WO2020083338A1

WO2020083338A1 - Use of combination of irinotecan, immune checkpoint inhibitor, and 5-fu in preparation of drugs for treating tumor diseases

Info

Publication number: WO2020083338A1
Application number: PCT/CN2019/113009
Authority: WO
Inventors: 曹永�; 王富济
Original assignee: 江苏恒瑞医药股份有限公司
Priority date: 2018-10-25
Filing date: 2019-10-24
Publication date: 2020-04-30
Also published as: TW202029964A; CN112543637A; CN112543637B

Abstract

Provided in the present application is use of a combination of Irinotecan, an immune checkpoint inhibitor, and 5-FU in preparation of drugs for preventing or treating tumor diseases. Specifically, provided in the present application is use of a combination of Irinotecan or a pharmaceutically acceptable salt thereof, a PD-L1 antibody or antigen binding segment, and 5-FU in preparation of drugs for preventing or treating tumor diseases.

Description

Use of irinotecan combined immune checkpoint inhibitor and 5-FU in the preparation of drugs for treating tumor diseases

Technical field

The present application belongs to the field of medicine, and relates to the use of irinotecan combined immune checkpoint inhibitors and 5-FU in the preparation of drugs for preventing or treating tumor diseases.

Background technique

Malignant tumors are major diseases that endanger people's lives and health. In recent years, with the rapid development of tumor biology and related disciplines, specific anti-tumor drugs targeting the targets of abnormal signaling systems in tumor cells have been the focus of new drug development. At the same time, the combination of multiple anti-tumor drugs for the treatment of tumor diseases is also a hotspot of scientific research.

Esophageal cancer refers to a type of malignant tumor that occurs in esophageal mucosal epithelium. It has the 8th highest morbidity and 6th mortality worldwide. More than 50% of esophageal cancers occur in China. In 2015, the number of new esophageal cancer patients in China was 479,700; from 2001 to 2011, the incidence of male tumors ranked fifth and the mortality rate ranked fourth. The incidence and mortality of females decreased year by year. trend. There is currently no accepted standard chemotherapy for first-line chemotherapy of advanced esophageal cancer. Most of them use platinum-based and / or fluorouracil-based regimens. The effective rate is 25% to 35%. Cisplatin combined with 5-FU is not shown in comparison with single-agent cisplatin Survival advantage. Therefore, exploring new first-line chemotherapy has become an urgent problem to be solved.

Irinotecan is a semi-synthetic derivative of camptothecin. Camptothecin can specifically bind to topoisomerase I, which induces reversible single-strand breaks, thereby unwinding the DNA double-stranded structure; irinotecan and its active metabolite SN-38 can bind to topoisomerase The I-DNA complex binds, thereby preventing reconnection of broken single strands. Existing research suggests that the cytotoxic effect of irinotecan is due to the interaction between the replicase and topoisomerase I-DNA-irinotecan (or SN-38) triple complex during DNA synthesis, resulting in DNA double Chain break. At present, irinotecan is mainly used clinically for the treatment of patients with advanced colorectal cancer.

5-fluorouracil (5-FU or KU) is a pyrimidine analog drug used in cancer treatment. It works by irreversibly inhibiting thymidylate synthase and belongs to antimetabolites. L. Assersohn et al. Disclosed a second-line clinical study of irinotecan combined with 5-fluorouracil in the treatment of advanced gastroesophageal cancer (Annals of Oncology 15: 64–69, 2004). The specific protocol is irinotecan 180mg / m ² + 5-fluorouracil 400mg / m ² + 5-fluorouracil 1200mg / m ^{2 was} continuously pumped for 48h, and was repeated every 2 weeks. Of the 38 patients with advanced gastroesophageal cancer, 39.5% had esophageal cancer, and 92.1% had histological type of adenocarcinoma. The total effective rate was 29%, mPFS was 3.7 months, and mOS was 6.4 months.

The PD-1 (programmed death receptor 1) antibody can specifically recognize and bind to PD-1 on the surface of lymphocytes, blocking the PD-1 / PD-L1 signaling pathway, thereby activating the immune killing effect of T cells on tumors, and mobilizing the body The immune system clears tumor cells in the body. PD-1 has two ligands, PD-L1 and PD-L2. PD-L1 is mainly expressed on T cells, B cells, macrophages and dendritic cells (dendritic cells, DC), and the expression on activated cells can be up-regulated after activation. PD-L1 suppresses the immune system by combining with PD-1 and B7-1, and many tumor cells and immune cells in the tumor tissue microenvironment express PD-L1. New research finds high PD-L1 detected in human tumor tissues such as breast cancer, lung cancer, gastric cancer, intestinal cancer, renal cancer, melanoma, non-small cell lung cancer, colon cancer, bladder cancer, ovarian cancer, pancreatic cancer and liver cancer Protein expression, and the expression level of PD-L1 are closely related to the clinical and prognosis of patients. Because PD-L1 acts as a second signaling pathway to inhibit T cell proliferation, blocking PD-L1 / PD-1 binding has become a very promising emerging target in the field of tumor immunotherapy. The combination of PD-1, PD-L1 antibody and other immune checkpoint inhibitors with other drugs is also a hot research area. At present, a series of anti-PD-L1 antibodies have been disclosed. Among them, WO2017084495 discloses a series of PD-L1 antibodies, which can effectively improve the effect of inhibiting tumorigenesis and development. The PD-L1 antibody code-named HRP00052 has excellent therapeutic effect.

A one-arm study from Japan (W. Joost Lesterhuis, et al, The Journal of Clinical Investigation, 2011, 128 (1): 3100-3108) reported the use of Nivolumab for transfluorouracils / platinum / yews The efficacy and safety of advanced esophageal squamous cell carcinoma patients with failed or intolerable drug therapy can be evaluated in 64 cases. The ORR after independent evaluation was 17%, of which 1 case obtained CR, and the median PFS and OS were 1.5 months and 2.3 months, respectively.

The present application provides the use of an irinotecan combined immune checkpoint inhibitor and 5-FU in the preparation of a medicament for preventing or treating tumor diseases, and shows a good tumor suppressing effect.

Summary of the invention

The present application provides the use of irinotecan or a pharmaceutically acceptable salt thereof in combination with an immune checkpoint inhibitor and 5-FU in the preparation of a medicament for preventing or treating tumor diseases.

In some embodiments, the irinotecan or a pharmaceutically acceptable salt thereof is administered in the form of liposomes. Preferred irinotecan liposomes contain irinotecan or a pharmaceutically acceptable salt thereof, neutral phospholipid and cholesterol, wherein the weight ratio of cholesterol to neutral phospholipid is 1: 3 to 5, preferably 1: 3.5 to 4.5 , Most preferably 1: 4.

The weight ratio of irinotecan or its pharmaceutically acceptable salt to neutral phospholipid is 1: 2-5, preferably 1: 2.5-4, most preferably 1: 4.

In a preferred embodiment, the neutral phospholipid is selected from one or more of hydrogenated soybean phosphatidylcholine, phosphatidylethanolamine, lecithin, cardiolipin, preferably hydrogenated soybean phospholipid.

In a preferred embodiment, the liposome further contains a lipid derivative of a hydrophilic polymer, preferably DSPE-PEG ₂₀₀₀ .

In certain embodiments, the immune checkpoint inhibitor is selected from PD-L1 antibodies or antigen-binding fragments.

In certain embodiments, any one of the PD-L1 antibodies or antigen-binding fragments is selected from the following CDR region sequences or mutant sequences thereof: antibody heavy chain variable region HCDR region sequence: SEQ ID NO: 1-3 ; And antibody light chain variable region LCDR region sequence: SEQ ID NO: 4-6;

details as follows:

HCDR1 is selected from:

SYWMH SEQ ID NO: 1

HCDR2 is selected from:

RI GPNSG FTSYNEKFKN SEQ ID NO: 2

HCDR3 is selected from:

GGSSYDYFDY SEQ ID NO: 3

LCDR1 is selected from:

RASESVSIHGTHLMH SEQ ID NO: 4

LCDR2 is selected from:

AASNLES SEQ ID NO: 5

LCDR3 is selected from:

QQSFEDPLT SEQ ID NO: 6.

In certain embodiments, the PD-L1 antibody or antigen-binding fragment comprises and amino acid sequence: SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity light chain variable region CDR sequences, and amino acids Sequence: SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 have at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98%, or 99% sequence identity of heavy chain variable region CDR sequences.

In certain embodiments, the PD-L1 antibody or antigen-binding fragment may be selected from murine antibodies, chimeric antibodies, humanized antibodies, human antibodies, and preferably humanized antibodies.

In certain embodiments, the PD-L1 antibody or antigen-binding fragment comprises and the amino acid sequence SEQ ID NO: 7 has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92 %, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity of the heavy chain variable region sequence, and the amino acid sequence SEQ ID NO: 8 has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity of the light chain variable region sequence.

In some embodiments, the PD-L1 antibody or antigen-binding fragment further comprises a heavy chain constant region of human IgG1, IgG2, IgG3 or IgG4 or a variant thereof, preferably a human heavy chain constant region of IgG2 or IgG4 It is more preferable to include an IgG4 heavy chain constant region that introduces F234A and L235A mutations; the humanized antibody light chain further includes a constant region of a human κ, λ chain, or a variant thereof.

In certain embodiments, the PD-L1 antibody or antigen-binding fragment comprises and the amino acid sequence SEQ ID NO: 9 has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92 %, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity of the heavy chain variable region sequence, and the amino acid sequence SEQ ID NO: 11 has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity of the light chain sequence.

In some embodiments, the heavy chain sequence of the PD-L1 antibody or antigen-binding fragment is SEQ ID NO: 9, and the light chain sequence is SEQ ID NO: 11.

SEQ ID NO: 7

SEQ ID NO: 8

Note: FR sequence is in italics; CDR sequence is underlined.

Heavy chain sequence

SEQ ID NO: 9

Heavy chain sequence encoding gene sequence

SEQ ID NO: 10

Light chain sequence

SEQ ID NO: 11

The light chain sequence encodes the gene sequence:

SEQ ID NO: 12

In certain embodiments, the tumor is selected from malignant tumors and benign tumors; the malignant tumor is selected from esophageal cancer. The esophageal cancer includes but is not limited to esophageal squamous cell carcinoma and esophageal adenocarcinoma.

In certain embodiments, the tumor is selected from advanced tumors, relapsed refractory tumors, failed and / or relapsed tumors after chemotherapy treatment, failed and / or relapsed tumors after radiation therapy, failed and / or targeted drug treatments and / or Relapsed tumor, failed immunotherapy and / or relapsed tumor.

In certain preferred embodiments, the tumor is selected from advanced esophageal cancer, recurrent esophageal cancer, and distant metastatic esophageal cancer.

In some embodiments, the pharmaceutically acceptable salt of irinotecan is the hydrochloride salt. Irinotecan and their liposomes are administered at a dose of 40-200 mg / m ² based on irinotecan hydrochloride, such as 40 mg / m ² , 41 mg / m ² , 42 mg / m ² , 43 mg / m ² , 44mg / m ² , 45mg / m ² , 46mg / m ² , 47mg / m ² , 48mg / m ² , 49mg / m ² , 50mg / m ² , 51mg / m ² , 52mg / m ² , 53mg / m ² , 54mg / m ² , 55mg / m ² , 56mg / m ² , 57mg / m ² , 58mg / m ² , 59mg / m ² , 60mg / m ² , 61mg / m ² , 62mg / m ² , 63mg / m ² , 64mg / m ² , 65mg / m ² , 66mg / m ² , 67mg / m ² , 68mg / m ² , 69mg / m ² , 70mg / m ² , 71mg / m ² , 72mg / m ² , 73mg / m ² , 74mg / m ² , 75mg / m ² , 76mg / m ² , 77mg / m ² , 78mg / m ² , 79mg / m ² , 80mg / m ² , 81mg / m ² , 82mg / m ² , 83mg / m ² , 84mg / m ² , 85mg / m ² , 86mg / m ² , 87mg / m ² , 88mg / m ² , 89mg / m ² , 90mg / m ² , 95mg / m ² , 100mg / m ² , 105mg / m ² , 110mg / m ² , 115mg / m ² , 120mg / m ² , 130mg / m ² , 140mg / m ² , 150mg / m ² , 160mg / m ² , 170mg / m ² , 180mg / m ² , 190mg / m ² , 200mg / m ² .

In certain embodiments, the dose of 5-fluorouracil 1000-3000mg / m ^2, may be ^{1000mg / m 2, 1100mg / m} 2, 1200mg / m 2, 1300mg / m 2, 1400mg / m 2 , 1500mg / m ² , 1600mg / m ² , 1700mg / m ² , 1800mg / m ² , 1900mg / m ² , 2000mg / m ² , 2100mg / m ² , 2200mg / m ² , 2300mg / m ² , 2400mg / m ² ^{, 2500mg / m 2, 2600mg /} m 2, 2700mg / m 2, 2800mg / m 2, 2900mg / m 2, 3000mg / m 2.

In certain embodiments, the dose of the immune checkpoint inhibitor is selected from 1-50 mg / kg, preferably from 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7mg / kg, 8mg / kg, 9mg / kg, 10mg / kg, 11mg / kg, 12mg / kg, 13mg / kg, 14mg / kg, 15mg / kg, 16mg / kg, 17mg / kg, 18mg / kg, 19mg / kg, 20mg / kg, 21mg / kg, 22mg / kg, 23mg / kg, 24mg / kg, 25mg / kg, 26mg / kg, 27mg / kg, 28mg / kg, 29mg / kg, 30mg / kg, 31mg / kg, 32mg / kg, 33mg / kg, 34mg / kg, 35mg / kg, 36mg / kg, 37mg / kg, 38mg / kg, 39mg / kg, 40mg / kg, 42mg / kg, 45mg / kg, 47mg / kg, 50mg / kg, more preferably 1 mg / kg, 3 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 30 mg / kg, 40 mg / kg.

In certain embodiments, the dose of the immune checkpoint inhibitor is selected from 50-3000 mg, preferably from 50 mg, 60 mg, 70 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 375 mg, 400 mg, 425mg, 450mg, 475mg, 500mg, 550mg, 600mg, 650mg, 700mg, 750mg, 800mg, 850mg, 900mg, 950mg, 1000mg, 1050mg, 1100mg, 1150mg, 1200mg, 1250mg, 1300mg, 1350mg, 1400mg, 1450mg, 1500mg, 1550mg, 1600mg, 1650mg, 1700mg, 1750mg, 1800mg, 1850mg, 1900mg, 1950mg, 2000mg, 2050mg, 2100mg, 2150mg, 2200mg, 2250mg, 2300mg, 2350mg, 2400mg, 2450mg, 2500mg, 2550mg, 2600mg, 2650mg, 2700mg, 2750mg, 2800mg, 2850mg, 2900mg, 2950mg, 3000mg.

The combined administration route described herein is selected from oral administration, parenteral administration, and transdermal administration. The parenteral administration includes but is not limited to intravenous injection, subcutaneous injection, and intramuscular injection.

The present application further relates to the use of irinotecan or a pharmaceutically acceptable salt thereof in combination with an immune checkpoint inhibitor and 5-FU in the preparation of a medicament for preventing or treating tumor diseases, wherein the frequency of administration of irinotecan or a pharmaceutically acceptable salt It can be once a day, twice a day, three times a day, once a week, once every two weeks, once every three weeks, once a month, once every five weeks, once every six weeks. The frequency of 5-FU administration may be once a day, twice a day, three times a day, once a week, once every two weeks, once every three weeks, and once a month. The frequency of administration of the immune checkpoint inhibitor can be once a day, twice a day, three times a day, once a week, once every two weeks, once every three weeks, once a month, once every five weeks, once every six weeks.

In certain embodiments, the frequency of administration of irinotecan or a pharmaceutically acceptable salt thereof is one administration cycle every 2 weeks, and the administration is once every cycle, with a dose of 60 mg / m ² or 80 mg / m ² ; 5- The administration frequency of FU is every 2 weeks as a dosing cycle, once every cycle, the dose is 2400mg / m ² ; the administration frequency of the immune checkpoint inhibitor is every 2 weeks as a dosing cycle, every cycle Once the medicine, the dose is 10mg / kg.

In certain embodiments, when irinotecan or its pharmaceutically acceptable salt, 5-FU or immune checkpoint inhibitor drugs are administered to patients intolerable due to toxic and side effects, etc., the dosage of the drug and the administration can be adjusted appropriately The frequency of medicine.

In the solution described in this application, the combination optionally further includes other components, including but not limited to other anti-tumor drugs.

The present application also provides a method of treating tumor diseases, which includes administering irinotecan or a pharmaceutically acceptable salt thereof, an immune checkpoint inhibitor, and 5-FU to a patient.

In certain embodiments, the patient has not previously received systemic anti-tumor therapy.

In certain embodiments, the patient has received neoadjuvant / adjuvant and radical concurrent chemoradiotherapy, and the time from the last chemotherapy to the time of relapse or progression exceeds 6 months.

The present application also relates to a pharmaceutical composition comprising irinotecan or a pharmaceutically acceptable salt thereof, an immune checkpoint inhibitor and 5-FU, and one or more pharmaceutically acceptable carriers, excipients, and diluents. The pharmaceutical composition can be made into any pharmaceutically acceptable dosage form. For example, it can be formulated as tablets, capsules, pills, granules, solutions, suspensions, syrups, injections (including injections, sterile powders for injection and concentrated solutions for injection), suppositories, inhalants or sprays Agent.

The pharmaceutical composition containing irinotecan or a pharmaceutically acceptable salt thereof, an immune checkpoint inhibitor and 5-FU described in this application may be administered alone or in combination with one or more therapeutic agents.

The present application also provides a pharmaceutical packaging box in which the pharmaceutical composition of irinotecan or a pharmaceutically acceptable salt thereof, an immune checkpoint inhibitor, and 5-FU described herein is packaged.

In this application, irinotecan or a pharmaceutically acceptable salt thereof, an immune checkpoint inhibitor and 5-FU are co-administered, thereby enhancing antitumor activity and improving the therapeutic effect of tumor diseases.

Detailed description of the invention

The three-letter codes and one-letter codes of amino acids used in this application are as described in J. biol.chem, 243, p3558 (1968).

The "antibody" described in this application refers to an immunoglobulin, which is a tetrapeptide chain structure formed by connecting two identical heavy chains and two identical light chains through interchain disulfide bonds. The immunoglobulin heavy chain constant region has different amino acid composition and arrangement order, so its antigenicity is also different. According to this, immunoglobulins can be divided into five categories, or isotypes called immunoglobulins, namely IgM, IgD, IgG, IgA, and IgE, and their corresponding heavy chains are μ chain, δ chain, and γ chain, respectively. , Α chain, and ε chain. The same class of Ig can be divided into different subclasses according to the differences in the amino acid composition of the hinge region and the number and position of heavy chain disulfide bonds. For example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4. The light chain is divided into a kappa chain or a lambda chain by different constant regions. Each of the five types of Ig can have a κ chain or a λ chain.

In the present application, the antibody light chain described in the present application may further comprise a light chain constant region, and the light chain constant region comprises a human or murine kappa, lambda chain or a variant thereof.

In the present application, the antibody heavy chain of the present application may further comprise a heavy chain constant region, which comprises human or murine IgG1, IgG2, IgG3, IgG4 or variants thereof.

The antibodies of the present application include murine antibodies, chimeric antibodies, and humanized antibodies, preferably humanized antibodies.

The term "murine antibody" in this application is a monoclonal antibody to human PD-L1 prepared according to the knowledge and skills in the art. During preparation, the test subjects are injected with PD-L1 antigen, and then hybridomas expressing antibodies with desired sequence or functional properties are isolated. In a preferred embodiment of the present application, the mouse-derived PD-L1 antibody or antigen-binding fragment thereof may further comprise a light chain constant region of a mouse-derived κ, λ chain or a variant thereof, or further comprise a mouse-derived IgG1 , IgG2, IgG3 or its heavy chain constant region.

The term "chimeric antibody" is an antibody obtained by fusing the variable region of a murine antibody and the constant region of a human antibody, which can reduce the immune response induced by the murine antibody. To build a chimeric antibody, first build a hybridoma that secretes murine specific monoclonal antibodies, then clone the variable region gene from the mouse hybridoma cells, and then clone the human antibody constant region gene as needed The gene and human constant region gene are connected into a chimeric gene, and then inserted into a human vector. Finally, the chimeric antibody molecule is expressed in a eukaryotic industrial system or a prokaryotic industrial system. In a preferred embodiment of the present application, the light chain of the antibody of the PD-L1 chimeric antibody further comprises a light chain constant region of human κ, λ chain or a variant thereof. The antibody heavy chain of the PD-L1 chimeric antibody further comprises a heavy chain constant region of human-derived IgG1, IgG2, IgG3, IgG4 or a variant thereof. The constant region of the human antibody may be selected from the heavy chain constant region of human IgG1, IgG2, IgG3, or IgG4 or a variant thereof, preferably comprising the human heavy chain constant region of IgG2 or IgG4, or no amino acid mutation after using ADCC (antibody-dependent cell-mediated cytotoxicity, antibody-dependent cell-mediated cytotoxicity) toxic IgG4.

The term "humanized antibody", also known as CDR-grafted antibody, refers to the transplantation of mouse CDR sequences into the framework of human antibody variable regions, that is, different types of human germlines Antibodies produced in antibody framework sequences. It can overcome the strong antibody variable antibody reaction induced by the chimeric antibody due to carrying a large amount of mouse protein components. Such framework sequences can be obtained from public DNA databases or published references that include germline antibody gene sequences. For example, the germline DNA sequences of human heavy chain and light chain variable region genes can be found in the "VBase" human germline sequence database (available on the Internet at www.mrccpe.com.ac.uk/vbase), and in Kabat, EA, etc. People, 1991 Sequences of Proteins of Immunological Interest, found in 5th edition. In order to avoid the decrease of the immunogenicity and the decrease of the activity, the framework sequence of the variable region of the human antibody can be subjected to minimum reverse mutation or back mutation to maintain the activity. The humanized antibodies of the present application also include humanized antibodies that have been further affinity-matured to CDRs by phage display.

The "antigen-binding fragment" described in this application refers to a Fab fragment having antigen-binding activity, a Fab 'fragment, an F (ab') 2 fragment, and an Fv fragment ScFv fragment that binds to human PD-L1. The Fv fragment contains the antibody heavy chain variable region and the light chain variable region, but has no constant region, and has the smallest antibody fragment with all antigen binding sites. Generally, Fv antibodies also contain a polypeptide linker between the VH and VL domains, and can form the structure required for antigen binding. Different linkers can also be used to connect the variable regions of two antibodies into one polypeptide chain, called single chain antibody (single chain antibody) or single chain Fv (sFv). The term "associated with PD-L1" in this application means that it can interact with human PD-L1. The term "antigen binding site" in the present application refers to a discontinuous three-dimensional site on the antigen recognized by the antibody or antigen-binding fragment of the present application.

"Identity" refers to the sequence similarity between two polynucleotide sequences or between two polypeptides. When the positions in the two compared sequences are occupied by the same base or amino acid monomer subunit, for example, if each position of two DNA molecules is occupied by adenine, then the molecules are identical at that position. The percentage of identity between the two sequences is a function of the number of matching or identical positions shared by the two sequences divided by the number of positions compared × 100. For example, when the sequences are optimally aligned, if 6 of the 10 positions in the two sequences match or are identical, then the two sequences are 60% identical. In general, comparisons are made when aligning two sequences to obtain the maximum percentage of identity.

"Administration" and "treatment" when applied to animals, humans, experimental subjects, cells, tissues, organs or biological fluids refer to exogenous drugs, therapeutic agents, diagnostic agents or compositions and animals, humans, subjects Subjects, cells, tissues, organs or biological fluids. "Administration" and "treatment" may refer to, for example, treatment, pharmacokinetics, diagnosis, research, and experimental methods. The treatment of cells includes contact of reagents with cells, and contact of reagents with fluids, wherein the fluids contact cells. "Administering" and "treating" also mean in vitro and ex vivo treatment of, for example, cells by an agent, diagnosis, binding composition, or by another cell. "Treatment" when applied to human, veterinary or research subjects refers to therapeutic treatment, prophylactic or preventative measures, research and diagnostic applications. "Treatment" means administration of a therapeutic agent for internal or external use to a patient, such as a composition containing any of the binding compounds of the present application, the patient has one or more symptoms of the disease, and the therapeutic agent is known to have Therapeutic effect. Generally, the therapeutic agent is administered in an amount effective to alleviate one or more disease symptoms in the treated patient or group to induce the regression of such symptoms or inhibit the development of such symptoms to the extent of any clinical measurement. The amount of therapeutic agent effective to relieve the symptoms of any particular disease (also referred to as a "therapeutically effective amount") can vary based on various factors, such as the patient's disease state, age, and body weight, and the ability of the drug to produce a desired therapeutic effect in the patient. It is possible to assess whether the symptoms of the disease have been alleviated by any clinical testing methods commonly used by doctors or other professional health care personnel to assess the severity or progression of the symptoms. Although embodiments of the present application (eg, treatment methods or preparations) may be ineffective in relieving the symptoms of each target disease, according to any statistical test methods known in the art such as Student's test, chi-square test, according to Mann and Whitney's U test, Kruskal-Wallis test (H test), Jonckheere-Terpstra test and Wilcoxon test determined that it should alleviate the target disease symptoms in a statistically significant number of patients.

The "combination" described in this application is a mode of administration, which means that at least one dose of irinotecan or a pharmaceutically acceptable salt thereof and at least one dose of an immune checkpoint inhibitor are administered within a certain period of time, and at least One dose of 5-FU, three of which show pharmacological effects. The time period may be within one administration cycle, preferably within 4 weeks, within 3 weeks, within 2 weeks, within 1 week, or within 24 hours, more preferably within 12 hours. Irinotecan or its pharmaceutically acceptable salt, immune checkpoint inhibitor, and 5-FU can be administered simultaneously or sequentially. This period includes treatments in which irinotecan or its pharmaceutically acceptable salts, immune checkpoint inhibitors, and 5-FU are administered through the same route of administration or different routes of administration. The combined administration method described herein is selected from simultaneous administration, independently formulated and co-administered, or independently formulated and sequentially administered.

"Effective amount" includes an amount sufficient to ameliorate or prevent the symptoms or conditions of medical diseases. An effective amount also means an amount sufficient to allow or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on factors such as the condition to be treated, the patient's general health, the route and dosage of the method of administration, and the severity of side effects. The effective amount may be the maximum dose or dosing regimen that avoids significant side effects or toxic effects.

The "toxic intolerance" mentioned in this application means that the adverse reactions caused by the drug cannot continue to be treated.

Progression-free survival (PFS): from the random start to the date when the objective progression of the previous adenocarcinoma was recorded for the first time or to any cause of death, whichever occurs first.

Overall survival (OS) refers to the period from random to any cause of death. For the subjects who were still alive at the last follow-up, the OS was deleted based on the time of the last follow-up. For the subjects who were lost to follow-up, the OS was counted as data deletion based on the last confirmed survival time before the loss of follow-up. The OS for data censoring is defined as the time from random grouping to censoring.

Objective response rate (ORR) refers to the proportion of patients whose tumor shrinkage reaches a certain level and remains for a certain period of time, including cases of CR and PR. Use solid tumor response evaluation criteria (RECIST 1.1 standard) to assess objective tumor response. Subjects must be accompanied by measurable tumor lesions at baseline. The evaluation criteria for efficacy are divided into complete response (CR), partial response (PR), stability (SD), and progression (PD) according to the RECIST 1.1 standard.

Duration of remission (DoR): the time from the first PR or CR to the first PD or death.

Disease Control Rate (DCR) refers to the percentage of confirmed complete remission, partial remission, and stable disease (≥8 weeks) in the number of patients with evaluable efficacy.

Target lesion assessment

Complete remission (CR): All target lesions disappear, and all pathological lymph nodes (including target nodules and non-target nodules) must be reduced in diameter to <10 mm.

Partial remission (PR): The sum of target lesion diameters is reduced by at least 30% from the baseline level.

Disease progression (PD): the minimum value of the sum of the diameters of all the measured target lesions in the entire experimental study is taken as the reference, and the diameter and the relative increase are at least 20% (if the baseline measurement is the smallest, the baseline value is used as the reference); otherwise In addition, the absolute value of the sum of diameters must be increased by at least 5 mm (the appearance of one or more new lesions is also regarded as disease progression).

Disease Stability (SD): The target lesions did not decrease in PR to the extent of the increase, nor did they reach the PD level. Between the two, the minimum sum of diameters can be used as a reference during the study.

Evaluation of non-target lesions

Complete remission (CR): All non-target lesions disappear and tumor markers return to normal levels. All lymph nodes are of non-pathological size (short diameter <10mm).

Incomplete remission / non-progression: the presence of one or more non-target lesions and / or the persistent presence of tumor marker levels above normal levels.

Disease progression: Existing non-target lesions show clear progression. Note: The appearance of one or more new lesions is also considered disease progression.

detailed description

Example 1: A clinical trial of first-line treatment of advanced esophageal squamous cell carcinoma with irinotecan hydrochloride liposomes combined with PD-L1 antibody and 5-FU

1. Test drugs

Irinotecan hydrochloride liposomes were prepared as irinotecan liposome lyophilized powder injections according to the method disclosed in CN103120645A according to the prescriptions in the table below, with a specification of 40 mg / piece.

Table 1: Irinotecan liposome prescription

Note: a 40mg dosage is based on irinotecan hydrochloride (C ₃₃ H ₃₈ N ₄ O ₆ · HCl) without crystal water, b is basically removed in the final product, c is basically removed in the final product, and d is converted into anhydrous diphosphate The amount of sodium hydrogen, the actual use is sodium dihydrogen phosphate monohydrate, because irinotecan hydrochloride has crystal water, and there is a loss of about 8% after feeding, the actual dosage is according to irinotecan hydrochloride: hydrogenated soybean phosphatidylcholine chol Alkali = 3:10 ratio feeding.

5-fluorouracil injection, specification: 250mg / piece; R & D unit: Shanghai Xudong Haipu Pharmaceutical Co., Ltd.

The PD-L1 antibody corresponds to the code name HRP00052, the heavy chain sequence is SEQ ID NO: 9, and the light chain sequence is SEQ ID NO: 11. Specification: 600mg / support.

2. Enrolled subjects

(1) Age 18-70 years old, both men and women;

(2) Unresectable locally advanced / relapse diagnosed by histology or cytology (cannot accept radical treatment such as radical radiochemotherapy or radical radiotherapy) or distant metastatic esophageal squamous cell carcinoma

(3) For patients who have not received systemic anti-tumor therapy or received neoadjuvant / adjuvant and radical concurrent radiotherapy and chemotherapy, the time from the last chemotherapy to the time of relapse or progression exceeds 6 months.

3. Method of administration

Qualified subjects were given irinotecan hydrochloride liposomes, HRP00052 antibody and 5-FU.

The dosage of irinotecan hydrochloride liposome is 60 kg / m ² or 80 kg / m ² per cycle, the dosage of 5-FU is 2400 mg / m ² per cycle, and the dosage of HRP00052 is 10 mg / kg per cycle. The dosing cycle of the three drugs is one every two weeks, and the dosing is on the first day of each cycle.

4. Test results

A total of 23 subjects were enrolled, of which 22 subjects completed more than one efficacy evaluation (8 subjects completed 1 efficacy evaluation, 11 subjects completed 2 efficacy evaluations, and 3 subjects completed 3 efficacy evaluations). 9 subjects PR, 9 subjects SD, 4 subjects PD, the effective rate is shown in Table 1. The overall incidence of adverse events, the incidence of drug-related ≥3 grade adverse events, and the incidence of drug-related adverse events> 10% are shown in Table 2, Table 3, and Table 4. Among them, the adverse events of irinotecan + 5FU group data are irinotecan preparations

Adverse reaction data recorded in the market drug insert.

Table 1 The efficacy of this study

The objective response rate of this study was 40.9%, which was higher than the objective response rate of previous irinotecan related studies, demonstrating the better antitumor activity of the combined medication.

Table 2 Incidence of overall adverse events

病例数(N＝23)Number of cases (N = 23)	例次Example	例数Number of cases	发生率Incidence
不良事件Adverse event	10461046	21twenty one	91.30％91.30%
CTCAE≥3级AECTCAE≥3 grade AE	1717	99	39.13％39.13%
严重不良事件Serious adverse event	55	33	13.04％13.04%
导致永久停药的AEAE leading to permanent withdrawal	00	00	0％0%
特别关注不良事件Special attention to adverse events	00	00	0％0%

Table 3 Incidence of ≥3 grade adverse events related to study drug

Table 4 The incidence of drug-related adverse events> 10%

Taken together, irinotecan liposomes combined with PD-L1 antibody and 5-FU first-line treatment of advanced esophageal squamous cell carcinoma not only show the anti-tumor activity of the drug, but also show good safety data.

Claims

Use of irinotecan or a pharmaceutically acceptable salt thereof in combination with an immune checkpoint inhibitor and 5-FU in the preparation of a medicament for preventing or treating tumor diseases.
The use according to claim 1, wherein the irinotecan or its pharmaceutically acceptable salt is administered in the form of liposomes.
The use according to claim 2, wherein the irinotecan liposome contains irinotecan or a pharmaceutically acceptable salt thereof, neutral phospholipid and cholesterol.
The use according to claim 3, wherein the weight ratio of cholesterol to neutral phospholipid is 1: 3 to 5, preferably 1: 3.5 to 4.5, most preferably 1: 4.
The use according to claim 3, wherein the weight ratio of irinotecan or a pharmaceutically acceptable salt thereof to neutral phospholipid is 1: 2 to 5, preferably 1: 2.5-4, most preferably 1: 4 .
The use according to claim 3, wherein the neutral phospholipid is one or more selected from hydrogenated soybean phosphatidylcholine, phosphatidylethanolamine, lecithin, cardiolipin, preferably hydrogenated soybean phospholipid.
The use according to claim 3, wherein the liposome further contains a lipid derivative of a hydrophilic polymer, preferably a cultured phosphatidylethanolamine DSPE-PEG 2000 .
The use according to claim 1, wherein the immune checkpoint inhibitor is selected from PD-L1 antibodies or antigen-binding fragments.
The use according to claim 8, wherein any one of the PD-L1 antibodies or antigen-binding fragments is selected from the following CDR region sequences or mutant sequences thereof: antibody heavy chain variable region HCDR region sequence: SEQ ID NO: 1-3; and antibody light chain variable region LCDR region sequence: SEQ ID NO: 4-6; specific as follows:

HCDR1 is selected from:

SYWMH SEQ ID NO: 1

HCDR2 is selected from:

RI GPNSG FTSYNEKFKN SEQ ID NO: 2

HCDR3 is selected from:

GGSSYDYFDY SEQ ID NO: 3

LCDR1 is selected from:

RASESVSIHGTHLMH SEQ ID NO: 4

LCDR2 is selected from:

AASNLES SEQ ID NO: 5

LCDR3 is selected from:

QQSFEDPLT SEQ ID NO: 6.
The use according to claim 9, wherein the sequence of the heavy chain variable region of the PD-L1 antibody or antigen-binding fragment is SEQ ID NO: 7, and the sequence of the light chain variable region is SEQ ID NO: 8.
The use according to claim 10, wherein the heavy chain sequence of the PD-L1 antibody or antigen-binding fragment is SEQ ID NO: 9, and the light chain sequence is SEQ ID NO: 11.
The use according to any one of claims 1 to 11, wherein the tumor disease is selected from malignant tumors and benign tumors; preferably esophageal cancer, more preferably esophageal squamous cell carcinoma and esophageal adenocarcinoma.
The use according to any one of claims 1-12, wherein the tumor is selected from advanced tumors, relapsed refractory tumors, failed and / or relapsed tumors after chemotherapy treatment, failed radiotherapy and / or relapsed tumors Targeted drug therapy failed and / or relapsed tumor, immunotherapy failed and / or relapsed tumor.
The use according to any one of claims 1 to 13, wherein the tumor is selected from advanced esophageal cancer, recurrent esophageal cancer, and distant metastatic esophageal cancer.
The use according to any one of claims 1-14, wherein the pharmaceutically acceptable salt of irinotecan is hydrochloride, and its dosage range is selected from 40-200 mg / m 2 based on irinotecan hydrochloride.
The use according to any one of claims 1-15, wherein the dose of 5-FU is 1000-3000 mg / m 2 .
The use according to any one of claims 1-16, wherein the dose of the immune checkpoint inhibitor is selected from 1-50 mg / kg.
The use according to any one of claims 1-17, wherein the dose of the immune checkpoint inhibitor is selected from 50-3000 mg.
A pharmaceutical composition comprising irinotecan or a pharmaceutically acceptable salt thereof, an immune checkpoint inhibitor and 5-FU according to claim 1, and one or more pharmaceutically acceptable excipients, diluents or carriers.