Use of a KRAS G12D inhibitor in combination with an anti-EGFR antibody in the manufacture of a medicament for treating a solid tumor

Abstract

The present disclosure relates to the use of a KRAS G12D inhibitor in combination with an anti-EGFR antibody in the manufacture of a medicament for the treatment of a solid tumor. In particular, the present disclosure relates to the use of a KRAS G12D inhibitor in combination with an anti-EGFR antibody, or an antigen-binding fragment thereof, in the manufacture of a medicament for the treatment of a solid tumor having a KRAS G12D mutation.

Description

Technical Field

[0001] This disclosure relates to the use of a KRAS G12D inhibitor in combination with an anti-EGFR antibody or its antigen-binding fragment in the preparation of a medicament for treating KRAS G12D-mutant solid tumors, and belongs to the pharmaceutical field. Background Technology

[0002] RAS (Rapid Acid Spectroscopy) genes are among the most frequently mutated oncogenes in tumors, with approximately 30% of human malignancies associated with RAS gene mutations. The RAS family includes KRAS, NRAS, and HRAS, with KRAS mutations being the most common, accounting for about 85%. KRAS mutations are common in solid tumors, showing high frequency in the three leading causes of cancer death in humans—lung cancer (17%), colorectal cancer (33%), and pancreatic cancer (61%). In KRAS gene mutations, 97% involve mutations at amino acid residue 12 or 13, with G12D being a significant mutation. Data analysis of European and American populations shows that G12D mutations account for 36%, 12%, and 4% of patients with pancreatic cancer, colorectal cancer, and non-small cell lung cancer, respectively.

[0003] Once activated, KRAS regulates various functions of cell proliferation, survival, migration, and metabolism through numerous downstream signaling pathways, including RAF-MEK-ERK, PI3K-AKT-mTOR, and TIAM1-RAc. Mutations in the KRAS gene result in a persistently activated protein, leading to continued activation of downstream signaling pathways and promoting tumorigenesis. Because the KRAS protein lacks traditional small-molecule binding sites and exhibits extremely high affinity for guanosine monophosphate (GMP), making it very difficult to inhibit, it has long been considered an untreatable drug target. However, given the importance and prevalence of abnormal KRAS activation in cancer progression, KRAS remains a highly anticipated target in drug development. Currently, apart from KRAS G12C inhibitors, there are still no effective KRAS inhibitors for other mutations, leaving most patients with KRAS mutations without a cure. G12D, as a mutant widely and highly expressed in various tumors, presents significant clinical potential for the development of inhibitors.

[0004] WO2022268051A provides a novel KRAS G12D inhibitor compound (Formula I) with good pharmaceutical activity.

[0005]

[0006] We hope to develop new methods or uses for the administration of KRAS G12D inhibitors in combination with other therapeutic agents, particularly in combination with anti-EGFR antibodies or their antigen-binding fragments. Summary of the Invention

[0007] The purpose of this disclosure is to provide the use of a KRAS G12D inhibitor in combination with an anti-EGFR antibody or an antigen-binding fragment thereof in the preparation of a medicament for treating KRAS G12D-mutant solid tumors.

[0008] In some embodiments, the KRAS G12D inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0009] In an optional implementation, the solid tumor described in this disclosure is selected from colorectal cancer and lung cancer.

[0010] In some implementations, the solid tumor is selected from advanced solid tumors.

[0011] In some implementations, the advanced solid tumor is selected from advanced solid tumors that have relapsed or progressed after standard treatment, or for which there is no standard treatment option, or for which standard treatment is not applicable at this stage.

[0012] In optional embodiments, the colorectal cancer described in this disclosure is selected from adenocarcinoma (e.g., special types of adenocarcinoma or non-special types of adenocarcinoma), adenosquamous carcinoma, squamous cell carcinoma, and undifferentiated carcinoma.

[0013] In an optional implementation, the colorectal cancer described in this disclosure is selected from adenocarcinoma.

[0014] In an optional implementation, the advanced solid tumor described in this disclosure is selected from recurrent or metastatic colorectal adenocarcinoma.

[0015] In optional embodiments, the specific types of adenocarcinoma described in this disclosure include, but are not limited to, mucinous adenocarcinoma, signet ring cell carcinoma, medullary carcinoma, serrated adenocarcinoma, micropapillary carcinoma, adenomatous adenocarcinoma, adenosquamous carcinoma, and carcinoma with sarcomatoid components.

[0016] In an optional implementation, the lung cancer described in this disclosure is selected from non-small cell lung cancer and small cell lung cancer.

[0017] In an optional implementation, the lung cancer described in this disclosure is selected from non-small cell lung cancer.

[0018] In an optional embodiment, the advanced solid tumor described in this disclosure is selected from recurrent or metastatic non-small cell lung cancer. In an optional embodiment, the KRAS G12D-mutated solid tumor described in this disclosure is KRAS G12D-mutated non-small cell lung cancer.

[0019] In an optional implementation, the KRAS G12D-mutated solid tumor described in this disclosure is advanced non-small cell lung cancer with KRAS G12D mutation.

[0020] In some embodiments, the dosage of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from 50-2000 mg; the frequency of administration is selected from once a week, once every 2 weeks, once every 3 weeks or once every 4 weeks.

[0021] In an optional embodiment, the dosage of the compound of formula (I) described in this disclosure or a pharmaceutically acceptable salt thereof may be 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg, 1600 mg, 1700 mg, 1800 mg, 1900 mg, 2000 mg, or any value between any two; the dosing frequency is selected from once a week, once every two weeks, or once every three weeks.

[0022] In an optional embodiment, the dosage of the compound of formula (I) described in this disclosure or a pharmaceutically acceptable salt thereof may be 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, 1000 mg, or 1200 mg, and the frequency of administration may be selected from once a week, once every two weeks, or once every three weeks.

[0023] In an optional embodiment, the dosage of the compound of formula (I) of this disclosure or a pharmaceutically acceptable salt thereof is 200-600 mg, and the frequency of administration is selected from once a week, specifically, for example, a dosage of 300 mg once a week, a dosage of 400 mg once a week, a dosage of 500 mg once a week, or a dosage of 600 mg once a week.

[0024] In an optional embodiment, the compound of formula (I) described in this disclosure or a pharmaceutically acceptable salt thereof is administered at a dose of 800 mg once every 2 weeks.

[0025] In an optional embodiment, the compound of formula (I) described in this disclosure or a pharmaceutically acceptable salt thereof is administered at a dose of 1000 mg once every 2 weeks.

[0026] In an optional embodiment, the compound of formula (I) described in this disclosure or a pharmaceutically acceptable salt thereof is administered at a dose of 1200 mg once every 2 weeks.

[0027] In an optional embodiment, the dosage of the compound of formula (I) of this disclosure or a pharmaceutically acceptable salt thereof is 400-600 mg D1 + 800-1200 mg D8, administered once every 3 weeks.

[0028] In some embodiments, the anti-EGFR antibody or its antigen-binding fragment described in this disclosure comprises LCDR1 with an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence differing from SEQ ID NO:1 by 1, 2, or 3 amino acids; LCDR2 with an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence differing from SEQ ID NO:2 by 1, 2, or 3 amino acids; LCDR3 with an amino acid sequence as shown in SEQ ID NO:3 or an amino acid sequence differing from SEQ ID NO:3 by 1, 2, or 3 amino acids; HCDR1 with an amino acid sequence as shown in SEQ ID NO:4 or an amino acid sequence differing from SEQ ID NO:4 by 1, 2, or 3 amino acids; HCDR2 with an amino acid sequence as shown in SEQ ID NO:5 or an amino acid sequence differing from SEQ ID NO:5 by 1, 2, or 3 amino acids; and HCDR3 with an amino acid sequence as shown in SEQ ID NO:6 or an amino acid sequence differing from SEQ ID NO:6 by 1, 2, or 3 amino acids.

[0029] In some embodiments, the anti-EGFR antibody or its antigen-binding fragment described in this disclosure comprises LCDR1 with the amino acid sequence shown in SEQ ID NO:1, LCDR2 with the amino acid sequence shown in SEQ ID NO:2, LCDR3 with the amino acid sequence shown in SEQ ID NO:3, HCDR1 with the amino acid sequence shown in SEQ ID NO:4, HCDR2 with the amino acid sequence shown in SEQ ID NO:5, and HCDR3 with the amino acid sequence shown in SEQ ID NO:6.

[0030] In some embodiments, the anti-EGFR antibody or antigen-binding fragment thereof described in this disclosure comprises a light chain variable region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the amino acid sequence shown in SEQ ID NO:7, and a heavy chain variable region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the amino acid sequence shown in SEQ ID NO:8.

[0031] In some embodiments, the anti-EGFR antibody or its antigen-binding fragment comprises a light chain variable region with an amino acid sequence as shown in SEQ ID NO:7, and a heavy chain variable region with an amino acid sequence as shown in SEQ ID NO:8.

[0032] In some embodiments, the anti-EGFR antibody or its antigen-binding fragment comprises a light chain having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the amino acid sequence shown in SEQ ID NO:9, and a heavy chain having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the amino acid sequence shown in SEQ ID NO:10.

[0033] In some embodiments, the anti-EGFR antibody comprises a light chain with an amino acid sequence as shown in SEQ ID NO:9 and a heavy chain with an amino acid sequence as shown in SEQ ID NO:10.

[0034] In some implementations, the anti-EGFR antibody is cetuximab or a biosimilar thereof.

[0035] In some embodiments, the anti-EGFR antibody or its antigen-binding fragment is administered at a dose selected from about 100 mg / m² based on body surface area. 2 Approximately 800 mg / m 2 For example, selected from approximately 200 mg / m 2 Approximately 500 mg / m 2 For example, approximately 100 mg / m³ 2 Approximately 150 mg / m 2 Approximately 200 mg / m 2 Approximately 250 mg / m 2 Approximately 300 mg / m 2 Approximately 350 mg / m 2 Approximately 400 mg / m 2 Approximately 450 mg / m 2 Approximately 500 mg / m 2 Approximately 550 mg / m 2 Approximately 600 mg / m 2 Approximately 650 mg / m 2 Approximately 700 mg / m 2 Approximately 750 mg / m 2 or approximately 800 mg / m 2 The administration frequency is once a week or once every two weeks.

[0036] In some embodiments, the anti-EGFR antibody or its antigen-binding fragment is administered at a dose of approximately 200 mg / m² based on body surface area. 2 Approximately 250 mg / m 2 Approximately 300 mg / m 2 Approximately 350 mg / m 2 or approximately 400 mg / m 2 The administration frequency is once a week.

[0037] In some embodiments, the initial dose of the anti-EGFR antibody or its antigen-binding fragment is approximately 400 mg / m² based on body surface area. 2 The weekly dosage thereafter is approximately 250 mg / m² based on body surface area. 2 The medication should be administered once a week.

[0038] In some embodiments, the initial dose of the anti-EGFR antibody or its antigen-binding fragment is approximately 400 mg / m² based on body surface area. 2 The weekly dosage thereafter is approximately 200 mg / m² based on body surface area. 2 The medication should be administered once a week.

[0039] In some embodiments, the initial dose of the anti-EGFR antibody or its antigen-binding fragment is approximately 400 mg / m² based on body surface area. 2 The weekly dosage thereafter is approximately 150 mg / m² based on body surface area. 2 The medication should be administered once a week.

[0040] In some embodiments, the anti-EGFR antibody or its antigen-binding fragment is administered at a dose of approximately 500 mg / m² based on body surface area. 2 The medication should be administered once every two weeks.

[0041] In some embodiments, the anti-EGFR antibody or its antigen-binding fragment is administered at a dose of approximately 400 mg / m² based on body surface area. 2 The medication should be administered once every two weeks.

[0042] In some embodiments, the anti-EGFR antibody or its antigen-binding fragment is administered at a dose of approximately 300 mg / m² based on body surface area. 2 The dosing frequency is once every 2 weeks.

[0043] In an optional embodiment, the dosage of the compound of formula (I) or a pharmaceutically acceptable salt thereof is 200-600 mg, and the frequency of administration is selected from once a week; the initial dose of the anti-EGFR antibody or its antigen-binding fragment is approximately 400 mg / m² based on body surface area. 2The weekly dosage thereafter is approximately 250 mg / m² based on body surface area. 2 The medication should be administered once a week.

[0044] In an optional embodiment, the compound of formula (I) of this disclosure or a pharmaceutically acceptable salt thereof is administered at a dose of 300 mg once weekly; the anti-EGFR antibody or its antigen-binding fragment is initially administered at a dose of approximately 400 mg / m² based on body surface area. 2 The weekly dosage thereafter is approximately 250 mg / m² based on body surface area. 2 The medication should be administered once a week.

[0045] In an optional embodiment, the compound of formula (I) of this disclosure or a pharmaceutically acceptable salt thereof is administered at a dose of 400 mg once weekly; the anti-EGFR antibody or its antigen-binding fragment is initially administered at a dose of approximately 400 mg / m² based on body surface area. 2 The weekly dosage thereafter is approximately 250 mg / m² based on body surface area. 2 The medication should be administered once a week.

[0046] In an optional embodiment, the dosage of the compound of formula (I) or a pharmaceutically acceptable salt thereof is 500 mg, administered once weekly; the initial dose of the anti-EGFR antibody or its antigen-binding fragment is approximately 400 mg / m² based on body surface area. 2 The weekly dosage thereafter is approximately 250 mg / m² based on body surface area. 2 The medication should be administered once a week.

[0047] In an optional embodiment, the compound of formula (I) of this disclosure or a pharmaceutically acceptable salt thereof is administered at a dose of 800 mg, 1000 mg, or 1200 mg, once every 2 weeks; the anti-EGFR antibody or its antigen-binding fragment is initially administered at a dose of approximately 400 mg / m² based on body surface area. 2 The weekly dosage thereafter is approximately 250 mg / m² based on body surface area. 2 The medication should be administered once a week.

[0048] In an optional embodiment, the dosage of the compound of formula (I) or a pharmaceutically acceptable salt thereof is 800 mg, administered once every 2 weeks; the initial dose of the anti-EGFR antibody or its antigen-binding fragment is approximately 400 mg / m² based on body surface area. 2 The weekly dosage thereafter is approximately 250 mg / m² based on body surface area. 2 The medication should be administered once a week.

[0049] In an optional embodiment, the dosage of the compound of formula (I) or a pharmaceutically acceptable salt thereof is 1200 mg, administered once every 2 weeks; the initial dose of the anti-EGFR antibody or its antigen-binding fragment is approximately 400 mg / m² based on body surface area. 2 The weekly dosage thereafter is approximately 250 mg / m² based on body surface area. 2 The medication should be administered once a week.

[0050] In an optional embodiment, the dosage of the compound of formula (I) of this disclosure or a pharmaceutically acceptable salt thereof is 200-600 mg QW, and the frequency of administration is selected from once a week; the dosage of the anti-EGFR antibody or its antigen-binding fragment is about 500 mg / m² based on body surface area. 2 The medication should be administered once every two weeks.

[0051] In an optional embodiment, the compound of formula (I) of this disclosure or a pharmaceutically acceptable salt thereof is administered at a dose of 300 mg once weekly; the anti-EGFR antibody or its antigen-binding fragment is administered at a dose of approximately 500 mg / m² based on body surface area. 2 The medication should be administered once every two weeks.

[0052] In an optional embodiment, the compound of formula (I) of this disclosure or a pharmaceutically acceptable salt thereof is administered at a dose of 400 mg once weekly; the anti-EGFR antibody or its antigen-binding fragment is administered at a dose of approximately 500 mg / m² based on body surface area. 2 The medication should be administered once every two weeks.

[0053] In an optional embodiment, the compound of formula (I) of this disclosure or a pharmaceutically acceptable salt thereof is administered at a dose of 500 mg once weekly; the anti-EGFR antibody or its antigen-binding fragment is administered at a dose of approximately 500 mg / m² based on body surface area. 2 The medication should be administered once every two weeks.

[0054] In an optional embodiment, the dosage of the compound of formula (I) or a pharmaceutically acceptable salt thereof is 800 mg, 1000 mg, or 1200 mg, administered once every two weeks; the dosage of the anti-EGFR antibody or its antigen-binding fragment is approximately 500 mg / m² based on body surface area. 2 The medication should be administered once every two weeks.

[0055] In an optional embodiment, the dosage of the compound of formula (I) or a pharmaceutically acceptable salt thereof is 800 mg, administered once every 2 weeks; the dosage of the anti-EGFR antibody or its antigen-binding fragment is approximately 500 mg / m² based on body surface area. 2 The medication should be administered once every two weeks.

[0056] In an optional embodiment, the compound of formula (I) of this disclosure or a pharmaceutically acceptable salt thereof is administered at a dose of 1000 mg once every 2 weeks; the anti-EGFR antibody or its antigen-binding fragment is administered at a dose of approximately 500 mg / m² based on body surface area. 2 The medication should be administered once every two weeks.

[0057] In an optional embodiment, the dosage of the compound of formula (I) or a pharmaceutically acceptable salt thereof is 1200 mg, administered once every 2 weeks; the dosage of the anti-EGFR antibody or its antigen-binding fragment is approximately 500 mg / m² based on body surface area. 2 The medication should be administered once every two weeks.

[0058] The amino acid sequences corresponding to the aforementioned sequence numbers are shown below:

[0059] CDR sequence of anti-EGFR antibody or its antigen-binding fragment

[0060] LCDR1 SEQ ID NO:1 RASQSIGTNIH LCDR2 SEQ ID NO:2 YASESIS LCDR3 SEQ ID NO:3 QQNNNWPTT HCDR1 SEQ ID NO:4 NYGVH HCDR2 SEQ ID NO:5 VIWSGGNTDYNTPFTS HCDR2 SEQ ID NO:6 ALTYYDYEFAY

[0061] SEQ ID NO:7

[0062] DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELK

[0063] SEQ ID NO:8

[0064] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSA

[0065] SEQ ID NO:9

[0066] DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0067] SEQ ID NO:10

[0068] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQG TLVTVSAASTKGPSVFPLAPSSKSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0069] In an optional embodiment, the compound of formula (I) described in this disclosure or a pharmaceutically acceptable salt thereof is in the form of a pharmaceutical composition, specifically, liposomes.

[0070] In an optional embodiment, the compound of formula (I) described in this disclosure or a pharmaceutically acceptable salt thereof is administered via intravenous injection.

[0071] This disclosure also provides a method for treating KRAS G12D-mutated solid tumors by administering to a subject a compound of the aforementioned formula (I) or a pharmaceutically acceptable salt thereof and an anti-EGFR antibody or an antigen-binding fragment thereof.

[0072] In an optional implementation, the compound of formula (I) described above, or a pharmaceutically acceptable salt thereof, and an anti-EGFR antibody or an antigen-binding fragment thereof are provided to the subject in a therapeutically effective amount.

[0073] In an optional implementation, the subject described in this disclosure is a patient.

[0074] Another aspect of this disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for treating KRAS G12D-mutated solid tumors, which is administered in combination with an anti-EGFR antibody or an antigen-binding fragment thereof.

[0075] Another aspect of this disclosure provides an anti-EGFR antibody or an antigen-binding fragment thereof for treating KRAS G12D-mutated solid tumors, which is administered in combination with a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0076] In an optional implementation, the duration of administration of the KRASG12D inhibitor described herein with the anti-EGFR antibody or its antigen-binding fragment is selected from 2 weeks, 4 weeks, 8 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks or longer, such as half a year, 1 year or 2 years.

[0077] The combined routes of administration described in this disclosure are selected from oral administration, parenteral administration, and transdermal administration, wherein parenteral administration includes, but is not limited to, intravenous injection, subcutaneous injection, and intramuscular injection.

[0078] The subjects described in this invention may be mammals, specifically healthy humans or patients.

[0079] The “recurrent or metastatic colorectal adenocarcinoma” described in this disclosure refers to patients who have failed previous treatment with oxaliplatin, 5-fluorouracil, or irinotecan, and those with dMMR (mismatch repair gene deletion) or MSI-H (microsatellite instability) who have failed anti-PD-1 / PD-L1 therapy and have not received EGFR antibody therapy such as cetuximab or panitumumab.

[0080] The "combination" described in this disclosure refers to a route of administration in which at least one dose of a KRAS G12D inhibitor and at least one dose of an anti-EGFR antibody or its antigen-binding fragment are administered over a specified time period, wherein the two drugs exert pharmacological effects simultaneously at at least one point in time or over a specified period. Because some drugs have long half-lives, the two drugs can be administered over a longer time interval while still achieving the effect of simultaneous pharmacological effects at a specific point in time or over a specified period. The time period or interval depends on the half-life of the drugs, and is preferably within 4 weeks, 3 weeks, 2 weeks, 1 week, or less than 24 hours, more preferably within 12 hours. The KRAS G12D inhibitor and the anti-EGFR antibody or its antigen-binding fragment can be administered simultaneously or sequentially. The combination includes administering the KRAS G12D inhibitor and the anti-EGFR antibody or its antigen-binding fragment via the same or different routes of administration. The route of administration for the combination described in this disclosure is selected from simultaneous administration, independently formulated and co-administered, or independently formulated and sequentially administered.

[0081] Progression-free survival (PFS): The time from randomization to the date of first recorded objective progression of the tumor or to death from any cause, whichever comes first.

[0082] Overall survival (OS) refers to the period from randomization to death from any cause. For subjects still alive at the last follow-up, the OS is defined as the time to the last follow-up, indicating data loss. For subjects lost to follow-up, the OS is defined as the time to the last confirmed survival prior to loss to follow-up, indicating data loss. OS with data censoring is defined as the time from randomization to censoring.

[0083] The objective response rate (ORR) refers to the proportion of patients whose tumors shrink to a certain extent and remain so for a certain period of time, including cases of complete response (CR) and partial response (PR). The RECIST 1.1 criteria for evaluating response in solid tumors are used to assess objective response. Subjects must have measurable tumor lesions at baseline. The efficacy assessment criteria, according to RECIST 1.1, are divided into complete response (CR), partial response (PR), stable disease (SD), and progressive disease (PD).

[0084] The Disease Control Rate (DCR) is the percentage of patients with confirmed complete remission, partial remission, and stable disease (≥8 weeks) among those with evaluable efficacy.

[0085] Complete remission (CR): All target lesions disappear and the short diameter of all pathological lymph nodes (including target nodules and non-target nodules) must be reduced to <10 mm.

[0086] Partial remission (PR): The sum of the diameters of the target lesions is reduced by at least 30% compared to the baseline level.

[0087] Disease progression (PD): The minimum sum of the diameters of all target lesions measured throughout the entire experimental study is used as a reference, with a relative increase of at least 20% in the sum of diameters (or the baseline value if the baseline measurement is the minimum); in addition, the absolute value of the sum of diameters must increase by at least 5 mm (the appearance of one or more new lesions is also considered disease progression).

[0088] Disease stability (SD): The degree of reduction in target lesions does not reach the PR level, nor does the degree of increase reach the PD level; it falls between the two. The minimum value of the sum of diameters can be used as a reference in studies.

[0089] The term "effective amount" or "effective therapeutic amount" as used in this disclosure includes an amount sufficient to improve or prevent symptoms or conditions of a medical condition. 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 overall health, the route and dosage of administration, and the severity of side effects. An effective amount may be the maximum dose or administration regimen that avoids significant side effects or toxicity.

[0090] The three-letter and single-letter codes for amino acids used in this disclosure are as described in J. biol. chem, 243, p3558 (1968).

[0091] The term "antibody" is used in the broadest sense, encompassing various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies; monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies), full-length antibodies, and antibody fragments (or antigen-binding fragments, or antigen-binding portions), as long as they exhibit the desired antigen-binding activity. Antibodies can refer to immunoglobulins, which are tetrapeptide chains composed of two identical heavy chains and two identical light chains linked by interchain disulfide bonds. The amino acid composition and sequence of the constant region of the immunoglobulin heavy chain differ, thus their antigenicity also differs. Based on this, immunoglobulins can be divided into five classes, or isotypes of immunoglobulins: IgM, IgD, IgG, IgA, and IgE, with their corresponding heavy chains being μ, δ, γ, α, and ε chains, respectively. Within the same class of Ig, based on differences in the amino acid composition of the hinge region and the number and position of disulfide bonds in the heavy chain, different subclasses can be distinguished; for example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4. Light chains are classified as κ chains or λ chains based on differences in their constant regions. Each of the five classes of Ig can have either a κ chain or a λ chain. The sequence of approximately 110 amino acids near the N-terminus of the antibody heavy and light chains varies considerably, forming the variable region (V region); the remaining amino acid sequences near the C-terminus are relatively stable, forming the constant region (C region). The variable region includes three hypervariable regions (CDRs) and four relatively conserved backbone regions (FRs). The three hypervariable regions determine the antibody's specificity and are also known as complementarity-determining regions (CDRs). Each light chain variable region (VL) and heavy chain variable region (VH) consists of three CDRs and four FRs, arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The three CDRs of the light chain refer to LCDR1, LCDR2, and LCDR3; the three CDRs of the heavy chain refer to HCDR1, HCDR2, and HCDR3.

[0092] The determination or definition of a contact ligand (CDR) can be accomplished by resolving the structure of the antibody and / or the structure of the antibody-ligand complex, thereby enabling the definitive depiction of the CDR and the identification of residues containing the antibody binding site. This can be achieved using any of the various techniques known to those skilled in the art, such as X-ray crystallography. A variety of analytical methods can be used to identify CDRs, including but not limited to the Kabat numbering system, the Chothia numbering system, the AbM numbering system, the IMGT numbering system, contact definitions, and conformational definitions.

[0093] The Kabat numbering system is the standard for numbering residues in antibodies and is commonly used to identify CDR regions (see, for example, Johnson & Wu, 2000, Nucleic Acids Res., 28: 214-8). The Chothia numbering system is similar to the Kabat system, but it takes into account the location of certain structural loop regions (see, for example, Chothia et al., 1986, J. Mol. Biol., 196: 901-17; Chothia et al., 1989, Nature, 342: 877-83). The AbM numbering system uses a computer program integration suite produced by the Oxford Molecular Group to model antibody structures (see, for example, Martin et al., 1989, ProcNatl Acad Sci (USA), 86: 9268-9272; "AbMTM, A Computer Program for Modeling Variable Regions of Antibodies," Oxford, UK; Oxford Molecular, Ltd). The AbM numbering system uses a combination of knowledge databases and a de novo approach to model the tertiary structure of antibodies from basic sequences (see those described by Samudrala et al., 1999, “Ab Initio Protein Structure Prediction Using a Combined Hierarchical Approach” in PROTEINS, Structural, Function and Genetics Suppl., 3:194-198). Contact definitions are based on the analysis of available complex crystal structures (see, for example, MacCallum et al., 1996, J. Mol. Biol., 5:732-45). In conformational definitions, the position of the CDR can be identified as a residue that contributes enthalpy to antigen binding (see, for example, Makabe et al., 2008, Journal of Biological Chemistry, 283:1156-1166). Other CDR boundary definitions may not strictly follow one of the above methods but still overlap at least partially with the Kabat CDR, although they may be shortened or lengthened depending on the prediction or experimental results that a particular residue or group of residues does not significantly affect antigen binding.

[0094] "Antigen-binding fragments" include single-chain antibodies (i.e., heavy or light chains); Fab, modified Fab, Fab', modified Fab', F(ab')2, Fv, Fab-Fv, Fab-dsFv, single-domain antibodies (e.g., VH, VL, or VHH), scFv, bivalent, trivalent, or tetravalent antibodies, Bis-scFv, diabody, tribody, tetrabody, and epitope-binding fragments of any of the above (see, for example, Holliger and Hudson, 2005, Nature Biotech. 23(9): 1126-1136; Adair and Lawson, 2005, Drug Design Reviews-Online 2(3), 209-217).

[0095] The terms "homology," "identity," or "sequence identity" refer to the sequence similarity between two polynucleotide sequences or two polypeptides. When positions in two compared sequences are occupied by the same nucleotide or amino acid monomer—for example, if every position in two DNA molecules is occupied by the same nucleotide—then the molecules are homologous at that position. The percentage of homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the number of positions compared multiplied by 100%. For example, at optimal sequence alignment, if six out of ten positions in two sequences match or are homologous, then the two sequences are 60% homologous. Generally, comparisons are made when the highest percentage of homology is obtained by aligning the two sequences.

[0096] The terms “cancer,” “cancerous,” “proliferative disorder,” and “tumor” are not mutually exclusive when used in this disclosure.

[0097] The terms “give,” “apply,” and “treat,” when applied to animals, humans, experimental subjects, cells, tissues, organs, or biological fluids, refer to the contact of an exogenous drug, therapeutic agent, diagnostic agent, or composition with an animal, human, subject, cell, tissue, organ, or biological fluid, such as in therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Cellular treatment includes contact between a reagent and a cell, as well as contact between a reagent and a fluid, wherein the fluid is in contact with the cell. “Give,” “apply,” and “treat” also mean treatment, such as of cells, by means of a reagent, diagnostic agent, conjugate composition, or by means of another cell in vitro and ex vivo. When applied to humans, veterinary, or research subjects, it refers to therapeutic treatment, preventative or prophylactic measures, research, and diagnostic applications.

[0098] The term "treatment" means administering a therapeutic agent, such as a fusion protein or insulin analog comprising any of the present disclosure, to a subject who has, is suspected of having, or is predisposed to having one or more diabetes or hyperglycemia-related diseases or their symptoms, and the therapeutic agent is known to have a therapeutic effect on these symptoms. Typically, a therapeutic agent is administered in a treated subject or population in an amount that effectively relieves symptoms of one or more diseases by preventing or delaying the onset of symptoms or complications, reducing symptoms or complications, or eliminating the disease, condition, or symptom to any clinically measurable degree. The amount of a therapeutic agent that effectively relieves symptoms of any specific disease (also referred to as a "therapeuticly effective amount") can vary depending on a variety of factors, such as the subject's disease state, age, and weight, and the drug's ability to produce the desired therapeutic effect in the subject. Whether the disease symptoms have been relieved can be evaluated using any clinical test that a physician or other healthcare professional typically uses to assess the severity or progression of the symptoms. Although the embodiments of this disclosure (e.g., treatment methods or products) may be ineffective in alleviating the symptoms of the target disease in a particular subject, they should alleviate the symptoms of the target disease in a statistically significant number of subjects, as determined by any statistical test known in the art, such as the Student t-test, chi-square test, U-test according to Mann and Whitney, Kruskal-Wallis test (H-test), Jonckheere-Terpstra test, and Wilcoxon test. The patients to be treated are mammals, and preferably humans.

[0099] The term "about" means that a value is within an acceptable margin of error for a specific value determined by a person skilled in the art, the value depending in part on how it is measured or determined (i.e., the limits of the measurement system). For example, "about" may mean within or above 1 standard deviation. Alternatively, "about" or "substantially contains" may mean a range of up to 20%, such as between 1% and 15%, between 1% and 10%, between 1% and 5%, between 0.5% and 5%, or between 0.5% and 1%.

[0100] The values ​​in this disclosure are measured by instruments or calculated after instrument measurement, and there is a certain degree of error. Generally speaking, plus or minus 10% is within the reasonable error range. Attached Figure Description

[0101] Figure 1 Tumor volume of the test substance in the GP2D xenograft model (Mean±SEM);

[0102] Figure 2 Relative tumor volume (Mean±SEM) of the test substance in the GP2D xenograft model;

[0103] Figure 3Tumor weight (Mean±SEM) of the test substance in the GP2D xenograft tumor model;

[0104] Figure 4 Weight of the test substance in the GP2D xenograft model (Mean±SEM). Detailed Implementation

[0105] The present disclosure is further described in detail through the following examples. These examples are for illustrative purposes only and are not intended to limit the scope of the present disclosure.

[0106] Example 1. Clinical trials (Phase IB and Phase II) of the compound shown in Formula (I) in combination with cetuximab for the treatment of subjects with advanced solid tumors with KRAS G12D mutations.

[0107] 1.1 Research Drugs

[0108] The compound injection solution shown in formula (I)

[0109] Dosage form: Injection

[0110] Specification: 10ml:30mg

[0111] Route of administration: Intravenous infusion

[0112] Manufacturer: Jiangsu Hengrui Medicine Co., Ltd.

[0113] cetuximab

[0114] Manufacturer: Merck Healthcare KGaA

[0115] 1.2 Research Objectives and End Points

[0116] Phase 1: (IB Period)

[0117] Primary study objective: To evaluate the safety and tolerability of the compound shown in formula (I) in combination with cetuximab in subjects with advanced solid tumors with KRAS G12D mutations, and to determine the recommended dose for phase II (RP2D).

[0118] Primary endpoint: Maximum tolerated dose (MTD) and RP2D of the compound represented by formula (I) in combination with cetuximab in subjects with advanced solid tumors with KRAS G12D mutations;

[0119] Secondary study objective: To evaluate the preliminary efficacy of the compound shown in formula (I) in combination with cetuximab in subjects with advanced solid tumors with KRAS G12D mutations;

[0120] Secondary endpoints: investigator-assessed objective response rate (ORR), disease control rate (DCR), duration of response (DoR), progression-free survival (PFS), and overall survival (OS) in subjects with advanced solid tumors with KRAS G12D mutations, in combination with the compound shown in Formula (I).

[0121] Phase Two: (Phase II)

[0122] Primary study objective: To evaluate the preliminary efficacy of the compound shown in formula (I) in combination with cetuximab in subjects with advanced solid tumors with KRAS G12D mutations;

[0123] Primary endpoint: Investigator-assessed ORR in subjects with advanced solid tumors with KRAS G12D mutations treated with the compound shown in formula (I) in combination with cetuximab.

[0124] Secondary study objective: To evaluate the efficacy and safety of the compound shown in formula (I) in combination with cetuximab in patients with advanced solid tumors with KRAS G12D mutations.

[0125] Secondary endpoints: investigator-assessed DCR, DoR, PFS, and OS.

[0126] 1.3 Research Design

[0127] Phase IB (dose escalation)

[0128] This phase aims to determine the tolerability of the compound shown in Formula (I) in combination with cetuximab in patients with advanced solid tumors harboring KRAS G12D mutations, to determine the RP2D, and to assess safety and preliminary efficacy. Dose escalation was performed using a Bayesian optimal interval (BOIN) design.

[0129] During the study, a Safety Monitoring Committee (SMC) composed of the principal investigator and sponsor representatives will be established to review safety and efficacy data generated during the study. The SMC will make decisions on study-related issues, including but not limited to dose escalation of combination regimens, choice of combination regimens, selection of efficacy expansion cohorts, expansion initiation time, and RP2D.

[0130] The combination therapy is pre-set with the following dosage group: starting dose (which may be determined by SMC) of 1200 mg of the compound shown in formula (I) every 2 weeks, combined with cetuximab 500 mg / m². 2 Q2W. SMC will determine whether to proceed with a lower dose group or other dosing frequencies based on the safety and tolerability of the initial dose combination therapy (e.g., compound shown in formula (I) is 800 mg Q2W or 500 mg, 400 mg QW, cetuximab may be adjusted to an initial dose of 400 mg / m²). 2Subsequent cycles 250mg / m 2 QW). The SMC will determine the RP2D dose for combination therapy based on the tolerability of the compound shown in formula (I) in combination with cetuximab. The SMC may consider preliminary safety and efficacy data to determine the starting dose, lower dose group, intermediate dose group, higher dose group, or explore other dosing frequencies. The final MTD and RP2D for this phase will be determined by the SMC through comprehensive evaluation. The DLT observation period is 28 days.

[0131] The compound represented by formula (I) is administered via intravenous infusion every two weeks. Cetuximab is administered via intravenous infusion every two weeks. Each cycle lasts 14 days.

[0132] Phase II (Therapeutic Effect Expansion)

[0133] This phase aims to evaluate the antitumor efficacy of the compound shown in Formula (I) in combination with cetuximab in patients with advanced solid tumors (non-small cell lung cancer, colorectal cancer) with KRAS G12D mutations. Approximately 30 patients are planned to be enrolled in each cohort.

[0134] The compound represented by formula (I) is administered intravenously at a dose determined in the dose escalation phase (up to 1-3 doses may be selected for expansion, as discussed by the SMC), once every two weeks. Cetuximab is administered intravenously once weekly or twice weekly. Combined therapy continues until the criteria for discontinuation of treatment are met, with each cycle lasting 14 days.

[0135] 1.4 Inclusion criteria

[0136] Patients with unresectable locally advanced or metastatic solid tumors with confirmed KRAS G12D mutations.

[0137] Phase IB enrolls patients with advanced solid tumors who have relapsed or progressed after standard treatment, or for whom there is no standard treatment regimen, or for whom standard treatment is not applicable at this stage; Phase II enrolls patients with relapsed or metastatic advanced solid tumors who have failed previous standard treatments (including but not limited to systemic chemotherapy, molecularly targeted therapy, immunotherapy, biotherapy, and other investigational therapies); patients with non-small cell lung cancer who have progressed after immunotherapy and platinum-based chemotherapy; patients with colorectal cancer who have progressed after oxaliplatin, irinotecan, or fluorouracil chemotherapy; and patients with ≤3 lines of treatment.

[0138] 1.5 Administration method and dosage

[0139] The compound shown in formula (I): Intravenous infusion, 1200mg Q2W, 800mg Q2W, 200-500mg QW, 400mg-600mg D1 + 800mg-1200mg D8 Q3W or other dose groups). The first infusion time should not be less than 60 minutes. If no infusion-related reaction occurs after the first administration, the duration of each subsequent infusion may be appropriately shortened.

[0140] Cetuximab: Intravenous infusion, 500 mg / m² 2 Once every 2 weeks or for the first use, use 400 mg / m². 2 250mg / m 2 Once a week.

[0141] 1.6 Evaluation of Tumor Response

[0142] Objective response rate (ORR), as well as progression-free survival (PFS), disease control rate (DCR), and duration of response (DoR) based on tumor response assessment, are evaluated using imaging methods. Each tumor imaging evaluation includes enhanced CT scans of the chest, abdomen, pelvis, and lesion sites. If there is an allergy to contrast agents used in enhanced CT scans, a plain chest CT scan plus a pelvic and abdominal MRI scan can be performed.

[0143] The results showed that the ORR for non-small cell lung cancer was 55.6%.

[0144] Example 2. Study on the inhibitory effect of compound (I) (drug A) alone or in combination with cetuximab (drug B) on the growth of human colon cancer GP2D cell xenograft tumors in nude mice.

[0145] 1. Experimental Objective

[0146] This study used a nude mouse xenograft model of human colon cancer GP2D cells to evaluate the antitumor activity of the test drugs administered alone or in combination.

[0147] 2. Animal and Drug Information

[0148] 2.1 Mice

[0149] Species and strain: BALB / c Nude mice; sex and age: female, 8-10 weeks old; weight: 18-20g, with a deviation of approximately ±20% of the mean weight; number of animals inoculated: 38; number of animals enrolled: 28; animal source: Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0150] 2.2 Drugs

[0151] Drug A Injection

[0152] Solvent: 0.9% Saline

[0153] Storage conditions for the drug solution: Store at 4℃, protected from light, and prepare and use immediately.

[0154] Preparation of the drug administration solution: Take 0.275 mL of drug A injection (3 mg / mL), add 5.225 mL of 0.9% Saline, mix until a homogeneous and clear solution is obtained, store at 4°C protected from light, and prepare fresh before use.

[0155] Drug B

[0156] Solvent: 0.9% Saline

[0157] Storage conditions for the drug solution: Store at 4℃, protected from light, and prepare fresh before use.

[0158] Preparation of the drug administration solution: Take 0.53 mL of drug B solution (7.55 mg / mL), add 3.47 mL of 0.9% Saline, mix until a homogeneous and clear solution is obtained, store at 4°C protected from light, and prepare fresh before use.

[0159] 2.3 Cell lines

[0160] The human colon cancer cell line GP2D was purchased from Nanjing Kebai Biotechnology Co., Ltd.

[0161] 2.4 Culture medium

[0162] MEM basal medium and fetal bovine serum (FBS) were purchased from GIBCO (Grand Island, NY, USA). Matrigel was purchased from Corning (Corning, NY, USA).

[0163] 3. Experimental Design

[0164] The experimental design is shown in Table 1 below:

[0165] Table 1. Experimental Design for In Vivo Efficacy Evaluation

[0166]

[0167] 4. Experimental Methods

[0168] 4.1 Model Establishment

[0169] GP2D cells were cultured in DMEM medium containing 10% FBS and maintained in a 37°C saturated humidity incubator with 5% CO2.

[0170] Logarithmic growth phase GP2D cells were collected and resuspended in DMEM basal medium containing 50% Matrigel, and the cell concentration was adjusted to 2 × 10⁶ cells / year. 7 / mL. Under aseptic conditions, 0.1 mL of cell suspension was injected subcutaneously into the right back of mice at an inoculation concentration of 2 × 10⁹ / mL. 6 / 0.1mL / mouse.

[0171] 4.2 Grouped Dosing

[0172] When the average tumor volume reaches 150 mm 3 When the tumor volume was around 10%, the animals were randomly grouped according to their tumor volume, so that the difference in tumor volume between the groups was less than 10% of the mean.

[0173] The day of grouping is designated as Day 0, and medication is initiated according to the animal's body weight. During the medication period, if an individual animal's body weight decreases by more than 15% compared to Day 0 (BWL ≥ 15%), medication will be discontinued until the animal's body weight recovers (BWL < 15%), at which point medication will be resumed.

[0174] 4.3 Weighing and Observation

[0175] Animal body weight and tumor volume were measured twice a week during the experiment. Tumor length and width were measured using a digital caliper, and tumor volume was estimated using the length and width measurements.

[0176] Clinical symptoms in animals were observed and recorded once daily during the experiment. Clinical observation included the animals' overall health status, abnormal weight, abnormal behavior, and other drug-related adverse reactions.

[0177] 4.4 Explanation of Experimental Endpoint

[0178] According to animal welfare regulations, if an individual experimental animal meets any of the following conditions during the experiment, that animal will be removed from the experimental group and euthanized: 1. The animal's body weight decreases by more than 20% compared to Day 0 (BWL ≥ 20%); 2. The animal exhibits severe adverse reactions, such as blindness or paralysis; 3. The tumor volume is greater than 2000 mm². 3 And the veterinarian confirms the impact on the animal's condition; 4. Open ulcers form on the surface of the tumor.

[0179] 4.5 Evaluation Indicators

[0180] The formula for calculating tumor volume (TV) is: 1 / 2 × a × b 2 , where a and b are the measured length and width of the tumor, respectively;

[0181] Tumor growth inhibition rate (%TGI) TV The calculation formula is: (1-TV) T / TV C )×100%, TV C The mean tumor volume of the negative control group, TV T This represents the average tumor volume in the treatment group;

[0182] The formula for calculating relative tumor volume (RTV) is: Vt / V0, where V0 is the tumor volume at the time of grouping and Vt is the tumor volume at each measurement.

[0183] Relative tumor proliferation rate (%T / C) RTV The calculation formula is: T RTV / C RTV ×100%, where T RTV For the treatment group RTV, C RTV This serves as the negative control group for RTV.

[0184] Tumor inhibition rate (%TGI) TW The calculation formula is: (TW) C -TW T ) / TW C ×100%, of which TW C The mean tumor weight of the negative control group, TW T The average tumor weight is for the treatment group.

[0185] The formula for calculating the percentage change in animal body weight (%BWC) is: (BW... t -BW0) / BW0×100%, where BW t BW0 represents the animal's weight at each measurement, while BW0 represents the animal's weight at the time of grouping.

[0186] According to the "Technical Guidelines for Non-Clinical Studies of Cytotoxic Antitumor Drugs" issued by the China National Medical Products Administration (NMPA) in November 2006, %T / C RTV A drug is considered effective if the mortality rate is ≤40% and statistical analysis shows a p-value <0.05. If the number of animal deaths related to the drug exceeds 20%, the drug dose is considered to be severely toxic.

[0187] 5. Statistical Analysis

[0188] In this study, all experimental data are expressed as mean ± SEM.

[0189] With time points as the X-axis, tumor volume (mm) 3 A tumor growth curve was plotted with time points on the X-axis and animal weight (g) on ​​the Y-axis; an animal weight change curve was plotted with time points on the X-axis and animal weight (g) on ​​the Y-axis. Two-tailed t-tests were used for comparisons between groups, with P < 0.05 considered statistically significant, and P < 0.01 and P < 0.001 considered highly statistically significant (Microsoft Excel 2016, Redmond, WA, USA).

[0190] 6. Experimental Results

[0191] Log-phase GP2D cells were collected and tumor cells were subcutaneously inoculated into the right back of nude mice. The actual inoculation was 2.0 × 10⁶ cells. 6 Cells / 0.1mL / mouse. A total of 38 animals were inoculated in this experiment, divided into two groups of 28. Drugs were administered immediately after grouping. Drug A was administered via tail vein once a week for 5 consecutive weeks (IV, QW×5); cetuximab was administered via tail vein once every 3 days for 10 consecutive weeks (IV, Q3D×10). On Day 30, the remaining animals were sacrificed, tumors were harvested, weighed, photographed, and the experiment concluded.

[0192] 6.1 Evaluation of the antitumor activity of the test substance

[0193] In this experiment, the effects of drug A injection, drug B alone or in combination, on tumor volume and tumor weight in human colon cancer GP2D tumor-bearing mice are shown in Table 1. Figure 1 , Figure 2 and Figure 3 .

[0194] On Day 30, the average tumor weight in the group treated with drug A injection alone was 0.389 ± 0.050 g, and the tumor inhibition rate (%TGI) was [missing value]. TW The tumor proliferation rate was 62.31%, which was significantly different from the mean tumor weight of the solvent control group (1.032±0.171g) (P<0.01); the relative tumor proliferation rate %T / C was... RTV The percentage was 41.51% (P<0.001).

[0195] The mean tumor weight in the drug B monotherapy group was 0.367 ± 0.032 g, % TGI TW The tumor weight was 64.44%, which was significantly different from the mean tumor weight of the solvent control group (P<0.01); the relative tumor proliferation rate %T / C RTV The percentage was 36.02% (P<0.001).

[0196] The average tumor weight in the group receiving both drug A injection and drug B in combination was 0.177 ± 0.036 g, % TGI TW The tumor weight was 82.85%, which was significantly different from the mean tumor weight of the solvent control group (P<0.001), and the relative tumor proliferation rate %T / C was [missing value]. RTV The mean tumor weight was 22.77% (P<0.001). There were highly significant differences in mean tumor weight compared to both the drug A injection alone group and the drug B injection alone group (P<0.001). There were also highly significant differences in relative tumor volume (RTV) compared to both the drug A injection alone group and the drug B injection alone group (P<0.001, P<0.01).

[0197] Experimental results showed that drug B administration, drug A injection, and the combined drug B administration group all effectively inhibited the growth and proliferation of human colon cancer GP2D cells in BALB / c Nude mice (%T / C). RTV ≤40%, P<0.001). Drug A injection alone significantly inhibited the growth and proliferation of human colon cancer GP2D cells in BALB / c Nude mice (P<0.001). The combined drug A and drug B injection group showed highly significant differences compared to the drug A injection alone group and the drug B alone group (P<0.001, P<0.01).

[0198] Table 2. Antitumor activity of test substances in GP2D xenograft models

[0199]

[0200] *:P<0.05compare with vehicle group; **:P<0.01compare with vehiclegroup; ***:P<0.001compare with vehicle group.##:P<0.01compare with group2; ###:P<0.001compare with group 2.$$:P<0.01compare with group 3;$$$:P<0.001compare with group 3.

[0201] 6.2 Effects of the test substance on the body weight of tumor-bearing animals

[0202] In this experiment, the effects of drug A injection, drug B alone or in combination, on the body weight of GP2D tumor-bearing mice are shown in Table 3. Figure 4 On Day 30, the average body weight of mice in the solvent control group, the drug A injection alone group, the drug B injection alone group, and the drug A injection and drug B combined group increased by 1.37%-7.30% (0.27g-1.46g) compared with the start of the experiment on Day 0. No drug-related animal deaths were observed during the experiment.

[0203] Table 3. Effects of the test substance on animal body weight in the GP2D xenograft model.

[0204]

[0205] 7. Experiment Summary

[0206] Experimental results showed that drug B alone, drug A injection, and drug B combined with other drugs effectively inhibited the growth and proliferation of human colon cancer GP2D cells in BALB / c Nude mice (%T / C). RTV ≤40%, P<0.001). Drug A injection alone significantly inhibited the growth and proliferation of human colon cancer GP2D in BALB / c Nude mice (P<0.001). The combined drug A and drug B injection group showed highly significant differences compared to the drug A injection alone group and the drug B alone group (P<0.001, P<0.01).

[0207] On Day 30, the average body weight of mice in the solvent control group and each drug-treated group increased by 1.37%–7.30% (0.27g–1.46g) compared to the start of the experiment on Day 0. No drug-related animal deaths were observed during the experiment.

[0208] In summary, administration of drug A injection at a dose of 1.5 mg / kg once weekly via tail vein for 5 consecutive weeks, combined with administration of drug B at a dose of 10 mg / kg once every 3 days via tail vein for 10 consecutive weeks, effectively inhibited the growth of human colon cancer GP2D cell xenografts in nude mice. Furthermore, there was a highly significant difference compared to the groups treated with drug A injection alone or drug B injection alone. During the experiment, no changes in animal body weight or adverse reactions related to the combined administration of drug A injection and drug B were observed, indicating that the tumor-bearing mice tolerated the dosages well under the experimental conditions.

Claims

1. Use of a KRAS G12D inhibitor in combination with an anti-EGFR antibody or an antigen-binding fragment thereof in the preparation of a medicament for treating KRAS G12D-mutant solid tumors, wherein the KRAS G12D inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

2. The use according to claim 1, wherein the solid tumor is selected from colorectal cancer and lung cancer.

3. The use according to any one of claims 1-2, wherein the solid tumor is selected from advanced solid tumors.

4. The use according to claim 3, wherein the advanced solid tumor is selected from advanced solid tumors that have relapsed or progressed after standard treatment, or for which there is no standard treatment option, or for which standard treatment is not applicable at this stage.

5. The use according to any one of claims 2-4, wherein the colorectal cancer is selected from adenocarcinoma, adenosquamous carcinoma, squamous cell carcinoma, undifferentiated carcinoma, preferably adenocarcinoma.

6. The use according to any one of claims 2-4, wherein the lung cancer is selected from non-small cell lung cancer and small cell lung cancer, preferably non-small cell lung cancer.

7. The use according to any one of claims 1-6, wherein the dosage of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from 50-2000 mg; the frequency of administration is selected from once a week, once every two weeks, once every three weeks, or once every four weeks; preferably, the dosage of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is selected from 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, 1000 mg, or 1200 mg, and the frequency of administration is selected from once a week or once every two weeks. Once or every 3 weeks; more preferably, the dosage of the compound of formula (I) or its pharmaceutically acceptable salt is selected from 200 mg, 300 mg, 400 mg, 500 mg, and the frequency of administration is selected from once a week; or the dosage of the compound of formula (I) or its pharmaceutically acceptable salt is 800 mg, 1000 mg, 1200 mg, and the frequency of administration is once every 2 weeks; most preferably, the dosage of the compound of formula (I) or its pharmaceutically acceptable salt is 1200 mg and the frequency of administration is once every 2 weeks.

8. The use according to any one of claims 1-7, wherein the anti-EGFR antibody or its antigen-binding fragment comprises LCDR1 with an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence differing from SEQ ID NO:1 by 1, 2, or 3 amino acids; LCDR2 with an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence differing from SEQ ID NO:2 by 1, 2, or 3 amino acids; LCDR3 with an amino acid sequence as shown in SEQ ID NO:3 or an amino acid sequence differing from SEQ ID NO:3 by 1, 2, or 3 amino acids; HCDR1 with an amino acid sequence as shown in SEQ ID NO:4 or an amino acid sequence differing from SEQ ID NO:4 by 1, 2, or 3 amino acids; HCDR2 with an amino acid sequence as shown in SEQ ID NO:5 or an amino acid sequence differing from SEQ ID NO:5 by 1, 2, or 3 amino acids; and HCDR3 with an amino acid sequence as shown in SEQ ID NO:6 or an amino acid sequence differing from SEQ ID NO:6 by 1, 2, or 3 amino acids; preferably, the anti-EGFR antibody or its antigen-binding fragment comprises an amino acid sequence as shown in SEQ ID NO:

1. LCDR1 as shown in NO:1, LCDR2 as shown in SEQ ID NO:2, LCDR3 as shown in SEQ ID NO:3, HCDR1 as shown in SEQ ID NO:4, HCDR2 as shown in SEQ ID NO:5, and HCDR3 as shown in SEQ ID NO:

6.

9. The use according to any one of claims 1-8, wherein the anti-EGFR antibody or its antigen-binding fragment comprises a light chain variable region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the amino acid sequence shown in SEQ ID NO:7, and a heavy chain variable region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the amino acid sequence shown in SEQ ID NO:8; preferably, the anti-EGFR antibody or its antigen-binding fragment 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.

10. The use according to any one of claims 1-9, wherein the anti-EGFR antibody or its antigen-binding fragment comprises a light chain having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the amino acid sequence shown in SEQ ID NO: 9, and a heavy chain having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the amino acid sequence shown in SEQ ID NO: 10; preferably, the anti-EGFR antibody comprises a light chain with the amino acid sequence shown in SEQ ID NO: 9, and a heavy chain with the amino acid sequence shown in SEQ ID NO: 10; most preferably, the anti-EGFR antibody is cetuximab or a biosimilar thereof.

11. The use according to any one of claims 1-10, wherein the anti-EGFR antibody or its antigen-binding fragment is administered at a dose of about 100 mg / m² based on body surface area. 2 Approximately 800 mg / m 2 For example, approximately 200 mg / m³ 2 Approximately 500 mg / m 2 For example, approximately 100 mg / m³ 2 Approximately 150 mg / m 2 Approximately 200 mg / m 2 Approximately 250 mg / m 2 Approximately 300 mg / m 2 Approximately 350 mg / m 2 Approximately 400 mg / m 2 Approximately 450 mg / m 2 Approximately 500 mg / m 2 Approximately 550 mg / m 2 Approximately 600 mg / m 2 Approximately 650 mg / m 2 Approximately 700 mg / m 2 Approximately 750 mg / m 2 or approximately 800 mg / m 2 The dosing frequency is once a week or once every two weeks; preferably, the anti-EGFR antibody or its antigen-binding fragment is administered at a dose of approximately 500 mg / m² based on body surface area. 2 The dosing frequency is once every 2 weeks; or, the initial dose of the anti-EGFR antibody or its antigen-binding fragment is approximately 400 mg / m² based on body surface area. 2 The weekly dosage thereafter is approximately 250 mg / m² based on body surface area. 2 The medication should be administered once a week.

12. The use according to any one of claims 1-11, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is in the form of a pharmaceutical composition; preferably, the pharmaceutical composition is a liposome.

13. The use according to any one of claims 1-12, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered via intravenous injection.

14. A compound of formula (I) or a pharmaceutically acceptable salt thereof for treating KRAS G12D-mutated solid tumors, administered in combination with an anti-EGFR antibody or an antigen-binding fragment thereof according to any one of claims 1-13.

15. An anti-EGFR antibody or an antigen-binding fragment thereof for treating KRAS G12D-mutated solid tumors, administered in combination with a compound of formula (I) according to any one of claims 1-13 or a pharmaceutically acceptable salt thereof.

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