WEE1 compounds for treatment of uterine serous cancer

Abstract

Disclosed herein are methods of identifying or selecting a subject that is sensitive to a WEE1 inhibitor (e.g., ZN-c3) and using a WEE1 inhibitor (e.g., ZN-c3) to treat or inhibit cancer, including non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer, or uterine serous cancer (USC).

Description

[0001] Incorporation by reference into priority application

[0002] Any and all applications for which foreign or domestic priority is claimed, such as identified in an application data sheet or request filed with this application, are hereby incorporated by reference under 37 CFR 1.57 and Rules 4.18 and 20.6, including U.S. Provisional Application No. 63 / 370,580, filed on August 5, 2022, which is hereby specifically incorporated by reference in its entirety. This application is related to PCT / US2022 / 024079, which claims priority to U.S. Provisional Application Serial No. 63 / 174,004, filed on April 12, 2021, and U.S. Provisional Application Serial No. 63 / 174,005, filed on April 12, 2021, all of which are hereby specifically incorporated by reference in their entirety. Background Art Technical Field

[0003] The present application generally relates to methods of identifying or selecting subjects or individuals that are sensitive to a WEE1 inhibitor (e.g., ZN-c3) and using a WEE1 inhibitor (e.g., ZN-c3) to treat or inhibit cancer, including non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer, or uterine serous carcinoma (USC).

[0004] describe

[0005] DNA is constantly exposed to environmental damage. Light, chemicals, stress, and cell replication cause single-strand or double-strand breaks along the DNA backbone. Typically, organisms defend against DNA damage by repair proteins that reattach or resynthesize damaged DNA. The correct working of these proteins is essential to life. Incorrect substitution of nucleotides into DNA can cause mutations (and other genetic changes, including but not limited to insertions, deletions, and frameshifts), genetic diseases, and loss of protein function. Complete loss of DNA repair can lead to cell death, tumor progression, and cancer.

[0006] Cell cycle checkpoints are important for correct DNA repair, ensuring that cells do not progress with cell replication before their genomic integrity is restored. WEE1 is a nuclear kinase that participates in the G2-M cell cycle checkpoint arrest for DNA repair prior to mitotic entry. Normal cells repair damaged DNA during G1 arrest. Cancer cells often have a defective G1-S checkpoint and rely on a functional G2-M checkpoint for DNA repair. WEE1 is overexpressed in various cancer types, and many inhibitors and / or degraders of WEE1 are known to those skilled in the art. See, for example, WO 2019 / 173082 and WO 2020 / 069105.

[0007] Uterine serous carcinoma (USC) is a highly aggressive type II endometrial cancer. See, e.g., Ferriss JS et al., Uterine serous carcinoma: key advances and novel treatment approaches. International Journal of Gynecologic Cancer 2021; 31: 1165-1174. A Phase II study of adavosertib in recurrent uterine serous carcinoma has been reported. See Liu JF et al., Phase II study of the WEE1 inhibitor adavosertib in recurrent uterine serous carcinoma. Journal of Clinical Oncology 2021; 39. However, existing treatments for USC and other cancers that are sensitive to WEE 1 inhibitors remain limited, and therefore additional therapies and methods for identifying or selecting individuals who will benefit from such therapies are urgently needed. Summary of the invention

[0008] Various embodiments described herein relate to methods of providing therapy (e.g., administering drug ZN-c3) to a subject or individual having cancer (e.g., a cancer sensitive to a WEE 1 inhibitor, such as non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer, or uterine serous carcinoma (USC)), and methods of identifying or selecting an individual or subject who will benefit from receiving such therapy. The methods described herein may include administering a therapeutically effective amount of a WEE 1 inhibitor (e.g., drug ZN-c3) or a pharmaceutically acceptable salt thereof to a subject or individual in need thereof, and may also include screening the subject or individual for regulation in a biological sample obtained from the subject or individual of a protein phosphatase 2 scaffold subunit alpha (PPP2R1A) protein or a gene encoding the PPP2R1A protein, such as a polymorphism that confers sensitivity to a WEE 1 inhibitor (e.g., drug ZN-c3), such as a polymorphism selected from P179R, S256F, or R183W, or a nucleic acid encoding a polymorphism selected from P179R, S256F, or R183W. Methods for identifying or selecting a subject or individual with sensitivity to a WEE 1 inhibitor (e.g., drug ZN-c3) are contemplated, whereby a biological sample from the subject or individual is analyzed to determine the presence or absence of a modulation in the protein phosphatase 2 scaffold subunit alpha (PPP2R1A) protein or a gene encoding the PPP2R1A protein, such as a polymorphism that confers sensitivity to a WEE 1 inhibitor (e.g., drug ZN-c3), such as a polymorphism selected from P179R, S256F, or R183W, or a nucleic acid encoding a polymorphism selected from P179R, S256F, or R183W. These and other embodiments are described in more detail below.

[0009] 1. A method for identifying or selecting a subject with sensitivity to drug ZN-c3, the method comprising: identifying in a biological sample obtained from the subject or individual a regulation in the protein phosphatase 2 scaffold subunit alpha (PPP2R1A) protein or a gene encoding the PPP2R1A protein, such as one or more polymorphisms that confer sensitivity to the drug ZN-c3, preferably a polymorphism selected from P179R, S256F or R183W or a nucleic acid encoding a polymorphism selected from P179R, S256F or R183W; and when the regulation, such as any one or more polymorphisms in the PPP2R1A gene or protein, is identified, selecting or identifying the subject as a subject with sensitivity to the drug ZN-c3.

[0010] 2. The method according to alternative 1, further comprising administering ZN-c3 to the subject when the modulation or polymorphism is identified in the biological sample.

[0011] 3. The method according to any one of Alternatives 1 or 2, wherein the subject has cancer.

[0012] 4. The method according to alternative 3, wherein the cancer comprises non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, uterine serous carcinoma (USC) or endometrial cancer.

[0013] 5. A method according to any one of Alternatives 2 to 4, wherein the regression or inhibition of the cancer is greater than 7%, such as 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more.

[0014] 6. The method according to any one of Alternatives 2 to 5, further comprising administering to the subject an agent or a combination of agents that inhibits the PPP2R1A gene or protein.

[0015] 7. The method according to alternative embodiment 6, wherein the agent or combination of agents comprises the PP2A catalytic inhibitor LB-100.

[0016] 8. A method according to any one of Alternatives 1 to 7, wherein the modulation of PPP2R1A comprises one or more gain of function mutations.

[0017] 9. A method according to any one of Alternatives 1 to 7, wherein the regulation of PPP2R1A comprises one or more loss of function mutations.

[0018] 10. The method according to any one of Alternatives 1 to 7, wherein the modulation of PPP2R1A comprises one or more mutations leading to overexpression of PPP2R1A.

[0019] 11. The method according to any one of Alternatives 1 to 7, wherein the modulation of PPP2R1A comprises one or more mutations that result in underexpression of PPP2R1A.

[0020] 12. A method for inhibiting, ameliorating or treating cancer or its sequelae in a subject, the method comprising: identifying regulation in the protein phosphatase 2 scaffold subunit alpha (PPP2R1A) protein or the gene encoding the PPP2R1A protein in a biological sample obtained from the subject, wherein the polymorphism confers sensitivity to the drug ZN-c3, such as a polymorphism selected from P179R, S256F or R183W or a nucleic acid encoding a polymorphism selected from P179R, S256F or R183W; and administering ZN-c3 to the subject when the polymorphism is identified in the biological sample.

[0021] 13. The method of alternative 12, wherein the cancer comprises non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, uterine serous carcinoma (USC), or endometrial cancer.

[0022] 14. A method according to alternative scheme 12 or 13, wherein the regression or inhibition of the cancer is greater than 7%, such as 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more.

[0023] 15. The method according to any one of Alternatives 12 to 14, wherein the subject is additionally provided with an agent or combination of agents that inhibits the PPP2R1A gene or protein.

[0024] 16. The method according to alternative 15, wherein the agent or combination of agents comprises the PP2A catalytic inhibitor LB-100.

[0025] 17. A method according to any one of Alternatives 12 to 16, wherein the modulation of PPP2R1A comprises one or more gain of function mutations.

[0026] 18. A method according to any one of Alternatives 12 to 16, wherein the modulation of PPP2R1A comprises one or more loss of function mutations.

[0027] 19. The method according to any one of Alternatives 12 to 16, wherein the modulation of PPP2R1A comprises one or more mutations leading to overexpression of PPP2R1A.

[0028] 20. The method of any one of Alternatives 12 to 16, wherein the modulation of PPP2R1A comprises one or more mutations that result in underexpression of PPP2R1A.

[0029] 21. A compound ZN-c3 for use in inhibiting, ameliorating or treating cancer or its sequelae in a subject identified as having a regulation in the protein phosphatase 2 scaffold subunit alpha (PPP2R1A) protein or a gene encoding the PPP2R1A protein in a biological sample obtained from the subject, such as a polymorphism that confers sensitivity to the drug ZN-c3, for example a polymorphism selected from P179R, S256F or R183W or a nucleic acid encoding a polymorphism selected from P179R, S256F or R183W.

[0030] 22. Compound ZN-c3 for use according to alternative 21, wherein the cancer comprises non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, uterine serous carcinoma (USC) or endometrial cancer.

[0031] 23. The compound ZN-c3 for use according to alternative scheme 21, wherein the regression or inhibition of the cancer is greater than 7%, such as 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more.

[0032] 24. Compound ZN-c3 for use according to any one of Alternatives 21 to 23, wherein the subject is additionally provided with an agent or a combination of agents that inhibits the PPP2R1A gene or protein.

[0033] 25. Compound ZN-c3 for use according to alternative 24, wherein the agent or combination of agents comprises the PP2A catalytic inhibitor LB-100.

[0034] 26. Compound ZN-c3 for use according to any of alternatives 21 to 25, wherein the modulation of PPP2R1A comprises one or more gain of function mutations.

[0035] 27. Compound ZN-c3 for use according to any one of alternatives 21 to 25, wherein the modulation of PPP2R1A comprises one or more loss of function mutations.

[0036] 28. Compound ZN-c3 for use according to any one of alternatives 21 to 25, wherein the modulation of PPP2R1A comprises one or more mutations leading to overexpression of PPP2R1A.

[0037] 29. Compound ZN-c3 for use according to any one of alternatives 21 to 25, wherein the modulation of PPP2R1A comprises one or more mutations leading to underexpression of PPP2R1A.

[0038] 30. A method for inhibiting, ameliorating or treating cancer or its sequelae in a subject, the method comprising: administering to the subject an agent or a combination of agents that inhibits the PPP2R1A gene or protein; and administering ZN-c3 to the subject.

[0039] 31. The method of alternative 30, wherein the agent or combination of agents that inhibit the PPP2R1A gene or protein is provided to the subject alone.

[0040] 32. The method of alternative 30 or 31, wherein the agent or combination of agents that inhibits the PPP2R1A gene or protein is provided to the subject prior to administration of ZN-C3.

[0041] 33. A method according to alternative embodiment 30, wherein the agent or combination of agents that inhibit the PPP2R1A gene or protein is provided to the subject within an interval of 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 11 minutes, 12 minutes, 13 minutes, 14 minutes, 15 minutes, 16 minutes, 17 minutes, 18 minutes, 19 minutes or 20 minutes.

[0042] 34. A method according to any one of Alternatives 30 to 33, wherein the cancer comprises non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, uterine serous carcinoma (USC), or endometrial cancer.

[0043] 35. A method according to any one of alternatives 30 to 34, wherein the regression or inhibition of the cancer is greater than 7%, such as 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more.

[0044] 36. The method of any one of Alternatives 30 to 35, wherein the agent or combination of agents comprises the PP2A catalytic inhibitor LB-100.

[0045] 37. A product combination for use in inhibiting, ameliorating or treating cancer or its sequelae in a subject, the product combination comprising compound ZN-c3 and an agent or a combination of agents that inhibits PPP2R1A gene or protein.

[0046] 38. The product combination for use according to alternative 37, wherein the cancer comprises non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, uterine serous carcinoma (USC) or endometrial cancer.

[0047] 39. A product combination for use according to any of alternatives 37 or 38, wherein the regression or inhibition of the cancer is greater than 7%, such as 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more.

[0048] 40. A product combination for use according to any one of Alternatives 37 to 39, wherein the agent or combination of agents comprises the PP2A catalytic inhibitor LB-100.

[0049] 41. The method according to any one of Alternatives 1 to 20 or the compound ZN-c3 for use according to any one of Alternatives 21 to 29, wherein the presence of the polymorphism is determined using next generation sequencing (NGS), sequencing, polymerase chain reaction (PCR), loop-mediated isothermal amplification, recombinase polymerase amplification or antibody detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 Depicted are the percentage cell density of A427 (a first non-small cell lung cancer cell line) cells after 72 hours of treatment with a 10-point dose response of ZN-c3, comparing cells transfected with siRNA against PPP2R1A to cells transfected with a scrambled control siRNA. Cell titer luminescence analysis was used to calculate the percentage cell density. Cells transfected with PPP2R1A siRNA were approximately 3.5-fold more sensitive to ZN-c3 (Table 1).

[0051] Figure 2Depicted is the mRNA expression of PPP2R1A in A427 cells treated with scrambled control siRNA or siRNA against PPP2R1A. mRNA is expressed as a percentage of the scrambled control.

[0052] Figure 3 Depicted are the percentage cell density of H1975 (a second non-small cell lung cancer cell line) cells after 72 hours of treatment with a 10-point dose response of ZN-c3, comparing cells transfected with siRNA against PPP2R1A to cells transfected with a scrambled control siRNA. Cell titer luminescence analysis was used to calculate the percentage cell density. Cells transfected with PPP2R1A siRNA had an approximately 3.7-fold increase in sensitivity to ZN-c3 (Table 2).

[0053] Figure 4 Depicted is the mRNA expression of PPP2R1A in H1975 cells treated with scrambled control siRNA or siRNA against PPP2R1A. mRNA is expressed as a percentage of the scrambled control.

[0054] Figure 5 Depicted are the percentage cell density of Hs578T (breast cancer cell line) cells after 72 hours of treatment with a 10-point dose response of ZN-c3, comparing cells transfected with siRNA against PPP2R1A to cells transfected with a scrambled control siRNA. Cell titer luminescence analysis was used to calculate the percentage cell density. Cells transfected with PPP2R1A siRNA had an approximately 2.4-fold increase in sensitivity to ZN-c3 (Table 3).

[0055] Figure 6 Depicted is the mRNA expression of PPP2R1A in Hs578T cells treated with scrambled control siRNA or siRNA against PPP2R1A. mRNA is expressed as a percentage of the scrambled control.

[0056] Figure 7 Depicted are the percentage cell density of SW620 (colorectal cancer cell line) cells after 72 hours of treatment with a 10-point dose response of ZN-c3, comparing cells transfected with siRNA against PPP2R1A to cells transfected with a scrambled control siRNA. Cell titer luminescence analysis was used to calculate the percentage cell density. Cells transfected with PPP2R1A siRNA had an approximately 1.9-fold increase in sensitivity to ZN-c3 (Table 4).

[0057] Figure 8 Depicted is the mRNA expression of PPP2R1A in SW620 cells treated with scrambled control siRNA or siRNA against PPP2R1A. mRNA is expressed as a percentage of the scrambled control.

[0058] Fig. 9 Depicted are the percentage cell density of SKOV3 (ovarian cancer cell line) cells after 72 hours of treatment with a 10-point dose response of ZN-c3, comparing cells transfected with siRNA against PPP2R1A to cells transfected with a scrambled control siRNA. Cell titer luminescence analysis was used to calculate the percentage cell density. Cells transfected with PPP2R1A siRNA were approximately 2.7-fold more sensitive to ZN-c3 (Table 5).

[0059] Fig.10 Depicted is the mRNA expression of PPP2R1A in SKOV3 cells treated with scrambled control siRNA or siRNA against PPP2R1A. mRNA is expressed as a percentage of the scrambled control.

[0060] Fig.11 Depicts the percentage cell density of HEC1B and HEC-59 (endometrial cancer cell line) cells after 72 hours of ZN-c3 treatment with 10-point dose response, comparing cells transfected with siRNA for PPP2R1A to cells transfected with scrambled control siRNA. HEC1B has a missense mutation (W257L; L429F) in PPP2R1A, making siRNA for PPP2R1A less effective in terms of sensitivity to ZN-c3. HEC-59 is PPP2R1A wild type. Cell titer luminescence analysis is used to calculate the percentage cell density. HEC1BPPP2R1A siRNA transfected cells increased sensitivity to ZN-c3 by about 1.4 times, while HEC-59PPP2R1A siRNA transfected cells increased sensitivity to ZN-c3 by about 2.5 times (Table 6).

[0061] Fig.12 Depicted are Western blot analyses of endometrial cancer cell lines HEC1B and HEC-59 with and without siRNA-mediated knockdown of PPP2R1A. Vinculin was used as a loading control.

[0062] Fig.13 Depicted are the percentage cell density of SKOV3 ovarian cancer cells after 72 hours of treatment with a 10-point dose response of ZN-c3, comparing cells transduced with a non-targeting guide RNA to cells with PPP2R1A knockout after transduction with a CRISPR guide RNA targeting PPP2R1A. Cellular titer luminescence analysis was used to calculate the percentage cell density. The sensitivity of PPP2R1A CRISPR knockout cells to ZN-c3 increased by approximately 1.7-fold (Table 7).

[0063] Fig.14Depicted is a Western blot analysis of the ovarian cancer cell line SKOV3 with and without CRISPR-mediated knockout of the PPP2R1A gene. β-Actin was used as a loading control.

[0064] Fig.15 Depicted are the percentage cell density of H1975 lung cancer cells after 72 hours of treatment with a 10-point dose response of ZN-c3, comparing cells transduced with non-targeting guide RNA to cells genetically knocked out PPP2R1A after transduction with CRISPR guide RNA targeting PPP2R1A. Cell titer luminescence analysis was used to calculate the percentage cell density. PPP2R1A CRISPR knockout cells were approximately 1.9-fold more sensitive to ZN-c3 (Table 8).

[0065] Fig.16 Depicted is a Western blot analysis of H1975 lung cancer cells with and without CRISPR-mediated knockout of PPP2R1A. β-Actin was used as a loading control.

[0066] Fig.17 Depicted are the percentage cell density of OV17R ovarian cancer cells after 72 hours of treatment with ZN-c3 at a 10-point dose response. OV17R has a S256F hotspot missense mutation in PPP2R1A. This cell line is highly sensitive to ZN-c3 treatment, and may be partially sensitive to PPP2R1A mutations (Table 9). Cell titer luminescence analysis was used to calculate the percentage cell density.

[0067] Fig.18 Depicted are the percentage inhibition of cell growth in HEC-59 endometrial cancer cells after treatment with ZN-c3 alone, PP2A catalytic inhibitor LB-100 alone, or a combination of ZN-c3+LB-100. The HEC-59 cell line is PPP2R1A wild type. Single agent treatment with either compound was not as effective as the combination therapy. Cell titer luminescence analysis was used to calculate the percentage inhibition. The dose of each compound is shown in Table 10.

[0068] Fig.19 Depicted are the percentage inhibition of cell growth in OV17R ovarian cancer cells after treatment with ZN-c3 alone, PP2A catalytic inhibitor LB-100 alone, or a combination of ZN-c3+LB-100. OV17R has the PPP2R1A missense mutation S256F. The percentage inhibition was calculated using a cell titer luminescence assay. The dose of each compound is shown in Table 11.

[0069] Fig. 20Depicted is the percentage inhibition of cell growth in HEC-59 endometrial cancer cells after siRNA-mediated knockdown of PPP2R1A, after treatment with ZN-c3 alone, PP2A catalytic inhibitor LB-100 alone, or a combination of ZN-c3+LB-100. Knockdown of PPP2R1A resulted in increased sensitivity to ZN-c3 for both single agent and combination therapy. Cell titer luminescence analysis was used to calculate the percentage inhibition. The doses of each compound are shown in Table 12.

[0070] Fig.21 Depicted is the percentage inhibition of cell growth in HEC1B endometrial cancer cells after siRNA-mediated knockdown of PPP2R1A, after treatment with ZN-c3 alone, PP2A catalytic inhibitor LB-100 alone, or a combination of ZN-c3+LB-100. Knockdown of PPP2R1A resulted in a slight increase in sensitivity, as shown above. The percentage inhibition was calculated using a cell titer luminescence assay. The dose of each compound is shown in Table 13. DETAILED DESCRIPTION

[0071] WEE1 is a tyrosine kinase, which is an important component of ATR-mediated G2 cell cycle checkpoint control, which prevents entry into mitosis in response to cell DNA damage. ATR phosphorylates and activates CHK1, which in turn activates WEE1, resulting in selective phosphorylation of cyclin-dependent kinase 1 (CDK1) at Tyr15, thereby stabilizing the CDK1-cyclin B complex and stopping cell cycle progression. This process confers a survival advantage by allowing tumor cells to obtain time for repairing damaged DNA before entering mitosis. The inhibition of WEE1 eliminates the G2 checkpoint, promotes cancer cells with DNA damage to enter unplanned mitosis, and experiences cell death via mitotic barriers. WEE1 inhibition and / or degradation have the potential to sensitize tumors to DNA damaging agents (such as cisplatin) and induce tumor cell death.

[0072] definition

[0073] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those of ordinary skill in the art. Unless otherwise indicated, all patents, applications, published applications, and other publications cited herein are incorporated by reference in their entirety. In the event that there are multiple definitions for a term herein, the definitions in that section shall prevail unless otherwise indicated.

[0074] The terms "WEE1 inhibition", "WEE1 inhibitors" and similar terms used herein refer to inhibiting the activity or function of WEE1 tyrosine kinase, for example by degrading WEE1 tyrosine kinase and / or by reducing the activity of WEE1 tyrosine kinase in mediating CDK1 phosphorylation. WEE1 inhibitors that act by degrading WEE1 tyrosine kinase may be referred to herein as WEE1 degraders.

[0075] As used herein, "subject" refers to an animal that is the object of treatment, observation, or experiment. The subject animal can be a mammal, such as, but not limited to, mice, rats, rabbits, guinea pigs, dogs, cats, sheep, goats, cattle, horses, primates (e.g., monkeys, chimpanzees, and apes), and in particular humans. In some embodiments, the subject can be a human. In some embodiments, the subject can be a child and / or an infant. In other embodiments, the subject can be an adult.

[0076] As used herein, the terms "treat" and "therapy" do not necessarily mean a complete cure or elimination of a disease or condition. Any degree of alleviation of any undesirable signs or symptoms of a disease or condition may be considered treatment and / or therapy. In addition, treatment may include actions that may worsen a subject's overall sense of well-being or appearance.

[0077] The terms "therapeutically effective amount" and "effective amount" are used to indicate the amount of an active compound (e.g., ZN-c3 or a pharmaceutically acceptable salt thereof) that elicits the indicated biological or pharmaceutical response. For example, a therapeutically effective amount of such a ZN-c3 compound, salt, or composition may be the amount required to prevent, alleviate, or ameliorate the symptoms of a disease or condition, or to prolong the survival of the subject being treated. The response may occur in a tissue, system, animal, or human, and includes alleviating the signs or symptoms of the disease or condition being treated. The determination of an effective amount is well within the capabilities of one skilled in the art, given the disclosure provided herein. The therapeutically effective amount (required to be a dose) of a ZN-c3 compound, salt, or composition herein will depend on the route of administration, the type of animal (including humans) being treated, and the physical characteristics of the particular animal under consideration. The dose may be modulated to achieve the desired effect, but the dose will depend on factors such as body weight, diet, concurrent medications, and other factors that will be recognized by those skilled in the medical field.

[0078] For example, an effective amount of a ZN-c3 compound, salt or composition may be an amount that results in the following effects: (a) reduction, alleviation or disappearance of one or more symptoms caused by USC, (b) reduction in tumor size, (c) elimination of tumors and / or (d) long-term disease stabilization (growth arrest) of tumors. For another example, an effective amount of a ZN-c3 compound, salt or composition may be an amount that results in reduced activity and / or phosphorylation (such as CDC2 phosphorylation) of WEE1. Reduction of WEE1 activity is known to those skilled in the art and can be determined by analyzing WEE1 intrinsic kinase activity and downstream substrate phosphorylation.

[0079] As used herein, the term "dosage regimen" refers to the manner in which the ZN-c3 compound is administered to a subject, including the route of administration, dosage, and dosing interval. A dosing regimen may include a "periodic" dosing phase, during which a specific dose (e.g., 300 mg) is administered at regular intervals (e.g., once daily) over a specific time period (e.g., three days). A dosing regimen may further include an "intermittent" dosing phase, during which one or more dosing parameters such as dosage and / or dosage interval are changed or altered. For example, an intermittent dosing phase may include a "rest" phase, during which the ZN-c3 compound is not administered or is administered at a reduced dose and / or less frequent.

[0080] It should be understood that the compounds described herein may be isotopically labeled. Substitution with isotopes such as deuterium may provide certain therapeutic advantages resulting from improved metabolic stability, such as, for example, increased half-life in vivo or reduced dosage requirements. Each chemical element as represented in the compound structure may include any isotope of the element. For example, in the compound structure, it may be explicitly disclosed or understood that hydrogen atoms are present in the compound. At any position of the compound where hydrogen atoms may be present, the hydrogen atoms may be any isotope of hydrogen, including but not limited to hydrogen-1 (protium) and hydrogen-2 (deuterium). Therefore, unless otherwise expressly specified in the context, the compounds mentioned herein encompass all possible isotopic forms.

[0081] Where a range of values ​​is provided, it should be understood that the upper and lower limits and every intermediate value between the upper and lower limits of the range are encompassed within the embodiments.

[0082] For the terms and phrases used in this application, especially in the appended claims and their variations, unless otherwise expressly stated, they should be understood as open-ended and not restrictive. For the aforementioned examples, the term "including" should be understood as "including but not limited to", "including but not limited to", etc.; as used herein, the term "including" is synonymous with "including", "containing" or "characterized as" and is inclusive or open-ended, and does not exclude additional unlisted elements or method steps; the term "having" should be interpreted as "at least having"; the term "including" should be interpreted as "including but not limited to"; the term "example" is used to provide an illustrative example of the item being discussed, rather than an exhaustive or restrictive list thereof; and the use of terms such as "preferably", "preferred", "desired" and "desired" and words of similar semantics should not be understood to imply that certain features are critical, necessary or even important for structure or function, but are intended only to highlight alternative or additional features that may or may not be used in a specific embodiment. In addition, the term "including" should be interpreted as synonymous with the phrase "at least having" or "at least including". When used in the context of a compound, composition, or device, the term "comprising" means that the compound, composition, or device includes at least the recited features or components, but may also include additional features or components.

[0083] For substantially any plural and / or singular terms used herein, a person skilled in the art may convert from the plural to the singular and / or from the singular to the plural, as long as it is appropriate to the context and / or application. For the sake of clarity, various singular / plural permutations may be explicitly stated herein. The indefinite article "a" or "an" does not exclude a plurality. The fact that certain measures are mentioned in mutually different dependent claims does not indicate that a combination of these measures cannot be used to make the advantages more prominent. Any figure marks in the claims should not be understood as limiting their scope.

[0084] In some embodiments, a method for selecting or identifying a subject with sensitivity to the drug ZN-c3 is provided. In some embodiments, the method comprises: identifying or detecting a regulation in the protein phosphatase 2 scaffold subunit alpha (PPP2R1A) protein or a gene encoding the PPP2R1A protein, for example, such regulation may be an overexpression or underexpression of the PPP2R1A protein or a transcript encoding the PPP2R1A protein, or may be a polymorphism in the PPP2R1A protein or a gene or transcript encoding the PPP2R1A protein. In some embodiments, a polymorphism that confers sensitivity to the drug ZN-c3 is identified or detected in the method. In some such embodiments, the polymorphism is selected from P179R, S256F or R183W or a nucleic acid encoding a polymorphism selected from P179R, S256F or R183W. The regulation in the above-mentioned PPP2R1A protein or the gene or transcript encoding the PPP2R1A protein is determined by analyzing a biological sample obtained from the subject suffering from cancer (such as non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer or uterine serous carcinoma (USC)). In some embodiments, the method includes selecting the subject as a subject with sensitivity to the drug ZN-c3 when any one or more of the aforementioned polymorphisms in the PPP2R1A gene or protein are identified. In some embodiments, once the subject is identified or selected as a subject with sensitivity to ZN-c3, ZN-c3 is provided to the identified or selected subject, for example, the subject is placed on a dosing regimen of ZN-c3.

[0085] In some embodiments, a method for inhibiting, improving or treating cancer or its sequelae in a subject is provided, the cancer or its sequelae are, for example, non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer or uterine serous carcinoma (USC). In some embodiments, the method includes identifying or detecting regulation in a protein phosphatase 2 scaffold subunit α (PPP2R1A) protein or a gene encoding the PPP2R1A protein, for example, such regulation may be overexpression or underexpression of a transcript of the PPP2R1A protein or a transcript encoding the PPP2R1A protein, or may be a polymorphism in a gene or transcript of the PPP2R1A protein or a PPP2R1A protein. In some embodiments, a polymorphism conferring sensitivity to drug ZN-c3 is identified or detected in the method. In some embodiments, in a biological sample obtained from the subject, a polymorphism is selected from P179R, S256F or R183W or a nucleic acid encoding a polymorphism selected from P179R, S256F or R183W. In some embodiments, when the polymorphism is identified in the biological sample, ZN-c3 is administered to the subject.

[0086] In some embodiments, compound ZN-c3 is provided for use in inhibiting, ameliorating or treating cancer or its sequelae in a subject identified as having regulation in a protein phosphatase 2 scaffold subunit alpha (PPP2R1A) protein or a gene encoding the PPP2R1A protein in a biological sample obtained from the subject, wherein the polymorphism confers sensitivity to drug ZN-c3, such as a polymorphism selected from any one or more of P179R, S256F or R183W or a nucleic acid encoding a polymorphism selected from any one or more of P179R, S256F or R183W. In some such uses, the cancer is, for example, non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer or uterine serous carcinoma (USC).

[0087] In some embodiments, a method of inhibiting, ameliorating or treating cancer or its sequelae in a subject is provided, the cancer or its sequelae, such as non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer or uterine serous carcinoma (USC). In some embodiments, the method comprises administering to the subject an agent or combination of agents that inhibits the PPP2R1A gene or protein, and administering ZN-c3 to the subject.

[0088] In some embodiments, a product combination for use in inhibiting, ameliorating or treating a subject's cancer or its sequelae (e.g., non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer, or uterine serous carcinoma (USC)) is provided, the product combination comprising compound ZN-c3 and an agent or a combination of agents that inhibit the PPP2R1A gene or protein.

[0089] In some embodiments, the above method includes administering ZN-c3 to the subject when a regulation in the PPP2R1A protein or a gene or transcript encoding the PPP2R1A protein (such as a polymorphism, preferably a polymorphism selected from any one or more of P179R, S256F or R183W or a polymorphism in a gene encoding a polymorphism selected from any one or more of P179R, S256F or R183W) is identified in the biological sample.

[0090] In some embodiments, the subject has cancer.

[0091] In some embodiments, the cancer comprises non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, or endometrial cancer.

[0092] In some embodiments, the regression or inhibition of the cancer is greater than 7%, such as 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more.

[0093] In some embodiments, the method further comprises administering to the subject an agent or combination of agents that inhibits the PPP2R1A gene or protein.

[0094] In some embodiments, the agent or combination of agents provided to the subject comprises the PP2A catalytic inhibitor LB-100.

[0095] In some embodiments, the subject is provided with the agent or combination of agents that inhibit the PPP2R1A gene or protein.

[0096] In some embodiments, the subject is provided with the agent or combination of agents that inhibits the PPP2R1A gene or protein prior to administration of ZN-C3.

[0097] In some embodiments, the agent or combination of agents that inhibit the PPP2R1A gene or protein is provided to the subject within an interval of 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 11 minutes, 12 minutes, 13 minutes, 14 minutes, 15 minutes, 16 minutes, 17 minutes, 18 minutes, 19 minutes, or 20 minutes.

[0098] In some embodiments, the presence of any one or more of the polymorphisms (e.g., a polymorphism in a gene encoding P179R, S256F, or R183W) is determined using next generation sequencing (NGS), sequencing, polymerase chain reaction (PCR), loop-mediated isothermal amplification, recombinase polymerase amplification, or antibody detection.

[0099] In some embodiments, modulation of PPP2R1A comprises one or more gain of function mutations.

[0100] In some embodiments, modulation of PPP2R1A comprises one or more loss of function mutations.

[0101] In some embodiments, modulation of PPP2R1A comprises one or more mutations that result in overexpression of PPP2R1A.

[0102] In some embodiments, modulation of PPP2R1A comprises one or more mutations that result in underexpression of PPP2R1A.

[0103] WEE1 Compound

[0104] ZN-c3 is a selective, orally bioavailable small molecule inhibitor of WEE1, a key component of the G2 / M cell cycle checkpoint that prevents cells from entering mitosis to allow repair of DNA damage. ZN-c3 exhibits significant in vitro antitumor activity in multiple cell lines and xenograft models.

[0105]

[0106] ZN-c3 can be prepared as described in WO 2019 / 173082 (see, e.g., Example 9B therein), which is hereby incorporated by reference herein and is particularly useful for the purpose of describing methods for preparing ZN-c3 and preparing salts thereof and pharmaceutical compositions thereof. If there is any difference between the chemical name and structure of ZN-c3, the structure should be given more weight.

[0107] Pharmaceutical composition

[0108] Some embodiments described herein relate to pharmaceutical compositions which may include an effective amount of a ZN-c3 compound, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, excipient, or a combination thereof.

[0109] The term "pharmaceutical composition" refers to a mixture of ZN-c3 and / or a pharmaceutically acceptable salt thereof with other chemical components (such as a diluent or carrier). Pharmaceutical compositions facilitate administration of compounds to an organism. Pharmaceutical compositions can also be obtained by reacting a compound with an inorganic acid or an organic acid (such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid). Pharmaceutical compositions will generally be formulated according to a specific intended route of administration.

[0110] The term "physiologically acceptable" defines a carrier, diluent or excipient that does not abrogate the biological activity and properties of the ZN-c3 compound and does not cause significant damage or injury to the animal to which the composition is intended to be delivered.

[0111] As used herein, "carrier" refers to a compound that facilitates the incorporation of a compound into cells or tissues. For example, but not limited to, dimethyl sulfoxide (DMSO) is a commonly used carrier that facilitates the uptake of many organic compounds into cells or tissues of a subject.

[0112] As used herein, "diluent" refers to an ingredient in a pharmaceutical composition that does not have significant pharmaceutical activity but may be necessary or desirable pharmaceutically. For example, a diluent can be used to increase the volume of a potent drug whose mass is too small to manufacture and / or administer. It can also be a liquid used to dissolve a drug to be administered by injection, ingestion, or inhalation. A common form of diluent in the art is a buffered aqueous solution, such as, but not limited to, phosphate-buffered saline that mimics the pH and isotonicity of human blood.

[0113] As used herein, "excipient" refers to a substantially inert substance added to a pharmaceutical composition to provide, but not limited to, bulk, consistency, stability, binding ability, lubrication, disintegration ability, etc. to the composition. For example, stabilizers such as antioxidants and metal chelators are excipients. In one embodiment, the pharmaceutical composition comprises an antioxidant and / or a metal chelator. A "diluent" is a type of excipient.

[0114] The pharmaceutical compositions described herein can be administered to human patients themselves, or administered to compositions wherein the pharmaceutical compositions are mixed with other active ingredients (such as in combination therapy), or mixed with carriers, diluents, excipients, or combinations thereof. The correct formulation depends on the selected route of administration. The techniques for preparing and administering the compounds described herein are known to those skilled in the art.

[0115] The pharmaceutical compositions disclosed herein can be manufactured in a manner known per se, for example, by conventional mixing, dissolving, granulating, dragee preparation, grinding, emulsifying, encapsulating, embedding or tableting processes. In addition, the ZN-c3 active ingredient is generally included in an amount effective to achieve its intended purpose and can be provided as a salt with a pharmaceutically compatible counterion.

[0116] A variety of art-recognized techniques for administering ZN-c3 compounds, salts, and / or compositions can be used, including but not limited to oral, rectal, pulmonary, topical, aerosol, injection, infusion, and parenteral delivery (including intramuscular, subcutaneous, intravenous, intramedullary, intrathecal, direct intraventricular, intraperitoneal, intranasal, and intraocular injection). In some embodiments, ZN-c3 or a pharmaceutically acceptable salt thereof can be administered orally.

[0117] The composition may (if desired) be present in a package or dispenser device, which may include one or more unit dosage forms containing the active ingredient. The package may, for example, include a metal or plastic foil, such as a blister package. The package or dispenser device may be accompanied by instructions for use. The package or dispenser may also be accompanied by a notice associated with the container form as specified by a government agency regulating the manufacture, use or sale of the drug, which reflects the agency's approval of the form of the drug for human or veterinary administration. For example, such notices may be labels approved by the U.S. Food and Drug Administration for prescription drugs or approved product inserts. Compositions comprising ZN-c3 and / or salts formulated in a compatible pharmaceutical carrier as described herein may also be prepared and placed in an appropriate container and labeled for use in treating the indicated disorder.

[0118] Uses and methods of treatment or inhibition

[0119] Some embodiments described herein relate to a method of treating or inhibiting cancer (e.g., non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer, or uterine serous carcinoma (USC)) in a subject, the method comprising administering to the subject a therapeutically effective amount of ZN-c3 or a pharmaceutically acceptable salt thereof, preferably according to a dosing regimen.

[0120] Some embodiments described herein relate to the use of a therapeutically effective amount of ZN-c3 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a subject having cancer (e.g., non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer, or uterine serous carcinoma (USC)), preferably according to a dosing regimen. Other embodiments relate to ZN-c3 for use in treating, inhibiting, or ameliorating a cancer (e.g., non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer, or uterine serous carcinoma (USC)) in a subject, comprising administering a therapeutically effective amount of ZN-c3 or a pharmaceutically acceptable salt thereof to the subject, preferably according to a dosing regimen. In various embodiments, the dosing regimen comprises orally administering ZN-c3 to the subject. In many of these embodiments, the subject is identified or selected as a subject with sensitivity to ZN-c3, and such sensitivity can be determined by analyzing a biological sample from the subject for modulation of the PPP2R1A protein or a gene encoding PPP2R1A, such as overexpression of the PPP2R1A protein or a transcript encoding the PPP2R1A protein or underexpression of the PPP2R1A protein or a transcript encoding the PPP2R1A protein. In some embodiments, the modulation of the PPP2R1A protein or a gene or transcript encoding the PPP2R1A protein involves a polymorphism in the PPP2R1A protein or a gene or transcript encoding the PPP2R1A protein, such as any one or more of P179R, S256F, or R183W, or a gene or transcript encoding a P179R, S256F, or R183W polymorphism.

[0121] [In various embodiments, the therapeutically effective amount of ZN-c3 administered to the subject is ≥300 mg per day. For example, in some embodiments, the therapeutically effective amount of ZN-c3 administered to the subject is about 300 mg per day. In other embodiments, the therapeutically effective amount of ZN-c3 administered to the subject is about 350 mg per day. In other embodiments, the therapeutically effective amount of ZN-c3 administered to the subject is about 200 mg per day. In some embodiments, the therapeutically effective amount of ZN-c3 administered to the subject is ≥300 mg, once a day (QD). In one embodiment, the therapeutically effective amount of ZN-c3 administered to the subject is about 300 mg, once a day (QD). In another embodiment, the therapeutically effective amount of ZN-c3 administered to the subject is about 350 mg, once a day (QD). In yet another embodiment, the therapeutically effective amount of ZN-c3 administered to the subject is about 200 mg, once a day (QD). In some embodiments, the therapeutically effective amount of ZN-c3 administered to a subject is ≥ 150 mg twice daily (BID). In one embodiment, the therapeutically effective amount of ZN-c3 administered to a subject is about 150 mg twice daily (BID). In another embodiment, the therapeutically effective amount of ZN-c3 administered to a subject is about 175 mg twice daily (BID). In another embodiment, the therapeutically effective amount of ZN-c3 administered to a subject is about 100 mg twice daily (BID).

[0122] In various embodiments, the dosing regimen includes a periodic dosing phase during which the dose and frequency of administering ZN-c3 to the subject are fixed for a period of time, such as 3 or more consecutive days. For example, in one embodiment, the periodic dosing phase includes a daily dose in the range of about 200 mg per day to about 350 mg per day, which is fixed for at least three consecutive days. In one embodiment, the daily dose is administered once a day (QD). In another embodiment, the daily dose is administered in divided doses provided twice, three times, or four times a day. In one embodiment, the periodic dosing phase includes administration once a day for at least three consecutive days.

[0123] In various embodiments, the dosing regimen includes an intermittent dosing phase during which the dose and frequency of administering ZN-c3 to the subject are changed. In one embodiment, the intermittent dosing phase includes at least one change in the amount of ZN-c3 administered to the subject daily. In one embodiment, the intermittent dosing phase includes at least one rest day, such as 1, 2, 3, 4, 5, 6 or 7 rest days. For example, in one embodiment, the dosing regimen includes a periodic phase during which ZN-c3 is orally administered to the subject once daily for five days, followed by an intermittent or rest phase during which ZN-c3 is not administered for two days. This dosing regimen can be referred to as 5 days of dosing / 2 days of resting, and can be repeated or cycled as often as needed depending on the specific circumstances.

[0124] In some embodiments, the subject is a human. In some embodiments, the subject has cancer, such as non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer, or uterine serous carcinoma (USC). In some embodiments, the USC is advanced USC. In some embodiments, the subject has recurrent USC.

[0125] In various embodiments, the subject has previously been treated for cancer, such as non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer, or uterine serous carcinoma (USC) by a prior treatment regimen. Various treatment regimens are known to those skilled in the art or are being actively developed. In one embodiment, the prior treatment regimen may include administering a drug, antibody, CAR T cell, surgery, and / or radiation prior to administering ZN-c3. For example, in various embodiments, the prior therapy may include at least one selected from carboplatin, paclitaxel, bevacizumab, trastuzumab, pamoate, lenvatinib, or doxorubicin.

[0126] The amount of a ZN-c3 compound or a pharmaceutically acceptable salt thereof that is effective in treating a particular case of cancer in a subject (e.g., non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, endometrial cancer, or uterine serous carcinoma (USC)) will likely vary not only with the details of the compound or salt selected, but also with the route of administration, the nature and / or symptoms of the disease or condition being treated, and the age and condition of the patient, and will ultimately be determined by the attending physician or clinician. In the case of administration of a pharmaceutically acceptable salt, the dose may be calculated as the free base. As will be appreciated by those skilled in the art, in certain circumstances it may be necessary to administer a ZN-c3 compound, salt, or composition in an amount exceeding or even far exceeding the dosage ranges described herein in order to effectively and aggressively treat particularly aggressive cases of USC.

[0127] The desired dose may conveniently be presented in a single dose or in divided doses administered at appropriate intervals (e.g., in two, three, four or more sub-doses per day). The sub-dose itself may be further divided, for example, into a number of discrete, loosely spaced administrations.

[0128] As will be apparent to those skilled in the art, the useful in vivo dose to be administered and the specific mode of administration will vary depending on the age, weight, severity of the affliction, species of mammal being treated, the specific ZN-c3 compound, salt or composition employed, and the specific use for which these therapies are employed. Determination of effective dose levels (i.e., dose levels necessary to achieve the desired results) can be accomplished by those skilled in the art using conventional methods, e.g., human clinical trials, in vivo studies, and in vitro studies. For example, useful doses of ZN-c3 or pharmaceutically acceptable salts thereof can be determined by comparing their in vitro and in vivo activities in animal models. Such comparisons can be accomplished by comparison with established drugs such as carboplatin and / or paclitaxel.

[0129] The dosage and interval can be adjusted individually to provide a plasma level of the active part sufficient to maintain the regulatory effect or minimum effective concentration (MEC). The MEC of each compound will vary, but can be estimated based on in vivo data and / or in vitro data. The dosage necessary to achieve the MEC will depend on individual characteristics and route of administration. However, HPLC determinations or bioassays can be used to determine the plasma concentration. The MEC value can also be used to determine the dose interval. The composition should be administered using a regimen that maintains the plasma level 10% to 90% higher than the MEC, preferably between 30% and 90%, most preferably between 50% and 90%. In the case of local administration or selective uptake, the effective local concentration of the drug may not be related to the plasma concentration.

[0130] It should be noted that in the event of toxicity or organ dysfunction, the attending physician will know how and when to terminate, interrupt or adjust the administration. Conversely, in the event that the clinical response is not sufficient (excluding toxicity), the attending physician will also know to adjust the treatment to a higher level. The magnitude of the administered dose in the management of the disorder of interest will vary according to the severity of the disease or condition to be treated and the route of administration. For example, the severity of the disease or condition can be assessed in part by standard prognostic assessment methods. In addition, the dosage and possible dosage frequency will also vary according to the age, weight and response of the individual patient. Programs comparable to those discussed above can be used in veterinary medicine.

[0131] The efficacy and toxicity of the ZN-c3 compounds, salts and compositions disclosed herein can be evaluated using known methods. For example, the toxicology of ZN-c3 can be established by determining its in vitro toxicity to cell lines, such as mammalian and preferably human cell lines. The results of such studies are generally predictive of toxicity in animals, such as mammals or more specifically humans. Alternatively, known methods can be used to determine the toxicity of a particular compound in an animal model, such as a mouse, rat, rabbit, dog or monkey. Several recognized methods, such as in vitro methods, animal models or human clinical trials, can be used to establish the efficacy of a particular compound. When selecting a model to determine efficacy, a skilled person can follow the prior art to select an appropriate model, dose, route of administration and / or regimen.

[0132] In some embodiments, the sensitivity of the subject to ZN-c3 is determined by analyzing the presence of one or more PPP2R1A polymorphisms (e.g., any one or more of P179R, S256F, or R183W or genes or transcripts encoding P179R, S256F, or R183W polymorphisms). Such polymorphisms can be identified, for example, by using a whole genome sequencing method (including next generation sequencing (NGS)). In some embodiments, the PPP2R1A polymorphism is identified using a method based on polymerase chain reaction (PCR). In some embodiments, PCR-based methods include allele-specific PCR (ASPCR), high resolution melting analysis (HRM), digital PCR, and low denaturation temperature co-amplification PCR (COLD-PCR) using Taqman probes. In some embodiments, mass spectrometry is used to identify PPP2R1A polymorphisms. In some embodiments, the mass spectrometry used can be matrix-assisted laser desorption / ionization time of flight (MALDI-TOF-MS). In some embodiments, the PPP2R1A polymorphism is identified using a small nucleotide polymorphism (SNP) array. In some embodiments, the PPP2R1A polymorphism is identified using a commercial SNaPshot multiplex kit. In some embodiments, the PPP2R1A polymorphism is identified using denaturing high performance liquid chromatography (DHPLC).

[0133] In some embodiments, primers used during NGS or PCR reactions are read by the target locus or its complement.

[0134] In some embodiments, the PPP2R1A mutation is a hotspot mutation. In some embodiments, the genomic locus of each codon where the hotspot is located is: S256: hg19: chr19: 52,716,321-52,716,323; R183: hg19: chr19: 52,715,981-52,715,983; and / or P179: hg19: chr19: 52,715,969-52,715,971. In some embodiments, these hotspot mutations are present in uterine cancer, ovarian cancer, breast cancer, lung cancer, colorectal cancer, prostate cancer, salivary gland cancer, or other cancers.

[0135] In some embodiments, the regulation in PPP2R1A is detected, such as the above-mentioned polymorphism. In some embodiments, the regulation includes one or more gain of function mutations in PPP2R1A. In some embodiments, the regulation includes one or more loss of function mutations in PPP2R1A. In some embodiments, the regulation includes one or more mutations that cause PPP2R1A to be overexpressed. In some embodiments, the regulation includes one or more mutations that cause PPP2R1A to be underexpressed.

[0136] Example 1 Dose response of cancer cell lines sensitized to ZN-c3 using PPP2R1A siRNA

[0137] The effect of PPP2R1A knockdown on affecting cancer cell sensitivity to ZN-c3 was evaluated. Various cancer cell lines were transfected with PPP2R1A siRNA or control scrambled siRNA and cells were treated with a 10-point dose response of ZN-c3 for 72 hours. Cell titer luminescence analysis was used to calculate the percentage of cell density. PPP2R1A siRNA targets the following bases Chr.19:52189802–52226425. siRNA transfection was initiated 48 hours prior to treatment with ZN-c3. ZN-c3 dose response evaluation included ZN-c3 doses of 10uM, 3.33uM, 1.11uM, 370.3nM, 123.4nM, 41.1nM, 13.7nM, 4.5nM, and 1.5nM.

[0138] The first non-small cell lung cancer cell line A427 was treated with PPP2R1A siRNA or control siRNA, and the sensitivity of the transfected cells to ZN-c3 was assessed by measuring the percentage decrease in cell density ( Figure 1 PPP2R1A siRNA-mediated knockdown of PPP2R1A was evaluated by comparing PPP2R1A expression under conditions treated with siCTRL and siPPPR12A ( Figure 2Knockdown of PPP2R1A by siRNA was successful, and A427 cells transduced with siPPP2R1A demonstrated a 3.5-fold increase in sensitivity to ZN-c3 (Table 1).

[0139] Table 1

[0140] siCTRL siPPP2R1A EC50 0.1957 0.05544

[0141] A second non-small cell lung cancer cell line, H1975, was treated with PPP2R1A siRNA or control siRNA, and the sensitivity of the transfected cells to ZN-c3 was assessed by measuring the percent decrease in cell density ( Figure 3 PPP2R1A siRNA-mediated knockdown of PPP2R1A was evaluated by comparing PPP2R1A expression under conditions treated with siCTRL and siPPPR12A ( Figure 4 Knockdown of PPP2R1A by siRNA was successful, and H1975 cells transduced with siPPP2R1A demonstrated a 3.7-fold increase in sensitivity to ZN-c3 (Table 2).

[0142] Table 2

[0143] siCTRL siPPP2R1A EC50 0.5595 0.1500

[0144] The breast cancer cell line Hs578T was treated with PPP2R1A siRNA or control siRNA, and the sensitivity of the transfected cells to ZN-c3 was assessed by measuring the percentage decrease in cell density ( Figure 5 PPP2R1A siRNA-mediated knockdown of PPP2R1A was evaluated by comparing PPP2R1A expression under conditions treated with siCTRL and siPPPR12A ( Figure 6 Knockdown of PPP2R1A by siRNA was successful, and Hs578T cells transduced with siPPP2R1A demonstrated a 2.4-fold increase in sensitivity to ZN-c3 (Table 3).

[0145] Table 3

[0146] siCTRL siPPP2R1A EC50 0.1699 0.07133

[0147] The colorectal cancer cell line SW620 was treated with PPP2R1A siRNA or control siRNA, and the sensitivity of the transfected cells to ZN-c3 was assessed by measuring the decrease in the percentage of cell density ( Figure 7PPP2R1A siRNA-mediated knockdown of PPP2R1A was evaluated by comparing PPP2R1A expression under conditions treated with siCTRL and siPPPR12A ( Figure 8 Knockdown of PPP2R1A by siRNA was successful, and SW620 cells transduced with siPPP2R1A demonstrated a 1.9-fold increase in sensitivity to ZN-c3 (Table 4).

[0148] Table 4

[0149] siCTRL siPPP2R1A EC50 0.4499 0.2364

[0150] The ovarian cell line SKOV3 was treated with PPP2R1A siRNA or control siRNA, and the sensitivity of the transfected cells to ZN-c3 was assessed by measuring the percent decrease in cell density ( Fig. 9 PPP2R1A siRNA-mediated knockdown of PPP2R1A was evaluated by comparing PPP2R1A expression under conditions treated with siCTRL and siPPPR12A ( Fig.10 Knockdown of PPP2R1A by siRNA was successful, and SKOV3 cells transduced with siPPP2R1A demonstrated a 2.7-fold increase in sensitivity to ZN-c3 (Table 5).

[0151] Table 5

[0152] siCTRL siPPP2R1A EC50 0.4804 0.1797

[0153] Endometrial cancer cell lines HEC1B and HEC-59 were treated with PPP2R1A siRNA or control siRNA, and the sensitivity of the transfected cells to ZN-c3 was assessed by measuring the percent decrease in cell density ( Fig.11 PPP2R1A siRNA-mediated knockdown of PPP2R1A was evaluated by comparing PPP2R1A protein expression under conditions treated with siCTRL and siPPPR12A by western blotting, using focal adhesion protein as a control ( Fig.12). Due to the missense mutations W257L and L429F in PPP2R1A in this cell line, knocking down PPP2R1A by siRNA is limited to HEC1B cells. However, HEC-59 cells have wild-type PPP2R1A, and knocking down PPP2R1A by siRNA in this cell line is successful. HEC1B cells transfected with PPP2R1A siRNA demonstrated a 1.4-fold increase in sensitivity to ZN-c3 (Table 6). HEC-59 cells transfected with PPP2R1A demonstrated a 2.5-fold increase in sensitivity to ZN-c3 (Table 6). Therefore, these results show how knocking down PPP2R1A plays a key role in sensitizing cancer cells to ZN-c3-mediated growth inhibition.

[0154] Table 6

[0155] EC50 siCTRL siPPP2R1A HEC1B 0.7072 0.4900 HEC-59 1.038 0.4357

[0156] Example 2 Dose response of cancer cell lines sensitized to ZN-c3 using CRISPR-mediated PPP2R1A knockout

[0157] After observing that siRNA-mediated knockdown of PPP2R1A sensitized cancer cell lines to ZN-c3 treatment, it was also interesting to evaluate whether CRISPR-mediated knockout of PPP2R1A produced similar effects. Various cancer cell lines were transfected with CRISPR guide RNA or non-targeted guide RNA targeting PPP2R1A, and cells were treated with ZN-c3 for 72 hours with a 10-point dose response. The 10-point dose response to ZN-c3 included the same dose as used in Example 1. As in Example 1, no medium replacement was performed during the 72-hour ZN-c3 dose response evaluation period. Less than 20 cell passages were completed between CRISPR-mediated knockout and ZN-c3 treatment. CRISPR was performed by targeting PPP2R1A with a PPP2R1A sgRNA sequence. Cell titer luminescence analysis was used to calculate the cell density percentage.

[0158] The ovarian cancer cell line SKOV3 was treated with CRISPR guide RNA targeting PPP2R1A or non-targeting guide RNA, and the sensitivity of the transfected cells to ZN-c3 was assessed by measuring the percentage reduction in cell density ( Fig.13 CRISPR-mediated knockdown of PPP2R1A was evaluated by comparing PPP2R1A protein expression under PPPR12A treatment conditions with non-targeting guide RNA and CRISPR-targeting guide RNA via Western blotting, with β-actin as a control ( Fig.14Knockout of PPP2R1A by CRISPR was successful, and SKOV3 cells transduced with guide RNA targeting PPP2R1A demonstrated a 1.7-fold increase in sensitivity to ZN-c3 (Table 7).

[0159] Table 7

[0160] sqN sgPPP2R1A EC50 0.3856 0.2239

[0161] The lung cancer cell line H1975 was treated with CRISPR guide RNA targeting PPP2R1A or non-targeting guide RNA, and the sensitivity of the transfected cells to ZN-c3 was assessed by measuring the percentage reduction in cell density ( Fig.15 CRISPR-mediated knockout of PPP2R1A was evaluated by comparing PPP2R1A expression under conditions treated with non-targeting guide RNA and CRISPR guide RNA targeting PPPR12A ( Fig.16 Knockout of PPP2R1A by CRISPR was successful, and H1975 cells transduced with guide RNA targeting PPP2R1A demonstrated a 1.9-fold increase in sensitivity to ZN-c3 (Table 8).

[0162] Table 8

[0163] sqN sgPPP2R1A EC50 0.5155 0.2752

[0164] The sensitivity of the ovarian cancer cell line OV17R to ZN-c3 was evaluated by measuring the decrease in the percentage of cell density ( Fig.17 OV17R cells have a S256F hotspot missense mutation in PPP2R1A, thus rendering these cells highly sensitive to ZN-c3 without the need for an additional mechanism for PPP2R1A knockdown or knockout (Table 9).

[0165] Table 9

[0166] OV17R EC50 0.1391

[0167] Example 3 Response of cancer cell lines to ZN-c3, LB-100 or ZN-c3+LB-100 treatment

[0168] After observing that both knockdown and knockout of PPP2R1A can sensitize cancer cells to Zn-C3, the effect of pharmacological inhibition of PPP2R1A by catalytic inhibitor LB-100 was evaluated. Various cancer cell lines were treated with ZN-c3 alone, PP2A catalytic inhibitor LB-100 alone, or a combination of ZN-c3+LB-100. As in Examples 1 and 2, the treatment period was 72 hours, and the percent inhibition was calculated using cell titer luminescence analysis.

[0169] The sensitivity of the endometrial cancer cell line HEC-59 to ZN-c3 alone, LB-100 alone, or the combination of ZN-c3 + LB-100 was evaluated ( Fig.18 ). The dosages of ZN-c3 and LB-100 are shown in Table 10. ZN-c3 treatment alone was able to achieve about 25% inhibition, while LB-100 treatment achieved about half of this value. However, combination therapy utilizing both ZN-c3 and LB-100 was significantly more effective and achieved almost 75% inhibition.

[0170] Table 10

[0171] Compound dose ZN-c3 400nM LB-100 1μM

[0172] The sensitivity of the ovarian cancer cell line OV17R to ZN-c3 alone, LB-100 alone, or the combination of ZN-c3+LB-100 was evaluated ( Fig.19 ). The dosages of ZN-c3 and LB-100 are shown in Table 11. ZN-c3 treatment alone was able to achieve about 30% inhibition, while LB-100 treatment achieved about 12% inhibition. Combination therapy with both ZN-c3 and LB-100 demonstrated a modest increase to about 40% inhibition compared to ZN-c3 alone. The small increase in percent inhibition observed in the combination therapy may be a result of the PPP2R1A missense mutation S256F present in OV17R, which is not present in other cancer lines tested with wild-type PPP2R1A.

[0173] Table 11

[0174] Compound dose ZN-c3 75nM LB-100 1μM

[0175] The sensitivity of the endometrial cancer cell line HEC-59 to ZN-c3 alone, LB-100 alone, or the combination of ZN-c3+LB-100 was evaluated after treatment with siCTRL or siPPP2R1A ( Fig. 20). The dosages of ZN-c3 and LB-100 are shown in Table 12. In cells treated with siCTRL, ZN-c3 treatment alone was able to achieve about 12% inhibition, while LB-100 treatment achieved about half of this value. However, in cells treated with siCTRL, the combination therapy using both ZN-c3 and LB-100 was significantly more effective and exceeded 25% inhibition. It is noteworthy that, in cells treated with siPPP2R1A, the percentage of inhibition achieved by ZN-c3 treatment alone and by the combination of ZN-c3 and LB-100 was approximately doubled compared to cells treated with siCTRL. In cells with PPP PPP2R1A knockdown, the percentage of inhibition of LB-100 alone was only slightly enhanced. These data demonstrate that, in the context of reduced PPP2R1A expression, the pairing of pharmacological inhibition (LB-100) with Zn-C3 treatment provides a strong combination approach for cancer treatment.

[0176] Table 12

[0177] Compound dose ZN-c3 300nM LB-100 1μM

[0178] The sensitivity of the endometrial cancer cell line HEC1B to ZN-c3 alone, LB-100 alone, or the combination of ZN-c3+LB-100 was also evaluated after treatment with siCTRL or siPPP2R1A ( Fig.21 ). The doses of ZN-c3 and LB-100 are shown in Table 13. In cells treated with siCTRL, ZN-c3 treatment alone was able to achieve approximately 30% inhibition, while LB-100 treatment achieved 25% inhibition. In siCTRL-treated cells, combination therapy with both ZN-c3 and LB-100 was more effective than either treatment alone and approached 50% inhibition. ZN-c3 treatment alone and the combination of ZN-c3 and LB-100 achieved approximately the same percentage of inhibition regardless of whether the cells had been treated with siPPP2R1A or siCTRL. This result is similar to the evaluation of HEC1B in the previous example, where missense mutations W257L and L429F within PPP2R1A in this cell line limited the induction of sensitivity to ZN-c3.

[0179] Table 13

[0180] Compound dose ZN-c3 200nM LB-100 1μM

[0181] Example 4 Clinical Trial of ZN-c3 in Patients with PPP2R1A Genetic Variants .

[0182] Having observed that genetic ablation and pharmacological inhibition of PPP2R1A sensitize cancer cell lines to ZN-c3, the next goal is to study the clinical response to ZN-c3 in human patients with hotspot mutations in PPP2R1A. As part of the clinical study, five USC patients with known previous genomic profiling information of PPP2R1A were evaluated. Three patients had the P179R variant of PPP2R1A, and two patients had the S256F variant (Table 14). The clinical response of all patients can be evaluated by at least one tumor assessment using computed tomography (CT) and / or magnetic resonance imaging (MRI) imaging. Four of the five patients (80%) showed clinical benefit from ZN-c3. The target response rate for patients with PPP2R1A mutations was 20%.

[0183] For Subject 1, a starting dose of 300 mg QD ZN-c3 was administered, the dose was reduced to 5 days on, 2 days off on Day 44, the dose was further reduced to 200 mg QD on Day 51, and the final dose was given on Day 230.

[0184] For Subject 2, a starting dose of 300 mg QD ZN-c3 was administered, the dose was reduced to 200 mg QD on Day 36, the dose was further reduced to 200 mg on 5 days and 2 days off on Day 50, and the final dose was given on Day 84.

[0185] For Subject 3, a starting dose of 300 mg QD ZN-c3 was administered, the dose was reduced to 200 mg QD on Day 71, and the final dose was given on Day 172.

[0186] For Subject 4, a starting dose of 300 mg QD ZN-c3 was administered with no dose reductions during the study, and the final dose was given on Day 64.

[0187] For Subject 5, a starting dose of 300 mg QD ZN-c3 was administered, the dose was reduced to 150 mg BID on Day 36, and the final dose was given on Day 62.

[0188] Table 14

[0189] Subject number PPP2R1A variants Best Overall Response 1 S256F cPR (-43%) 2 P179R SD (-26%) 3 P179R SD (-12%) 4 P179R SD (-7%) 5 S256F PD

[0190] cPR = confirmed partial response. SD = stable disease. PD = progressive disease

[0191] In addition, although the foregoing has been described in some detail by way of illustration and example for the purpose of clarity and understanding, it will be appreciated by those skilled in the art that many and various modifications may be made without departing from the essence of the present disclosure. Therefore, it should be clearly understood that the forms disclosed herein are merely illustrative and are not intended to limit the scope of the present disclosure, but also encompass all modifications and alternatives that are consistent with the true scope and essence of the present disclosure.

Claims

1. A method for identifying or selecting a subject who is sensitive to the drug ZN-c3, the method comprising: identifying, in a biological sample obtained from said subject or individual, a regulation in a protein phosphatase 2 scaffold subunit alpha (PPP2R1A) protein or a gene encoding said PPP2R1A protein, such as one or more polymorphisms conferring sensitivity to said drug ZN-c3, preferably a polymorphism selected from P179R, S256F or R183W or a nucleic acid encoding a polymorphism selected from P179R, S256F or R183W; and When the regulation is identified, such as any one or more of the polymorphisms in the PPP2R1A gene or protein, the subject is selected or identified as a subject having sensitivity to the drug ZN-c3.

2. The method of claim 1, further comprising administering ZN-c3 to the subject when the modulation or polymorphism is identified in the biological sample.

3. The method according to any one of claims 1 or 2, wherein the subject has cancer.

4. The method of claim 3, wherein the cancer comprises non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, uterine serous carcinoma (USC), or endometrial cancer.

5. The method of any one of claims 2 to 4, wherein the regression or inhibition of the cancer is greater than 7%, such as 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more.

6. The method according to any one of claims 2 to 5, further comprising administering to the subject an agent or combination of agents that inhibits the PPP2R1A gene or protein.

7. The method of claim 6, wherein the agent or combination of agents comprises the PP2A catalytic inhibitor LB-100.

8. The method according to any one of claims 1 to 7, wherein the modulation of PPP2R1A comprises one or more gain of function mutations.

9. The method of any one of claims 1 to 7, wherein the modulation of PPP2R1A comprises one or more loss of function mutations.

10. The method according to any one of claims 1 to 7, wherein the modulation of PPP2R1A comprises one or more mutations leading to overexpression of PPP2R1A.

11. The method according to any one of claims 1 to 7, wherein the modulation of PPP2R1A comprises one or more mutations leading to under-expression of PPP2R1A.

12. A method for inhibiting, improving or treating cancer or its sequelae in a subject, the method comprising: identifying, in a biological sample obtained from the subject, regulation in a protein phosphatase 2 scaffold subunit alpha (PPP2R1A) protein or a gene encoding the PPP2R1A protein, wherein the polymorphism confers sensitivity to drug ZN-c3, such as a polymorphism selected from P179R, S256F or R183W or a nucleic acid encoding a polymorphism selected from P179R, S256F or R183W; and When the polymorphism is identified in the biological sample, ZN-c3 is administered to the subject.

13. The method of claim 12, wherein the cancer comprises non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, uterine serous carcinoma (USC), or endometrial cancer.

14. The method of claim 12 or 13, wherein the regression or inhibition of the cancer is greater than 7%, such as 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more.

15. The method according to any one of claims 12 to 14, wherein the subject is additionally provided with an agent or combination of agents that inhibits the PPP2R1A gene or protein.

16. The method of claim 15, wherein the agent or combination of agents comprises the PP2A catalytic inhibitor LB-100.

17. The method of any one of claims 12 to 16, wherein the modulation of PPP2R1A comprises one or more gain of function mutations.

18. The method of any one of claims 12 to 16, wherein the modulation of PPP2R1A comprises one or more loss of function mutations.

19. The method of any one of claims 12 to 16, wherein the modulation of PPP2R1A comprises one or more mutations leading to overexpression of PPP2R1A.

20. The method of any one of claims 12 to 16, wherein the modulation of PPP2R1A comprises one or more mutations resulting in under-expression of PPP2R1A.

21. A compound ZN-c3 for use in inhibiting, ameliorating or treating cancer or its sequelae in a subject identified as having a regulation in the protein phosphatase 2 scaffold subunit alpha (PPP2R1A) protein or a gene encoding the PPP2R1A protein in a biological sample obtained from the subject, such as a polymorphism that confers sensitivity to the drug ZN-c3, for example a polymorphism selected from P179R, S256F or R183W or a nucleic acid encoding a polymorphism selected from P179R, S256F or R183W.

22. The compound ZN-c3 for use according to claim 21, wherein the cancer comprises non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, uterine serous carcinoma (USC) or endometrial cancer.

23. The compound ZN-c3 for use according to claim 21, wherein the regression or inhibition of the cancer is greater than 7%, such as 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more.

24. Compound ZN-c3 for use according to any one of claims 21 to 23, wherein the subject is additionally provided with an agent or a combination of agents that inhibits the PPP2R1A gene or protein.

25. Compound ZN-c3 for use according to claim 24, wherein the agent or combination of agents comprises the PP2A catalytic inhibitor LB-100.

26. Compound ZN-c3 for use according to any one of claims 21 to 25, wherein the modulation of PPP2R1A comprises one or more gain of function mutations.

27. Compound ZN-c3 for use according to any one of claims 21 to 25, wherein the modulation of PPP2R1A comprises one or more loss of function mutations.

28. Compound ZN-c3 for use according to any one of claims 21 to 25, wherein the modulation of PPP2R1A comprises one or more mutations leading to overexpression of PPP2R1A.

29. Compound ZN-c3 for use according to any one of claims 21 to 25, wherein the modulation of PPP2R1A comprises one or more mutations leading to under-expression of PPP2R1A.

30. A method for inhibiting, ameliorating or treating cancer or its sequelae in a subject, the method comprising: administering to the subject an agent or a combination of agents that inhibits the PPP2R1A gene or protein; as well as ZN-c3 is administered to the subject.

31. The method of claim 30, wherein the agent or a combination of agents that inhibit the PPP2R1A gene or protein is provided to the subject alone.

32. The method of claim 30 or 31, wherein the agent or combination of agents that inhibits the PPP2R1A gene or protein is provided to the subject prior to administration of ZN-C3.

33. The method of claim 30, wherein the agent or combination of agents that inhibit the PPP2R1A gene or protein is provided to the subject within an interval of 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 11 minutes, 12 minutes, 13 minutes, 14 minutes, 15 minutes, 16 minutes, 17 minutes, 18 minutes, 19 minutes or 20 minutes.

34. The method of any one of claims 30 to 33, wherein the cancer comprises non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, uterine serous carcinoma (USC), or endometrial cancer.

35. The method of any one of claims 30 to 34, wherein the regression or inhibition of the cancer is greater than 7%, such as 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more.

36. The method of any one of claims 30 to 35, wherein the agent or combination of agents comprises the PP2A catalytic inhibitor LB-100.

37. A product combination for use in inhibiting, ameliorating or treating cancer or its sequelae in a subject, the product combination comprising compound ZN-c3 and an agent or a combination of agents that inhibits PPP2R1A gene or protein.

38. A product combination for use according to claim 37, wherein the cancer comprises non-small cell lung cancer, breast cancer, colorectal cancer, ovarian cancer, uterine serous carcinoma (USC) or endometrial cancer.

39. A product combination for use according to any one of claims 37 or 38, wherein the regression or inhibition of the cancer is greater than 7%, such as 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50% or more.

40. A product combination for use according to any one of claims 37 to 39, wherein the agent or combination of agents comprises the PP2A catalytic inhibitor LB-100.

41. The method according to any one of claims 1 to 20 or compound ZN-c3 for use according to any one of claims 21 to 29, wherein the presence of the polymorphism is determined using next generation sequencing (NGS), sequencing, polymerase chain reaction (PCR), loop-mediated isothermal amplification, recombinase polymerase amplification or antibody detection.

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