Combination of WEE1 inhibitor and PKMYT1 inhibitor

Abstract

The present invention provides the use of a combination of a WEE1 inhibitor and a PKMYT1 inhibitor for the treatment of cancer.

Description

Technical Field

[0001] This invention generally relates to combinations of WEE1 inhibitors and PKMYT1 inhibitors. More specifically, this invention relates to the use of combinations of WEE1 inhibitors and PKMYT1 inhibitors in the treatment of cancer in patients in need. Background Technology

[0002] DNA is constantly subjected to both endogenous damage (e.g., stalled replication forks, reactive oxygen species) and exogenous damage (UV radiation, ionizing radiation, chemicals) that can lead to DNA damage. Therefore, cells have developed complex mechanisms to combat these harmful events, which would otherwise compromise genome integrity and lead to genomically unstable diseases such as cancer. These mechanisms are collectively known as the DNA damage response (DDR). A portion of the overall DDR is the activation of various checkpoint pathways that regulate specific DNA repair mechanisms at different stages throughout the cell cycle, including the G1, S, G2, and mitotic checkpoints. Most cancer cells lose their G1 checkpoint due to p53 mutations and therefore rely on the G2 checkpoint for necessary DNA damage correction before attempting to enter mitosis and divide into two daughter cells.

[0003] DNA damage repair inhibitors have been proposed as potentially useful treatment methods. Nevertheless, improved cancer treatment options remain needed. This invention aims to provide these improved cancer therapies. Specifically, the goal is to provide cancer therapies that exhibit superior therapeutic efficacy and / or reduced side effects. Summary of the Invention

[0004] The present invention provides drug combinations, drugs for combination therapy, pharmaceutical compositions and kits, as set forth in the appended claims and further described in the following embodiments.

[0005] This invention relates to combinations of WEE1 inhibitors and PKMYT1 inhibitors. It should be understood that any reference to WEE1 inhibitor or PKMYT1 inhibitor also includes pharmaceutically acceptable salts of the inhibitor, even if not so explicitly mentioned.

[0006] The present invention further relates to a combination of a WEE1 inhibitor and a PKMYT1 inhibitor, which is used as a pharmaceutical agent.

[0007] The present invention further relates to a combination of WEE1 inhibitors and PKMYT1 inhibitors for the treatment of cancer in patients in need.

[0008] The present invention further relates to a WEE1 inhibitor for treating cancer in patients in need, wherein the WEE1 inhibitor is administered in combination with a PKMYT1 inhibitor.

[0009] The present invention further relates to a PKMYT1 inhibitor for treating cancer in patients in need, wherein the PKMYT1 inhibitor is administered in combination with a WEE1 inhibitor.

[0010] In some aspects of the present invention, the WEE1 inhibitor is a compound of formula (I). (I), Or its pharmaceutically acceptable salt.

[0011] In some aspects of the present invention, the WEE1 inhibitor is administered as follows: a. From day 1 to day 3 of the 21-day cycle, b. On days 1 to 3 and days 8 to 10 of the 21-day cycle, c. On days 1 to 3, 8 to 10, and 15 to 17 of the 21-day cycle, d. From day 1 to day 5 of the 21-day cycle, e. Days 1 to 5 and days 8 to 12 within a 21-day cycle, f. During days 1 to 5, 8 to 12, and 15 to 19 of the 21-day cycle, g. From day 1 to day 14 of the 21-day cycle, h. Daily within a 21-day cycle, i. On days 1 to 5, 8 to 10, and 15 to 17 of the 28-day cycle, j. From day 1 to day 5 of the 28-day cycle, k. On days 1 to 5 and days 8 to 10 of the 28-day cycle, 1. On days 1 to 5 and days 8 to 12 of the 28-day cycle, m. During days 1 to 5, 8 to 12, and 15 to 19 of the 28-day cycle, n. Days 1 to 3, 8 to 10, and 15 to 17 of a 28-day cycle. o. On days 1 to 3 and days 8 to 10 of a 28-day cycle, or p. Days 1 to 3 of a 28-day cycle.

[0012] In some aspects of the present invention, the WEE1 inhibitor is administered as follows: a. A dose of about 30 to about 1000 mg per WEE1 inhibitor treatment day, preferably in the range of about 30 to about 720 mg per WEE1 inhibitor treatment day, and / or b. Orally, and / or c. On the day of WEE1 inhibitor treatment, in a single dose, and / or d. At approximately the same time on each WEE1 inhibitor treatment day.

[0013] In some aspects of this invention, the PKMYT1 inhibitor is administered as follows: a. A number of consecutive or non-consecutive 7 or more days in a 21-day cycle, such as 14 or more consecutive or non-consecutive days or days per day, and / or b. On days 1 to 3, 8 to 10, and 15 to 17 of the 21-day cycle, and / or c. On the same day as the WEE1 inhibitor treatment date, preferably on the same day as defined in any of (a) to (p) of the above-described 21-day or 28-day cycle for the WEE1 inhibitor, and / or d. On a different day from the treatment day of the WEE1 inhibitor in a 21-day or 28-day cycle, wherein the treatment day of the WEE1 inhibitor in a 21-day or 28-day cycle may be, in particular, the WEE1 inhibitor treatment day specified herein, for example, under items (a) to (p) above, and / or e. To administer PKMYT1 inhibitors and WEE1 inhibitors in an alternating manner, wherein one of the drugs is administered for x days and then paused for y days, and the other drug is paused during the x days and administered during the y days, followed by another period of x days and y days, etc., wherein x and y are independently selected from 1, 2, 3, 4, 5, 6 and 7, such as 1, 2, 3 or 4, wherein in some respects x and y are selected such that x + y = 7, for example, an administration regimen consisting of three days of administration of WEE1 inhibitor followed by four days of administration of PKMYT1 inhibitor, etc. (or vice versa); in other respects x and y may be the same and both are jointly selected from 1, 2, 3, 4, 5, 6 and 7, especially 1, 2 or 3, such as three days of administration of WEE1 inhibitor followed by three days of administration of PKMYT1 inhibitor, etc.

[0014] In some aspects of this invention, the PKMYT1 inhibitor is administered as follows: a. On each day of PKMYT1 inhibitor treatment, at a dose ranging from about 60 mg to about 400 mg, or from about 100 mg to about 300 mg, or at a dose of about 240 mg, and / or b. Once or twice daily on each PKMYT1 inhibitor treatment day, and / or c. Administer once or twice daily at a dose selected from approximately 30 to approximately 100 mg, or approximately 60 to approximately 100 mg, or approximately 80 to approximately 100 mg per administration, and / or d. Administer a dose ranging from approximately 80 to approximately 100 mg twice daily for each PKMYT1 inhibitor treatment day, and / or, e. 240 mg daily on each day of PKMYT1 inhibitor treatment, and / or f. via oral communication.

[0015] In some aspects of the invention, the combination, WEE1 inhibitor and / or PKMYT1 inhibitor are administered over 1, 2, 3, 4, 5, 6 or more 21-day cycles or over 1, 2, 3, 4, 5, 6 or more 28-day cycles.

[0016] In some aspects of the invention, the cancer is ovarian cancer; primary peritoneal cancer; fallopian tube cancer; epithelial ovarian cancer (EOC), including serous EOC, endometrioid EOC, or clear cell EOC; gastric cancer; colorectal cancer; bladder cancer; cervical cancer; esophageal cancer; small cell lung cancer (SCLC); non-small cell lung cancer (NSCLC); breast cancer; soft tissue sarcoma; glioma; or endometrial cancer, including serous uterine carcinoma (USC).

[0017] In some aspects of the invention, the cancer is characterized by CCNE1 overexpression and / or CCNE1 amplification.

[0018] In some aspects of the invention, the cancer is characterized by ATRX deficiency, FBXW7 inactivation mutation or PPP2R1A inactivation mutation.

[0019] In some aspects of this invention, the cancer is metastatic.

[0020] In some aspects of the invention, metastatic cancer includes brain metastases.

[0021] In some aspects of the present invention, the PKMYT1 inhibitor is a compound of formula (II). (II), Or its pharmaceutically acceptable salt.

[0022] The present invention further relates to a pharmaceutical composition comprising, as further specified herein, a combination thereof, or comprising a WEE1 inhibitor as specified herein or a PKMYT1 inhibitor as specified herein, for use in treating cancer in patients in need, wherein said use is as described herein.

[0023] The present invention further relates to a kit comprising a combination as further specified herein, wherein the WEE1 inhibitor and the PKMYT1 inhibitor are provided in separate dosage forms and / or containers, which may be used to treat cancer in patients in need, wherein the use is as described herein. Attached Figure Description

[0024] Figure 1The results obtained in Example 1 are depicted in the bar graph, showing the relative PKMYT1 expression in A549 cells transfected with 25 nM, 50 nM, or 100 nM PKMYT1 siRNA or out-of-order siRNA.

[0025] Figure 2 The results obtained in Example 2 are depicted as follows: (A) a representative bright-field image of A549WT cells or A549 WEE1 KO cells 72 hours after transfection with 50 nM PKMYT1 siRNA or disordered siRNA, and (B) a figure showing the quantification of live cells per well by flow cytometry at 72 hours and 96 hours after transfection.

[0026] Figure 3 The results obtained in Example 3 (a cell line-derived xenograft model of ovarian cancer) are depicted: changes in tumor volume over time in various treatment groups (left panel) and changes in average body weight of animals over time (right panel).

[0027] Figure 4 Depicting the results obtained in Example 4 (a cell line-derived xenograft model of breast cancer): The figure shows the changes in tumor volume over time in various treatment groups.

[0028] Figure 5 Depicting the results obtained in Example 5 (PDX model of breast cancer): The figure shows the changes in tumor volume over time in various treatment groups.

[0029] Figure 6 Depicting the results obtained in Example 5 (PDX model of ovarian cancer): The figure shows the changes in tumor volume over time in various treatment groups. Detailed Implementation

[0030] definition

[0031] To facilitate a better understanding of the invention, certain terms are specifically defined below. Unless explicitly defined elsewhere in this document, all other technical and scientific terms used herein have the meanings commonly understood by those skilled in the art.

[0032] As used herein, including in the appended claims, the singular forms of words such as “a”, “an” and “the”, unless the context clearly indicates otherwise, include their corresponding multiple references.

[0033] It should be understood that whenever an aspect is described in this document using the language "contains", other similar aspects described using the terms "composed of" and / or "mainly composed of" are also provided.

[0034] As used herein, the term "and / or" as used in phrases such as "A and / or B" is intended to include "A and B", "A or B", and "A and B". Similarly, the term "and / or" as used in phrases such as "A, B and / or C" is intended to cover each of the following: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0035] As used herein, “WEE1 inhibitor” refers to a compound that inhibits the activity of the WEE1 kinase, for example, with an IC50 < 10 nM in ADP-GLO kinase assays or an IC50 < 100 nM in enzyme profiling. Compounds of formula (I) in this article are examples of WEE1 inhibitors. As used herein, “WEE1 inhibitor” generally exhibits selectivity, meaning that it does not inhibit PKMYT1 to the extent that the compound can also be identified as a PKMYT1 inhibitor according to the following definition.

[0036] In some respects, a “brain-penetrating” WEE1 inhibitor refers to a WEE1 inhibitor capable of crossing the blood-brain barrier (whether or not it is damaged) and distributing into the brain. In some respects, the ability of a compound to cross the blood-brain barrier and distribute into the brain is reflected by its brain-to-plasma concentration ratio exceeding a certain threshold. For example, compared to AZD1775 and Zn-c3, compounds of formula (I) exhibit a better ability to cross the blood-brain barrier and distribute into the brain, as demonstrated by their higher brain-to-plasma concentration ratio (see, for example, Example 1 of WO 2024 / 170480). Therefore, compounds of formula (I) can be referred to as brain-penetrating WEE1 inhibitors.

[0037] As used herein, the term "biomarker," sometimes also referred to as "biomarker material," is a measurable indicator of a biological state or condition. A biomarker is a molecule whose activity, conformation, localization, and / or abundance can be objectively measured and serves as an indicator of normal biological processes, pathogenic processes, or a predictor of pharmacological response to therapeutic treatment. Biomarkers can be specific cells, molecules, genes, gene products, proteins, enzymes, or hormones. They contribute to the understanding of disease mechanisms and the effectiveness of drug treatments. Biomarkers can be measured and evaluated using blood, urine, cells, or tissues.

[0038] It should be noted that for some biomarkers (such as CCNE1, FBXW7, or PPP2R1A) used in this invention, the genes and the resulting gene products (e.g., mRNA and / or proteins) use the same names or abbreviations. Those skilled in the art will understand that amplification, mutation, or deletion of a given biomarker refers to the biomarker gene, while references to expression levels (e.g., reduced expression or overexpression) refer to the corresponding biomarker gene product.

[0039] As used in this article, the term “PKMYT1” or “Myt1” refers to membrane-associated tyrosine and threonine-specific cdc2 inhibitory kinase (Myt1) (gene name PKMYT1).

[0040] As used herein, the term "PKMYT1 inhibitor" or "Myt1 inhibitor" refers to a compound that, upon contacting the enzyme Myt1, reduces the activity of Myt1, whether in vitro, in cell culture, or in animals, such that the measured Myt1 IC50 is 100 nM or less (e.g., 30 nM or less, 10 nM or less, or 3 nM or less) and can be as low as 100 pM or 10 pM. More preferably, the Myt1 IC50 is less than 20 nM (e.g., 1 nM to 20 nM). The Myt1 IC50 value can be determined by PKMYT1 cell-based activity assays as described by J. Szychowski et al. in J. Med. Chem. 2022, 65(15), 10251-10284, doi: 10.1021 / acs.jmedchem.2c00552. PKMYT1 inhibitors can also be dual inhibitors, inhibiting both WEE1 and PKMYT1.

[0041] The terms “CCNE1” and “cyclin E1” are used interchangeably in this document. It refers to G1 / S-specific cyclin E1 (gene name: CCNE1). Cells overexpressing CCNE1 are those exhibiting higher CCNE1 activity than cells normally expressing CCNE1. CCNE1 amplification is present in cells expressing at least 3 copies, compared to diploid normal cells with 2 copies. CCNE1-overexpressing cells can be, for example, cells expressing CCNE1 amplification. Therefore, cells expressing more than 3 copies of CCNE1 may be overexpressing CCNE1. CCNE1 expression levels can also be affected by transcriptional alterations or CCNE1 protein stability, regardless of copy number changes. CCNE1 overexpression can be measured, for example, by identifying the expression levels of gene products in tumor cells or tissues (e.g., CCNE1 mRNA transcript count or CCNE1 protein content) and comparing them to the expression levels in healthy (non-circulating) cells or tissues or tumor tissues where CCNE1 overexpression is not yet known. For example, tumor samples subjected to immunohistochemical (IHC) testing and intensity scoring, in which at least 25% of tumor cells show moderate (intensity score 2 or higher) or strong (intensity score 3 or higher) staining of CCNE1, can be considered as CCNE1 overexpressing tumors (equivalent to an H score ≥50). Methods for reproducible semi-quantitative protein detection in tissues using IHC and pathologist-based scoring are described, for example, in Meyerholz and Beck, 2018, Principles and approaches for reproducible scoring of tissue stains in research, Laboratory Investigation, Vol. 98, No. 7, pp. 844-855, ISSN 0023-6837, https: / / doi.org / 10.1038 / s41374-018-0057-0 (https: / / www.sciencedirect.com / science / article / pii / S0023683722010194).

[0042] The term "FBXW7" is used herein to refer to a protein 7 gene, transcript, or protein containing an F-box / WD repeat sequence. An FBXW7 mutant gene, sometimes referred to herein as an FBXW7 gene with an inactivating mutation, is a gene that cannot produce functional FBXW7 protein or produces a reduced amount of FBXW7 protein in the cell.

[0043] As used herein, the term "PPP2R1A" refers to the α-isoform of the 65 kDa regulatory subunit A of serine / threonine protein phosphatase 2A, an enzyme encoded by the PPP2R1A gene in the human body. The PPP2R1A gene encodes the constant regulatory subunit of protein phosphatase 2. Protein phosphatase 2 is one of four major serine / threonine phosphatases and is involved in the negative regulation of cell growth and division. It consists of a common heteromeric core enzyme composed of a catalytic subunit associated with multiple regulatory subunits and a constant regulatory subunit. The constant regulatory subunit A acts as a backbone molecule to coordinate the assembly of the catalytic subunit and the variable regulatory subunit B. This gene encodes the α-isoform of the constant regulatory subunit A. A mutant PPP2R1A gene, sometimes referred to herein as a PPP2R1A gene with an inactivating mutation, is a gene that fails to produce functional PPP2R1A protein or produces a reduced amount of PPP2R1A protein in cells.

[0044] Gene mutations or amplifications can be assessed, for example, by next-generation sequencing (NGS), and the reduction or overexpression of a given gene product can be assessed by NGS, qPCR, Western blotting, or IHC techniques.

[0045] "Applying at approximately the same time" means applying at the same time as the application time on the reference date or at a time that deviates from the application time on the reference date by no more than approximately one hour.

[0046] For example, a 21-day “treatment cycle” refers to a period of time, for example, 21 consecutive days, during which the drug of interest is administered on one or more days of the treatment cycle. As in this invention, a combination of at least two drugs of interest is administered, preferably in a coordinated manner such that the treatment cycles for both drugs begin on the same day. However, it is also conceivable to administer one drug according to a first treatment cycle and another drug according to a second treatment cycle, wherein the duration and / or start dates of the two treatment cycles may differ from each other.

[0047] As used herein, the term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic acid (including inorganic or organic acids). Examples of acceptable salts derived from acids include quaternary salts, acetates, carbonates, carbamates, sulfonates, strong inorganic acids, and the like. Generally, pharmaceutically acceptable salts can be used to modify the solubility or hydrolytic characteristics of compounds, or in sustained-release formulations. It should be understood that, unless otherwise stated, as used herein, the reference to WEE1 inhibitors or PKMYT1 inhibitors also includes pharmaceutically acceptable salts.

[0048] As used herein, the term “subject” means any animal (e.g., a mammal) that will become a recipient of a particular therapy, including but not limited to humans, non-human primates, rodents, and similar animals. Generally, the terms “subject” and “patient” are used interchangeably in this document with respect to human subjects.

[0049] As used herein, “combination” means the presence of a combination of at least a WEE1 inhibitor and a PKMYT1 inhibitor. The combination may be provided in the same pharmaceutical composition, or may be provided separately in, for example, two separate pharmaceutical compositions. Similarly, any reference to “combination” administration should be understood as administration of the combination to the same patient such that the combination may be administered together or separately from each other. If administered separately from each other, administration includes simultaneous (parallel) and sequential administration in any order. For example, a WEE1 inhibitor and a PKMYT1 inhibitor may be administered in combination by first administering the WEE1 inhibitor and then subsequently administering the PKMYT1 inhibitor, or vice versa.

[0050] As used herein, terms such as “treating,” “treatment,” “to treat,” “alleviating,” or “to alleviate” refer to therapeutic measures of curing, slowing, reducing, or halting or reversing the progression or severity of a diagnosed pathological condition, symptom, or disease. Therefore, those requiring treatment include those already diagnosed with or suspected of having the condition. In some respects, the cancer of an individual is considered successfully “treated” according to the method of the invention if the patient exhibits one or more of the following: a reduction in the number of cancer cells or the complete absence of cancer cells; a reduction in tumor size or burden; inhibition or absence of cancer cell infiltration into surrounding organs; inhibition or absence of tumor metastasis; inhibition or absence of tumor growth; relief of one or more symptoms associated with a specific cancer; a reduction in morbidity and mortality; an improvement in quality of life; a reduction in tumorigenicity, frequency, or capacity for tumorigenicity; a reduction in the number or frequency of cancer stem cells in the tumor; differentiation of tumorigenic cells into a non-tumorigenic state; and an increased chance of complete response (CR), partial response (PR), disease control (e.g., CR, PR, stable disease SD), longer survival without progression, and longer disease-free survival, a reduced chance of disease progression (PD), and an increased time before progression. For a patient group or population, successful treatment may produce endpoints such as improved overall response rate (ORR), best overall response (BOR), duration of response (DOR), disease control rate (DCR), progression-free survival (PFS), overall survival (OS), time to progression (TTP), or any combination thereof.

[0051] As used in this article, when referring to cancer or patients with cancer, the terms “relapse,” “to relapse,” “recurrence,” and “to recur” mean a worsening of the disease and / or its signs and symptoms after a period of improvement, stability, or absence of the disease.

[0052] As used in this article, when referring to cancer or a patient with cancer, the terms “progression” or “to progress” mean that the cancer has worsened due to the growth of existing lesions and / or due to the appearance of new lesions.

[0053] As used herein, the term "WEE1 inhibitor treatment day" refers to the day on which the WEE1 inhibitor is administered according to the present invention.

[0054] As used herein, the term "PKMYT1 inhibitor treatment day" refers to the day on which a PKMYT1 inhibitor is administered according to the present invention.

[0055] As used herein, the term "WEE1-PKMYT1 combination therapy day" refers to the day on which the WEE1 inhibitor and PKMYT1 inhibitor are administered according to the present invention.

[0056] As used herein, the term “therapeutic effective dose” refers to the amount of a drug that effectively “treats” a subject’s or patient’s disease or condition. In the case of cancer, a therapeutically effective dose of a drug may reduce the number of cancer cells; reduce tumor size or burden; inhibit (i.e., to some extent slow and, in some respects, prevent) the infiltration of cancer cells into surrounding organs; inhibit (i.e., to some extent slow and, in some respects, prevent) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more cancer-related symptoms to some extent; and / or induce a favorable response, such as an improved overall response rate (ORR), best overall response (BOR), duration of response (DOR), disease control rate (DCR), progression-free survival (PFS), overall survival (OS), complete response (CR) rate, partial response (PR) rate, or, in some cases, stable disease (SD) rate, reduced disease progression (PD), increased time to tumor progression (TTP), or any combination thereof. See the definition of “treatment” in this article.

[0057] In some respects, the term "therapeutic effective amount" used for a given drug in monotherapy, such as "therapeutic effective amount of WEE1," "therapeutic effective amount of a PKMYT1 inhibitor," or "therapeutic effective amount of combination therapy," refers to the amount by which the drug or combination of drugs effectively "treats" the disease or condition of the subject or patient while maintaining an acceptable profile of safety. In some respects, when a combination of a WEE1 inhibitor and a PKMYT1 inhibitor is administered, the corresponding therapeutic effective amount may be referred to as "therapeutic effective amount of the WEE1-PKMYT1 combination therapy." In the case where the WEE1 inhibitor is a compound of formula (I), when administered as monotherapy, the "therapeutic effective amount of WEE1" for a compound of formula (I) may differ from the "therapeutic effective amount of the WEE1-PKMYT1 combination therapy." In the case of cancer, a treatment with an effective dose of a drug can reduce the number of cancer cells; reduce tumor size or burden; inhibit (i.e., slow down to some extent and stop in some aspect) the infiltration of cancer cells into surrounding organs; inhibit (i.e., slow down to some extent and stop in some aspect) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more cancer-related symptoms to some extent; and / or produce a favorable response, such as improved overall response rate (ORR), best overall response (BOR), duration of response (DOR), disease control rate (DCR), progression-free survival (PFS), overall survival (OS), complete response (CR) rate, partial response (PR) rate, or in some cases, stable disease (SD) rate, reduced disease progression (PD), increased time to progression (TTP), or any combination thereof. See the definition of “treatment” in this article.

[0058] As used in this article, “overall survival” (OS) in clinical trials refers to the time from patient enrollment, first treatment administration, or randomization to death from any cause or review of the last known survival date. Improved OS includes an increase in life expectancy compared to an initially (naive) or untreated individual or patient. Overall survival refers to a patient’s survival for a defined period of time, such as one year, five years, etc., from the time of randomization or first treatment.

[0059] As used herein, the terms “pharmaceutical formulation” or “pharmaceutical composition” refer to an article which is in a form that allows the bioactivity of the active ingredient to be effective and which does not contain any other components that would have unacceptable toxicity to a subject to be administered the formulation.

[0060] As used herein, the term "oral dosage form" refers to any form of pharmaceutical composition suitable for oral administration.

[0061] It should be understood that any reference to WEE1 inhibitors or PKMYT1 inhibitors also includes their pharmaceutically acceptable salts, even if not so explicitly mentioned.

[0062] Information introduced in this disclosure by the phrases “in some respects,” “in a particular respect,” or similar wording should be understood to allow it to be freely combined with any other information provided elsewhere in this disclosure, regardless of whether such information is also introduced by the phrases “in some respects,” “in a particular respect,” or any other wording. However, exceptions to this general rule apply to situations where the information to be combined is inconsistent or even contradictory or mutually exclusive, such that the resulting combined information would be inconsistent, ambiguous, and / or technically meaningless.

[0063] combination

[0064] This invention relates to a combination of WEE1 inhibitors and PKMYT1 inhibitors.

[0065] This invention further relates to a combination of a WEE1 inhibitor and a PKMYT1 inhibitor, used as a pharmaceutical agent. This invention also relates to a method of using a combination of a WEE1 inhibitor and a PKMYT1 inhibitor as a pharmaceutical agent. This use or method may comprise administering a therapeutically effective amount of a WEE1 inhibitor, a therapeutically effective amount of a PKMYT1 inhibitor, or a therapeutically effective amount of both a WEE1 inhibitor and a PKMYT1 inhibitor.

[0066] This invention relates to a combination of a WEE1 inhibitor or a pharmaceutically acceptable salt thereof with a PKMYT1 inhibitor or a pharmaceutically acceptable salt thereof for the treatment of cancer in patients in need, or for the preparation of an agent for the treatment of cancer in patients in need.

[0067] Any disclosure relating to the WEE1 inhibitors in this article should be understood as a combination of any disclosure relating to the PKMYT1 inhibitors in this article.

[0068] In some aspects of the invention, the WEE1 inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0069] In some aspects of the invention, the WEE1 inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof, and the PKMYT1 inhibitor is a compound of formula (II) or a pharmaceutically acceptable salt thereof.

[0070] use

[0071] This invention relates to a WEE1 inhibitor for treating cancer in patients in need, wherein the WEE1 inhibitor is administered in combination with a PKMYT1 inhibitor.

[0072] This invention relates to PKMYT1 inhibitors for the treatment of cancer in patients in need, wherein the PKMYT1 inhibitor is administered in combination with a WEE1 inhibitor.

[0073] method

[0074] This invention also relates to methods of treating cancer in patients in need using combinations, WEE1 inhibitors, or PKMYT1 inhibitors, as well as methods of combination therapy described herein. Any disclosure of treating cancer with combinations, using WEE1 inhibitors, or using PKMYT1 inhibitors is to be understood as relating to methods for treating cancer, and vice versa.

[0075] It should be understood that any reference to a compound or combination used herein also covers methods of using that compound. For example, any reference to the use of a compound or combination for the treatment of cancer should be understood to also cover methods of using that compound or combination to treat cancer, or methods of treating cancer with that compound or combination.

[0076] Specific compounds

[0077] In some aspects of the present invention, the WEE1 inhibitor is a compound of formula (I). (I), Or its pharmaceutically acceptable salt.

[0078] Compound (I) is a selective WEE1 inhibitor.

[0079] In some respects, a WEE1 inhibitor is any compound described in patent applications WO2018090939, WO2022155202, WO2022256680, WO2013126656, WO2020192581, and WO2008153207, each of which is incorporated herein by reference in its entirety. Specifically, a WEE1 inhibitor may be one of the compounds in the following formula, or a pharmaceutically acceptable salt thereof: Also known as AZD-1775 (adavosertib); Also known as Zn-C3 (azenosertib); Also known as IMP7068 (as described in WO2018090939); Also known as STC-8123 (as described in WO2022155202); Also known as ATRN-W1051 (as described in WO2022256680); Also known as PD0407824; Also known as CJM061; Also known as SC0191; Also known as PD0166285; (as described in WO2013126656), or (As described in WO2008153207).

[0080] In some aspects, the WEE1 inhibitor is compound SY-4835 or any compound described in patent application WO2020192581, especially the compounds disclosed in claims 3 and 6 of this document and the compound of Example 59 of CN111718348 identified as SY-4835 in Q. Ye et al., Bioorganic & Medicinal Chemistry, 2023, 87, 117312, https: / / doi.org / 10.1016 / j.bmc.2023.117312.

[0081] In some aspects of the present invention, the PKMYT1 inhibitor is a compound of formula (II). (II), Or a pharmaceutically acceptable salt thereof. Compounds of formula (II) may also be called lunresertib or RP-6306.

[0082] In some respects, PKMYT1 inhibitors are any compounds described by J. Szychowski et al. in J. Med. Chem. 2022, 65(15), 10251-10284, doi: 10.1021 / acs.jmedchem.2c00552 and patent application WO2021 / 195781A1, each of which is incorporated herein by reference in its entirety. In other respects, PKMYT1 inhibitors are any compounds described in patent applications WO2023 / 022659 A1 (and particularly the inhibitors described in its "therapeutic" portion and claim 39), WO 2023 / 155892 A, WO 2023 / 155871 A, WO 2023 / 155870 A, WO2024 / 112853 A1, or WO 2024 / 137671 A1, each of which is incorporated herein by reference in its entirety. In other respects, PKMYT1 inhibitors are AST-NS2301 of Allist Pharmaceuticals, ACR-2316 of Acrivon Therapeutics, EVT-0002518 of Evariste, or XL495 of Exelixis.

[0083] In some aspects, PKMYT1 inhibitors are as described in claim 1 of WO2021 / 195781 A1, as follows: A compound of the following formula

[0084] Or a pharmaceutically acceptable salt thereof, wherein X, Y, and Z are each independently N or CR. 2 ; R 1 and each R 2 Independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 ynyl, optionally substituted C3-8 cycloalkyl, optionally substituted C3-8 cycloalkenyl, optionally substituted C2-9 heterocyclic, optionally substituted C2-9 heterocyclic C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, halogen, cyano, -N(R) 7 )2、-OR 7 - C(O)N(R) 8 )2、-SO2N(R 8 )2、-SO2R 7 A or -Q-R7B; or R 1With one and R 1 Adjacent R 2 Combining to form optionally substituted C3-6 alkylene groups; R 3 and R 4 Each of them is independently a optionally substituted C1-6 alkyl or halogen; R 5 For H or -N(R) 7 )2; R 6 -C(O)NH(R) 8 -C(O)R 7A or -SO2R 7A ; Each R 7 Independently hydrogen, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl C1-6 alkyl, optionally substituted C3-8 cycloalkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclic, optionally substituted C1-9 heteroaryl, optionally substituted C1-9 heteroaryl C1-6 alkyl, or -SO2R 7A ; or two Rs 7 The group, together with the atoms attached to both, forms an optionally substituted C2-9 heterocyclic group; Each R 7A Independently, it is an optionally substituted C1-6 alkyl, an optionally substituted C3-8 cycloalkyl, or an optionally substituted C6-10 aryl; Each R 7B Independently hydroxyl, optionally substituted C1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C2-9 heterocyclic, optionally substituted C1-9 heteroaryl, -N(R 7 )2、-C(O)N(R 8 )2、-SO2N(R 8 )2、-SO2R 7A Or optionally substituted alkoxy groups; Each R 8 Independently, it is hydrogen, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkoxyalkyl, optionally substituted C6-10 arylC1-6 alkyl, optionally substituted C6-10 aryl, optionally substituted C3-8 cycloalkyl, or optionally substituted C1-9 heteroaryl; or two Rs. 8 Together with the atoms to which it is attached, it forms an optionally substituted C2-9 heterocyclic group; Q can be a substituted C1-6 alkylene group, a substituted C2-6 alkenyl group, a substituted C2-6 ynynyl group, a substituted C3-8 cycloalkylene group, a substituted C3-8 cycloalkenyl group, a substituted C6-10 aryl group, a substituted C2-9 heterocyclic group, or a substituted C1-9 heteroaryl group.

[0085] Compound (I) is a WEE1 inhibitor. Compound (II) is a PKMYT1 inhibitor. In some aspects, N-oxides or N-oxide derivatives of compounds of formula (I) and / or formula (II) may be used.

[0086] In certain aspects of the combinations, uses, and methods according to the invention, the WEE1 inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof. All other information provided herein should be understood to apply particularly to aspects where the WEE1 inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0087] In other specific aspects of the combinations, uses, and methods according to the invention, the WEE1 inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof, and the PKMYT1 inhibitor is a compound of formula (II) or a pharmaceutically acceptable salt thereof. All other information provided herein should be understood to apply particularly to the aspects where the WEE1 inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof and the PKMYT1 inhibitor is a compound of formula (II) or a pharmaceutically acceptable salt thereof.

[0088] Dosage, timing and route of administration

[0089] As used herein, the term “about” describes a deviation of ±10% from the indicated value. Individual values ​​should be understood as describing a range between any of the described values.

[0090] The dosage instructions provided herein should be understood as characterizing the mass of the free base compound. If a pharmaceutically acceptable salt of the corresponding compound is used, the dosage instructions may need to be adjusted according to the molecular weight of the compound in that salt form.

[0091] In some aspects of the combinations, uses or methods according to the invention, the combinations, WEE1 inhibitors and / or PKMYT1 inhibitors described herein are administered orally.

[0092] In some respects, WEE1 inhibitors are administered according to a regimen selected from the following: a. From day 1 to day 3 of the 21-day cycle, b. On days 1 to 3 and days 8 to 10 of the 21-day cycle, c. On days 1 to 3, 8 to 10, and 15 to 17 of the 21-day cycle, d. From day 1 to day 5 of the 21-day cycle, e. Days 1 to 5 and days 8 to 12 within a 21-day cycle, f. During days 1 to 5, 8 to 12, and 15 to 19 of the 21-day cycle, g. From day 1 to day 14 of the 21-day cycle, h. Daily within a 21-day cycle, i. On days 1 to 5, 8 to 10, and 15 to 17 of the 28-day cycle, j. From day 1 to day 5 of the 28-day cycle, k. On days 1 to 5 and days 8 to 10 of the 28-day cycle, 1. On days 1 to 5 and days 8 to 12 of the 28-day cycle, m. During days 1 to 5, 8 to 12, and 15 to 19 of the 28-day cycle, n. Days 1 to 3, 8 to 10, and 15 to 17 of a 28-day cycle. o. On days 1 to 3 and days 8 to 10 of a 28-day cycle, or p. Days 1 to 3 of a 28-day cycle.

[0093] In some aspects of the combinations, uses, or methods according to the invention, the WEE1 inhibitor is administered as follows: a. A dose of about 30 to about 1000 mg per WEE1 inhibitor treatment day, preferably in the range of about 30 to about 720 mg per WEE1 inhibitor treatment day, and / or b. Daily, and / or c. On the day of WEE1 inhibitor treatment, in a single dose, and / or d. At approximately the same time on each WEE1 inhibitor treatment day.

[0094] In some aspects, WEE1 inhibitors are administered at doses ranging from about 30 to about 1000 mg per WEE1 inhibitor treatment day, preferably from about 30 to about 720 mg per WEE1 inhibitor treatment day, and in some aspects from about 90 to about 720 mg per WEE1 inhibitor treatment day or from about 150 to about 720 mg per WEE1 inhibitor treatment day, such as at a dose of about 200 mg or about 150 mg per WEE1 inhibitor treatment day. For example, WEE1 inhibitors may be administered in a single dose on the WEE1 inhibitor treatment day.

[0095] In some respects, the WEE1 inhibitor is administered at approximately the same time on each WEE1 inhibitor treatment day, such as at the same time on each WEE1 inhibitor treatment day ± about 60 minutes, preferably ± 60 minutes.

[0096] In some aspects, the WEE1 inhibitor and / or PKMYT1 inhibitor are administered after fasting, preferably 4 hours after fasting. In the same or other aspects, the combination, the WEE1 inhibitor and / or PKMYT1 inhibitor may be administered after fasting, preferably 2 hours after fasting.

[0097] In some aspects of the combination, use, or method according to the invention, on the WEE1 inhibitor treatment day, the WEE1 inhibitor is administered before the PKMYT1 inhibitor is administered. For example, on the WEE1 inhibitor treatment day, PKMYT1 inhibitor administration is initiated approximately 5 minutes to 30 minutes after the WEE1 inhibitor administration, and less than 1 hour to 4 hours after the WEE1 inhibitor administration. In some aspects, the time interval between the WEE1 inhibitor administration and the PKMYT1 inhibitor administration can be any interval from 5 minutes to 240 minutes, 15 minutes to 180 minutes, or 30 minutes to 120 minutes. In other aspects, the two drugs can be administered substantially simultaneously; for example, the time interval between the WEE1 inhibitor administration and the PKMYT1 inhibitor administration can be 5 minutes to 30 minutes or 5 minutes to 15 minutes.

[0098] In some aspects of the combination, use, or method according to the invention, on the WEE1 inhibitor treatment day, the PKMYT1 inhibitor is administered before the WEE1 inhibitor administration. For example, on the WEE1 inhibitor treatment day, the WEE1 inhibitor is administered approximately 5 minutes, such as 30 minutes, and less than 1 hour, such as 4 hours, after the start of PKMYT1 inhibitor administration. In some aspects, the time interval between the WEE1 inhibitor administration and the PKMYT1 inhibitor administration can be any period from 5 minutes to 240 minutes, 15 minutes to 180 minutes, or 30 minutes to 120 minutes. In other aspects, the two drugs can be administered substantially simultaneously; for example, the time interval between the PKMYT1 inhibitor administration and the WEE1 inhibitor administration can be 5 minutes to 30 minutes or 5 minutes to 15 minutes.

[0099] In some aspects of the combination, use or method according to the invention, the simultaneous administration of WEE1 inhibitors and PKMYT1 inhibitors may be in the form of separate pharmaceutical compositions or in the form of a single pharmaceutical composition comprising the two agents.

[0100] In some aspects of the combination, use or method according to the invention, the combination, WEE1 inhibitor and / or PKMYT1 inhibitor are administered over 1, 2, 3, 4, 5, 6 or more cycles.

[0101] In some aspects of the combinations, uses, or methods according to the invention, the combinations, WEE1 inhibitors, and / or PKMYT1 inhibitors are administered within 1, 2, 3, 4, 5, 6, or more 21-day or 28-day cycles. Typically, there is no interruption between any consecutive cycles; that is, the day following the previous 21-day or 28-day cycle may be the first day of the next consecutive cycle.

[0102] In some aspects of the combinations, uses, or methods according to the present invention, the PKMYT1 inhibitor is administered as follows: a. A number of consecutive or non-consecutive 7 or more days in a 21-day cycle, such as 14 or more consecutive or non-consecutive days or days per day, and / or b. On days 1 to 3, 8 to 10, and 15 to 17 of the 21-day cycle, and / or c. On the same day as the WEE1 inhibitor treatment, preferably on the same day as defined in any of the regimen aspects (a) to (p) described herein with respect to the WEE1 inhibitor administration regimen, and / or d. On a different day from the treatment day of the WEE1 inhibitor in a 21-day or 28-day cycle, wherein the treatment day of the WEE1 inhibitor in a 21-day or 28-day cycle may be, in particular, the WEE1 inhibitor treatment day specified herein, for example, under items (a) to (p) above, and / or e. To administer PKMYT1 inhibitors and WEE1 inhibitors in an alternating manner, wherein one of the drugs is administered for x days and then paused for y days, and the other drug is paused during the x days and administered during the y days, followed by another period of x days and y days, etc., wherein x and y are independently selected from 1, 2, 3, 4, 5, 6 and 7, such as 1, 2, 3 or 4, wherein in some respects x and y are selected such that x + y = 7, for example, an administration regimen consisting of three days of administration of WEE1 inhibitor followed by four days of administration of PKMYT1 inhibitor, etc. (or vice versa); in other respects x and y may be the same and both are jointly selected from 1, 2, 3, 4, 5, 6 and 7, especially 1, 2 or 3, such as three days of administration of WEE1 inhibitor followed by three days of administration of PKMYT1 inhibitor, etc.

[0103] In some aspects of the combinations, uses, or methods according to the present invention, the PKMYT1 inhibitor is administered as follows: a. On each day of PKMYT1 inhibitor treatment, at a dose ranging from about 60 mg to about 400 mg, or from about 100 mg to about 300 mg, or at a dose of about 240 mg, and / or b. Once or twice daily on each PKMYT1 inhibitor treatment day, and / or c. Administer once or twice daily at a dose selected from approximately 30 to approximately 100 mg, or approximately 60 to approximately 100 mg, or approximately 80 to approximately 100 mg per administration, and / or d. Administer a dose ranging from approximately 80 to approximately 100 mg twice daily for each PKMYT1 inhibitor treatment day, and / or, e. 240 mg daily on each day of PKMYT1 inhibitor treatment, and / or f. via oral communication.

[0104] In some aspects of the combination, use or method according to the invention, the PKMYT1 inhibitor is administered orally twice daily at a dose of about 40 mg, about 60 mg, about 80 mg or about 100 mg per administration.

[0105] In one particular aspect, WEE1 inhibitors and PKMYT1 inhibitors are administered according to one of the following options: Both A.WEE1 inhibitors and PKMYT1 inhibitors were administered on days 1-3, 8-10, and 15-17 of a 21-day cycle. Both B.WEE1 inhibitors and PKMYT1 inhibitors were administered on days 1-3, 8-10, 15-17, and 22-24 of a 28-day cycle. C.WEE1 inhibitors are administered daily, and PKMYT1 inhibitors are administered on days 1-3, 8-10, and 15-17 of a 21-day cycle. D.WEE1 inhibitors are administered daily, and PKMYT1 inhibitors are administered on days 1-3, 8-10, 15-17, and 22-24 of a 28-day cycle. E.WEE1 inhibitors were administered on days 1-3 and 8-10 of a 21-day cycle, and PKMYT1 inhibitors were administered on days 1-3, 8-10, and 15-17 of a 21-day cycle, or Both F.WEE1 inhibitors and PKMYT1 inhibitors were administered on days 1-3 and 15-17 of a 28-day cycle.

[0106] In each of the above options A through F, the WEE1 inhibitor and the PKMYT1 inhibitor are each administered once or twice per administration day. The administration of the WEE1 inhibitor and the PKMYT1 inhibitor once or twice per administration day can be chosen independently. For example, in a particular aspect related to each of the above options A through F, the WEE1 inhibitor is administered once per WEE1 inhibitor treatment day and the PKMYT1 inhibitor is administered twice per PKMYT1 inhibitor treatment day.

[0107] In another specific aspect, both WEE1 inhibitors and PKMYT1 inhibitors are administered daily. The duration of this administration is not specifically limited, except in the event of serious adverse effects. It may be, for example, 1, 2, 3, 4, 5, 6 or more 21-day cycles, or 1, 2, 3, 4, 5, 6 or more 28-day cycles, or any other duration.

[0108] In some aspects, the combination, WEE1 inhibitor, and / or PKMYT1 inhibitor are administered on days 1, 2, and 3 of a 21-day cycle. The combination, WEE1 inhibitor, and / or PKMYT1 inhibitor may also be administered on days 1, 2, 3, 8, 9, and 10 of a 21-day cycle.

[0109] In some cases, WEE1 inhibitors are administered daily and PKMYT1 inhibitors are administered on days 1 through 3, days 8 through 10, and days 15 through 17 of a 21-day cycle.

[0110] In some cases, WEE1 inhibitors were administered on days 1 through 3 and days 8 through 10 of a 21-day cycle, and PKMYT1 inhibitors were administered on days 1 through 3, days 8 through 10, and days 15 through 17 of a 21-day cycle.

[0111] In some aspects of the combination, use or method according to the invention, the administration of the WEE1 inhibitor is performed separately from, simultaneously with, concurrently with or sequentially with the administration of the PKMYT1 inhibitor.

[0112] In the combinations, uses, and methods of the present invention, the WEE1 inhibitor may be administered at a dose ranging from about 30 to about 1000 mg of free base per WEE1 inhibitor treatment day, preferably from about 30 to about 720 mg of free base. In some aspects, the compound of formula (I) may preferably be administered at a dose ranging from about 90 to about 720 mg of free base or from about 150 to about 720 mg of free base per WEE1 inhibitor treatment day.

[0113] For example, WEE1 inhibitors can be administered at doses of approximately 30, 60, 75, 90, 100, 120, 130, 150, 200, 220, 250, 260, 300, 320, 350, 360, 400, 420, 450, 460, 500, 520, 550, 600, 620, 650, 700, 720, 750, 800, 820, 850, 900, 920, 950, or 1000 mg of free base per WEE1 inhibitor treatment day. Specifically, WEE1 inhibitors can be administered at doses of 30, 60, 75, 90, 100, 120, 130, 150, 200, 220, 250, 260, 300, 320, 350, 360, 400, 420, 450, 460, 500, 520, 550, 600, 620, 650, 700, 720, 750, 800, 820, 850, 900, 920, 950, or 1000 mg of free base per WEE1 inhibitor treatment day.

[0114] In a more specific aspect, WEE1 inhibitors may be administered at doses of approximately 30, 60, 75, 90, 100, 120, 150, 200, 250, 260, 300, 350, 360, 400, 450, 460, 500, 520, 550, or 720 mg of free base per WEE1 inhibitor treatment day. For example, WEE1 inhibitors may be administered at doses of 30, 60, 75, 90, 100, 120, 150, 200, 250, 260, 300, 350, 360, 400, 450, 460, 500, 520, 550, or 720 mg of free base per WEE1 inhibitor treatment day.

[0115] In another specific aspect, WEE1 inhibitors may be administered at doses of approximately 100, 150, 200, 250, 300, 350, 400, 450, 500, 520, 550, 600, 620, 650, 700, 720, 750, 800, 820, 850, 900, 920, 950, or 1000 mg of free base per WEE1 inhibitor treatment day. For example, compounds of formula (I) may be administered at doses of approximately 100, 150, 200, 300, 400, or 520 mg of free base per WEE1 inhibitor treatment day.

[0116] In another aspect of the invention, the WEE1 inhibitor can be administered at a dose ranging from 100 to 1000 mg of free base per WEE1 inhibitor treatment day, preferably from 150 to 720 mg of free base. For example, the compound of formula (I) can be administered at a dose of 100, 150, 200, 250, 300, 350, 400, 450, 500, 520, 550, 600, 620, 650, 700, 720, 750, 800, 820, 850, 900, 920, 950, or 1000 mg of free base per WEE1 inhibitor treatment day. Specifically, the compound of formula (I) can be administered at a dose of 100, 150, 200, 300, 400, 520, or 720 mg of free base per WEE1 inhibitor treatment day.

[0117] In some cases, an effective dose of WEE1 or a combination of WEE1 and effective doses can be administered.

[0118] In some aspects of the invention, the WEE1 inhibitor is administered orally.

[0119] In some aspects of the invention, the WEE1 inhibitor is administered as a single dose (QD) per WEE1 inhibitor treatment day or as a twice-dose (BID) per WEE1 inhibitor treatment day.

[0120] In some aspects, the WEE1 inhibitor is administered on days 1, 2, and 3 of a 21-day cycle. In other aspects of the invention, the WEE1 inhibitor is administered on days 1, 2, 3, 8, 9, and 10 of a 21-day cycle. In some aspects, if the WEE1 inhibitor, particularly the compound of formula (I), is administered on days 1, 2, 3, 8, 9, and 10 of a 21-day cycle, it is optionally possible to administer the WEE1 inhibitor, particularly the compound of formula (I), only on days 1, 2, and 3 of subsequent 21-day cycles. This variation in administration dates can be advantageous, especially when the patient experiences side effects after administration on days 1, 2, 3, 8, 9, and 10. Similar variations in administration dates can be considered, such as administering the WEE1 inhibitor daily during the first 21-day cycle or more 21-day cycles, and subsequently administering the WEE1 inhibitor on a reduced number of days in subsequent 21-day cycles.

[0121] In some aspects, the WEE1 inhibitor may be administered at the dose indicated above for each WEE1 inhibitor treatment day on days 1, 2, and 3 of a 21-day cycle. In other aspects, the WEE1 inhibitor may be administered at the dose indicated above for each WEE1 inhibitor treatment day on days 1, 2, 3, 8, 9, and 10 of a 21-day cycle.

[0122] In these respects, WEE1 inhibitors can be administered in 1, 2, 3, 4, 5, 6 or more cycles. Typically, there is no interruption between any consecutive cycles; that is, the day after the previous 21-day cycle can be the first day of the next consecutive cycle. In some respects, treatment with a WEE1 inhibitor such as a compound of formula (I) can continue until disease progression, unacceptable toxicity, patient decision to discontinue, physician's decision to discontinue, initiation of subsequent anti-tumor therapy, completion of the clinical trial, or death.

[0123] In some aspects of the invention, the WEE1 inhibitor is administered, for example, at approximately the same time on each WEE1 inhibitor treatment day within a given cycle, for example, at the same time ± about 60 minutes, preferably ± 60 minutes, on each WEE1 inhibitor treatment day. In a more specific aspect, the compound of formula (I) is administered in the morning, for example, between 5 a.m. and noon.

[0124] In some aspects of the use or method according to the invention, the WEE1 inhibitor compound is administered after fasting, preferably after a 4-hour fasting period. In the same or other aspects, fasting may follow the administration of the WEE1 inhibitor, preferably for 2 hours.

[0125] In an alternative aspect of the use and method according to the invention, the WEE1 inhibitor is administered while the patient is eating.

[0126] In yet another alternative aspect of the use or method according to the invention, the administration of the WEE1 inhibitor is independent of the patient’s food status.

[0127] In some aspects of the method of the present invention, PKMYT1 inhibitors are used in therapeutically effective amounts for intended purposes.

[0128] In some cases, a therapeutically effective dose of a PKMYT1 inhibitor or a combination of therapeutically effective doses of PKMYT1 inhibitors may be administered.

[0129] In some respects, PKMYT1 inhibitors may be administered on the same day as WEE1 inhibitors, such as on the same day in a 21-day cycle or a 28-day cycle, or at different times, as defined below.

[0130] PKMYT1 inhibitors can be administered to patients in a single dose or multiple doses. Multiple doses can be administered at intervals ranging from 1 to 24 hours, 1 to 7 days, 1 to 4 weeks, or 1 to 12 months between administrations. PKMYT1 inhibitors can be administered according to a time schedule. In other respects, PKMYT1 inhibitors can be administered without a predetermined time schedule. PKMYT1 inhibitors can be administered, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times daily, every 2nd, 3rd, 4th, 5th, or 6th day, 1, 2, 3, 4th, 5th, 6th, or 7 times weekly, 1, 2, 3, 4, 5, or 6 times monthly, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times annually. In some respects, it is possible to administer a specific dosing regimen specially adjusted according to the individual patient's needs and the professional judgment of the person administering or supervising the administration of the composition.

[0131] An effective dose of a PKMYT1 inhibitor may be, for example, a total daily dose ranging from 0.05 mg to 3000 mg. More specific dose ranges may include, for example, 10 to 1000 mg (e.g., 50-800 mg). In some cases, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg of a PKMYT1 inhibitor may be administered. Alternatively, the dose may be calculated using the patient's weight. Of course, the attending physician will determine the appropriate dose based on the patient's condition, age, sex, weight, etc.

[0132] There are no particular restrictions on the duration of multiple doses of PKMYT1 inhibitors administered to patients. For example, PKMYT1 inhibitors can be administered over periods of 1–7 days, 1–12 weeks, or 1–3 months. The duration of administration can also be longer, such as 4–11 months or 1–30 years. PKMYT1 inhibitors can be administered to patients during symptom flare-ups. Generally, the amount of PKMYT1 inhibitor administered can vary during the administration period. When PKMYT1 inhibitors are administered daily, administration can be, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times daily.

[0133] In some respects, PKMYT1 inhibitors can be administered daily as a single dose (QD) or as a twice-daily dose (BID).

[0134] In some respects, PKMYT1 inhibitors can be administered in an intermittent or weekly intermittent regimen as a single dose (QD) per PKMYT1 inhibitor treatment day or as a twice-dose (BID) per PKMYT1 inhibitor treatment day.

[0135] In some respects, PKMYT1 inhibitors can be administered at doses of approximately 5 to approximately 600 mg of free base per PKMYT1 inhibitor treatment day, such as approximately 5, approximately 15, approximately 30, approximately 60, approximately 75, approximately 80 mg, approximately 90, approximately 100, approximately 120, approximately 140, approximately 150, approximately 200, approximately 240, approximately 250, approximately 260, approximately 300, approximately 350, approximately 360, approximately 400, approximately 450, approximately 460, approximately 500, approximately 520, approximately 550, or approximately 600 mg per PKMYT1 inhibitor treatment day. In other respects, PKMYT1 inhibitors can be administered at doses of 60 mg, 80 mg, 100 mg, 140 mg, or 240 mg of free base per PKMYT1 inhibitor treatment day.

[0136] In other respects, PKMYT1 inhibitors can be administered daily in the form of a single dose (QD) of 5 mg to 600 mg, preferably 240 mg, of free base per PKMYT1 inhibitor treatment day.

[0137] In some respects, PKMYT1 inhibitors can be administered daily in the form of a twice-daily dose (BID) of 5 mg to 600 mg, preferably 60 mg or 80 mg, of free base per PKMYT1 inhibitor treatment day.

[0138] In some respects, PKMYT1 inhibitors can be administered intermittently weekly in the form of a twice-daily dose (BID) of 5 mg to 600 mg, preferably 60 mg to 140 mg, of free base per PKMYT1 inhibitor treatment day.

[0139] In some respects, PKMYT1 inhibitors can be administered intermittently in the form of a twice-daily dose (BID) of 5 mg to 600 mg, preferably 80 mg to 100 mg, of free base per PKMYT1 inhibitor treatment day.

[0140] In any aspect of the combinations described herein, the WEE1 inhibitor may be a compound of formula (I). In any aspect of the combinations described herein, the PKMYT1 inhibitor may be a compound of formula (II). Specifically, in some aspects, the combination of compounds of formula (I) and (II) demonstrates synergistic effects and an acceptable safety profile in the treatment of cancer.

[0141] It should be understood that any of the specific compounds, administration, dosage, regimens, and indications described herein can be used in any combination. For example, this disclosure provides combinations, uses, and methods in which a WEE1 inhibitor is administered at a dose ranging from about 30 to about 1000 mg per WEE1 inhibitor treatment day, preferably from about 30 to about 720 mg per WEE1 inhibitor treatment day, and a PKMYT1 inhibitor is administered at a dose ranging from about 100 mg to about 400 mg daily, such as from about 200 mg to about 300 mg, and more specifically about 240 mg. Therefore, it should be understood that any of the administration doses, durations, and routes of administration of the WEE1 inhibitor or PKMYT1 inhibitor described herein can be combined with the administration doses, durations, and routes of administration of the corresponding other.

[0142] For example, any administration of a WEE1 inhibitor in a 21-day cycle can be combined with administration of a PKMYT1 inhibitor in the same 21-day cycle or in different cycles.

[0143] In any aspect of the combinations herein, the WEE1 inhibitor may be a compound of formula (I). In any aspect of the combinations herein, the PKMYT1 inhibitor may be a compound of formula (II). It should also be understood that any designation for the dose, duration, or administration of the WEE1 inhibitor may apply to the compound of formula (I). Similarly, any designation for the dose, duration, or administration of the PKMYT1 inhibitor may apply to the compound of formula (II). Compounds of formula (I) and formula (II) may therefore be preferably used at the doses, durations, and routes of administration described herein.

[0144] It should be further understood that any indications regarding the administration, dosage, duration of treatment, or indications of WEE1 inhibitors in this article apply particularly to compounds of formula (I).

[0145] It should be further understood that any instructions in this article regarding the administration, dosage, duration of treatment or indications for PKMYT1 inhibitors apply particularly to compounds of formula (II).

[0146] For example, in some aspects of the combination described herein, the WEE1 inhibitor is a compound of formula (I) administered at a dose ranging from about 100 mg to about 260 mg per treatment day, such as at a dose of about 100 mg, about 120 mg, about 150 mg, about 200 mg, or about 260 mg once daily. In these aspects, the PKMYT1 inhibitor may be administered at a dose ranging from about 40 mg to about 100 mg on days 1 to 3, 8 to 10, and 15 to 17 of a 21-day cycle, such as at a dose of about 40 mg, about 60 mg, about 80 mg, or about 100 mg twice daily, preferably at a dose of 60 mg or 80 mg twice daily. Both the compound of formula (I) and the PKMYT1 inhibitor may be administered orally. The PKMYT1 inhibitor may target cancers characterized by CCNE1 overexpression and / or amplification, FBXW7 gene inactivation mutations, and / or PPP2R1A inactivation mutations. PKMYT1 inhibitors can be compounds of formula (II).

[0147] In some aspects of the combination described herein, the WEE1 inhibitor is a compound of formula (I) administered once daily on days 1 to 3 and days 8 to 10 of a 21-day cycle at doses ranging from about 200 mg to about 720 mg per treatment day, such as about 200 mg, about 300 mg, about 400 mg, about 520 mg, or about 720 mg. In these aspects, the PKMYT1 inhibitor may be administered on days 1 to 3, days 8 to 10, and days 15 to 17 of a 21-day cycle at doses ranging from about 40 mg to about 100 mg, such as twice daily at doses of about 40 mg, about 60 mg, about 80 mg, or about 100 mg, preferably twice daily at doses of 60 mg or 80 mg. Both the compound of formula (I) and the PKMYT1 inhibitor may be administered orally. PKMYT1 inhibitors can target cancers characterized by CCNE1 overexpression and / or amplification, FBXW7 gene inactivation mutations, and / or PPP2R1A inactivation mutations. PKMYT1 inhibitors can be compounds of formula (II).

[0148] Radiation therapy

[0149] In some aspects of the combination, use or method according to the invention, the WEE1 inhibitor is used in combination with the PKMYT1 inhibitor and radiotherapy.

[0150] The radiotherapy used can be any suitable type of radiotherapy, such as 3D conformal radiotherapy (3DCRT), intensity-modulated radiotherapy (IMRT), focal radiotherapy, or fractionated radiotherapy.

[0151] In some respects, radiotherapy is fractionated radiotherapy.

[0152] In some respects, the radiation therapy is fractionated radiation therapy and is administered in fractions of 1 Gy to 100 Gy, preferably 2 Gy to 80 Gy, more preferably 10 Gy to 60 Gy, and even more preferably 60 Gy over five consecutive days per week until the total radiation dose has been administered.

[0153] In some respects, radiotherapy is used in combination with compounds of formula (I) and formula (II).

[0154] Indications

[0155] In some aspects of the combinations, uses, or methods according to the invention, cancer is treated, including ovarian cancer; primary peritoneal cancer; fallopian tube cancer; epithelial ovarian cancer (EOC), including serous EOC, endometrioid EOC, or clear cell EOC; gastric cancer; colorectal cancer; bladder cancer; cervical cancer; esophageal cancer; small cell lung cancer (SCLC); non-small cell lung cancer (NSCLC); breast cancer; soft tissue sarcoma; glioma; or endometrial cancer, including uterine serous carcinoma (USC).

[0156] In some respects, cancer is characterized by CCNE1 overexpression and / or amplification.

[0157] In some aspects of the invention, a subject is treated with cancer, wherein a therapeutically effective amount of the combination, WEE1 inhibitor, and / or PKMYT1 inhibitor is administered to the subject in need, wherein the cancer has been previously identified as cancer overexpressing CCNE1.

[0158] In some aspects of the invention, a subject is treated with cancer, wherein the subject is administered a therapeutically effective amount of a combination of a WEE1 inhibitor and / or a PKMYT1 inhibitor, wherein the cancer is a cancer that overexpresses CCNE1.

[0159] In some aspects of the invention, the combination, WEE1 inhibitor, and / or PKMYT1 inhibitor induce cell death in cancer cells overexpressing CCNE1, comprising contacting the cells with an effective amount of the combination, WEE1 inhibitor, and / or PKMYT1 inhibitor.

[0160] In some cases, cancers that overexpress CCNE1 include uterine cancer, ovarian cancer, breast cancer, stomach cancer, esophageal cancer, lung cancer, or endometrial cancer.

[0161] In some aspects of the invention, treating a subject's cancer includes administering a therapeutically effective amount of the combination, a WEE1 inhibitor, and / or a PKMYT1 inhibitor to the subject in need, wherein the cancer has been previously identified as having an inactivating mutation of the FBXW7 gene.

[0162] In some aspects of the invention, treating a subject's cancer includes administering to a subject in need a therapeutically effective amount of the combination, a WEE1 inhibitor, and / or a PKMYT1 inhibitor, wherein the cancer has an inactivating mutation in the FBXW7 gene.

[0163] In some aspects of the invention, inducing cell death in FBXW7-mutant cancer cells includes contacting the cells with effective amounts of the combination, a WEE1 inhibitor, and / or a PKMYT1 inhibitor. In some aspects, the FBXW7-mutant cancer is uterine cancer, colorectal cancer, breast cancer, lung cancer, or esophageal cancer.

[0164] In some respects, cancer is characterized by ATRX deficiency or PPP2R1A inactivation mutations.

[0165] In some aspects of the combinations, uses, or methods according to the invention, the cancer is newly diagnosed.

[0166] In some aspects of the combinations, uses, or methods according to the invention, cancer has progressed or recurred after initial or prior cancer treatment.

[0167] In some aspects of the combinations, uses, or methods according to the invention, initial or prior cancer treatment includes the administration of radiotherapy.

[0168] In some aspects of the combinations, uses, or methods according to the invention, the cancer is metastatic.

[0169] In some aspects of the combinations, uses, or methods according to the invention, metastatic cancer includes brain metastases.

[0170] Pharmaceutical Compositions and Kits

[0171] This invention also relates to pharmaceutical compositions comprising the aforementioned combinations, WEE1 inhibitors as described herein, or PKMYT1 inhibitors as described herein, and their use in the preparation of medicaments for treating cancer in patients of need. Unless otherwise specified, the following indications also apply to pharmaceutical compositions comprising only a WEE1 inhibitor, pharmaceutical compositions comprising only a PKMYT1 inhibitor, and pharmaceutical compositions comprising both a WEE1 inhibitor and a PKMYT1 inhibitor. The possible presence of a WEE1 inhibitor or a PKMYT1 inhibitor, or both, is collectively referred to as the “active ingredient.”

[0172] The pharmaceutical compositions of the present invention, suitable for oral administration (oral dosage forms), can be in solid or liquid form. Suitable solid oral dosage forms include capsules (which may comprise powders, granules, and / or microtablets and may be soft or hard capsules), tablets, powders, or granules, and the like, each containing a predetermined amount of the active ingredient. Suitable liquid oral dosage forms include solutions, emulsions, or suspensions. The pharmaceutical compositions of the present invention can also be in sustained-release formulations.

[0173] Any inert component commonly used as a carrier or diluent may be used as a pharmaceutically acceptable excipient in the solid oral formulations of the present invention, such as gums, starches, sugars, cellulose materials, acrylates, or mixtures thereof. Preferred diluents include, for example, microcrystalline cellulose and anhydrous lactose. The composition may further comprise a disintegrant (e.g., croscarmellose sodium, sodium glycolate starch) and a lubricant (e.g., magnesium stearate), and may additionally comprise one or more additives selected from: binders (e.g., hydroxypropyl cellulose), flow aids (e.g., silica), buffers (e.g., citric acid), surfactants (e.g., Tween 80), solubilizers (e.g., cyclodextrin), plasticizers (e.g., triacetin), emulsifiers (e.g., sodium lauryl sulfate), stabilizers (e.g., povidone, ascorbic acid), viscosity increasers (e.g., hydroxypropyl methylcellulose), sweeteners (e.g., sucrose), film-forming agents (e.g., cellulose-based systems, polymers), colorants (e.g., iron oxide), flavoring agents, or any combination thereof.

[0174] The oral pharmaceutical compositions of the present invention can be prepared by any of the pharmaceutical methods, but all methods include the step of binding the active ingredient with a carrier constituting one or more essential components. Generally, the composition is prepared by uniformly and tightly mixing the active ingredient with a liquid carrier or a finely powdered solid carrier, or both, and subsequently shaping the product into the desired form, if necessary. For example, tablets can be prepared by compression or molding, optionally with one or more excipients. Compressed tablets can be prepared by compressing the active ingredient in a free-flowing form (such as powder or granules) in a suitable machine, optionally mixed with a binder, lubricant, inert diluent, surfactant, or dispersant. Molded tablets can be manufactured by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0175] WEE1 inhibitors

[0176] Pharmaceutical compositions containing WEE1 inhibitors may, for example, have unit doses of an active ingredient (free base) selected from the range of 10 mg to 250 mg or 20 mg to 200 mg, such as 20, 30, 60, 80, 100, 130, 140, 150, 200, and 240 mg. Moderate concentrations can be readily manufactured to cover alternative dose levels.

[0177] For example, WEE1 inhibitors are formulated in capsules, such as hard gelatin capsules containing microtablets, each microtablet containing 10 mg of the active ingredient. In some aspects, WEE1 inhibitors are compounds of formula (I). The following capsule concentrations (strengths) are available: 20, 30, 60, 100, 130, 150, and 200 mg of the active ingredient (free base). Medium capsule concentrations can be readily manufactured to cover alternative dose levels.

[0178] In some aspects, solid oral dosage forms are packaged in blister packs and / or bottles. In some aspects, one, two, three, four, five, or six units of the WEE1 inhibitor are administered with each intake. Preferably, one or two units of the WEE1 inhibitor are administered with each intake.

[0179] For example, when formulated as capsules containing microtablets, the excipients in each microtablet may include microcrystalline cellulose, anhydrous lactose, hydroxypropyl cellulose, sodium glycolate starch, anhydrous colloidal silica, and magnesium stearate. Alternative excipients may be used in oral dosage forms.

[0180] PKMYT1 inhibitors

[0181] Pharmaceutical compositions containing PKMYT1 inhibitors may, for example, have unit doses of the active ingredient (free base) selected from the range of 10 mg to 250 mg or 20 mg to 200 mg, such as 20, 30, 60, 80, 100, 130, 140, 150, 200, and 240 mg. Moderate concentrations can be readily manufactured to cover alternative dose levels.

[0182] Reagent test kit

[0183] This invention also relates to kits comprising the aforementioned combinations, WEE1 inhibitors for use, or PKMYT1 inhibitors for use, for treating cancer in patients in need, or for preparing pharmaceutical agents for treating cancer in patients in need. In some aspects, the kit comprises a WEE1 inhibitor, specifically a compound of formula (I), and a PKMYT1 inhibitor, specifically a compound of formula (II), and instructions for use in treating cancer, specifically according to the uses and methods described herein. Optionally, in certain aspects, the kit may comprise a WEE1 inhibitor, specifically a compound of formula (I), and a PKMYT1 inhibitor, specifically a compound of formula (II), in individual dosage forms or containers.

[0184]

[0185] In some aspects of the invention, the combination of WEE1 inhibitors and PKMYT1 inhibitors exhibits synergistic effects in treating cancer, such as synthetic lethality.

[0186] In vitro studies have shown that the combination of WEE1 inhibitors and PKMYT1 inhibitors significantly increases the induction of cell death (synthetic lethality). In vivo, this combination effectively and significantly improves tumor growth inhibition and can even induce complete regression. The combination is also well-tolerated in vivo.

[0187] The synthetic lethal effects of the combinations disclosed herein are particularly applicable to the treatment of diseases or conditions, such as cancer, and especially those that depend on the activity of membrane-associated tyrosine and threonine-specific cdc2 inhibitory kinase (Myt1) (gene name PKMYT1) (e.g., cancers with CCNE1 amplification / overexpression or FBXW7 mutations).

[0188] The aspects of this disclosure may be further defined and illustrated with reference to the following non-limiting examples. Those skilled in the art will appreciate that many modifications or alterations may be made to, for example, the materials and methods described, without departing from the scope of this disclosure.

[0189] Experimental Section

[0190] Example 1: Efficiently knocking down PKMYT1 using siRNA

[0191] A549 cells (lung cancer cell line) were reverse-transfected with various concentrations of DharmaFECT 1 and PKMYT1 siRNA pools for 48 hours, followed by PKMYT1 mRNA analysis by RT-qPCR. PKMYT1 gene expression was reported as a percentage of relative PKMYT1 expression and knockdown relative to the housekeeping gene GAPDH and to cells transfected with matched concentrations of scrambled siRNA (negative control). N = 2 transfections. Values ​​are shown as mean + / - SD.

[0192] The results are plotted in Figure 1 In A549 cells, DharmaFECT combined with PKMYT1 siRNA at doses ranging from 25 to 100 nM mediated over 90% PKMYT1 knockdown.

[0193] Cell confluence was similar across siRNA doses 48 hours post-transfection. A dose of 50 nM siRNA was chosen for Example 2.

[0194] Example 2: Knockdown of synthetic lethality in WEE1 gene knockout (KO) cells by PKMYT1 siRNA

[0195] A549 cells (WT or WEE1 KO) were transfected in reverse at 50 nM using DharmaFECT 1 and PKMYT1 siRNA pools or randomized siRNA. (A) Representative bright-field images of cells 72 hours after transfection (data at 96 hours are similar and not shown in the figure) and (B) Quantification of viable cells per well by flow cytometry at 72 and 96 hours after transfection.

[0196] The results are plotted in Figure 2 In A549WT cells, transfection with PKMYT1 siRNA did reduce cell viability at 72 and 96 hours, but viable cells remained. Transfection with PKMYT1 siRNA in A549 WEE1 KO cells resulted in synthetic lethality, leading to undetectable viable cells.

[0197] Example 3: In vivo synthetic lethality of combined PKMYT1 inhibitor and WEE1 inhibitor (ovarian cancer cell line-derived xenograft model)

[0198] Balb / c nude mice were fixed and subcutaneously inoculated in 0.2 mL of PBS solution (mixed with matrix gel, 1:1) at a concentration of 1 × 10⁶ cells / mL in the right ventral region. 7 OVCAR-3 ovarian tumor cells were used for tumor development. When the average tumor size reached approximately 150-200 mm... 3Animals were randomly assigned to four study groups using a "matched distribution" / "stratified" method (StudyDirector™ software, version 3.1.399.19). The date of randomization was labeled Day 0. Morbidity and mortality were assessed daily after tumor cell inoculation. Individual mortality rates and observed clinical signs were meticulously recorded. Tumor volume was measured twice weekly in two dimensions using calipers, and the volume was expressed in mm. 3 This means that the formula is: V=(L×W×W) / 2, where V is the tumor volume, L is the tumor length (the longest tumor size), and W is the tumor width (the longest tumor size perpendicular to L).

[0199] After being grouped according to the study design, patients were immediately treated with the vehicle, compound (I) (30 mg / kg QD) (referred to as [compound (I)]), RP6306 (10 mg / kg or 20 mg / kg BID), and combinations thereof for 37 days.

[0200] Tumor growth inhibition (TGI): TGI% is an indicator of antitumor activity and is expressed as: TGI (%) = 100 × (1 - T / C). T and C are the mean tumor volume (or weight) of the treatment group and the control group, respectively, on a given day.

[0201] The results are plotted in Figure 3 In the experimental animals, treatment with the test formulation (I) compound was tolerable, and no significant treatment-related weight loss or adverse clinical signs were observed in mice after combination with RP-6306. On day 37 after randomization, the mean tumor volume in the load-treated group (Group 1) reached 1387.86 mm³. Monotherapy with 30 mg / kg of the formulation (I) compound (Group 2) produced only a mild antitumor response, with a TGI of 17.18% compared to the control group (p>0.05). Treatment with 20 mg / kg and 10 mg / kg of RP-6306 (Groups 3 and 4) produced mild but not significant antitumor activity, with TGIs of 38.9% and 42.26%, respectively (p>0.05). When compound (I) was combined with RP-6306, improved and superior antitumor activity was observed in groups 5 and 6 (RP6306 20 mg / kg and 10 mg / kg, respectively), where all tumors were designated as completely regressed (TGI 94.23% and 82.54% (p<0.01); all below the lower limit of quantitation). Furthermore, the combination of compound (I) and RP6306 (at either dose) induced significant tumor growth inhibition compared to administration of either single agent as monotherapy (p<0.01).

[0202] Example 4: In vivo efficacy of compound (I) in combination with RP6306 in a cell line-derived xenograft model of breast cancer.

[0203] Balb / c nude mice were fixed and subcutaneously inoculated in 0.2 mL of PBS solution (mixed with matrix gel, 1:1) at a concentration of 4 × 10⁶ cells / mL in the right ventral region. 6 One SUM149PT breast tumor cell was used for tumor development. When the average tumor size reached approximately 190 mm... 3 Animals were randomly assigned to four study groups using a "matched distribution" / "stratified" method (StudyDirector™ software, version 3.1.399.19). The date of randomization was labeled Day 0. Morbidity and mortality were assessed daily after tumor cell inoculation. Individual animal mortality rates and observed clinical signs were meticulously recorded. Tumor volume was measured twice weekly in two dimensions using calipers, and the volume was expressed in mm. 3 This means that the formula is: V=(L×W×W) / 2, where V is the tumor volume, L is the tumor length (the longest tumor size), and W is the tumor width (the longest tumor size perpendicular to L).

[0204] After being grouped according to the study design, patients were immediately treated with the carrier, compound (I) (30 mg / kg QD) (referred to as [compound (I) (Cpd of formula (I))]), RP6306 (20 mg / kg BID), and combination thereof for 28 days.

[0205] Tumor growth inhibition (TGI): TGI% is an indicator of antitumor activity and is expressed as: TGI (%) = 100 × (1 - T / C). T and C are the mean tumor volume (or weight) of the treatment group and the control group, respectively, on a given day.

[0206] The results are plotted in Figure 4 In the experimental animals, treatment with compound (I) was tolerable, and no significant treatment-related weight loss or adverse clinical signs were observed in mice after combination with RP-6306. On day 28 after randomization, the mean tumor volume in the load-treated group (Group 1) reached 1471 mm. 3Monotherapy with compound (I) at 30 mg / kg (Group 2) produced only a mild antitumor response, with a TGI of 20.67% compared to the control group (p<0.05). Treatment with RP-6306 at 20 mg / kg (Group 3) did not produce any significant antitumor activity, with a TGI of -10.17% (p>0.05). Improved and superior antitumor activity was observed in Group 4 following the combination of compound (I) and RP-6306, where all tumors regressed by day 18 compared to the initial tumor volume, and the TGI was 100.71% by day 28 (p<0.01). Furthermore, the combination of compound (I) and RP6306 induced significant tumor growth inhibition compared to administration of either drug as monotherapy (p<0.01).

[0207] Example 5: In vivo efficacy of compound (I) in combination with RP6306 in patient-derived xenograft (PDX) models of breast and ovarian cancer.

[0208] NCG mice were fixed and subcutaneously inoculated with 3×3×3 mm patient-derived breast or ovarian tumor cell fragments in streptomy gel in the right ventral region for tumor development. When the average tumor size of the breast cancer PDX reached approximately 185 mm... 3 Or the average tumor size of the ovarian PDX reached approximately 150 mm. 3 Animals were randomly assigned to four study groups using a "matched distribution" or "stratified" method. The date of randomization was designated as day 0. Morbidity and mortality were assessed daily after tumor cell inoculation. Individual animal mortality rates and observed clinical signs were meticulously recorded. Tumor volume was measured twice weekly in two dimensions using calipers, and the volume was expressed in mm. 3 This means that the formula is: V=(L×W×W) / 2, where V is the tumor volume, L is the tumor length (the longest tumor size), and W is the tumor width (the longest tumor size perpendicular to L).

[0209] After being grouped according to the study design, patients were immediately treated with the carrier, compound (I) (30 mg / kg QD) (referred to as [compound (I) (Cpd of formula (I))]), RP6306 (20 mg / kg BID), and combination thereof for 28 days.

[0210] Tumor growth inhibition (TGI): TGI% is an indicator of antitumor activity and is expressed as: TGI (%) = 100 × (1 - T / C). T and C are the mean tumor volume (or weight) of the treatment group and the control group, respectively, on a given day.

[0211] LD1 model of breast cancer: The LD1 model used was characterized by CCNE1 amplification (26 copies) and high CCNE1 mRNA expression (139 transcripts per million as determined by RNAseq).

[0212] The results are plotted in Figure 5 In the experimental animals, treatment with compound (I) was tolerable, and no significant treatment-related weight loss or adverse clinical signs were observed in mice after combination with RP-6306. On day 35 after randomization, the mean tumor volume in the load-treated group (Group 1) reached 1284 mm. 3 Monotherapy with compound (I) at 30 mg / kg (Group 2) produced only a mild antitumor response, with a TGI of 20% compared to the control group (p<0.05). Treatment with RP-6306 at 20 mg / kg (Group 3) did not produce any significant antitumor activity. Improved and superior antitumor activity was observed in Group 4 following the combination of compound (I) and RP-6306, in which all tumors achieved complete regression (no detectable tumors) by day 38, and this regression continued until the study was completed on day 52 (p<0.01). Furthermore, the combination of compound (I) and RP6306 induced significant tumor growth inhibition compared to administration of either single agent as monotherapy (p<0.01).

[0213] Ovarian PDX280 model: The results are plotted in Figure 6 In the experimental animals, treatment with compound (I) was tolerable, and no significant treatment-related weight loss or adverse clinical signs were observed in mice after combination with RP-6306. On day 28 after randomization, the mean tumor volume in the load-treated group (Group 1) reached 471.5 mm. 3 Monotherapy with compound (I) at 30 mg / kg (Group 2) produced only a mild antitumor response, with a TGI of 2.8% compared to the control group (p>0.05). Treatment with RP-6306 at 20 mg / kg (Group 3) did not produce any significant antitumor activity, with a TGI of -2.5% (p>0.05). Improved and superior antitumor activity was observed in Group 4 following the combination of compound (I) and RP-6306, with 7 out of 8 tumors designated as completely regressed by day 28 compared to the initial tumor volume, and a TGI of 183% compared to the load-bearing control (p<0.01). Furthermore, the combination of compound (I) and RP6306 induced significant tumor growth inhibition compared to administration of either drug as monotherapy (p<0.01).

Claims

1. A combination of a WEE1 inhibitor and a PKMYT1 inhibitor.

2. The combination as described in claim 1, which is used as a pharmaceutical agent.

3. The combination as described in claim 1 or 2, applied to treat cancer in patients in need.

4. A WEE1 inhibitor for treating cancer in patients in need, wherein the WEE1 inhibitor is administered in combination with a PKMYT1 inhibitor.

5. A PKMYT1 inhibitor for the treatment of cancer in patients in need, wherein the PKMYT1 inhibitor is administered in combination with a WEE1 inhibitor.

6. The combination of any one of claims 1 to 3, the WEE1 inhibitor of claim 4, or the PKMYT1 inhibitor of claim 5, wherein the WEE1 inhibitor is a compound of formula (I). (I), or a pharmaceutically acceptable salt thereof.

7. A combination of the applications of claim 3 or 6, a WEE1 inhibitor of the application of claim 4 or 6, or a PKMYT1 inhibitor of the application of claim 5 or 6, wherein the WEE1 inhibitor is administered as follows: a. From day 1 to day 3 of the 21-day cycle, b. On days 1 to 3 and days 8 to 10 of the 21-day cycle, c. On days 1 to 3, 8 to 10, and 15 to 17 of the 21-day cycle, d. From day 1 to day 5 of the 21-day cycle, e. Days 1 to 5 and days 8 to 12 within a 21-day cycle, f. During days 1 to 5, 8 to 12, and 15 to 19 of the 21-day cycle, g. From day 1 to day 14 of the 21-day cycle, h. Daily within a 21-day cycle, i. During days 1 to 5, 8 to 10, and 15 to 17 of the 28-day cycle, j. From day 1 to day 5 of the 28-day cycle, k. On days 1 to 5 and days 8 to 10 of the 28-day cycle, 1. On days 1 to 5 and days 8 to 12 of the 28-day cycle, m. During days 1 to 5, 8 to 12, and 15 to 19 of the 28-day cycle, n. Days 1 to 3, 8 to 10, and 15 to 17 of a 28-day cycle. o. On days 1 to 3 and days 8 to 10 of a 28-day cycle, or p. Days 1 to 3 of a 28-day cycle.

8. A combination of the applications of any one of claims 3, 6, or 7, a WEE1 inhibitor of the application of any one of claims 4, 6, or 7, or a PKMYT1 inhibitor of the application of any one of claims 5, 6, or 7, wherein the WEE1 inhibitor is administered as follows: a. A dose of about 30 to about 1000 mg per WEE1 inhibitor treatment day, preferably in the range of about 30 to about 720 mg per WEE1 inhibitor treatment day, and / or b. Orally, and / or c. On the day of WEE1 inhibitor treatment, administer a single dose, and / or d. The treatment days for each WEE1 inhibitor were at approximately the same time.

9. A combination of the applications of any one of claims 3, 6, 7, or 8, a WEE1 inhibitor of the application of any one of claims 4, 6, 7, or 8, or a PKMYT1 inhibitor of the application of any one of claims 5, 6, 7, or 8, wherein the PKMYT1 inhibitor is administered as follows: a. A number of consecutive or non-consecutive 7 or more days in a 21-day cycle, such as a number of consecutive or non-consecutive 14 or more days or days per day, and / or b. On days 1 to 3, 8 to 10, and 15 to 17 of the 21-day cycle, and / or c. On the same day as the WEE1 inhibitor treatment, preferably on the same day as defined in claim 7, and / or d. On a different day from the treatment date of the WEE1 inhibitor in a 21-day or 28-day cycle, and / or e. The PKMYT1 inhibitor and the WEE1 inhibitor are administered alternately, wherein one of the drugs is administered for x days and then paused for y days, and the other drug is paused during said x days and administered during said y days, followed by another period of x days and y days, etc., wherein x and y are independently selected from 1, 2, 3, 4, 5, 6 and 7.

10. A combination of the applications of any one of claims 3 to 9, a WEE1 inhibitor of the application of any one of claims 4 to 9, or a PKMYT1 inhibitor of the application of any one of claims 5 to 9, wherein the PKMYT1 inhibitor is administered as follows: a. Each day of PKMYT1 inhibitor treatment, at a dose ranging from about 60 mg to about 400 mg, or from about 100 mg to about 300 mg, or at a dose of about 240 mg, and / or b. Once or twice daily on each PKMYT1 inhibitor treatment day, and / or c. Administered once or twice daily at a dose selected from approximately 30 to approximately 100 mg, or approximately 60 to approximately 100 mg, or approximately 80 to approximately 100 mg per administration, and / or d. Administer a dose ranging from approximately 80 to approximately 100 mg twice daily for each PKMYT1 inhibitor treatment day, and / or, e. 240 mg daily on each day of PKMYT1 inhibitor treatment, and / or f. via oral communication.

11. A combination of the applications of any one of claims 7 to 10, a WEE1 inhibitor of the application of any one of claims 7 to 10, or a PKMYT1 inhibitor of the application of any one of claims 7 to 10, wherein the WEE1 inhibitor and the PKMYT1 inhibitor are administered according to one of the following options: A. Both the WEE1 inhibitor and the PKMYT1 inhibitor were administered on days 1-3, 8-10, and 15-17 of a 21-day cycle. B. Both the WEE1 inhibitor and the PKMYT1 inhibitor were administered on days 1-3, 8-10, 15-17, and 22-24 of a 28-day cycle. C. The WEE1 inhibitor is administered daily, and the PKMYT1 inhibitor is administered on days 1-3, 8-10, and 15-17 of a 21-day cycle. D. The WEE1 inhibitor is administered daily, and the PKMYT1 inhibitor is administered on days 1-3, 8-10, 15-17, and 22-24 of a 28-day cycle. E. The WEE1 inhibitor was administered on days 1-3 and 8-10 of a 21-day cycle, and the PKMYT1 inhibitor was administered on days 1-3, 8-10, and 15-17 of a 21-day cycle, or F. Both the WEE1 inhibitor and the PKMYT1 inhibitor were administered on days 1-3 and 15-17 of a 28-day cycle.

12. A combination of the applications of any one of claims 7 to 11, a WEE1 inhibitor of the application of any one of claims 7 to 11, or a PKMYT1 inhibitor of the application of any one of claims 7 to 11, wherein the combination, the WEE1 inhibitor, and / or the PKMYT1 inhibitor are administered over 1, 2, 3, 4, 5, 6, or more 21-day cycles or over 1, 2, 3, 4, 5, 6, or more 28-day cycles.

13. A combination of the applications of any one of claims 3 or 6 to 12, a WEE1 inhibitor of the application of any one of claims 4 or 6 to 12, or a PKMYT1 inhibitor of the application of any one of claims 5 or 6 to 12, wherein the cancer is ovarian cancer; primary peritoneal cancer; fallopian tube cancer; epithelial ovarian cancer (EOC), including serous EOC, endometrioid EOC, or clear cell EOC; gastric cancer; colorectal cancer; bladder cancer; cervical cancer; esophageal cancer; small cell lung cancer (SCLC); non-small cell lung cancer (NSCLC); breast cancer; soft tissue sarcoma; glioma; or endometrial cancer, including serous uterine carcinoma (USC).

14. A combination of the applications of any one of claims 3 or 6 to 13, a WEE1 inhibitor of the application of any one of claims 4 or 6 to 13, or a PKMYT1 inhibitor of the application of any one of claims 5 or 6 to 13, wherein the cancer is characterized by CCNE1 overexpression and / or CCNE1 amplification.

15. A combination of the applications of any one of claims 3 or 6 to 13, a WEE1 inhibitor of the application of any one of claims 4 or 6 to 13, or a PKMYT1 inhibitor of the application of any one of claims 5 or 6 to 13, wherein the cancer is characterized by ATRX deficiency, FBXW7 inactivating mutation, or PPP2R1A inactivating mutation.

16. A combination of the applications of any one of claims 3 to 15, a WEE1 inhibitor of the application of any one of claims 4 to 15, or a PKMYT1 inhibitor of the application of any one of claims 5 to 15, wherein the cancer is metastatic.

17. A combination of the applications of claim 16, a WEE1 inhibitor of the application of claim 15, or a PKMYT1 inhibitor of the application of claim 15, wherein the metastatic cancer comprises brain metastases.

18. A combination of the applications of any one of claims 1, 2, 3, or 6 to 17, a WEE1 inhibitor of the application of any one of claims 4, 6 to 17, or a PKMYT1 inhibitor of the application of any one of claims 5, 6 to 17, wherein the PKMYT1 inhibitor is a compound of formula (II). (II), or a pharmaceutically acceptable salt thereof.

19. A pharmaceutical composition comprising any one of claims 1 to 3 and / or a combination further specified in claims 6 and / or 18, or comprising a WEE1 inhibitor specified in one or more of claims 4, 6 or 18 or a PKMYT1 inhibitor specified in one or more of claims 5, 6 or 18, for the treatment of cancer in a patient in need, wherein the application is described in any one of claims 3 to 5 or 7 to 17.

20. A kit comprising a combination as specified in any one of claims 1 to 3 and / or further specified in claims 6 and / or 18, wherein the WEE1 inhibitor and the PKMYT1 inhibitor are provided in separate dosage forms and / or containers, optionally for use in treating cancer in a patient in need, wherein the use is described in any one of claims 3 to 5 or 7 to 17.

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