chinese medicine was taken orally intermittently for conditioning. The efficacy evaluation by CT after chemotherapy was stable. Enhanced CT on May 25, 2020 showed: a small nodule in the lingular segment of the upper lobe of the left lung, about 0.4 cm, enlarged compared with before. Abdominal MRI on June 2, 2020 showed: multiple metastatic tumors in the liver, enlarged compared with before, and the largest one was located in the lower segment of the right posterior lobe of the liver. There were also scattered small abnormal signal nodules in the liver, increased compared with before, indicating tumor progression.

[0295] On June 19, 2020, oral administration of Anlotinib Hydrochloride Capsules 10 mg once a day was started, taken continuously for 2 weeks and stopped for 1 week. 14C12H1L1: administered once every 3 weeks, 200 mg / time, by intravenous infusion. Each 3-week period was one treatment cycle until disease progression or intolerance.

[0296] Pathology report: Biopsy pathology on February 28, 2018: neuroendocrine tumor with necrosis, low Ki-67 proliferation index, consistent with NET G2, Ki-67 (15%), SSTR2 (3+), PD-L (+, 5%).

[0297] Dosage: Anlotinib Hydrochloride: 10 mg, 14C12H1L1: 200 mg

[0298] Efficacy evaluation:

[0299] Screening period: Target lesion: 42 mm; Non-target lesions: Multiple liver metastases, lung metastases, rectal lesions.

[0300] After administration in the 2nd cycle: Target lesion: 40 mm; Non-target lesions: NCR / NPD.

[0301] After administration in the 4th cycle: Target lesion: 38 mm; Non-target lesions: NCR / NPD.

[0302] 3. Patient: c01007 - Poorly differentiated cholangiocarcinoma / biliary tract cancer

[0303] Past medical history: 65-year-old male. MRI on May 31, 2019: A mass at the junction of the left and right lobes of the liver, suspected to be malignant cholangiocarcinoma. Hepatic segment V + partial segment IV + cholecystectomy was performed on June 11, 2019. The postoperative pathological diagnosis on June 12, 2019 was poorly differentiated cholangiocarcinoma. From August 20, 2019 to October 26, 2019, 3 cycles of oxaliplatin combined with capecitabine chemotherapy regimen were administered. Specific regimen: Oxaliplatin 190 mg, capecitabine 1000 g bid po d1 - 14, q21d. Enhanced chest, abdomen and pelvis CT on July 20, 2020 showed: Multiple small nodules in the liver, multiple enlarged lymph nodes in the abdominal cavity and retroperitoneum, multiple bone changes in the spine and pelvis.

[0304] On August 7, 2020, treatment with oral anlotinib hydrochloride capsules 10 mg once daily was started, taken continuously for 2 weeks and stopped for 1 week. 14C12H1L1: Administered once every 3 weeks, 200 mg / time, by intravenous infusion. Each 3-week period is one treatment cycle until disease progression or intolerance.

[0305] Pathology report: Postoperative pathology on June 12, 2019: Poorly differentiated cholangiocarcinoma (partially showing spindle cell morphology, about 30%), with focal necrosis and a large amount of lymphocyte infiltration. The maximum tumor diameter was 3.6 cm, and vascular tumor thrombus was visible. The tumor did not involve the liver capsule. The hepatic basal resection margin was cancer-free. The portal area of the surrounding liver tissue was accompanied by significant inflammatory cell infiltration. Gallbladder adenomyoma, with active epithelial hyperplasia and glandular cystic dilation. Metastatic carcinoma in lymph nodes 2 / 4, without involvement outside the lymph node capsule. Gastric left vascular paracardial lymph node 1 / 1, lymph nodes in groups 12 and 13 1 / 1, lymph nodes in groups 7, 8, 9 0 / 1, lymph nodes in group 12P 0 / 1, staging: PT2N1. Immunohistochemical results showed: CK18(2+), CK19(3+), CK7(focal 1+), GPC3(-), Hepatpocyte(-), AFP(-), CA19.9(1+), CD34(vascular endothelium+), Ki-67(+, 60%). In-situ hybridization results showed: EBER(-).

[0306] Dosage: Anlotinib Hydrochloride: 10 mg, 14C12H1L1: 200 mg

[0307] Efficacy evaluation:

[0308] Screening period: Target lesion: 68 mm; Non-target lesions: Multiple lymph node metastases; Liver metastasis.

[0309] After administration in the 2nd cycle: Target lesion: 53 mm; Non-target lesions: NCR / NPD.

[0310] 4. Patient: c01010 - Invasive urothelial carcinoma

[0311] Medical History: A 69-year-old male underwent a CT scan in May 2012, which revealed: 1. Hyperdensity of the lower left ureter with left hydronephrosis and dilatation of the mid- and upper ureter, suggesting a high likelihood of a mass. 2. Poor perfusion and excretion of the left kidney. 3. Multiple cysts in the left kidney. Ureteral tumor was considered an option, but urine cytology revealed suspicious cancer cells. He underwent a full-length left ureterectomy and left nephrectomy in May 2012. Postoperative pathology revealed left ureteral transitional cell carcinoma. Biopsy on March 17, 2013, revealed a papillary urothelial tumor of low malignant potential. On March 21, 2013, he underwent transurethral resection of the bladder tumor. Postoperative pathology revealed grade II transitional cell carcinoma. On March 1, 2014, he underwent transurethral resection of the bladder tumor. Pathology revealed grade II transitional cell carcinoma. Postoperative intravesical chemotherapy with pirarubicin was administered. On May 17, 2014, pathology revealed a small amount of severely degenerated, compressed, and deformed tumor tissue. On May 24, 2014, a transurethral resection of the bladder tumor was performed, followed by intravesical chemotherapy with pirarubicin. On September 7, 2016, a CT scan revealed multiple lesions on the posterior bladder wall. Cystoscopic pathology (in the trigone) suggested cystitis glandularis. On February 20, 2017, a CT scan revealed postoperative changes in the left kidney, including right hydronephrosis, right ureteral dilatation, and a space-occupying lesion on the posterior bladder wall, involving the right ureteral bladder entrance. On February 27, 2017, a transurethral resection of the bladder tumor with right DJ tube placement was performed. Postoperative pathology revealed high-grade invasive urothelial carcinoma in the right ureteral opening, trigone, and posterior bladder wall. A whole-body bone scan on May 22, 2019, concluded that multiple bone metastases were present throughout the body. From June 19, 2019, to December 14, 2019, the patient received six cycles of systemic chemotherapy with gemcitabine and docetaxel, followed by two cycles of oral maintenance chemotherapy with tegafur, and zoledronic acid for bone metastasis. The best outcome was SD. On January 4, 2020, imaging studies showed stable disease, and blood tumor markers increased compared to the previous period, suggesting tumor progression. From January 7, 2020, to January 14, 2020, the patient received one cycle of chemotherapy with gemcitabine and docetaxel, along with zoledronic acid for bone metastasis. On the sixth day of chemotherapy, a routine blood test revealed leukopenia and thrombocytopenia, leading to a supplementary diagnosis of bone marrow suppression. Chemotherapy was discontinued, and treatment was initiated to increase leukocyte and platelet counts. From February 13, 2020, to July 28, 2020, the patient received 120 mg of oral tegafur on days 2-14, along with four cycles of zoledronic acid. The best outcome was SD. CT report from July 13, 2020: Postoperative changes after left ureteral tumor surgery and right abdominal fistula surgery. Multiple right kidney cysts are likely, similar to previous findings. Multiple small lymph nodes are present in both groins, similar to previous findings. Multiple cords and micronodules are present in both lungs, similar to previous findings. Emphysema is present. Multiple calcifications are present in the aortic and coronary artery walls. Bilateral pleural thickening is present. Abnormal bone density on the scan planes suggests the possibility of metastasis.

[0312] On September 1, 2020, treatment with oral anlotinib hydrochloride capsules at a dose of 12 mg once daily was initiated. It was administered for 2 consecutive weeks followed by a 1-week break. For 14C12H1L1, it was administered once every 3 weeks at a dose of 200 mg per time by intravenous infusion. Each 3-week period was considered one treatment cycle until disease progression or intolerance occurred.

[0313] Pathology report: Postoperative pathology on July 27, 2017: (right ureteral orifice, trigone of bladder, posterior wall of bladder) were all high-grade invasive urothelial carcinoma.

[0314] Dosage: Anlotinib hydrochloride: 12 mg, 14C12H1L1: 200 mg

[0315] Efficacy evaluation:

[0316] Screening period: Target lesions: 10 mm; Non-target lesions: bone metastasis lesions, pulmonary nodules.

[0317] After the second cycle of administration: Target lesions: 7.4 mm; Non-target lesions: NCR / NPD.

[0318] 5. Patient: c01014 - moderately differentiated adenocarcinoma of the distal common bile duct / cholangiocarcinoma

[0319] Past medical history: A 72-year-old female. In December 2017, abdominal B-ultrasound and laboratory tests showed a mass in the lower bile duct and obstructive jaundice. PTCD for jaundice reduction was performed on December 27, 2017. After puncture and drainage, liver function and jaundice indicators improved significantly. "Pancreatoduodenectomy" was performed on January 9, 2018, with a smooth operation process and good postoperative recovery. Postoperative pathology on January 10, 2018 showed moderately differentiated adenocarcinoma of the distal common bile duct, and no postoperative chemotherapy or radiotherapy was given. In March 2020, upper abdominal discomfort recurred, and CT showed a mass in the gallbladder fossa. From April 15, 2020 to July 29, 2020, 10 cycles of nab-paclitaxel + capecitabine were given systemically. The specific regimen was: nab-paclitaxel 100 mg on day 1 every 21 days + capecitabine 2.5 g on days 1 - 7 every 21 days, without serious adverse reactions. The best efficacy was SD. Enhanced CT of the whole abdomen and pelvis on August 13, 2020 showed: 1. Changes after "cholangiocarcinoma resection and cholecystectomy": Multiple lymph nodes of varying sizes beside the superior mesenteric artery, considered metastatic, enlarged compared with the previous film, with necrosis in the lesions, please follow up in combination with clinical findings. 2. Circumferential thickening and enhancement of the common hepatic duct wall: Inflammatory lesion. 3. Small cyst in the right lobe of the liver. 4. Encapsulated fluid collection in front of the right renal anterior fascia, smaller compared with the previous film. 5. Hemangioma of the first lumbar vertebra. CT indicated progression.

[0320] On September 18, 2020, treatment with oral Anlotinib Hydrochloride Capsules at a dose of 12 mg once daily was initiated, administered for 2 consecutive weeks followed by a 1-week break. 14C12H1L1: Administered once every 3 weeks, 200 mg per dose, by intravenous infusion. Each 3-week period was considered one treatment cycle, continuing until disease progression or intolerance. The patient had good overall tolerance during medication, and no grade III or above adverse events occurred, allowing continued medication.

[0321] Pathology report: Postoperative pathology on January 10, 2018: Moderately differentiated adenocarcinoma at the terminal common bile duct, tumor size approximately 1.5x1x0.8 cm; carcinoma invaded the entire thickness of the bile duct wall and extended to the pancreas and duodenal wall; no definite intravascular cancer thrombus was seen, but nerve invasion was observed; cancer was found in the peripancreatic lymph nodes (1 / 4); no cancer was found in the perigastric lymph nodes (0 / 7); no cancer was seen at the resection margin; no cancer was found in the lymph nodes of group 8 (0 / 1); no cancer was found in the gallbladder; the pathological stage of the tumor was pT3N1. Immunohistochemical results: CDX-2 (-), cK19 (+), CK20 (+), CK7 (+), MUC-1 (-), MUC-2 (-), KI-67 (hot area 80% +).

[0322] Dosage: Anlotinib Hydrochloride: 12 mg, 14C12H1L1: 200 mg

[0323] Efficacy evaluation:

[0324] Screening period: Target lesions: 69 mm; Non-target lesions: None.

[0325] After administration in the 2nd cycle: Target lesions: 59 mm; Non-target lesions: None.

[0326] 6. Patient: c01013 - Metastatic colorectal cancer

[0327] Previous medical history: A 27-year-old female. On March 12, 2019, the patient had abdominal pain and diarrhea for one and a half months and fever for one week. Pathological diagnosis: (Sigmoid colon) adenocarcinoma. On March 21, 2019, adhesiolysis, anterior resection of rectal cancer, and pelvic abscess drainage were performed, with good recovery; postoperative pathological diagnosis on March 27, 2019: (Rectal) ulcerative mucinous adenocarcinoma invaded the extraperitoneal soft tissue, no nerve invasion or intravascular cancer thrombus was seen; CT plain scan on July 28, 2020: Multiple small nodules in both lungs, a small cyst in the right lobe of the liver, and a pelvic mass. Pelvic MRI on July 31, 2020: 1. Pelvic space-occupying lesion 2. Pelvic effusion. On August 6, 2020, laparotomy + adhesiolysis + left ovarian cystectomy + pelvic mass biopsy were performed, and postoperative pathology indicated metastatic colorectal cancer.

[0328] On September 11, 2020, treatment with oral anlotinib hydrochloride capsules at a dose of 12 mg once daily was initiated. The treatment was administered for 2 weeks followed by a 1-week break. 14C12H1L1 was administered once every 3 weeks at a dose of 200 mg per administration by intravenous infusion. Each 3-week period was considered one treatment cycle, and the treatment continued until disease progression or intolerance. The patient had good overall tolerance during the medication period and could continue the treatment.

[0329] Pathology report: Postoperative pathology on August 6, 2020: (Pelvic mass) metastatic colorectal cancer. Immunohistochemistry showed CK7 and Pax-8 positive, suggesting Lynch syndrome. Examination of the uterus and ovaries was recommended. Immunohistochemistry results: CK20(+), CK7 (focal +), PAX-8 (focal +), Ki-67 (+, 80%), CDX2(+), Villin(+), ER(++), PR(++).

[0330] Dosage: Anlotinib hydrochloride: 12 mg, 14C12H1L1: 200 mg

[0331] Efficacy evaluation:

[0332] Screening period: Target lesions: 75 mm; Non-target lesions: None.

[0333] Unplanned on September 27, 2020: Target lesions: 67 mm; Newly developed lesion: Right adnexal mass 42 mm; Overall response assessment: IUPD.

[0334] After administration in the second cycle: Target lesions: 49 mm; Newly developed lesion: Right adnexal mass 42 mm; Overall response assessment: IUPD.

[0335] 7. Patient: S06001 / C06001 - moderately differentiated cholangiocarcinoma / biliary tract cancer

[0336] In September 2017, the patient developed right upper abdominal pain without obvious cause. On February 23, 2018, a chest CT showed multiple nodules in both lungs, considered to be metastatic tumors, and a space-occupying lesion in the liver. On February 24, 2018, an upper abdominal CT showed findings consistent with a giant mass in the right lobe of the liver with intrahepatic, intraperitoneal, and retroperitoneal lymph node metastases. On March 8, 2018, the pathological results showed: moderately differentiated adenocarcinoma (right lobe of the liver). Combined with immunohistochemistry, it was consistent with moderately differentiated cholangiocarcinoma. Subsequently, the patient received gemcitabine + oxaliplatin chemotherapy multiple times. On May 5, 2019, a contrast-enhanced CT of the chest + whole abdomen showed: 1. Multiple nodular lesions in both lungs, all considered to be metastatic tumors; 2. Fibrous foci in the lower lobes of both lungs; 3. Findings consistent with liver tumors with multiple retroperitoneal lymph node metastases; cancer thrombus in the right branch of the portal vein; tortuous and thickened gastroesophageal varices. On May 13, 2019, the patient underwent transcatheter arterial chemoembolization (drug-loaded microspheres) + splenic artery embolization under local anesthesia in the catheterization laboratory. The patient was discharged without obvious discomfort after the operation. On June 12, 2019, a plain chest + contrast-enhanced CT of the whole abdomen showed: 1. Findings consistent with multiple metastatic tumors in both lungs, similar to the previous ones; 2. Fibrous foci in the lower lobes of both lungs; 3. Findings consistent with changes after liver tumor treatment, obvious necrosis of the lesions, scattered small enhanced areas; cancer thrombus in the right branch of the portal vein; 4. Varices around the cardia and left ovary; 5. Splenic infarction; 6. Considered uterine fibroids. On April 20, 2020, a CT showed: 1. Findings consistent with multiple metastatic tumors in both lungs, worse than the previous film; 2. Fibrous foci in the lower lobes of both lungs; 3. Findings consistent with changes after liver tumor treatment, the range seemed to increase compared with the previous film, multiple intraperitoneal and retroperitoneal lymph node metastases; cancer thrombus in the right branch of the portal vein; 4. Varices around the cardia; 5. Splenic infarction. On April 22, 2020, the patient underwent bronchial artery chemoinfusion + transcatheter arterial chemoembolization (drug-loaded microspheres), and the patient recovered well after the operation.

[0337] Dosage: Anlotinib Hydrochloride: 10 mg, 14C12H1L1: 200 mg

[0338] First medication time: Starting from June 18, 2020, the patient took anlotinib hydrochloride capsules 12 mg orally once a day for 2 weeks and then stopped for 1 week. 14C12H1L1 was administered once every 3 weeks, 200 mg per time, by intravenous infusion. Each 3-week period was one treatment cycle until disease progression or intolerance. The patient had good overall tolerance during the medication period and could continue the medication.

[0339] Efficacy evaluation: Screening period: Target lesions: 214.2 mm; Non-target lesions: Multiple metastases in the lungs, peritoneum, and liver

[0340] After the second cycle of drug administration: Target lesions: 183.5 mm; Non-target lesions: Present, no progression

[0341] After the fourth cycle of drug administration: Target lesions: 173.3 mm; Non-target lesions: Present, no progression

[0342] After the 6th cycle of administration: Target lesion: 180.7 mm; Non-target lesions: Present, no progression

[0343] 8. Patient: S06002 / C06002 - Gastric cancer

[0344] One year ago, the patient had anorexia without obvious cause, felt fatigued, accompanied by belching, acid reflux and heartburn, without symptoms such as nausea and vomiting. Gastroscopy in May 2019 showed huge irregular elevations on the anterior and posterior walls and the lesser curvature of the gastric antrum, and the pathology suggested adenocarcinoma. Gastroscopy on August 5, 2019 showed: Gastric cancer, reflux esophagitis. Chest, upper abdomen and lower abdomen CT (August 5, 2019) showed: Gastric cancer with multiple lymph node metastases: Micronodular foci in the left lung: Multiple high-density foci in the bone. Abdominal ultrasound showed: Multiple cholesterol crystals in the gallbladder: Solid protrusion on the left lateral wall of the bladder. Echocardiogram showed: Degenerative changes of the aortic valve, reduced left ventricular diastolic function. For treatment, "palliative gastrectomy" was performed. During the operation, no obvious metastatic foci were seen in the liver, spleen, abdominal cavity and pelvis. The cancer mass was located in the lesser curvature of the gastric body, about 8*7*6 cm in size, penetrating the serosa layer. Multiple enlarged lymph nodes were seen in groups 2, 4, 6, 7, and 8. The cancer mass was densely adhered to the pancreas and part of the transverse colon mesentery. Lymph nodes in groups 1, 2, 3, 4, 5, 6, 7, 8, and 11 were removed during the operation. The operation process was smooth. Blood transfusion was performed during the operation. The postoperative pathological results showed: Ulcerative moderately and poorly differentiated adenocarcinoma in the gastric antrum. Immunohistochemical results: Ulcerative moderately and poorly differentiated adenocarcinoma in the gastric antrum. Tumor size: 3*2.8*0.8 cm. The cancer tissue invaded the submucosa of the serosa layer. No obvious cancer thrombus was seen in the blood vessels. No cancer tissue was seen at the upper and lower surgical margins and the greater omentum. Cancer metastasis was seen in 1 / 4 of the lymph nodes on the greater curvature side, and no cancer metastasis was seen in 0 / 12 of the lymph nodes on the lesser curvature side. The patient had an upper abdomen and lower abdomen MRI examination on May 26, 2020 due to "abdominal discomfort" (May 27, 2020), which showed: 1. Space-occupying lesions in the abdominal cavity and retroperitoneum, considering possible lymphatic metastasis; 2. Cyst in the left lobe of the liver. Gastroscopy examination on May 28, 2020 showed: 1. Squamous epithelial papillomatous hyperplasia of the esophagus; 2. Severe reflux esophagitis; 3. Residual gastritis; 4. Bile reflux; 5. Gastric retention. Past history: The patient had a history of "diabetes" for more than 16 years, regularly took hypoglycemic drugs, and the blood sugar was well controlled. Had a history of "hypertension" for more than 10 years, and the self-administered drugs were well controlled. Had a history of "premature beats", took traditional Chinese medicine for conditioning, and had no obvious symptoms usually.

[0345] Dosage: Anlotinib hydrochloride: 10 mg, 14C12H1L1: 200 mg

[0346] First administration time: Starting from June 19, 2020, oral administration of 12 mg of anlotinib hydrochloride capsules once a day for 2 consecutive weeks, followed by a 1-week break. 14C12H1L1: Administered once every 3 weeks, 200 mg per dose, by intravenous infusion. Each 3-week period is one treatment cycle until disease progression or intolerance. The patient had good overall tolerance during medication and could continue taking the drug.

[0347] Efficacy evaluation: Screening period: Target lesion: 25.5 mm; Non-target lesion: Metastasis in the descending part of the duodenum

[0348] After the second cycle of administration: Target lesion: 19.6 mm; Non-target lesion: NCR / NPD

[0349] After the fourth cycle of administration: Target lesion: 19.6 mm; Non-target lesion: NCR / NPD

[0350] After the sixth cycle of administration: Target lesion: 19.3 mm; Non-target lesion: NCR / NPD

[0351] 9. Patient: S06004 / C06003 - Biliary tract cancer

[0352] The patient reported abdominal distension after drinking alcohol more than 2 years ago, accompanied by symptoms such as poor tolerance, acid reflux, belching, and diarrhea. Enhanced MR showed: Abnormal shape and signal in the middle and lower segments of the common bile duct, consistent with cholangiocarcinoma and biliary system dilation, and multiple hepatic cysts. Laparoscopic pancreatoduodenectomy was performed on June 7, 2018. The postoperative pathological report on June 12, 2018 showed: (Left half of the liver) Intrahepatic cholangiocarcinoma (tumor area: 4.5 * 3 cm), moderately differentiated cancer cells, adjacent to the liver capsule, no cancer thrombus in the blood vessels was detected, nerve invasion was detected, and no cancer cells were detected at the liver resection margin and the hepatic duct resection margin. Another sample (liver tissue biopsy) showed cholangiocarcinoma in the examined tissue, and no cancer was found in the other sample of the gallbladder. Then, oral administration of tegafur was started in November 2019. An abdominal plain CT scan was performed on March 29, 2020, and the results showed: 1. Findings on postoperative review of cholangiocarcinoma, with ascites. 2. No obvious abnormalities were found on the chest CT plain scan. The patient reported stopping tegafur on their own. An abdominal enhanced CT scan was performed on June 23, 2020, and the results showed: 1. Findings on postoperative review of cholangiocarcinoma. 2. Consistent with peritoneal metastasis and ascites.

[0353] Dosage: Anlotinib hydrochloride: 10 mg, 14C12H1L1: 200 mg

[0354] First administration time: Starting from July 31, 2020, oral administration of 12 mg of anlotinib hydrochloride capsules once a day for 2 consecutive weeks, followed by a 1-week break. 14C12H1L1: Administered once every 3 weeks, 200 mg per dose, by intravenous infusion. Each 3-week period is one treatment cycle until disease progression or intolerance. The patient had good overall tolerance during medication and could continue taking the drug.

[0355] Efficacy evaluation: Screening period: Target lesion: 37.5 mm; Non-target lesions: peritoneal cavity, retroperitoneal metastasis

[0356] After the second cycle of administration: target lesions: 37.2 mm; non-target lesions: NCR / NPD

[0357] 10. Patient: S06005 / C06004 - Cholangiocarcinoma / Bile tract cancer

[0358] One year prior, the patient had experienced recurrent upper abdominal pain accompanied by icteric skin and sclera, dark tea-colored urine, and no significant fever, chills, or diarrhea. An upper abdominal contrast-enhanced MRI performed on April 10, 2019, showed an enlarged gallbladder with thickened gallbladder walls, extensive dilatation of the intrahepatic and extrahepatic bile ducts, and localized stenosis of the left and right hepatic ducts and the proximal common hepatic duct. This was consistent with a space-occupying lesion in the portal area (left and right hepatic ducts and the proximal common hepatic duct), and enlarged lymph nodes in the portal area and portal space, consistent with cholangiocarcinoma and cholecystitis. On April 12, 2019, under general anesthesia with endotracheal intubation, a radical resection of hilar cholangiocarcinoma and right hemihepatectomy was performed successfully. Pathological results were released on April 18, 2019: (left hepatic duct) cholangiocarcinoma, with moderately to poorly differentiated cancer cells invading the muscularis and serosa. The right hepatic duct had submucosal invasion but not the muscularis and serosa. The gallbladder and right hemiliver tissue were not involved. The resection margins of the common hepatic duct and gallbladder were negative. One lymph node was found in the left hepatic duct, but no cancer cells were found. The patient also had chronic cholecystitis and cholecystoadenia. Six cycles of oral S-1a were administered from May 11, 2019, to March 2020. On April 19, 2020, an enhanced abdominal CT scan revealed the following: 1. Postoperative changes in the right hepatic lobe, with low-density lesions in the left hepatic lobe, suggesting metastasis; 2. Postoperative cholecystectomy; 3. Prostatic hyperplasia. On April 26, 2020, an upper (or lower) abdominal MRI with plain scan and enhanced liver examination revealed the following: 1. Postoperative changes in the portal bile duct; multiple intrahepatic nodules, suggesting metastasis. Abnormally enhancing lesions at the margin of the left inner lobe, suggesting a high likelihood of metastasis. 2. Possible accessory spleen. The patient self-administered capecitabine for one cycle from June 15 to July 12, 2020. A chest and abdominal examination, performed on July 21, 2020, revealed the following: 1. A plain lung CT scan revealed no significant abnormalities; 2. Postoperative changes in the right hepatic lobe, with multiple liver masses, suggesting metastasis, which were increased in number and size compared to previous examinations; and possible portal vein tumor thrombosis. 3. Postoperative cholecystectomy; 4. Prostatic hyperplasia.

[0359] Dosage: Anlotinib hydrochloride: 10mg, 14C12H1L1: 200mg

[0360] First administration time: Starting from August 11, 2020, orally take 12 mg of Anlotinib Hydrochloride Capsules once a day for treatment, take continuously for 2 weeks and stop for 1 week, 14C12H1L1: administer once every 3 weeks, 200 mg / time, by intravenous infusion. Each 3-week period is one treatment cycle until disease progression or intolerance. The patient had good overall tolerance during medication and could continue taking the medicine.

[0361] Efficacy evaluation: Screening period: Target lesion: 42.8 mm; Non-target lesions: Multiple liver metastases, mesenteric lymph node metastases

[0362] After the second cycle of administration: Target lesion: 41.7 mm; Non-target lesions: iUPD

[0363] 11. Patient: S06007 / C06006 - Diffuse infiltrating adenocarcinoma of bile duct / Biliary tract cancer

[0364] On September 7, 2017, the results of upper abdominal plain MR examination showed: Consistent with the MR and MRCP manifestations of a mass at the lower end of the common bile duct and biliary tract obstruction and dilation. The results of lung plain scan + enhanced CT of the whole abdomen showed: 1. A few fibrotic nodules in both lungs. 2. A little interstitial change in both lower lungs. 3. Masses in the common bile duct and cystic duct with dilation of intrahepatic and extrahepatic bile ducts and pancreatic duct. 4. Chronic cholecystitis. 5. Considered a small liver cyst. 6. Thrombosis of the celiac trunk abdominal wall. Laparoscopic pancreatoduodenectomy was performed smoothly. The postoperative pathology showed: Diffuse infiltrating adenocarcinoma of bile duct, poorly differentiated, invading the serosa, negative surgical margin of bile duct, cauliflower-like papillary adenocarcinoma of pancreatic duct, moderately and poorly differentiated, invading the muscular layer of duodenum, one cancer nodule seen, lymph nodes 0 / 31. Starting from November 2, 2017 after the operation, Tegafur 20 mg was given orally bid for chemotherapy. After discontinuing orally for two cycles due to intolerance, it was stopped in March 2019.

[0365] Dosage: Anlotinib Hydrochloride: 12 mg, 14C12H1L1: 200 mg

[0366] First administration time: Starting from September 9, 2020, orally take 12 mg of Anlotinib Hydrochloride Capsules once a day for treatment, take continuously for 2 weeks and stop for 1 week, 14C12H1L1: administer once every 3 weeks, 200 mg / time, by intravenous infusion. Each 3-week period is one treatment cycle until disease progression or intolerance. The patient had good overall tolerance during medication and could continue taking the medicine.

[0367] Efficacy evaluation: Screening period: Target lesion: 38.9 mm; Non-target lesions: Liver, peritoneum, supraclavicular lymph node metastases

[0368] After the second cycle of administration: Target lesion: 38.7 mm; Non-target lesions: iUPD

[0369] 12. Patient: S06008 / C06007 - Intrahepatic cholangiocarcinoma

[0370] One year ago, an abdominal ultrasound during a physical examination revealed a liver space-occupying lesion. MR examination showed: a space-occupying lesion in the left lobe of the liver, considered a neoplastic lesion; a small hemangioma in the right lobe of the lung. The enhanced CT of the upper abdomen showed: a space-occupying lesion in the left lobe of the liver, considered likely to be malignant, with a high possibility of being cholangiogenic; a small hemangioma in the lower segment of the right posterior lobe of the liver. On July 30, 2019, laparoscopic left hepatectomy was performed under general anesthesia. The operation was successful. The postoperative pathology showed: moderately to poorly differentiated adenocarcinoma, consistent with intrahepatic cholangiocarcinoma, about 5.5×4.5×4 cm in size, the cancer invaded the surrounding liver tissue, focally involved the liver tissue, and focally involved the liver capsule; cancer was visible at the surgical margin; nerve invasion was visible, and multiple satellite nodules were visible in the surrounding liver tissue. Immunohistochemical results: PMS2(+), MSH2(+), MSH6(+), MLH1(+), PD-1 (tumor cells -, lymphocytes about 2% +), CK7(+), CK20 (individually +), CK19 (weak +), P53 (wild type), Ki-67(50%). From September 2019 to February 2020 after the operation, oxaliplatin + gemcitabine chemotherapy was given. The specific chemotherapy cycle is unknown. Then, on February 10, 2020, the CT imaging results of the lungs / mediastinum / chest wall showed: multiple lesions in both lungs, and the lesion in the upper lobe of the right lung increased compared with before. Localized thickening of the bilateral pleura. Postoperative changes in the left lobe of the liver. Considering that the metastasis increased compared with before, the chemotherapy regimen was changed to paclitaxel + tegafur in February 2020. During this period, paclitaxel was discontinued, but oral tegafur chemotherapy was given. The specific dates and chemotherapy cycles are unknown. Tegafur was stopped on August 12, 2020.

[0371] Dosage: Anlotinib hydrochloride: 12 mg, 14C12H1L1: 200 mg

[0372] First administration time: Starting from September 16, 2020, anlotinib hydrochloride capsules were orally administered once a day at a dose of 12 mg for 2 weeks and then stopped for 1 week. 14C12H1L1: administered once every 3 weeks, 200 mg per dose, by intravenous infusion. Each 3-week period is one treatment cycle until disease progression or intolerance. The patient had good overall tolerance during the medication and could continue the medication.

[0373] Efficacy evaluation: Screening period: Target lesion: 71.3 mm; Non-target lesion: hilar lymph node metastasis

[0374] After the second cycle of administration: Target lesion: 44.4 mm; Non-target lesion: NCR / NPD

[0375] According to the content disclosed in the present application, although the compositions and methods of the present application have been described according to the preferred embodiments, for those skilled in the art, without departing from the concept, spirit and scope of the present application, changes can be made to the compositions and / or methods described herein and the steps or the order of the steps of the methods.

[0376] The disclosures of all documents cited herein are incorporated herein by reference to the extent that they provide exemplary, procedural, and other details to supplement the content described herein. Sequence Listing <110> Chia Tai Tianqing Pharmaceutical Group Co., Ltd. <120> Pharmaceutical combination of quinoline derivatives and PD-1 monoclonal antibody <130> 2020.11.27 <160> 8 <170> SIPOSequenceListing 1.0 <210> 1 <211> 6 <212> PRT <213> Synthetic sequence <400> 1 Gln Asp Ile Asn Thr Tyr 1 5 <210> 2 <211> 3 <212> PRT <213> Synthetic sequence <400> 2 Arg Ala Asn 1 <210> 3 <211> 9 <212> PRT <213> Synthetic sequence <400> 3 Leu Gln Tyr Asp Glu Phe Pro Leu Thr 1 5 <210> 4 <211> 8 <212> PRT <213> Synthetic sequence <400> 4 Gly Phe Ala Phe Ser Ser Tyr Asp 1 5 <210> 5 <211> 8 <212> PRT <213> Synthetic sequence <400> 5 Ile Ser Gly Gly Gly Arg Tyr Thr 1 5 <210> 6 <211> 11 <212> PRT <213> Synthetic sequence <400> 6 Ala Asn Arg Tyr Gly Glu Ala Trp Phe Ala Tyr 1 5 10 <210> 7 <211> 107 <212> PRT <213> Synthetic sequence <400> 7 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Met Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Phe Thr Cys Arg Ala Ser Gln Asp Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Thr Leu Ile 35 40 45 Tyr Arg Ala Asn Arg Leu Val Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Met Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Glu Phe Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 8 <211> 118 <212> PRT <213> Synthetic sequence <400> 8 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 Ala Phe Ser Ser Tyr 20 25 30 Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Asp Trp Val 35 40 45 Ala Thr Ile Ser Gly Gly Gly Arg Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Asn Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Asn Arg Tyr Gly Glu Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115

Claims

1. Use of a human PD-1 antibody and a tyrosine kinase inhibitor in the preparation of a medicament for treating cholangiocarcinoma, in, wherein the human PD-1 antibody comprises a light chain and a heavy chain, the light chain comprises light chain complementarity-determining regions LCDR1, LCDR2 and LCDR3 having amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 respectively, and the heavy chain comprises heavy chain complementarity-determining regions HCDR1, HCDR2 and HCDR3 having amino acid sequences shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively, wherein the tyrosine kinase inhibitor is a compound of formula I or a pharmaceutically acceptable salt thereof, 2. The use according to claim , wherein the pharmaceutically acceptable salt of the compound of formula I is the hydrochloride salt of 1-[[[4-(4-fluoro-2-methyl-1H-indol-5-yl)oxy-6-methoxyquinolin-7-yl]oxy]methyl]cyclopropanamine.

3. The use according to claim 1, wherein the pharmaceutically acceptable salt of the compound of formula I is the dihydrochloride salt of 1-[[[4-(4-fluoro-2-methyl-1H-indol-5-yl)oxy-6-methoxyquinolin-7-yl]oxy]methyl]cyclopropanamine.

4. The use according to claim 1, wherein the human PD-1 antibody comprises a light chain variable region having an amino acid sequence shown in SEQ ID NO:7 and a heavy chain variable region having an amino acid sequence shown in SEQ ID NO:

8.

5. The use according to claim 1, wherein the human PD-1 antibody is 14C12H1L1.

6. Use of a human PD-1 antibody in the preparation of a medicament for the combined treatment of cholangiocarcinoma with a tyrosine kinase inhibitor, Among them, wherein the human PD-1 antibody comprises a light chain and a heavy chain, the light chain comprises light chain complementarity-determining regions LCDR1, LCDR2 and LCDR3 having amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 respectively, and the heavy chain comprises heavy chain complementarity-determining regions HCDR1, HCDR2 and HCDR3 having amino acid sequences shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively, wherein the tyrosine kinase inhibitor is a compound of formula I or a pharmaceutically acceptable salt thereof, 7. The use according to claim 6, wherein the pharmaceutically acceptable salt of the compound of formula I is the hydrochloride salt of 1-[[[4-(4-fluoro-2-methyl-1H-indol-5-yl)oxy-6-methoxyquinolin-7-yl]oxy]methyl]cyclopropanamine.

8. The use according to claim 6, wherein the pharmaceutically acceptable salt of the compound of formula I is the dihydrochloride salt of 1-[[[4-(4-fluoro-2-methyl-1H-indol-5-yl)oxy-6-methoxyquinolin-7-yl]oxy]methyl]cyclopropanamine.

9. The use according to claim 6, wherein the human PD-1 antibody comprises a light chain variable region having the amino acid sequence as shown in SEQ ID NO: 7 and a heavy chain variable region as shown in SEQ ID NO:

8.

10. The use according to claim 6, wherein the human PD-1 antibody is 14C12H1L1.

11. The use according to any one of claims 1-10, wherein the human PD-1 antibody and the compound of formula I or a pharmaceutically acceptable salt thereof are each in the form of a pharmaceutical composition.

12. The use according to any one of claims 1-10, wherein the biliary tract cancer includes intrahepatic cholangiocarcinoma, extrahepatic cholangiocarcinoma, and gallbladder cancer.

13. The use according to any one of claims 1-10, wherein the biliary tract cancer is inoperable or metastatic biliary tract cancer, or biliary tract cancer that has failed first-line or more than first-line chemotherapy.

14. The use according to any one of claims 1-10, wherein the human PD-1 antibody is administered at a frequency of once every 2 weeks or once every 3 weeks.

15. The use according to any one of claims 1-10, wherein the human PD-1 antibody is administered in one or more uniform doses selected from 200 mg.

16. The use according to claim 5 or 10, wherein, The hydrochloride salt of the compound of formula I is administered once a day, 12 mg or 10 mg each time, orally for 2 consecutive weeks and then stopped for 1 week, and 14C12H1L1 is administered once every 3 weeks, 200 mg each time.

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