Combining PLK1 inhibitors with antiangiogenic agents to treat metastatic cancer

Abstract

Provided are methods, compositions, and kits for treating metastatic cancer in a subject, which can include administering to an antiangiogenic treatment-naive subject a treatment that includes inhibiting angiogenesis and a PLK1 inhibitor (e.g., onvansertib) in a manner sufficient to reduce or inhibit the progression of metastatic cancer.

Description

[Technical Field]

[0001] Related Applications This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Patent Application No. 63 / 405,466, filed September 11, 2022, and U.S. Provisional Patent Application No. 63 / 515,831, filed July 26, 2023, the contents of which are incorporated by reference herein in their entirety for all purposes.

[0002] The present disclosure relates generally to the field of cancer treatment. [Background technology]

[0003] Polo-like kinase 1 (PLK1) is a serine / threonine kinase and the best-characterized member of a family of five closely related regulatory proteins. PLK1 is a master regulator of mitosis, controlling cell entry and progression into mitosis. PLK1 performs several essential functions throughout the mitotic (M) phase of the cell cycle, including regulating centrosome maturation and spindle assembly, removing cohesin from chromosome arms, inactivating anaphase-promoting complex / cyclosome (APC / C) inhibitors, and regulating mitotic exit and cytokinesis. PLK1 plays a critical role in centrosome function and bipolar spindle assembly. PLK1 regulates the interaction of kinetochores with spindle microtubules, which is required for successful separation and segregation of chromatids into the appropriate mother and daughter cells. PLK1 also functions as a negative regulator of p53 family members, resulting in the ubiquitination and subsequent degradation of p53 / TP53, inhibition of p73 / TP73-mediated apoptosis-inducing functions, and phosphorylation / degradation of bora, a cofactor of Aurora kinase A. PLK1 localizes to centrosomes, kinetochores, and the central spindle during various stages of mitosis. PLK1 is aberrantly overexpressed in various human cancers and correlates with cell proliferation and poor prognosis. Summary of the Invention

[0004] The most advanced stage of cancer is stage IV, defined by the cancer's spread from where it first began to distant parts of the body (e.g., metastasis). Angiogenesis is required for metastasis to occur, and several antiangiogenic agents have been approved for human use to treat metastatic cancer. However, according to the Centers for Disease Control and Prevention, cancer still accounts for approximately one in five deaths in the United States. More effective treatments for advanced (e.g., metastatic) cancer are needed.

[0005] Disclosed herein are methods of treating cancer, including metastatic cancer. The methods for treating cancer, in some embodiments, comprise administering a PLK1 inhibitor and an anti-angiogenic agent to a subject with metastatic cancer, thereby reducing or inhibiting the progression of the metastatic cancer, wherein the subject has not previously been treated with angiogenesis inhibitors. The methods for treating cancer, in some embodiments, can comprise administering a PLK1 inhibitor and an anti-angiogenic agent to a subject, thereby reducing or inhibiting the progression of the metastatic cancer, wherein the subject has not previously been treated with angiogenesis inhibitors, or wherein the subject has not previously been treated with angiogenesis inhibitors. The methods for treating cancer, in some embodiments, can comprise administering a PLK1 inhibitor and an anti-angiogenic agent to a subject, thereby reducing or inhibiting the progression of the metastatic cancer, wherein the subject is known to have not previously been treated with angiogenesis inhibitors, or wherein the subject is known to have not previously been treated with angiogenesis inhibitors. A method for treating cancer, in some embodiments, can include identifying a subject with metastatic cancer and who has not received prior cancer treatment, and administering to the subject a PLK1 inhibitor and an anti-angiogenic agent, thereby reducing or inhibiting the progression of the metastatic cancer. The method, in some embodiments, includes identifying a subject with metastatic cancer and who has not received prior treatment, including inhibition of angiogenesis, and administering to the subject a PLK1 inhibitor and an anti-angiogenic agent, thereby reducing or inhibiting the progression of the metastatic cancer.

[0006] In some embodiments, the method includes administering to the subject chemotherapy, a PLK1 inhibitor, and an anti-angiogenic agent. In some embodiments, the subject has not previously received chemotherapy. In some embodiments, the subject has not previously received chemotherapy for the metastatic cancer. The chemotherapy can include treatment with FOLFIRI, abiraterone, FOLFOX, an anti-EGFR agent, a KRAS-directed inhibitor, gemcitabine, Abraxane, nanoliposomal irinotecan, 5-FU, FOLFIRINOX, FOLFOXIRI, or a combination thereof. In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent synergistically reduces or inhibits metastatic cancer progression compared to PLK1 inhibitor treatment alone, anti-angiogenic agent treatment alone, and / or the additive effects of PLK1 inhibitor treatment alone and anti-angiogenic agent treatment alone. In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent improves one or more therapeutic effects in a subject compared to a control or baseline. In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent reduces the oncogenic allele burden in a subject compared to a subject that has previously undergone treatment that includes inhibiting angiogenesis.

[0007] Also disclosed herein are methods of improving objective response rate (ORR), progression-free survival (PFS), or both in the treatment of metastatic cancer. The methods, in some embodiments, comprise administering a PLK1 inhibitor and an anti-angiogenic agent to a subject with metastatic cancer, thereby improving the ORR and / or PFS in the subject, where the subject has not previously been treated for cancer or where the subject has not previously been treated with angiogenesis inhibition. The methods, in some embodiments, comprise administering a PLK1 inhibitor and an anti-angiogenic agent to a subject with metastatic cancer, thereby improving the ORR and / or PFS in the subject, where the subject is known to have not previously been treated for cancer or where the subject is known to have not previously been treated with angiogenesis inhibition. In some embodiments, the methods comprise identifying a subject with metastatic cancer who has not previously been treated for cancer. In some embodiments, the methods comprise identifying a subject with metastatic cancer who has not previously been treated for cancer. In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent synergistically improves the subject's ORR and / or PFS compared to PLK1 inhibitor treatment alone, anti-angiogenic agent treatment alone, and / or the additive effects of PLK1 inhibitor treatment alone and anti-angiogenic agent treatment alone. In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent improves one or more therapeutic effects in the subject compared to a control or baseline, optionally including tumor size from metastatic cancer, objective response rate (ORR), duration of response, time to response, progression-free survival (PFS), overall survival (OS), disease control rate (DCR), oncogenic allele burden, or a combination thereof. In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent improves the subject's ORR, improves the subject's PFS, improves the subject's OS, improves the subject's DCR, reduces the subject's oncogenic allele burden, or a combination thereof, compared to a subject who has received a previous treatment that includes inhibition of angiogenesis. In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent improves the subject's ORR, PFS, or both by at least 50% compared to subjects who have received prior treatment that includes inhibition of angiogenesis.In some embodiments, administering a PLK1 inhibitor and an anti-angiogenic agent to a subject results in a difference in ORR in the subject of 50%, 55%, 65%, 70%, 75%, or more, or a value or range between any two of these values. In some embodiments, administering a PLK1 inhibitor and an anti-angiogenic agent to a subject results in an mPFS in the subject of 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, or more, or a value or range between any two of these values. As described herein, administering a PLK1 inhibitor and an anti-angiogenic agent to a subject can include administering to the subject a PLK1 inhibitor, an anti-angiogenic agent, and one or more anti-cancer therapeutics or therapies (e.g., one or more chemotherapeutics).

[0008] Non-limiting examples of metastatic cancers include metastatic colorectal cancer, metastatic bladder cancer, metastatic breast cancer, metastatic kidney cancer, metastatic lung cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic prostate cancer, metastatic gastric cancer, metastatic thyroid cancer, metastatic uterine cancer, metastatic kidney cancer, metastatic cervical cancer, metastatic recurrent glioblastoma, or a combination thereof. In some embodiments, the PLK1 inhibitor is selective and / or specific for PLK1. In some embodiments, the PLK1 inhibitor is onvansertib, BI2536, volasertib (BI 6727), GSK461364, AZD1775, CYC140, HMN-176, HMN-214, rigosertib (ON-01910), MLN0905, TKM-080301, TAK-960, or Ro3280. In some embodiments, the subject has not previously been treated with an angiogenesis inhibitor, and optionally, the angiogenesis inhibitor is the same as the antiangiogenesis agent. In some embodiments, the antiangiogenesis agent is bevacizumab. In some embodiments, the angiogenesis inhibitor is bevacizumab.

[0009] The PLK1 inhibitor and the anti-angiogenic agent can be administered simultaneously or sequentially. In some embodiments, the PLK1 inhibitor is administered prior to the administration of the anti-angiogenic agent, optionally prior to the administration of the anti-angiogenic agent on all days that the subject is administered the PLK1 inhibitor and the anti-angiogenic agent. In some embodiments, the PLK1 inhibitor is administered about 30 minutes to about 5 hours prior to the administration of the anti-angiogenic agent on a given day. In some embodiments, the PLK1 inhibitor is administered orally and the anti-angiogenic agent is administered intravenously or orally. In some embodiments, the anti-angiogenic agent and the PLK1 inhibitor are each administered to the subject at least twice or at least five times per week in one cycle. In some embodiments, the anti-angiogenic agent, the PLK1 inhibitor, or both are administered in cycles of at least 7 days, optionally with each treatment cycle being at least about 21 days, and further optionally with each treatment cycle being about 21 days to about 28 days.

[0010] In some embodiments, the PLK1 inhibitor is administered for at least 4 days of the cycle. In some embodiments, the PLK1 inhibitor is not administered for at least 1 day of the cycle. In some embodiments, the antiangiogenic agent is administered daily, weekly, every other week, every 3 weeks, every 4 weeks, or monthly. In some embodiments, the subject receives at least 2 cycles of antiangiogenic agent and PLK1 inhibitor administration. In some embodiments, the antiangiogenic agent is bevacizumab and the PLK1 inhibitor is onvansertib. Onvansertib is administered at a dose of, for example, 12 mg / m 2 ~90mg / m 2 Bevacizumab can be administered, for example, at about 1 mg / kg to 20 mg / kg, and optionally, bevacizumab is administered at about 5 mg / kg, about 7.5 mg / kg, about 10 mg / kg, or about 15 mg / kg. In some embodiments, the subject has had at least one prior cancer treatment, optionally without the use of an anti-angiogenic agent, a PLK1 inhibitor, or both. In some embodiments, one or more subjects have had a prior cancer remission. The prior remission can be a complete remission (CR) or a partial remission (PR).

[0011] In some embodiments, the method further includes one or more of: (1) determining the cancer status of the one or more subjects, (2) determining the responsiveness of the one or more subjects to PLK1 inhibitor treatment, and (3) administering one or more cancer therapeutic agents or therapies to the one or more subjects. The one or more subjects may be human. In some embodiments, reducing or inhibiting the progression of cancer comprises inhibiting the growth of one or more tumors in one or more subjects and / or reducing the number of cancer cells detected in one or more subjects by at least about 25%, 30%, 40%, 50%, 60%, or 70% compared to untreated subjects. In some embodiments, reducing or inhibiting the progression of cancer comprises inhibiting the growth of one or more tumors in one or more subjects and / or reducing the number of cancer cells detected in one or more subjects by at least about 25%, 30%, 40%, 50%, 60%, or 70% compared to the one or more subjects prior to administration of the PLK1 inhibitor and the anti-angiogenic agent. In some embodiments, the growth of at least one of the one or more tumors in one or more subjects is reduced by at least about 25%, 30%, 40%, 50%, 60%, or 70% after one or more cycles of treatment. In some embodiments, the size / volume of at least one of one or more tumors in one or more subjects is reduced by at least about 25%, 30%, 40%, 50%, 60%, or 70% after one or more cycles of treatment.

[0012] In some embodiments, the one or more cancer therapeutic agents or therapies include FOLFIRI, abiraterone, FOLFOX, an anti-EGFR agent, a KRAS-directed inhibitor, gemcitabine, Abraxane, nanoliposomal irinotecan, 5-FU, or a combination thereof, where the anti-EGFR agent is optionally cetuximab, and the KRAS-directed inhibitor is optionally a G12C inhibitor, a G12D inhibitor, or a combination thereof. In some embodiments, determining the responsiveness of one or more subjects comprises determining whether the subject is a responder to treatment, whether one or more subjects is in or heading for complete recovery (CR), or whether one or more subjects is in or heading for partial remission (PR). In some embodiments, determining the responsiveness of a subject comprises determining the subject's objective response rate (ORR), duration of response, time to response, progression-free survival (PFS), overall survival (OS), disease control rate (DCR), oncogenic allele burden, or a combination thereof. In some embodiments, determining the responsiveness of one or more subjects comprises determining whether one or more subjects have a partial response to treatment, whether the subjects have a complete response to treatment, whether the subjects have stable disease (SD), or whether the subjects have progressive disease (PD).

[0013] Also disclosed herein are kits, which in some embodiments include a PLK1 inhibitor and a manual providing instructions for administering the PLK1 inhibitor together with an anti-angiogenic agent to a subject with metastatic cancer, where the subject has not previously been treated for cancer, or the subject has not previously been treated for angiogenesis inhibition. In some embodiments, the kit includes a PLK1 inhibitor and a manual providing instructions for administering the PLK1 inhibitor together with an anti-angiogenic agent to a subject with metastatic cancer, where the subject is known to have not previously been treated for cancer, or the subject is known to have not previously been treated for angiogenesis inhibition.

[0014] In some embodiments, the instructions include instructions regarding co-administration of the PLK1 inhibitor and the anti-angiogenic agent. In some embodiments, the instructions include instructions regarding sequential administration of the PLK1 inhibitor and the anti-angiogenic agent. In some embodiments, the instructions include (1) instructions regarding oral administration of the PLK1 inhibitor, (2) instructions regarding oral administration of the anti-angiogenic agent, (3) instructions regarding intravenous administration of the anti-angiogenic agent, or any combination thereof. In some embodiments, the instructions include a statement that the subject has no prior treatment history that includes administration of an angiogenesis inhibitor, and optionally, the angiogenesis inhibitor is the same as the anti-angiogenic agent. In some embodiments, the instructions include a statement regarding administration of each of the anti-angiogenic agent and the PLK1 inhibitor to the subject at least twice or at least five times per week in one cycle. In some embodiments, the instructions include administering the antiangiogenic agent, the PLK1 inhibitor, or both in cycles of at least 7 days, optionally with each treatment cycle being at least about 21 days, and further optionally with each treatment cycle being about 21 to about 28 days. In some embodiments, the instructions include administering the PLK1 inhibitor for at least 4 days of the cycle. In some embodiments, the instructions include not administering the PLK1 inhibitor for at least 1 day of the cycle. In some embodiments, the instructions include administering the antiangiogenic agent daily, weekly, biweekly, every three weeks, every four weeks, or monthly. In some embodiments, the instructions include administering at least two cycles of the antiangiogenic agent and the PLK1 inhibitor. The antiangiogenic agent can be bevacizumab. The PLK1 inhibitor can be onvansertib. In some embodiments, the instructions include administering onvansertib at 12 mg / m 2 ~90mg / m 2 In some embodiments, the kit further comprises an anti-angiogenic agent. [Brief explanation of the drawings]

[0015] [Figure 1]FIG. 1 shows exemplary Kaplan-Meier survival curves for cancer patients treated with the methods and compositions provided herein, with and without prior anti-angiogenic therapy. [Figure 2] Figure 2 shows the number of events and the number of censored (non-reporting) patients for the studies described herein. [Figure 3] Figure 3 shows the objective response rates for cohorts of patients treated using the methods and compositions provided herein, separated into patients with and without prior antiangiogenic therapy. The boxed numbers are the total number of patients in the indicated group. [Figure 4A-4B] Figures 4A-4B show the best radiographic response and duration of response for 66 evaluable patients (as of June 16, 2023). *Radiographic response was determined according to RECIST 1.1. The waterfall plot and table represent interim data as of June 16, 2023, from ongoing clinical trials and unlocked databases. Patients 02-008 and 07-029 were classified as bevacizumab-naive in the data from July 25, 2022, but are now considered bevacizumab-exposed. mDOR CI: "-" indicates not reached. After external review of tumor measurements completed on May 12, 2023, patients 02-028 and 04-038 were determined to have a definite PR. [Figure 5A-5B] Figures 5A-5B are swimmer plots showing responses for 66 evaluable patients (as of June 16, 2023). *Swimmer plots / tables represent interim data as of June 16, 2023 from ongoing trials and unlocked databases. After external review of tumor measurements completed on May 12, 2023, patients 02-028 and 04-038 were determined to have a definite PR. [Figure 6] Figure 6 shows a plot showing progression-free survival for 66 evaluable patients (as of June 16, 2023). *Onvansertib mPFS is interim data as of June 16, 2023 from ongoing trials and unlocked databases. [Figure 7]Figure 7 shows a plot showing the transformation of tumor size from baseline. *Spider plot represents interim data as of June 16, 2023 from ongoing trials and unlocked databases. [Figure 8] Figure 8 shows the change in tumor volume in KRAS mutant xenograft models treated with vehicle, onvansertib, bevacizumab, or the combination of onvansertib and bevacizumab. 8-9 mice per group. Mean ± SEM is shown. An unpaired t-test was used to test for differences in tumor volume change between the combination treatment and the most effective control group on the last day of treatment. *p<0.05, ***p<0.001, ****p<0.0001. [Figure 9] FIG. 9 shows changes in tumor vascularization in KRAS mutant xenograft models upon treatment with vehicle, onvansertib, bevacizumab, or the combination of onvansertib and bevacizumab. [Figure 10] Figure 10 illustrates a non-limiting method that can be used to identify potential mechanisms of treatment resistance in KRAS mutant mCRC patients exposed to bev. DETAILED DESCRIPTION OF THE INVENTION

[0016] In the following detailed description, reference is made to the accompanying drawings, which form a part hereof. In the drawings, like numerals generally identify like elements unless context dictates otherwise. The illustrative embodiments described in the detailed description, drawings, and claims are not meant to be limiting. Other embodiments may be utilized, and other changes may be made, without departing from the spirit or scope of the subject matter presented herein. It will be readily understood that aspects of the present disclosure, as generally described herein and illustrated in the drawings, can be arranged, substituted, combined, separated, and designed in a wide variety of different configurations, all of which are expressly contemplated herein and make a part of this disclosure. All patents, published patent applications, other publications, and sequences from GenBank and other databases mentioned herein are incorporated by reference in their entirety for relevant art.

[0017] Disclosed herein are methods of treating cancer. In some embodiments, the methods include administering a PLK1 inhibitor and an anti-angiogenic agent to a subject with metastatic cancer, thereby reducing or inhibiting the progression of the metastatic cancer, wherein the subject has not previously been treated with angiogenesis inhibition, or the subject has not previously been treated for cancer (e.g., for metastatic cancer). In some embodiments, the methods can include administering a PLK1 inhibitor and an anti-angiogenic agent to a subject, thereby reducing or inhibiting the progression of the metastatic cancer, wherein the subject is known to have not previously been treated for cancer, or the subject is known to have not previously been treated for angiogenesis inhibition. In some embodiments, the methods for treating cancer can include identifying a subject with metastatic cancer who has not previously been treated for cancer, and administering a PLK1 inhibitor and an anti-angiogenic agent to the subject, thereby reducing or inhibiting the progression of the metastatic cancer. In some embodiments, the method includes identifying a subject with metastatic cancer who has not previously received a treatment involving angiogenesis inhibition, and administering to the subject a PLK1 inhibitor and an anti-angiogenic agent, thereby reducing or inhibiting the progression of the metastatic cancer. Any of the methods disclosed herein can be used as a first-line treatment for metastatic cancer, a second-line treatment for metastatic cancer, or both. To receive second-line cancer treatment, a subject will have previously received at least one cancer treatment that did not respond, stopped working, and / or had intolerable side effects. As disclosed herein, in some embodiments, a subject treated by a method disclosed herein has failed or is intolerant to treatment (e.g., chemotherapy, including oxaliplatin-based chemotherapy). In some embodiments, the chemotherapy is a fluoropyrimidine and oxaliplatin chemotherapy.

[0018] Disclosed herein are methods for improving objective response rate (ORR), progression-free survival (PFS), or both in the treatment of metastatic cancer. The methods, in some embodiments, comprise administering a PLK1 inhibitor and an anti-angiogenic agent to a subject suffering from metastatic cancer, thereby improving the ORR and / or PFS of the subject, wherein the subject has not previously been treated for cancer (e.g., for metastatic cancer) or the subject has not previously been treated for angiogenesis inhibition. The methods, in some embodiments, comprise administering a PLK1 inhibitor and an anti-angiogenic agent to a subject suffering from metastatic cancer, thereby improving the ORR and / or PFS of the subject, wherein the subject is known to have not previously been treated for cancer (e.g., for metastatic cancer) or the subject is known to have not previously been treated for angiogenesis inhibition. These methods can be used as first-line treatments for metastatic cancer, second-line treatments for metastatic cancer, or both. Disclosed herein are compositions and kits for treating cancer. In some embodiments, the kit comprises a PLK1 inhibitor and a manual providing instructions for administering the PLK1 inhibitor in combination with an anti-angiogenic agent to a subject to treat metastatic cancer, wherein the subject has not previously undergone treatment involving the inhibition of angiogenesis, or the subject has not previously undergone cancer treatment (e.g., treatment for metastatic cancer).

[0019] definition Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. See, e.g., Singleton et al., Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley & Sons (New York, NY 1994); Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press (Cold Spring Harbor, NY 1989). For purposes of this disclosure, the following terms are defined below.

[0020] As used herein, "subject" refers to an animal that is the object of treatment, observation, or experiment. "Animal" includes cold-blooded and warm-blooded vertebrates and invertebrates, such as fish, shellfish, reptiles, and especially mammals. "Mammals" include, but are not limited to, mice, rats, rabbits, guinea pigs, dogs, cats, sheep, goats, cows, horses, primates, such as monkeys, chimpanzees, and apes, especially humans. As used herein, a "patient" refers to a subject being treated by a medical professional, such as a physician or veterinarian, to attempt to cure, or at least ameliorate the effects of, a particular disease or disorder, or to prevent the disease or disorder from occurring in the first place. The patient may be an animal. In some embodiments, the patient is a mammal. In some embodiments, the patient is human.

[0021] As used herein, "administration" or "administering" refers to a method of providing a vertebrate with a dose of a pharmaceutically active ingredient. As used herein, "dose" refers to the total amount of an active ingredient (e.g., a PLK1 inhibitor (e.g., onvansertib) or an anti-angiogenic agent (e.g., bevacizumab)). As used herein, "unit dose" refers to the amount of therapeutic agent administered to a patient in one administration. As used herein, the term "daily dose" or "daily dosage" refers to the total amount of a pharmaceutical composition or therapeutic agent taken within a 24-hour period. As used herein, the term "delivery" refers to approaches, formulations, techniques, and systems for transporting a pharmaceutical composition or therapeutic agent into a patient's body as needed to safely achieve its desired therapeutic effect. In some embodiments, the composition or agent is formulated to deliver an effective amount of the composition or agent to the patient's bloodstream. As used herein, "formulated" or "formulation" refers to the process of combining different chemical entities, including one or more pharmaceutically active ingredients, into a dosage form. In some embodiments, two or more pharmaceutically active ingredients may be co-formulated into a single dosage form or combined dosage unit, or may be formulated separately and then combined into a combined dosage unit. A sustained release formulation is a formulation designed to slowly release a therapeutic agent in the body over an extended period of time, while an immediate release formulation is a formulation designed to rapidly release a therapeutic agent in the body over a short period of time.

[0022] As used herein, the term "pharmaceutically acceptable" indicates that the indicated material does not possess properties that would cause a reasonably prudent medical practitioner, taking into account the disease or condition being treated and the respective route of administration, to avoid administering the material to a patient. For example, such materials are generally required to be essentially sterile. As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulant, that is involved in carrying or transporting any supplement or composition, or its components, from one organ or body part to another, or delivering a drug to an affected tissue or tissue adjacent to the affected tissue. Carriers or excipients can be used to prepare compositions. Carriers or excipients can be selected to facilitate administration of a drug or prodrug. Examples of carriers include calcium carbonate, calcium phosphate, various sugars such as lactose, glucose, and sucrose, or starch species, cellulose derivatives, gelatin, vegetable oils, polyethylene glycol, and physiologically compatible solvents. Examples of physiologically compatible solvents include water for injection (WFI), saline, and sterile glucose solutions.

[0023] As used herein, the term "pharmaceutically acceptable salt" refers to any acid or base addition salt whose counterion is non-toxic to patients in pharmaceutical dosages of the salt. Numerous pharmaceutically acceptable salts are well known in the pharmaceutical art. When pharmaceutically acceptable salts of the compounds of the present disclosure are utilized in these compositions, the salts are preferably derived from inorganic or organic acids and bases. Such acid salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, and methanesulfonate. , 2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenyl-propionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, undecanoate, hydrohalides (e.g., chloride and hydrobromide), sulfate, phosphate, nitrate, sulfamate, malonate, salicylate, methylene-bis-b-hydroxynaphthoate, gentisate, isethionate, di-p-toluoyltartrate, ethanesulfonate, cyclohexylsulfamate, quinic acid, and the like. Pharmaceutically acceptable base addition salts include, but are not limited to, those derived from alkali or alkaline earth metal bases or conventional organic bases such as triethylamine, pyridine, piperidine, morpholine, N-methylmorpholine, and the like; for example, ammonium salts, alkali metal salts (such as sodium and potassium salts), alkaline earth metal salts (such as calcium and magnesium salts), salts with organic bases (such as dicyclohexylamine salts, N-methyl-D-glucamine), and salts with amino acids such as arginine and lysine.

[0024] As used herein, the terms "partial response," "partial remission," or "PR" refer to an improvement in a cancerous condition in response to treatment, as measured, for example, by tumor size and / or cancer marker levels. In some embodiments, "partial response" means that the size or level of a tumor or a blood marker indicative of a tumor has decreased by about 50% in response to treatment. The treatment can be any treatment for cancer, including, but not limited to, chemotherapy, radiation therapy, hormone therapy, surgery, cell or bone marrow transplantation, and immunotherapy. Tumor size can be detected by clinical or radiological means. Markers indicative of a tumor can be detected by means well known to those skilled in the art, such as ELISA or other antibody-based tests.

[0025] As used herein, the terms "complete response," "complete remission," "complete recovery," or "CR" refer to the disappearance of a cancerous condition, as measured, for example, by tumor size and / or cancer marker levels, following treatment, including, but not limited to, chemotherapy, radiation therapy, hormone therapy, surgery, cell or bone marrow transplantation, and immunotherapy. The presence of a tumor can be detected by clinical or radiological means. Tumor-indicating markers can be detected by means well known to those skilled in the art, such as ELISA or other antibody-based tests. However, a "complete response" does not necessarily mean that the cancer has been cured. Recurrence may occur after a complete response. A complete response of target lesions includes the disappearance of all target lesions and the reduction of pathological lymph nodes (whether target or non-target) to less than 10 mm in their short axis. A complete response of non-target lesions includes the disappearance of all non-target lesions and the normalization of tumor marker levels (all lymph nodes are non-pathological (short axis less than 10 mm)). If tumor markers were initially above the upper limit of normal, they must normalize for patients to be considered in clinical complete response for non-target lesions. Duration of overall CR is measured from the time the metrics first met CR to the first date of objectively documented disease progression or death from any cause. Participants without reported events will be censored at the last disease assessment.

[0026] As used herein, the term "stable disease" or "SD" means neither a sufficient shrinkage to qualify for PR nor a sufficient increase to qualify for progressive disease (PD) based on the smallest total diameter during the study. Duration of stable disease is measured from the start of treatment until criteria for progression are met, based on the smallest measurement recorded since the start of treatment, including the baseline measurement. As used herein, the term "disease progression" or "PD" with respect to target lesions means an increase of at least 20% in the sum of the diameters of the target lesions, based on the smallest sum during the study (including the baseline sum if this was the first value during the study). In addition to the 20% relative increase, an absolute increase in the sum of at least 5 mm must also be demonstrated (Note: the appearance of one or more new lesions is also considered progression). When disease progression or PD refers to non-target lesions, it means the appearance of one or more new lesions and / or overt progression of existing non-target lesions. Overt progression should not usually supersede the status of target lesions. It should represent a change in overall disease state, not an increase in a single lesion.

[0027] As used herein, the term "overall survival" or "OS" means the time from randomization (or enrollment) to death from any cause. Surviving participants are censored at the date of their last known survival. As used herein, the term "progression-free survival" or "PFS" means the time from randomization (or enrollment) to progression or death from any cause, whichever occurs first. Participants who remain alive without progression are censored at the date of their last disease assessment. As used herein, the term "best overall response" refers to the best response recorded from the start of treatment until disease progression / relapse (for disease progression, the lowest measurement recorded since treatment initiation is used as the reference). A patient's best response assignment depends on the achievement of both the measurement and confirmation criteria. Duration of overall response is measured from the time the measurement criteria meets CR or PR (whichever is recorded first) to the first date of objectively documented relapse or disease progression (for disease progression, the lowest measurement recorded since treatment initiation is used as the reference, or death from any cause; participants with no reported events are censored at the last disease assessment).

[0028] As used herein, the term "hydrate" refers to a complex formed by the combination of water molecules with solute molecules or ions. As used herein, the term "solvate" refers to a complex formed by the combination of solvent molecules with solute molecules or ions. The solvent may be an organic compound, an inorganic compound, or a mixture of both. Solvates are meant to include hydrates, hemihydrates, channel hydrates, and the like. Examples of solvents include, but are not limited to, methanol, N,N-dimethylformamide, tetrahydrofuran, dimethyl sulfoxide, and water. As used herein, a "therapeutically effective amount" or a "pharmaceutically effective amount" refers to an amount of a therapeutic agent that has a therapeutic effect. A dose of a pharmaceutically active ingredient that is therapeutically useful when administered alone or in combination with one or more additional therapeutic agents is a therapeutically effective amount. Thus, as used herein, a therapeutically effective amount refers to an amount of a therapeutic agent that produces a desired therapeutic effect as determined by the results of clinical trials and / or model animal studies. The therapeutically effective amount will vary depending on the compound, the disease, disorder, or condition and its severity, as well as the age, weight, etc., of the mammal being treated. The dose can conveniently be administered, for example, in divided doses up to four times daily or in sustained-release form.

[0029] As used herein, the terms "treat," "treatment," or "treating" refer to the administration of a therapeutic agent or pharmaceutical composition to a subject for prophylactic and / or therapeutic purposes. The term "prophylactic treatment" refers to treating a subject who does not yet exhibit symptoms of a disease or condition, but who is susceptible to or at risk for a particular disease or condition, whereby the treatment reduces the likelihood that the patient will develop the disease or condition. The term "therapeutic treatment" refers to administering treatment to a subject who already has a disease or condition. As used herein, a "therapeutic effect" relieves to some extent one or more symptoms of a disease or disorder. For example, a therapeutic effect can be observed by a reduction in subjective discomfort reported by the subject (e.g., a reduction in discomfort as documented in a self-administered patient questionnaire). As used herein, "prophylaxis," "prevent," "preventing," and grammatical variations thereof used herein refer to the prophylactic treatment of a subclinical condition in a subject, e.g., a mammal (including a human), to reduce the likelihood of the occurrence of a clinical condition. The method may partially or completely delay or prevent the onset or recurrence of a disorder or condition and / or one or more symptoms associated therewith, or may prevent a subject from acquiring or re-acquiring a disorder or condition, or may reduce the risk of a subject acquiring or re-acquiring a disorder or condition or one or more symptoms associated therewith. Subjects are selected for prophylactic therapy based on factors known to increase the risk of suffering from a clinical condition compared to the general population. "Prophylactic" therapy can be divided into (a) primary prevention and (b) secondary prevention. Primary prevention is defined as treatment in subjects who have not yet exhibited clinical disease, and secondary prevention is defined as the prevention of the secondary occurrence of the same or a similar clinical condition.

[0030] As used herein, the terms "partial response" and "partial remission" can refer to an improvement in a cancerous condition in response to treatment, for example, as measured by tumor size and / or cancer marker levels. In some embodiments, "partial response" means that the size or level of a tumor or a blood marker indicative of a tumor has decreased by about 50% in response to treatment. The treatment can be any treatment for cancer, including, but not limited to, chemotherapy, radiation therapy, hormone therapy, surgery, cell or bone marrow transplantation, and immunotherapy. Tumor size can be detected by clinical or radiological means. Markers indicative of a tumor can be detected by means well known to those skilled in the art, such as ELISA or other antibody-based tests. As used herein, the terms "complete response" or "complete remission" mean that the cancerous state, as measured by, for example, tumor size and / or cancer marker levels, has disappeared after treatment, including, but not limited to, chemotherapy, radiation therapy, hormone therapy, surgery, cell or bone marrow transplantation, and immunotherapy. The presence and / or size of a tumor can be detected by clinical or radiological means. Markers indicative of a tumor can be detected by means well known to those skilled in the art, such as ELISA or other antibody-based tests. However, a "complete response" does not necessarily indicate that the cancer has been cured, as recurrence may occur after a complete response.

[0031] cancer The methods, compositions, and kits disclosed herein can be used to treat cancer, including metastatic cancer. In some embodiments, a method for treating cancer comprises administering to a subject in need thereof (e.g., a subject with metastatic cancer) an antiangiogenic agent (e.g., bevacizumab) and a PLK1 inhibitor (e.g., onvansertib), or a pharmaceutically acceptable salt, solvate, stereoisomer, or prodrug thereof. In some embodiments, the subject is naive to treatment to inhibit angiogenesis (e.g., the subject has not been treated with bevacizumab alone, or the subject has not been treated with bevacizumab in combination with one or more anticancer agents or one or more anticancer therapies (e.g., chemotherapy)). In some embodiments, the subject is naive to cancer treatment. The subject can, for example, be a subject with no prior cancer diagnosis. In some embodiments, the subject is naive to treatment with bevacizumab and / or chemotherapy.

[0032] The methods, compositions, and kits disclosed herein can be used for various types of cancer. The cancer can be a solid tumor, a liquid tumor, or a combination thereof. In some embodiments, the cancer is a solid tumor, including but not limited to melanoma, renal cell carcinoma, lung cancer, bladder cancer, breast cancer, cervical cancer, colon cancer, gallbladder cancer, laryngeal cancer, liver cancer, thyroid cancer, stomach cancer, salivary gland cancer, prostate cancer, pancreatic cancer, Merkel cell carcinoma, brain and central nervous system cancer, and any combination thereof. In some embodiments, the cancer is a liquid tumor. In some embodiments, the cancer is a hematological cancer. Non-limiting examples of hematological cancers include diffuse large B-cell lymphoma ("DLBCL"), Hodgkin's lymphoma ("HL"), non-Hodgkin's lymphoma ("NHL"), follicular lymphoma ("FL"), acute myeloid leukemia ("AML"), and multiple myeloma ("MM"). Additionally, the diseases or conditions provided herein include refractory or recurrent malignant tumors whose growth can be inhibited using the methods and compositions disclosed herein.

[0033] In some embodiments, the cancer is a metastatic cancer. As used herein, "metastatic cancer" may refer to a case in which a cancer spreads (metastasizes) from its original site to another part of the body. Virtually all cancers have this ability to spread. Whether metastasis occurs depends on the complex interplay of many tumor cell factors, including the type of cancer, the maturity (degree of differentiation) of tumor cells, the location and duration of the cancer's presence, and other factors that are not fully understood. As used herein, the term "metastasis" refers to the formation of secondary tumor foci that gradually grow from the primary lesion to discontinuous sites. The metastatic process is a multistep mechanism in which metastatic cancer cells escape from the primary tumor, enter the circulation, invade distant tissue sites, and grow into macroscopic tumors at the target site. The metastatic cancer can be metastatic colorectal cancer, metastatic bladder cancer, metastatic breast cancer, metastatic kidney cancer, metastatic lung cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic prostate cancer, metastatic gastric cancer, metastatic thyroid cancer, metastatic uterine cancer, metastatic kidney cancer, metastatic cervical cancer, metastatic recurrent glioblastoma, or a combination thereof.

[0034] PLK1 inhibitors Polo-like kinases (PLKs) are a family of five highly conserved serine / threonine protein kinases. PLK1 is a master regulator of mitosis and is involved in several stages of the cell cycle, including mitotic entry, centrosome maturation, bipolar spindle formation, chromosome segregation, and cytokinesis. PLK1 has been shown to be overexpressed in solid tumors and hematological malignancies, including AML. PLK1 inhibition induces G2-M arrest and subsequent apoptosis in cancer cells and has emerged as a promising targeted therapy. Several PLK inhibitors are being investigated in clinical trials. In a randomized phase II study of previously untreated AML patients for whom induction therapy was inadequate, intravenous administration of the pan-PLK inhibitor volasertib (BI6727) in combination with LDAC significantly improved overall survival compared with LDAC alone. A subsequent randomized phase III study failed to confirm the benefit of the combination and reported an increased risk of serious infections. PLK1 promotes HR during double-strand DNA break (DSB) repair. PLK1 phosphorylates Rad51 and BRCA1, promoting their recruitment to DSB sites and thereby facilitating HR-mediated DNA repair. PLK1 inhibitors can be selective and / or specific for PLK1. The PLK1 inhibitor can be a dihydropteridinone, a pyridopyrimidine, an aminopyrimidine, a substituted thiazolidinone, a pteridine derivative, a dihydroimidazo[1,5-f]pteridine, a meta-substituted thiazolidinone, a benzylstyryl sulfone analog, a stilbene derivative, or any combination thereof. The PLK1 inhibitor can be onvansertib, BI2536, volasertib (BI 6727), GSK461364, AZD1775, CYC140, HMN-176, HMN-214, rigosertib (ON-01910), MLN0905, TKM-080301, TAK-960, or Ro3280.

[0035] Onvansertib (also known as PCM-075, NMS-1286937, NMS-937, "compound of formula (I)" in U.S. Pat. No. 8,927,530, and the IUPAC name 1-(2-hydroxyethyl)-8-{[5-(4-methylpiperazin-1-yl)-2-(trifluoromethoxy)phenyl]amino}-4,5-dihydro-1H-pyrazolo[4,3-h]quinazoline-3-carboxamide) is a selective, ATP-competitive PLK1 inhibitor. Biochemical assays demonstrated high specificity for PLK1 among a panel of 296 kinases, including other PLK members. Onvansertib has demonstrated potent antitumor activity in vitro and in vivo in both solid and hematologic malignancy models. Onvansertib is the first orally administered PLK1-specific ATP-competitive inhibitor to enter clinical trials and has demonstrated antitumor activity in various preclinical models. Onvansertib inhibited cell proliferation at nanomolar concentrations in AML cell lines and tumor growth in xenograft models of AML. Onvansertib also significantly augmented the antitumor activity of cytarabine in disseminated AML models. [ka] Onvansertib

[0036] Onvansertib exhibits high potency in proliferation assays with low nanomolar activity against multiple cell lines derived from both solid and hematologic tumors. It potently induces mitotic cell cycle arrest and subsequent apoptosis in cancer cell lines and inhibits xenograft tumor growth in mice at well-tolerated doses after oral administration via a distinct PLK1-mediated mechanism of action. Furthermore, onvansertib has demonstrated activity in combination with approved cytotoxic agents, such as irinotecan, enhancing tumor regression in HT29 human colon adenocarcinoma xenografts compared with each agent alone, and in combination with cytarabine prolonged survival in disseminated AML animal models. Onvansertib possesses favorable pharmacological parameters and good oral bioavailability in rodents and non-rodents. Its antitumor activity has been demonstrated in various preclinical models using various dosing regimens, potentially providing flexibility in dosing schedules and meriting clinical investigation. Onvansertib has several advantages over volasertib (BI6727, another PLK1 inhibitor), including higher potency and specificity for the PLK1 isoenzyme, and oral bioavailability.

[0037] A Phase I, first-in-human, dose-escalation study of onvansertib in patients with advanced / metastatic solid tumors identified neutropenia and thrombocytopenia as the primary dose-limiting toxicities. These hematologic toxicities were expected based on the drug's mechanism of action and were reversible, resolving within 3 weeks. The half-life of onvansertib was estimated to be 20-30 hours. Onvansertib's oral bioavailability and short half-life offer the opportunity for a simple, controlled, and flexible dosing schedule, potentially minimizing toxicity and improving the therapeutic window. To identify biomarkers associated with clinical response, pharmacodynamic and biomarker studies, including baseline genomic profiling, serial monitoring of mutant allele fractions in plasma, and the degree of PLK1 inhibition in circulating blast cells, have been conducted and are described in WO 2021 / 146322, the contents of which are incorporated herein by reference in their entirety.

[0038] The cancer treatments of the present disclosure can include administering a PLK1 inhibitor (e.g., onvansertib) to a subject with cancer for a desired period of time in one, two, or more cycles. The desired period of each cycle can independently be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more days. A cycle can be, for example, at least 20, 21, 22, 23, 24, 25, or more days in length. For example, a single treatment cycle can include administering a PLK1 inhibitor (e.g., onvansertib) for 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more days in one cycle (e.g., a cycle of at least 21 days (e.g., 21-28 days)). In some embodiments, treatment may involve administering a PLK1 inhibitor (e.g., onvansertib) for 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 days, or a range between any two of these values, in one cycle (e.g., a cycle of at least 21 days (e.g., 21-28 days)). Administration of the PLK1 inhibitor (e.g., onvansertib) in a single cycle of treatment may be continuous or separated by one or more periods (e.g., a 1- or 2-day rest period). In some embodiments, treatment involves administering the PLK1 inhibitor (e.g., onvansertib) for 5 days in a 21- to 28-day cycle. In some embodiments, the duration of administration of the PLK1 inhibitor in one cycle may be different from the duration of administration of the PLK1 inhibitor in one or more other cycles. For example, the PLK1 inhibitor can be administered to a subject for 10 days in the first cycle (e.g., days 1-5 of the first 14 days and days 1-5 of the subsequent 14 days of a 28-day cycle) and for 14 days in the second cycle (e.g., days 1-7 of the first 14 days and days 1-7 of the subsequent 14 days of a 28-day cycle). The length of each cycle can vary. For example, cycle 1 can be 28 days, and cycle 2 can be 21 days.

[0039] The cancer treatments disclosed herein include a PLK1 inhibitor (e.g., onvansertib) administered at a dose of 12 mg / m 2 ~90mg / m 2 For example, treatment may involve administering a PLK1 inhibitor (e.g., onvansertib) at or about such an amount, e.g., as a daily dose. 2 , 10 mg / m 2 , 12 mg / m 2 , 14 mg / m 2 , 15 mg / m 2 , 16 mg / m 2 , 18 mg / m 2 , 20 mg / m 2 , 23 mg / m 2 , 27 mg / m 2 , 30 mg / m 2 , 35 mg / m 2 , 40 mg / m 2 , 45 mg / m 2 , 50 mg / m 2 , 55 mg / m 2 , 60 mg / m 2 , 65 mg / m 2 , 70 mg / m 2 , 80 mg / m 2 , 85 mg / m 2 , 90 mg / m 2 , a range between any two of these values, or 8 mg / m 2 ~90mg / m 2 In some embodiments, the daily dose of the PLK1 inhibitor (e.g., onvansertib) can be adjusted (e.g., increased or decreased within a range) for the subject during treatment or during a single cycle of treatment (e.g., the first, second, third, and subsequent cycles). In some embodiments, the daily dose of the PLK1 inhibitor (e.g., onvansertib) is 12 mg / m or more. 2 , 15 mg / m 2 , 18 mg / m 2 , or 24 mg / m 2 In some embodiments, the daily dose of the PLK1 inhibitor (e.g., onvansertib) is 15 mg / m 2The daily dose of the PLK1 inhibitor (e.g., onvansertib) in each treatment cycle can vary. For example, the daily dose of the PLK1 inhibitor (e.g., onvansertib) in the first cycle is 12 mg / m 2 and the daily dose of the PLK1 inhibitor (e.g., onvansertib) in the second cycle can be, for example, 15 mg / m 2 In some embodiments, the daily dose of the PLK1 inhibitor (e.g., onvansertib) for the second cycle can then be increased to, for example, 18 mg / m 2 Without being bound by any particular theory, the mg / m 2 Dosages are considered to be based on body surface area (BSA) equivalent to a fixed dose ranging from 20 to 45 mg.

[0040] The maximum concentration (C) of a PLK1 inhibitor in the blood of a subject when the PLK1 inhibitor (e.g., onvansertib) is administered as a single agent or in combination with one or more additional cancer therapeutic agents (e.g., FOLFIRI and bevacizumab) max ) (during or after treatment) can be about 100 nmol / L to about 1500 nmol / L. For example, the C of a PLK1 inhibitor in the blood of a subject when the PLK1 inhibitor (e.g., onvansertib) is administered as a single agent or in combination with one or more additional cancer therapeutic agents (e.g., FOLFIRI and bevacizumab) can be max can be or can be about 100 nmol / L, 200 nmol / L, 300 nmol / L, 400 nmol / L, 500 nmol / L, 600 nmol / L, 700 nmol / L, 800 nmol / L, 900 nmol / L, 1000 nmol / L, 1100 nmol / L, 1200 nmol / L, 1300 nmol / L, 1400 nmol / L, 1500 nmol / L, a range between any two of these values, or any value between 200 nmol / L and 1500 nmol / L. The area under the curve (AUC) of a plot of the concentration of the PLK1 inhibitor in the blood of a subject versus time (e.g., AUC during the first 24 hours after administration) when the PLK1 inhibitor (e.g., onvansertib) is administered as a single agent or in combination with one or more additional cancer therapeutic agents (e.g., FOLFIRI and bevacizumab) is administered. 0-24 ) can be from about 1,000 nmol / L h to about 400,000 nmol / L h. For example, when a PLK1 inhibitor (e.g., onvansertib) is administered as a single agent or in combination with one or more additional cancer therapeutic agents (e.g., FOLFIRI and bevacizumab), the AUC (e.g., AUC during the first 24 hours after administration) of a plot of the PLK1 inhibitor concentration in the subject's blood against time can be used. 0-24 ) can be or be about 1000 nmol / L-h, 5000 nmol / L-h, 10,000 nmol / L-h, 15,000 nmol / L-h, 20,000 nmol / L-h, 25,000 nmol / L-h, 30,000 nmol / L-h, 35,000 nmol / L-h, 40,000 nmol / L-h, a range between any two of these values, or any value between 1000 nmol / L-h and 400,000 nmol / L-h.

[0041] The time to reach maximum concentration of a PLK1 inhibitor in the blood of a subject (T) when the PLK1 inhibitor (e.g., onvansertib) is administered as a single agent or in combination with one or more additional cancer therapeutic agents (e.g., FOLFIRI and bevacizumab) max ) can be about 1 hour to about 5 hours. For example, when a PLK1 inhibitor (e.g., onvansertib) is administered as a single agent or in combination with one or more additional cancer therapeutic agents (e.g., FOLFIRI and bevacizumab), the time to reach the maximum concentration of the PLK1 inhibitor in the blood of a subject (T max ) can be or be about 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 4.5 hours, 5 hours, a range between any two of these values, or any value from 1 hour to 5 hours. The elimination half-life (T) of a PLK1 inhibitor (e.g., onvansertib) in the blood of a subject when the PLK1 inhibitor is administered as a single agent or in combination with one or more additional cancer therapeutic agents (e.g., FOLFIRI and bevacizumab) is measured. 1 / 2 ) can be about 10 hours to about 60 hours. For example, when a PLK1 inhibitor (e.g., onvansertib) is administered as a single agent or in combination with one or more additional cancer therapeutic agents (e.g., FOLFIRI and bevacizumab), the elimination half-life (T) of the PLK1 inhibitor in the blood of a subject can be measured. 1 / 2 ) can be or be about 10 hours, 15 hours, 20 hours, 25 hours, 30 hours, 35 hours, 40 hours, 45 hours, 50 hours, 55 hours, 60 hours, a range between any two of these values, or any value from 10 hours to 60 hours.

[0042] Antiangiogenic agents Angiogenesis, known as the inappropriate formation of new blood vessels, is generally required for cancer metastasis. "Angiogenesis" is often observed in tumors, where endothelial cells secrete a group of growth factors that are mitogenic to the endothelium, causing endothelial cell elongation and proliferation, resulting in the generation of new blood vessels. Inhibition of angiogenesis has been shown to cause tumor regression in animal models and to be effective in treating metastatic cancer in humans. The terms "antiangiogenic agent" and "angiogenesis inhibitor" shall have their ordinary meanings and may be used interchangeably herein to refer to any agent capable of inhibiting angiogenesis. Some antiangiogenic agents have been approved for use in humans for the treatment of cancer. In some embodiments of the methods disclosed herein, the subject has not previously undergone treatment, including administration of an angiogenesis inhibitor.

[0043] The angiogenesis inhibitor and / or anti-angiogenic agent can inhibit VEGF-A, VEGFR-1, VEGFR-2, VEGFR-3, EGFR, HER2, PDGFR family proteins, RAF, Kit (or c-Kit), FLT3, CSF-1R, RET, Abl, Itk, LcK, c-FMS, FGFR family proteins, c-Met, PlGF, TNF-α, IFN, IL, bFGF, mTOR, or any combination thereof. Antiangiogenic agents include afatinib (Giotrif®), axitinib (Inlyta®), bevacizumab (Avastin®), cabozantinib (Cometriq®), cetuximab (Erbitux®), erlotinib (Tarceva®), everolimus (Afinitor®), gefitinib (Iressa®), imatinib (Gleevec®), lapatinib (Tykerb®), lenalidomide (Revirimid®), lenvatinib mesylate (Lenvima®), necitumumab (Portrazza®), and others. The medicament for treating rheumatoid arthritis may be steroid therapy (Tregs), neratinib (Nerlynx®), panitumumab (Vectibix®), pazopanib (Votrient®), pertuzumab (Perjeta®), ramucirumab (Cyramza®), regorafenib (Stivarga®), sorafenib (Nexavar®), sunitinib (Sutent®), thalidomide (Sinovir, Thalomid®), trastuzumab (Ontruzant®), vandetanib (Caprelsa®), or Ziv-Aflibercept (Zaltrap®).Antiangiogenic agents include afatinib (Giotrif®), axitinib (Inlyta®), bevacizumab (Avastin®), cabozantinib (Cometriq®), cetuximab (Erbitux®), erlotinib (Tarceva®), everolimus (Afinitor®), gefitinib (Iressa®), imatinib (Gleevec®), lapatinib (Tykerb®), lenalidomide (Revlimid®), lenvatinib mesylate (Lenvima®), necitumumab (Portrazza®), and rituximab (Tykerb®). The medicament for treating rheumatoid arthritis may be steroid therapy, such as steroid therapy with steroids like steroids like steroids like steroids, or steroids like steroids like steroids like steroids. The medicament for treating rheumatoid arthritis may be steroids like steroids like steroids, such as steroids like steroids like steroids, or steroids like steroids like steroids. The medicament for treating rheumatoid arthritis may be steroids like steroids like steroids, such as steroids like steroids, or steroids like steroids. The medicament for treating rheumatoid arthritis may be steroids like steroids like steroids, such as steroids like steroids, or steroids like steroids. The medicament for treating rheumatoid arthritis may be steroids like steroids like steroids, such as steroids like steroids, or steroids like steroids.

[0044] Bevacizumab (also referred to herein as "bev") can be used in combination with chemotherapy in both first-line and second-line therapy for treating cancer. Other antiangiogenic agents, such as ramucirumab and aflibercept, can be used in second-line therapy. Prior to the currently disclosed methods, the overall survival (OS) and median progression-free survival (mPFS) benefits in second-line therapy have been shown to be independent of whether bevacizumab was administered in the first line (e.g., the subject had received prior therapy involving angiogenesis inhibition). For example, a study examining OS and PFS in patients with and without prior bevacizumab treatment found that OS was 13.9 months in bevacizumab-naïve patients and 12.5 months in those with prior bevacizumab treatment. Regarding PFS, mPFS was 6.9 months in bevacizumab-naïve patients and 6.7 months in those with prior bevacizumab treatment. Antiangiogenic therapy incrementally improves response rates in both bevacizumab-experienced and bevacizumab-naive patients, with objective response rates ranging from 5% to 13% in bevacizumab-experienced patients compared with approximately 25% in bevacizumab-naive patients.

[0045] As disclosed herein, combination therapy of an antiangiogenic agent and a PLK1 inhibitor (including onvansertib) can surprisingly significantly enhance efficacy against metastatic cancer (e.g., metastatic colorectal cancer, metastatic bladder cancer, metastatic breast cancer, metastatic kidney cancer, metastatic lung cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic prostate cancer, metastatic gastric cancer, metastatic thyroid cancer, metastatic uterine cancer, metastatic kidney cancer, metastatic cervical cancer, metastatic recurrent glioblastoma, or a combination thereof) and can result in tumor regression and cancer-free survival in subjects who have not received prior treatment, including angiogenesis inhibition. As a result, the tumor regression and cancer-free survival / survival rates achieved by this combination can demonstrate surprising synergy (i.e., greater than additive, superior to the cumulative anti-tumor effects achieved by the antiangiogenic agent and the PLK1 inhibitor, respectively). The PLK1 inhibitor can be onvansertib. Provided herein are methods, compositions, and kits for treating metastatic cancer in a subject (e.g., a human patient suffering from metastatic cancer). Provided herein are methods, compositions, and kits for treating cancer in a subject (e.g., a human patient suffering from cancer), where the subject has not received prior treatment involving the inhibition of angiogenesis. The method includes administering to the patient an anti-angiogenic agent and a PLK1 inhibitor in a manner sufficient to inhibit the progression of the cancer. For example, the anti-angiogenic agent and the PLK1 inhibitor can be administered simultaneously, separately, or sequentially to a subject with cancer.

[0046] In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent synergistically reduces or inhibits the progression of metastatic cancer compared to the additive effects of PLK1 inhibitor treatment alone, anti-angiogenic agent treatment alone, and / or PLK1 inhibitor treatment alone and anti-angiogenic agent treatment alone. The inhibition or reduction of cancer progression that can be achieved by the methods disclosed herein using a PLK1 inhibitor (e.g., onvansertib) and an anti-angiogenic agent (e.g., bevacizumab) is enhanced or synergistic, rather than merely additive (i.e., the inhibition is greater than the combined inhibition of progression caused by the anti-angiogenic agent alone and the PLK1 inhibitor alone). The enhanced or synergistic efficacy or inhibition of the combination of an anti-angiogenic agent and a PLK1 inhibitor of the present disclosure can vary in different embodiments. In some embodiments, the enhanced or synergistic efficacy or inhibition of the combination of an anti-angiogenic agent and a PLK1 inhibitor disclosed herein is greater than or equal to 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 210%, 220%, 230%, 240%, 250%, 260%, 270%, 280%, 290%, 300%, 310%, 320%, 330%, 340%, 350%, 360%, 370%, 380%, 390%, 410%, 420%, 430%, 440%, 450%, 460%, 470%, 480%, 490%, 500%, 510%, 520%, 530%, 540%, 550%, 560%, 570%, 580%, 590%, 610%, 620%, 630%, 640%, 650%, 660%, 670%, 680%, 690%, 700%, 710%, 720%, 730%, 740%, 750%, 760%, 770%, 780%, 790%, 800%, 810%, 820%, 830%, 840%, 850%, 860%, 870%, 880%, 890%, 900%, 950%, 9 , 150%, 160%, 170%, 180%, 190%, 200%, 210%, 220%, 230%, 240%, 250%, 260%, 270%, 280%, 290%, 300%, or a number or range between any two of these values, or about such a percentage higher, or at least such a percentage higher, or at most such a percentage higher, or at most such a percentage higher.

[0047] The molar ratio of the PLK1 inhibitor (e.g., onvansertib) to the antiangiogenic agent (e.g., bevacizumab) can be, for example, about 1:200, 1:100, 1:90, 1:80, 1:70, 1:60, 1:50, 1:40, 1:30, 1:20, 1:10, 1:1, 10:1, 20:1, 30:1, 40:1, 50:1, 100:1, 1000:1, 2000:1, or 5000:1, or a number or range between any two of these values. In some embodiments, the enhanced or synergistic efficacy or inhibition of cancer progression caused by the combination of an antiangiogenic agent and a PLK1 inhibitor (e.g., onvansertib) is 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 250%, 300%, or a number or range between any two of these values, is greater, greater by about such a percentage, greater by at least such a percentage, greater by at least about such a percentage, greater by at most such a percentage, or greater by at most such a percentage than the combined inhibition of progression caused by the antiangiogenic agent and the PLK1 inhibitor (e.g., onvansertib) alone. For example, the combination of an antiangiogenic agent and a PLK1 inhibitor may result in 50%, 60%, 70%, 80%, 90% or more inhibition of cancer progression (50%, 40%, 30%, 20%, 10% or less cancer cell viability), whereas under the same conditions, the combined inhibition of the antiangiogenic agent alone and the PLK1 inhibitor alone may result in 10%, 20%, 25%, 30% or less inhibition of cancer progression (90%, 80%, 75%, 70% or more cancer cell viability). Thus, the enhanced or synergistic efficacy or inhibition of cancer progression caused by the combination of an antiangiogenic agent and a PLK1 inhibitor may be, for example, 50%, 60%, 70%, 80%, 90%, 100% or more greater than the combined inhibition of progression caused by the antiangiogenic agent alone and the PLK1 inhibitor alone. In some embodiments, the antiangiogenic agent is bevacizumab and the PLK1 inhibitor is onvansertib.

[0048] The antiangiogenic agent and the PLK1 inhibitor can be administered to a patient in any manner believed to be effective for treating cancer. The antiangiogenic agent can be administered together with the PLK1 inhibitor or separately. When administered separately, the antiangiogenic agent can be administered before or after the PLK1 inhibitor, or in a different administration cycle. The PLK1 inhibitor can be administered orally. The antiangiogenic agent can be administered intravenously or orally. The PLK1 inhibitor and the antiangiogenic agent can be administered simultaneously or sequentially. In some embodiments, it may be advantageous to administer the PLK1 inhibitor (e.g., onvansertib) to a subject before the antiangiogenic agent (e.g., bevacizumab), for example, on one or more days, or each day, that the PLK1 inhibitor and the antiangiogenic agent are administered to the subject. The time interval between the administration of the PLK1 inhibitor and the administration of the antiangiogenic agent can be, for example, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, a range between any two of these values, or any value between 30 minutes and 12 hours. In some embodiments, both the PLK1 inhibitor (e.g., onvansertib) and the anti-angiogenic agent (e.g., bevacizumab) are administered to the subject on, or at least about, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the days of a cycle (e.g., each cycle during combination treatment), optionally with the PLK1 inhibitor administered to the subject before the anti-angiogenic agent on each of the days on which both agents are administered, e.g., the PLK1 inhibitor is administered 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, or a range between any two of these values, or any value between 30 minutes and 12 hours, before administration of the anti-angiogenic agent.

[0049] The antiangiogenic agent and the PLK1 inhibitor can each be administered according to any schedule, such as, for example, once or more times daily or weekly; once, twice, three times, four times, five times, six times, or seven times weekly (daily); or once or more weeks. In some embodiments, the antiangiogenic agent and the PLK1 inhibitor are each administered to a patient in cycles of at least two times per week. In other embodiments, the antiangiogenic agent and the PLK1 inhibitor are each administered to a patient in cycles of at least five times per week. In some embodiments, the PLK1 inhibitor is administered daily, and the antiangiogenic agent is administered daily, weekly, every other week, every four weeks, every five weeks, or monthly. In further embodiments, the patient receives at least two cycles of administration. The patient can receive one or more cycles, for example, two, three, three, four, five, or more cycles of administration. The administration of two adjacent cycles can be consecutive, i.e., there can be no break between the last day of the first cycle and the first day of the second cycle. In some embodiments, two adjacent cycles of administration are separated by a drug holiday, i.e., an interval between the last day of the first cycle and the first day of the second cycle. The drug holiday (i.e., interval) can be, or can be, 1 day, 2 days, 3 days, 5 days, 7 days, 10 days, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or a value or range between any two of these values. In some embodiments, a patient receives three or four cycles of administration, each cycle comprising at least five doses per week (e.g., five days per week). Each cycle of a multi-cycle administration can have the same or different dosing schedule. For example, one cycle of a multi-cycle administration can include five consecutive days of daily administration of a PLK1 inhibitor and an anti-angiogenic agent per week, followed by two days of drug holiday per week for four weeks, while one or more other cycles of the same multi-cycle administration can include 28 consecutive days of daily administration of a PLK1 inhibitor and an anti-angiogenic agent per week for four weeks. The PLK1 inhibitor can be administered for at least four days of the cycle. In some embodiments, the PLK1 inhibitor is not administered on at least one day of the cycle.

[0050] The antiangiogenic agent can be administered to a patient at any suitable dose, for example, about, at least, or at most 0.1 mg / kg, 1 mg / kg, 5 mg / kg, 10 mg / kg, 20 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 200 mg / kg, 300 mg / kg, 400 mg / kg, 500 mg / kg, 600 mg / kg, 700 mg / kg, 800 mg / kg, 900 mg / kg, 1000 mg / kg, 1500 mg / kg, 2000 mg / kg, or a value between any two of these values. Weight-based dosage units (mg / kg) can be converted to other units (e.g., mg / m) using a conversion table, such as a body surface area (BSA) conversion table, as will be understood by those skilled in the art. 2 ) can be converted to a 200 mg / kg dose. The antiangiogenic agent (e.g., bevacizumab) can be administered at about 1 mg / kg to 20 mg / kg. In some embodiments, the antiangiogenic agent is bevacizumab and is administered at a dose of about, at least, or at most 1 mg / kg, 5 mg / kg, 10 mg / kg, 20 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 200 mg / kg, 300 mg / kg, 400 mg / kg, 500 mg / kg, 600 mg / kg, 700 mg / kg, 800 mg / kg, 900 mg / kg, 1000 mg / kg, or a number between any two of these values. Bevacizumab can be administered at about 5 mg / kg, about 7.5 mg / kg, about 10 mg / kg, or about 15 mg / kg.

[0051] The antiangiogenic agent can be administered to a patient once daily, twice daily, or three times daily. The antiangiogenic agent can be administered daily, weekly, every other week, every three weeks, every four weeks, or monthly. In some embodiments, the antiangiogenic agent is administered in a 7-56 day cycle that is daily, weekly, every other week, every three weeks, every four weeks, or monthly. In some embodiments, the antiangiogenic agent is administered in a 7-day, 8-day, 9-day, 10-day, 11-day, 12-day, 13-day, 14-day, 15-day, 16-day, 17-day, 18-day, 19-day, 20-day, 21-day, 22-day, 23-day, 24-day, 25-day, 26-day, 27-day, 28-day, 29-day, 30-day, 32-day, 35-day, 42-day, 49-day, or 56-day cycle. In some embodiments, the antiangiogenic agent is administered on days 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 32, 35, 42, 49, or 56 of a cycle. In some embodiments, the antiangiogenic agent is administered on day 1, 2, 3, 4, 5, 6, 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, 51, 52, 52, 53, 54, 55, and / or 56.In some embodiments, the antiangiogenic agent is not administered on days 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 52, 53, 54, 55, and / or 56.

[0052] In some embodiments, the subject has not previously received treatment involving the administration of an angiogenesis inhibitor. The angiogenesis inhibitor can be the same as the anti-angiogenic agent. Any currently known or hereafter discovered agent capable of inhibiting angiogenesis (e.g., anti-angiogenic), including anti-angiogenic agents that inhibit the activity of one or more growth factors or other proteins that promote angiogenesis, can be used in these methods. For example, the angiogenesis inhibitor and / or anti-angiogenic agent can inhibit vascular endothelial growth factor family proteins, such as VEGF-A, VEGFR-1, VEGFR-2, VEGFR-3, and PlGF, or the signaling pathways of such proteins. Other proteins and pathways that can be targeted by antiangiogenic agents include, but are not limited to, epidermal growth factor receptor (EGFR), human epidermal growth factor receptor 2 (HER2), platelet-derived growth factor receptor (PDGFR) family proteins, RAF kinase; tyrosine kinases including Kit (or c-Kit), RET, Abl, Itk, LcK, c-Met, and FLT3; colony-stimulating factor 1 receptor (CSF-1R) and c-FMS, fibroblast growth factor receptor (FGFR) family proteins, tumor necrosis factor alpha (TNF-α), interferon (IFN), interleukin (IL), basic fibroblast growth factor (bFGF), mammalian target of rapamycin (mTOR), or any combination thereof. Antiangiogenic agents can be small molecules, antibodies or fragments thereof, aptamers, RNA (e.g., miRNA), or any other agent capable of inhibiting angiogenesis. In some embodiments, the antiangiogenic or angiogenesis inhibitor is bevacizumab (eg, Avastin®).

[0053] Similarly, any now known or hereafter discovered PLK1 inhibitor can be used in these methods, including PLK1 inhibitors that are selective for PLK1 and PLK1 inhibitors that also inhibit the activity of other proteins. In some embodiments, the PLK1 inhibitor is a dihydropteridinone, a pyridopyrimidine, an aminopyrimidine, a substituted thiazolidinone, a pteridine derivative, a dihydroimidazo[1,5-f]pteridine, a meta-substituted thiazolidinone, a benzylstyryl sulfone analog, a stilbene derivative, or a combination thereof. In some of these embodiments, the PLK1 inhibitor is onvansertib, BI2536, volasertib (BI 6727), GSK461364, AZD1775, CYC140, HMN-176, HMN-214, rigosertib (ON-01910), MLN0905, TKM-080301, TAK-960, or Ro3280. The PLK1 inhibitor can be onvansertib. In these embodiments, onvansertib is administered at any suitable dose, for example, 12 mg / m 2 Less than 24 mg / m 2 or less than 24 mg / m 2 In some embodiments, onvansertib is administered to patients at a higher dose than about 12 mg / m 2 , about 15mg / m 2 , or about 18 mg / m 2 In some embodiments, onvansertib is administered to a patient daily. In further embodiments, onvansertib is administered in cycles of 3-10 days of daily administration and 2-16 days without administration of onvansertib. In some embodiments, onvansertib is administered to a patient at least 5 times per week in one cycle. A patient may receive 2, 3, or 4 cycles. In some embodiments, a patient receives 4 cycles of at least 5 days of daily administration of onvansertib and 1-2 days without administration of onvansertib.

[0054] In some embodiments, a PLK1 inhibitor alone or in combination with an anti-angiogenic agent is administered to a patient after one or more cycles of administration followed by a drug holiday. Drug holiday, as used herein, refers to a period during which a patient stops taking the PLK1 inhibitor and / or anti-angiogenic agent. The drug holiday can last from several days to several months. In some embodiments, the drug holiday can be 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, or any value or range between any two of these values. As will be understood by those skilled in the art, the dosage and timing of the antiangiogenic agent and PLK1 inhibitor may vary depending on the type (species, sex, age, weight, etc.) and condition of the subject being treated, as well as the severity of the disease or condition being treated. The antiangiogenic agent and PLK1 inhibitor may be formulated into a single pharmaceutical composition or two separate pharmaceutical compositions. The active ingredient may also be encapsulated in microcapsules prepared, for example, by coacervation techniques or interspecific polymerization, such as hydroxymethylcellulose or gelatin microcapsules and poly(methylmethacylate) microcapsules, respectively, in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or in macroemulsions. The methods, compositions, kits, and systems disclosed herein can be applied to various types of subjects. For example, the subject can be undergoing cancer treatment, in remission from cancer, having undergone one or more cancer treatments, or suspected of having cancer. The subject can have stage I cancer, stage II cancer, stage III cancer, and / or stage IV cancer. In some embodiments, the subject has stage IV cancer. In some embodiments, the subject has metastatic cancer. In some embodiments, the subject has not received prior treatment involving the inhibition of angiogenesis.

[0055] The treatment of the present disclosure may include administration of a PLK1 inhibitor (onvansertib) for a desired period during a cycle. Administration of the PLK1 inhibitor (and / or anti-angiogenic agent) may be daily, or there may be drug holidays between administration days. The drug holidays may be, for example, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, or more days. Administration may be once, twice, three times, four times, or more times on the days on which the PLK1 inhibitor (and / or anti-angiogenic agent) is administered to the patient. Administration may be, for example, once every 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days. The desired period of time can vary in length, for example, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, or more. Each treatment cycle can be of various lengths, for example, at least 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, or more. For example, one cycle of treatment can include administration of a PLK1 inhibitor (e.g., onvansertib) and / or an anti-angiogenic agent for 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or more days in one cycle (e.g., a cycle of at least 21 days (e.g., 21-28 days)). In some embodiments, treatment may involve administration of a PLK1 inhibitor (e.g., onvansertib) and / or an anti-angiogenic agent for, or at least for, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 days, or a range between any two of these values, in one cycle (e.g., a cycle of at least 21 days (e.g., 21-28 days)). Administration of the PLK1 inhibitor (e.g., onvansertib) and / or anti-angiogenic agent in one cycle of treatment may be continuous or separated by one or more periods (e.g., one or two days off).In some embodiments, treatment involves administration of a PLK1 inhibitor (e.g., onvansertib) for 5 days in a 21-28 day cycle.

[0056] In some embodiments, a PLK1 inhibitor (e.g., onvansertib) is administered to a subject in need thereof for 20 days (e.g., days 1-10 and days 15-24) in a 28-day cycle. The 20 days can be, for example, 10 consecutive days (e.g., days 1-10) of continuous daily administration and another 10 consecutive days (e.g., days 15-24) of continuous daily administration, or four sets of 5 days (e.g., days 1-5, days 8-12, days 15-19, and days 22-26) of continuous daily administration. In some embodiments, for example, if a patient is identified as having poor tolerance to a PLK1 inhibitor, the PLK1 inhibitor is administered to a subject in need thereof for 10 days (e.g., days 1-5 and days 15-19) in a 28-day cycle. The 10 days can be, for example, 10 consecutive daily administrations (e.g., days 1-10) or two consecutive daily administrations for each 5-day period (e.g., days 1-5 and days 15-19). In some embodiments, the PLK1 inhibitor is administered to a subject in need thereof every day throughout the entire cycle (e.g., every day for 28 days in a 28-day cycle). Depending on the need for inhibition / reversal of cancer progression in the subject, the subject can receive one, two, three, four, five, six, or more cycles of treatment. In the case of combined treatment, the administration cycle, administration schedule, and / or dosage of the antiangiogenic agent and the PLK1 inhibitor can be the same or different. In the case of combined treatment, the administration cycle, administration schedule, and / or dosage of the antiangiogenic agent can be adjusted depending on the administration cycle, administration schedule, and / or dosage of the PLK1 inhibitor. For example, an antiangiogenic agent (e.g., bevacizumab) can be administered in four 7-day cycles (e.g., daily administration on days 1-5, rest on days 6-7, repeated for 4 weeks) corresponding to the 28-day cycle of administration of a PLK1 inhibitor (e.g., onvansertib).

[0057] Treatment can be, for example, at a daily dose of 6 mg / m 2 ~90mg / m 2For example, treatment may include administration of a PLK1 inhibitor (e.g., onvansertib) at or about 6 mg / m 2 , 8 mg / m 2 , 10 mg / m 2 , 12 mg / m 2 , 14 mg / m 2 , 16 mg / m 2 , 18 mg / m 2 , 20 mg / m 2 , 23 mg / m 2 , 27 mg / m 2 , 30 mg / m 2 , 35 mg / m 2 , 40 mg / m 2 , 45 mg / m 2 , 50 mg / m 2 , 55 mg / m 2 , 60 mg / m 2 , 65 mg / m 2 , 70 mg / m 2 , 80 mg / m 2 , 85 mg / m 2 , 90 mg / m 2 , a value or range between any two of these values, or 8 mg / m 2 ~90mg / m 2 This may include daily administration at or about any value of 12 mg / m 2 or 15 mg / m 3 or 16 mg / m 4 or 18 mg / m 5 or 19 mg / m 6 or 20 mg / m 2 or 20 mg / m 3 or 20 mg / m 4 or 20 mg / m 5 or 20 mg / m 6 or 20 mg / m 2 or 20 mg / m 3 ... 2 In some embodiments, the PLK1 inhibitor (e.g., onvansertib) is administered at 15 mg / m for 10 days (e.g., days 1-5 and days 15-19) in a 28-day cycle. 2 In some embodiments, the PLK1 inhibitor (e.g., onvansertib) is administered at 8 mg / m daily (e.g., days 11-28) during a 28-day cycle.2 or 10 mg / m 2 In some embodiments, the daily dose of the PLK1 inhibitor (e.g., onvansertib) can be adjusted (e.g., increased or decreased within a range) for the subject during treatment, or during a cycle (e.g., the first, second, third, and subsequent cycles) of treatment. In some embodiments, the PLK1 inhibitor is administered at a dose of 12 mg / m 2 or about 12 mg / m 2 In some embodiments, the PLK1 inhibitor is administered at a dose of 15 mg / m 2 or about 15 mg / m 2 In some embodiments, the PLK1 inhibitor is administered at a dose of 18 mg / m 2 or approximately 18 mg / m 2 It is administered at .

[0058] The maximum concentration (C) of a PLK1 inhibitor in the blood of a subject when the PLK1 inhibitor (e.g., onvansertib) is administered alone or in combination with an antiangiogenic agent. max ) (during or after treatment) can be about 100 nmol / L to about 1500 nmol / L. For example, when a PLK1 inhibitor is administered alone or in combination with an anti-angiogenic agent, the C max can be or can be about 100 nmol / L, 200 nmol / L, 300 nmol / L, 400 nmol / L, 500 nmol / L, 600 nmol / L, 700 nmol / L, 800 nmol / L, 900 nmol / L, 1000 nmol / L, 1100 nmol / L, 1200 nmol / L, 1300 nmol / L, 1400 nmol / L, 1500 nmol / L, a range between any two of these values, or any value between 200 nmol / L and 1500 nmol / L. The area under the curve (AUC) of a plot of the concentration of the PLK1 inhibitor in the blood of a subject versus time (e.g., AUC during the first 24 hours after administration) when the PLK1 inhibitor (e.g., onvansertib) is administered alone or in combination with an anti-angiogenic agent. 0-24) can be about 1,000 nmol / L-h to about 400,000 nmol / L-h. For example, when a PLK1 inhibitor is administered alone or in combination with an anti-angiogenic agent, the AUC (e.g., AUC during the first 24 hours after administration) of a plot of the PLK1 inhibitor concentration in the subject's blood versus time can be used. 0-24 ) can be or be about 1000 nmol / L.hour, 5000 nmol / L.hour, 10,000 nmol / L.hour, 15,000 nmol / L.hour, 20,000 nmol / L.hour, 25,000 nmol / L.hour, 30,000 nmol / L.hour, 35,000 nmol / L.hour, 40,000 nmol / L.hour, a range between any two of these values, or any value from 1000 nmol / L.hour to 400,000 nmol / L.hour.

[0059] The time to reach the maximum concentration of a PLK1 inhibitor in the blood of a subject (T) when the PLK1 inhibitor (e.g., onvansertib) is administered alone or in combination with an anti-angiogenic agent. max ) can be about 1 hour to about 5 hours. For example, when a PLK1 inhibitor is administered alone or in combination with an anti-angiogenic agent, the time until the maximum concentration of the PLK1 inhibitor in the blood of a subject (T max ) can be or be about 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 4.5 hours, 5 hours, a range between any two of these values, or any value from 1 hour to 5 hours. The elimination half-life (T) of a PLK1 inhibitor (e.g., onvansertib) in the blood of a subject when the PLK1 inhibitor is administered alone or in combination with an anti-angiogenic agent. 1 / 2 ) can be about 10 hours to about 60 hours. For example, when a PLK1 inhibitor is administered alone or in combination with an anti-angiogenic agent, the elimination half-life (T 1 / 2 ) can be or be about 10 hours, 15 hours, 20 hours, 25 hours, 30 hours, 35 hours, 40 hours, 45 hours, 50 hours, 55 hours, 60 hours, a range between any two of these values, or any value from 10 hours to 60 hours.

[0060] As described herein, the methods, compositions, and kits described herein can reduce or inhibit the progression of cancer. In some embodiments, the reduction or inhibition comprises an inhibition of the growth of one or more tumors in the subject and / or a reduction in the number of cancer cells detected in the subject by at least about 25%, 30%, 40%, 50%, 60%, or 70%, compared to an untreated subject. In some embodiments, the reduction or inhibition comprises an inhibition of the growth of one or more tumors in the subject and / or a reduction in the number of cancer cells detected in the subject by at least about 25%, 30%, 40%, 50%, 60%, or 70%, compared to the subject prior to administration of the PLK1 inhibitor and the anti-angiogenic agent. The growth of at least one of the subject's one or more tumors can be reduced by, for example, at least about 25%, 30%, 40%, 50%, 60%, or 70% after one or more cycles of treatment. In some embodiments, the size / volume of at least one of the subject's one or more tumors is reduced by at least about 25%, 30%, 40%, 50%, 60%, or 70% after one or more cycles of treatment.

[0061] Additional cancer medications or therapies The methods, compositions, and kits disclosed herein can be used to treat cancer. In some embodiments, a method for treating cancer (e.g., mCRC) comprises administering an anti-angiogenic agent and a PLK1 inhibitor (e.g., onvansertib) to a subject (e.g., patient) in need thereof. The method may comprise administering a therapeutically effective amount of the anti-angiogenic agent and a therapeutically effective amount of the PLK1 inhibitor. The treatment may comprise administering at least one additional cancer therapeutic agent or cancer therapy. In some embodiments, the subject has not previously been treated with angiogenesis inhibition. In some embodiments, the subject has not previously been treated to inhibit angiogenesis (e.g., the subject has not previously been treated with bevacizumab alone, or the subject has not previously been treated with bevacizumab in combination with one or more anti-cancer agents or one or more anti-cancer therapies (e.g., chemotherapy)). In some embodiments, the subject has not previously been treated for cancer. The subject may, for example, not have a prior cancer diagnosis. In some embodiments, the subject has not previously been treated with bevacizumab and / or chemotherapy.

[0062] Non-limiting examples of additional cancer therapeutic agents or cancer therapies include surgery, chemotherapy, radiation therapy (including external beam, stereotactic radiation therapy, intraoperative radiation therapy, and brachytherapy), bone marrow transplant, immunotherapy, targeted drug therapy, cryoablation, or radiofrequency ablation. When the cancer is colorectal cancer, examples of treatments include surgery, radiofrequency ablation, cryoablation, radiation therapy, chemotherapy (including medications including capecitabine, 5-fluorouracil (5-FU), irinotecan, oxaliplatin, trifluridine / tipiracil), targeted therapy (including antiangiogenic therapy using, for example, bevacizumab, regorafenib, ziv-aflibercept, or ramucirumab; e.g., pembrolizumab, nivolumab, or ipilimumab; and immunotherapy using PLK1 inhibitors). The additional cancer therapeutic agent or cancer therapy can be chemotherapy, such as FOLFIRI, FOLFOX, XELOX (CAPOX), FOLFOXIRI, or a combination thereof. Chemotherapy regimens using fluorouracil are the standard treatment for advanced colorectal cancer. Fluorouracil is a pyrimidine analog and antimetabolite that incorporates into DNA molecules, halting synthesis and thereby preventing cancer cell replication. Examples of these regimens include FOLFOX and FOLFIRI. The additional cancer therapeutic agent or cancer therapy can include a therapeutically effective amount of FOLFIRI. FOLFIRI is a chemotherapy cocktail containing leucovorin (folinic acid), fluorouracil, and irinotecan hydrochloride. Leucovorin is a vitamin B derivative, and this combination enhances the cytotoxicity of fluorouracil. Irinotecan is a topoisomerase inhibitor that prevents DNA from uncoiling and replicating. FOLFIRI is often combined with other therapeutic agents (eg, bevacizumab) to improve efficacy and response rates.

[0063] FOLFOX is a chemotherapy regimen for the treatment of colorectal cancer consisting of folinic acid, fluorouracil, and oxaliplatin (ELOXATIN). FOLFOX can be divided into other subtypes, such as FOLFOX-4, FOLFOX-6, and FOLFOX-7, depending on how these three drugs are administered. FOLFOX is typically used to treat colorectal cancer. FOLFOX can also be used to treat pancreatic cancer and certain other cancers. FOLFOX is generally used as adjuvant therapy (in addition to primary therapy) for advanced cancers. However, FOLFOX can also be used as first-line therapy for colorectal adenocarcinoma, the most common type of colon cancer. In this combination, oxaliplatin exhibits synergistic effects with fluorouracil, with little overlap in toxicity. For decades, fluorouracil was the only drug with demonstrated activity against colorectal cancer, commonly in combination with leucovorin. Oxaliplatin and capecitabine are both relatively new drugs used to treat colorectal cancer. These drugs have been shown to act synergistically both in vivo and in vitro. The oxaliplatin-capecitabine combination chemotherapy, known as Xerox, has demonstrated high activity in metastatic colorectal cancer (mCRC). Capecitabine has demonstrated high efficacy as a first-line treatment for mCRC. Capecitabine is an oral fluoropyrimidine rationally designed to generate FU preferentially at tumor sites through a three-step enzymatic process that exploits the significantly higher activity of thymidine phosphorylase (TP) in tumors compared with healthy tissue. Compared with leucovorin and fluorouracil combination therapy (e.g., FOLFIRI and FOLFOX), capecitabine has fewer side effects, such as diarrhea, stomatitis, nausea, alopecia, and neutropenia, and is less likely to result in neutropenic fever / sepsis and associated hospitalization. However, hand-foot syndrome (HFS) is more common with capecitabine administration. Oxaliplatin is a third-generation cisplatin analogue and an organoplatinum complex. It is generally classified as an alkylating agent, although it lacks the ability to actually attach alkyl groups to DNA. Oxaliplatin is an essential component of various fluorouracil regimens (e.g., FOLFOX), which are the standard of care for metastatic and node-positive colorectal cancer.

[0064] Another combination of oxaliplatin and fluorouracil is known as FOLFOXIRI, which contains leucovorin, fluorouracil, oxaliplatin, and irinotecan. FOLFOXIRI is also a chemotherapy regimen approved for the treatment of advanced colorectal cancer, often in combination with bevacizumab. The additional cancer therapeutic agent can include FOLFIRI, bevacizumab, abiraterone, FOLFOX, an anti-EGFR agent, a KRAS-directed inhibitor, gemcitabine, Abraxane, nanoliposomal irinotecan, 5-FU, or a combination thereof. The PLK1 inhibitor and the cancer therapeutic agent or therapy can be administered simultaneously or sequentially. In some embodiments, the additional cancer therapeutic agent or therapy includes FOLFIRI, abiraterone, FOLFOX, an anti-EGFR agent, a KRAS-directed inhibitor, gemcitabine, Abraxane, nanoliposomal irinotecan, 5-FU, or a combination thereof. In some embodiments, the anti-EGFR agent is optionally cetuximab. In some embodiments, the KRAS-directed inhibitor is optionally a G12C inhibitor, a G12D inhibitor, or a combination thereof. In some embodiments, the additional cancer therapy is FOLFOX (leucovorin, fluorouracil, and oxaliplatin), FOLFIRI (leucovorin, fluorouracil, and irinotecan), or a combination thereof.

[0065] Methods for predicting / determining treatment efficacy and cancer status Disclosed herein are methods for treating cancer. In some embodiments, the methods include administering a PLK1 inhibitor and an anti-angiogenic agent to a subject with metastatic cancer, thereby reducing or inhibiting the progression of the metastatic cancer, wherein the subject has not received prior treatment involving angiogenesis inhibition. The methods described herein, in combination with the anti-angiogenic agent, are expected to be effective in treating various metastatic cancers, such as metastatic colorectal cancer, metastatic bladder cancer, metastatic breast cancer, metastatic kidney cancer, metastatic lung cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic prostate cancer, metastatic gastric cancer, metastatic thyroid cancer, metastatic uterine cancer, metastatic kidney cancer, metastatic cervical cancer, metastatic recurrent glioblastoma, or a combination thereof.

[0066] The method may include one or more of: (1) determining the subject's cancer status; (2) determining the subject's responsiveness to PLK1 inhibitor treatment; and (3) administering one or more cancer therapeutic agents or therapies for the cancer. In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent improves one or more therapeutic effects in the treated subject compared to a control or baseline. The one or more therapeutic effects can include tumor size from metastatic cancer, objective response rate (ORR), duration of response, time to response, progression-free survival (PFS), overall survival (OS), disease control rate (DCR), oncogenic allele burden, or a combination thereof. In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent improves the subject's ORR, improves the treated subject's PFS, improves the treated subject's OS, improves the treated subject's DCR, reduces the treated subject's oncogenic allele burden, or a combination thereof, compared to a subject who has previously received a therapy that includes inhibition of angiogenesis.

[0067] Disclosed herein are methods of improving objective response rate (ORR), progression-free survival (PFS), or both in a subject with metastatic cancer. In some embodiments, the methods involve administering to the subject a PLK1 inhibitor and an anti-angiogenic agent, thereby improving the ORR and / or PFS in the subject, wherein the subject has not previously received treatment involving inhibition of angiogenesis. In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent synergistically improves ORR and / or PFS in the treated subject compared to the additive effects of PLK1 inhibitor treatment alone, anti-angiogenic agent treatment alone, and / or PLK1 inhibitor treatment alone and anti-angiogenic agent treatment alone.

[0068] Tumor assessment and assessment of tumor burden and treatment efficacy can be performed based on RECIST criteria (Therasse et al. 2000), New Guidelines to Evaluate the Response to Treatment in Solid Tumors, Journal of National Melanoma Institute, Vol. 92; 205-16, and in some embodiments, are performed within the methods disclosed herein according to the revised RECIST guidelines (version 1.1) (Eisenhauer et al. 2009, New response evaluation criteria in solid tumors: revised RECIST guideline (version 1.1). Eur J Melanoma, 45(2):228-47.), which is incorporated herein by reference in its entirety. In some embodiments, the treatment results in a sustained response in the subject after cessation of treatment. "Sustained response" may refer to a sustained effect on tumor growth reduction after cessation of treatment. For example, the size of the tumor may remain the same or may become smaller compared to the size at the beginning of the administration phase. In some embodiments, the sustained response is at least as long as the treatment period, or at least 1.5, 2.0, 2.5, or 3.0 times the treatment period.

[0069] The treatment methods disclosed herein may result in partial or complete response. As used herein, "complete response" or "CR" refers to the disappearance of all target lesions; "partial response" or "PR" refers to at least a 30% reduction in the sum of the longest diameters (SLD) of target lesions, based on the baseline SLD; "stable disease" or "SD" may refer to the target lesions neither shrinking enough to qualify for PR nor increasing enough to qualify for PD, based on the smallest SLD since the start of treatment. As used herein, "objective response rate" (ORR) may refer to the sum of the complete response (CR) rate and the partial response (PR) rate. The treatment methods disclosed herein may result in an increase in progression-free survival (PFS) and overall survival (OS) of subjects to whom the combination therapy is administered. As used herein, "progression-free survival" (PFS) refers to the length of time during and after treatment that the disease being treated (e.g., cancer) does not worsen. Progression-free survival may include the amount of time a patient has a complete response or partial response, as well as the amount of time a patient has stable disease. As used herein, "overall survival" refers to the percentage of subjects in a group who are likely to be alive after a certain period of time. As used herein, "disease control rate" refers to complete response (CR) + partial response (PR) + stable disease (SD).

[0070] In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent improves the subject's ORR by 50%, by about such a percentage, by at least such a percentage, or by at least about such a percentage (e.g., 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 1 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or range between any two of these values) improvement. In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent improves the subject's PFS by 50%, by about such a percentage, by at least such a percentage, or by at least about such a percentage (e.g., 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 1 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or range between any two of these values) improvement.

[0071] In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent to a subject who has not received prior treatment involving inhibition of angiogenesis may result in an ORR of 69%, about such a percentage, at least such a percentage, or at least about such a percentage (e.g., 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or a number or range between any two of these values). In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent to a subject who has not previously received treatment involving inhibition of angiogenesis results in an ORR of about 23% or less (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, or a number or range between any two of these values). In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent to a subject who has not previously received a treatment involving inhibition of angiogenesis is continued for 13.5 months, about such a period, at least such a period, or at least about such a period (e.g., 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13 years, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13 years, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13 years, 14 years, 15 years ... , 13 years, 14 years, 15 years, 16 years, 17 years, 18 years, 19 years, 20 years, 21 years, 22 years, 23 years, 24 years, 25 years, 26 years, 27 years, 28 years, 29 years, 30 years, 31 years, 32 years, 33 years, 34 years, 35 years, 36 years, 37 years, 38 years, 39 years, 40 years, 41 years, 42 years, 43 years, 44 years, 45 years, 46 years, 47 years, 48 ​​years, 49 years, 50 years, 55 years, 60 years, 65 years, or more than 65 years). In some embodiments, administration of a PLK1 inhibitor and an anti-angiogenic agent to a subject who has not received prior treatment, including inhibition of angiogenesis, may result in a median PFS of about 7.8 months or less (e.g., 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, or a number or range between any two of these values).

[0072] The term "tumor growth inhibition" or "tumor growth reduction" can refer to causing a reduction or complete cessation of tumor growth and / or causing a regression in tumor size (e.g., diameter and / or volume). The term "tumor volume" or "tumor size" can refer to the total size of the tumor, which can include the tumor itself and, if applicable, affected lymph nodes. The presence or absence of a tumor and tumor size can be determined by various methods known in the art, such as by measuring tumor dimensions using calipers, computed tomography (CT) or magnetic resonance imaging (MRI) scans, mammography, and X-rays. Volume can be calculated using equations based on, for example, the z-axis diameter or standard shapes such as a sphere, ellipsoid, or cube. Tumor size can be assessed at any time before, during, or after at least one cycle of treatment with onvansertib and / or an antiangiogenic agent. Tumor size can be assessed at an initial time point and at one or more additional time points. In some embodiments, tumor size can be assessed in a subject and, e.g., an untreated subject, at comparable time points (e.g., an initial time point and one or more additional time points). Tumor growth can be determined, e.g., by measuring tumor size at an initial time point and measuring tumor size at one or more additional time points. In some embodiments, increased inhibition of tumor growth in a subject (e.g., improved ORR) indicates that the subject is responsive to the cancer treatment.

[0073] The inhibition of growth of at least one of the one or more tumors in a subject can be at least, or at least about, 1.1-fold, greater than 1.2-fold, greater than 1.3-fold, greater than 1.4-fold, greater than 1.5-fold, greater than 1.6-fold, greater than 1.7-fold, greater than 1.8-fold, greater than 1.9-fold, greater than 2-fold, or a number or range between or exceeding any two of these values, of the inhibition of growth caused by onvansertib and an anti-angiogenic agent in a subject who has been previously treated for inhibition of angiogenesis after one or more cycles of treatment. Inhibition of growth of at least one of the one or more tumors in a subject may be increased by, about, at least, or at least about 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or a value or range between any two of these values, relative to a subject that has been previously treated to inhibit angiogenesis. Growth of at least one of the one or more tumors in a subject may be inhibited by, about, at least, or at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or a value or range between any two of these values, relative to an untreated subject or a subject that has been previously treated to inhibit angiogenesis, relative to an untreated subject or a subject that has been previously treated to inhibit angiogenesis. Growth of at least one of the one or more tumors in a subject may be inhibited by, about, at least, or at least about 70%, 75%, 80%, 85%, 90%, 95%, 100%, or a value or range between any two of these values, relative to an untreated subject after one or more cycles of treatment. The subject may be tumor-free after one or more cycles of treatment.

[0074] The inhibition of growth of at least one of the one or more tumors in the subject may, after one or more cycles of treatment, exceed, exceed about, exceed at least, or exceed at least about 1.1-fold, greater than 1.2-fold, greater than 1.3-fold, greater than 1.4-fold, greater than 1.5-fold, greater than 1.6-fold, greater than 1.7-fold, greater than 1.8-fold, greater than 1.9-fold, greater than 2-fold, or a number or range between any two of these values, or a greater multiple. Inhibition of growth of at least one of the one or more tumors in a subject may be increased by, about, at least, or at least about 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or a value or range between any two of these values, after one or more cycles of treatment, compared to a subject treated with onvansertib alone or the antiangiogenic agent alone. Inhibition of growth of at least one of the one or more tumors in a subject may be inhibited by, about, at least, or at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, or a value or range between any two of these values, after one or more cycles of treatment, compared to an untreated subject. Growth of at least one of the one or more tumors in a subject can be inhibited by, about, at least, or at least about 70%, 75%, 80%, 85%, 90%, 95%, 100%, or a value or range between any two of these values, relative to an untreated subject after one or more cycles of treatment. The subject can be tumor-free after one or more cycles of treatment.

[0075] In some embodiments, the first time point is before or immediately before the combination treatment, and at least one of the one or more additional time points is at or after the completion of at least one cycle of the combination treatment. In some embodiments, the cycle of the combination treatment is the first cycle of the combination treatment. In some embodiments, the first time point is before or immediately before the first cycle of the combination treatment, and the one or more additional time points is at or after the completion of a second cycle of the combination treatment. In some embodiments, the first cycle of combination treatment immediately precedes the second cycle of combination treatment. In some embodiments, the method includes continuing the combination treatment on the subject if the subject is shown to be responsive to the combination treatment. In some embodiments, the method includes discontinuing the combination treatment on the subject and / or initiating a different combination treatment on the subject if the subject is not shown to be responsive to the combination treatment.

[0076] The first time point can be before or immediately before the combination treatment, and the one or more additional time points are at the end of or after at least one cycle of the combination treatment, optionally, the cycle of the combination treatment is the first cycle of the combination treatment. In some embodiments, the first time point is before or immediately before the first cycle of the combination treatment, and the one or more additional time points are at the end of or after the second cycle of the combination treatment, optionally, the first cycle of the combination treatment is immediately before the second cycle of the combination treatment. Determining a subject's responsiveness may include determining whether the subject is a responder to the treatment, whether the subject is in or heading toward complete remission (CR), or whether the subject is in or heading toward partial remission (PR). Determining a subject's responsiveness may include determining the subject's objective response rate (ORR), duration of response, time to response, progression-free survival (PFS), overall survival (OS), disease control rate (DCR), oncogenic allele burden, or a combination thereof. Determining a subject's responsiveness may include determining whether the subject is in partial response to the treatment, whether the subject is in complete response to the treatment, whether the subject is in stable disease (SD), or whether the subject is in progressive disease (PD). In some embodiments, the method includes initiating additional treatment in the subject if the subject's cancer is shown to have recurred. The additional treatment may be the same or different from the current or prior concomitant treatment.

[0077] Another method for evaluating the cancer status of a subject includes determining ECOG status.As used herein, ECOG status refers to Eastern Cooperative Oncology Group (ECOG) Performance Status (Oken M, et al. Toxicity and response criteria of the Eastern Cooperative Oncology Group. Am J Clin Oncol 1982;5(6):649-655), as follows: 0: fully active, can perform all pre-disease activities without limitation; 1: limited physically strenuous activity, but ambulatory, and can perform light or sedentary work, such as light housework and office work; 2: ambulatory and can perform all self-care, but cannot perform work activities, and moves more than 50% of the time when awake; 3: can only perform limited self-care, and is bed or chair-bound more than 50% of the time when awake; 4: completely disabled, cannot perform self-care, and is completely bed or chair-bound, and 5: death. As described herein, the patient achieves a complete or partial response after treatment with an antiangiogenic agent and a PLK1 inhibitor. In some embodiments, the patient may achieve a complete response. In some embodiments, the patient may achieve a partial response. In some embodiments, the patient is antiangiogenic-naive.

[0078] Disclosed herein are methods, compositions, kits, and systems for predicting / determining the clinical outcome of the disclosed cancer combination treatments, monitoring the combination treatments, predicting / determining a subject's responsiveness to the combination treatments, determining the cancer status in a subject, and improving the outcome of the combination treatments. These methods, compositions, kits, and systems can be used to guide combination treatments, provide combination treatment recommendations, and reduce or avoid unnecessary and ineffective combination treatments for patients. In some embodiments, the combination treatments disclosed herein can improve a subject's oncogenic allele dosage. As used herein, "allele dosage" refers to the ratio of mutant (e.g., oncogenic) alleles to wild-type alleles in a clinical sample used for genotyping. In some embodiments, the sample may contain circulating tumor DNA (ctDNA). ctDNA can be analyzed to predict / determine the clinical outcome of cancer treatment, monitor cancer treatment, predict / determine a subject's responsiveness to cancer treatment, determine the state of cancer in a subject, improve the outcome of cancer treatment, guide cancer treatment, provide treatment recommendations, and / or reduce or avoid ineffective combination treatments. Such analysis of ctDNA is described in WO2021146322, the contents of which are incorporated herein by reference in their entirety.

[0079] Methods of determining a subject's responsiveness to a combination therapy comprising an antiangiogenic agent and a PLK1 inhibitor of the present disclosure include, for example, analyzing ctDNA of a subject with cancer (which can include, where the subject is undergoing treatment and / or has previously undergone combination treatment), thereby determining the subject's responsiveness to the combination therapy. In some embodiments, determining the subject's responsiveness includes determining whether the subject is a responder to the treatment, whether the subject is in or heading toward a CR, or whether the subject is in or heading toward a partial response (PR). For example, analyzing the ctDNA can include detecting a variant allele frequency of ctDNA in a first sample obtained from the subject at a first time point, detecting a variant allele frequency of ctDNA obtained from the subject at one or more additional time points in one or more additional samples, and determining a difference in the variant allele frequency of ctDNA between the first sample and at least one of the one or more additional samples, wherein a decrease in the variant allele frequency in at least one of the additional samples relative to the first sample indicates that the subject is responsive to the cancer treatment.

[0080] In some embodiments, the first time point is before or immediately before the combination treatment, and at least one of the one or more additional time points is at or after the completion of at least one cycle of the combination treatment. In some embodiments, the cycle of the combination treatment is the first cycle of the combination treatment. In some embodiments, the first time point is before or immediately before the first cycle of the combination treatment, and the one or more additional time points is at or after the completion of a second cycle of the combination treatment. In some embodiments, the first cycle of combination treatment immediately precedes the second cycle of combination treatment. In some embodiments, the method includes continuing the combination treatment on the subject if the subject is shown to be responsive to the combination treatment. In some embodiments, the method includes discontinuing the combination treatment on the subject and / or initiating a different combination treatment on the subject if the subject is not shown to be responsive to the combination treatment.

[0081] Disclosed herein is a method for determining the cancer status of a subject, comprising analyzing the subject's ctDNA, thereby determining the cancer status of the subject. The subject may be currently receiving a combination treatment comprising an antiangiogenic agent and a PLK1 inhibitor of the present disclosure, a subject who has previously received a combination treatment of the present disclosure, and / or a subject in cancer remission. A subject in cancer remission may be in complete remission (CR) or partial remission (PR). In some embodiments, analyzing the ctDNA comprises detecting a mutant allele frequency of the ctDNA. In some embodiments, analyzing the ctDNA comprises detecting a mutant allele frequency of ctDNA obtained from the subject at a first time point in a first sample, detecting a mutant allele frequency of ctDNA obtained from the subject at one or more additional time points in one or more additional samples, and determining a difference in the mutant allele frequency of ctDNA between the first sample and at least one of the one or more additional samples, wherein an increase in the mutant allele frequency in the additional sample relative to the first sample indicates that the subject is at risk of cancer recurrence or has cancer recurrence.

[0082] In some embodiments, the first time point is before or immediately before the combination treatment, and the one or more additional time points are at the end of or after at least one cycle of the combination treatment, optionally, this cycle of the combination treatment. In some embodiments, the first time point is before or immediately before the first cycle of the combination treatment, and the one or more additional time points are at the end of or after a second cycle of the combination treatment, optionally, the first cycle of the combination treatment is immediately before the second cycle of the combination treatment. In some embodiments, the method includes initiating an additional treatment in the subject if the subject is shown to have a recurrence of cancer, which may be the same or different from the current or prior concomitant treatment.

[0083] The mutant allele frequency of ctDNA can be determined, for example, by the count of all mutations of ctDNA in each of the first sample and one or more additional samples, or by the average mutant allele frequency in each of the first sample and one or more additional samples. In some embodiments, the mutant allele frequency is the mutant allele frequency (MAF) of driver mutations in cancer (e.g., metastatic colorectal cancer, metastatic bladder cancer, metastatic breast cancer, metastatic kidney cancer, metastatic lung cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic prostate cancer, metastatic gastric cancer, metastatic thyroid cancer, metastatic uterine cancer, metastatic kidney cancer, metastatic cervical cancer, metastatic recurrent glioblastoma, or combinations thereof). In some embodiments, the mutant allele frequency is the MAF of one or more driver mutations in cancer (e.g., metastatic colorectal cancer, metastatic bladder cancer, metastatic breast cancer, metastatic kidney cancer, metastatic lung cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic prostate cancer, metastatic gastric cancer, metastatic thyroid cancer, metastatic uterine cancer, metastatic kidney cancer, metastatic cervical cancer, metastatic recurrent glioblastoma, or combinations thereof). In some embodiments, Log2(C1 / C0) < MAF threshold indicates a decrease in ctDNA MAF. C0 is the ctDNA MAF of the first sample, and C1 is the ctDNA MAF of one of the additional samples. In some embodiments, the MAF threshold is from 0.01 to -0.10, or about such a value. In some embodiments, the MAF threshold is 0.06, or about such a value. In some embodiments, the MAF threshold is 0.05, or about such a value. In some embodiments, the first sample comprises ctDNA of the subject before treatment, and one of the additional samples comprises ctDNA of the subject after treatment.

[0084] Driver mutations were detected in the following 75 genes: ABL1, ANKRD26, ASXL1, ATRX, BCOR, BCORL1, BRAF, BTK, CALR, CBL, CBLB, CBLC, CCND2, CDC25C, CDKN2A, CEBPA, CSF3R, CUX1, CXCR4, DCK, DDX41, DHX15, DNMT3A, ETNK1, ETV6, EZH2, FBXW7, FLT3, GATA1, GATA2, GNAS, HRAS, IDH1, IDH2, IKZF1, JAK2, JAK3, KDM6A, and KI. In some embodiments, at least one of the one or more driver mutations is a mutation in one of the 75 genes. In some embodiments, the one or more driver mutations is a mutation in one of the 75 genes. The driver mutation or at least one of the one or more driver mutations can be in a gene selected from TP53, ASXL1, DNMT3A, NRAS, SRSF2, TET2, SF3B1, FLT3, FLT3 ITD, IDH2, NPM1, RUNX1, CDKN2A, KRAS, STAG2, CALR, CBL, CSF3R, DDX41, GATA2, JAK2, PHF6, and SETBP1. In some embodiments, the driver mutation or at least one of the one or more driver mutations is in a gene selected from the group consisting of DNMT3A, TET2, NPM1, SRSF2, NRAS, CDKN2A, SF3B1, FLT3, ASXL1, SRSF2, IDH2, NRAS, and SF3B1. In some embodiments, the method further comprises determining a mutant allele frequency in one or more of the subject's ctDNA, PBMCs, and BMMCs.

[0085] ctDNA can be analyzed, for example, using polymerase chain reaction (PCR), next-generation sequencing (NGS), and / or droplet digital PCR (ddPCR). The samples disclosed herein can be derived, for example, from a subject's whole blood, a subject's plasma, a subject's serum, or a combination thereof. In some embodiments, ctDNA is derived from a subject's whole blood, a subject's plasma, a subject's serum, or a combination thereof. In some embodiments, the method includes analyzing the subject's ctDNA before treatment. In some embodiments, the treatment includes one or more cycles, and the ctDNA is analyzed before, during, and after each cycle of treatment. Each treatment cycle can be at least 21 days. In some embodiments, each treatment cycle is about 21 days to about 28 days. In some embodiments, the subject is human.

[0086] Disclosed herein is a method for improving a cancer treatment outcome, which may include detecting a mutant allele frequency in ctDNA obtained from the subject at a first time point in a first sample before the subject receives a combination treatment comprising an anti-angiogenic agent of the present disclosure and a PLK1 inhibitor; detecting a mutant allele frequency in ctDNA obtained from the subject at one or more additional time points in one or more additional samples after the subject receives the combination treatment; determining a difference in the mutant allele frequency in ctDNA between the first sample and at least one of the one or more additional samples, wherein a decrease in oncogenic allele load and / or mutant allele frequency in at least one of the additional samples relative to the first sample indicates that the subject is responsive to the combination treatment; and, if the subject is shown to be responsive to the combination treatment, continuing the combination treatment on the subject, or, if the subject is not shown to be responsive to the combination treatment, discontinuing the combination treatment on the subject and / or initiating a different cancer treatment on the subject.

[0087] Also included as disclosed herein are methods of treating cancer. The methods may include administering a combination treatment comprising an anti-angiogenic agent and a PLK1 inhibitor of the present disclosure to a subject in need thereof; determining a reduction in oncogenic allele burden and / or mutant allele frequency in a second sample from the subject obtained at a second time point after the subject has received the combination treatment relative to a first sample from the subject obtained at a first time point before the subject has received the combination treatment; and continuing the combination treatment. In some embodiments, the subject is a newly diagnosed cancer patient, e.g., a subject who has not previously been treated for cancer before the combination treatment. In some embodiments, the subject has previously been treated for cancer and is in remission for the cancer, e.g., the subject is in complete remission (CR) or partial remission (PR) after previously receiving the combination treatment. In some embodiments, the previous treatment does not include use of an anti-angiogenic agent, a PLK1 inhibitor, or both.

[0088] The first time point can be, for example, before or immediately before the combination treatment. At least one of the one or more additional time points can be, for example, at the end of or after at least one cycle of the combination treatment. In some embodiments, the cycle of the combination treatment is the first cycle of the combination treatment. In some embodiments, the first time point is before or immediately before the first cycle of the combination treatment, and the one or more additional time points are at the end of or after the second cycle of the combination treatment. In some embodiments, the first cycle of the combination treatment is immediately before the second cycle of the combination treatment. The mutant allele frequency of ctDNA can be determined, for example, by the count of all mutations of ctDNA in each of the first sample and one or more additional samples, and / or by the average mutant allele frequency in each of the first sample and one or more additional samples. In some embodiments, the mutant allele frequency is the mutant allele frequency (MAF) of a driver mutation of cancer (e.g., metastatic colorectal cancer, metastatic bladder cancer, metastatic breast cancer, metastatic kidney cancer, metastatic lung cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic prostate cancer, metastatic gastric cancer, metastatic thyroid cancer, metastatic uterine cancer, metastatic kidney cancer, metastatic cervical cancer, metastatic recurrent glioblastoma, or a combination thereof). In some embodiments, the mutant allele frequency is the MAF of one or more driver mutations of cancer (e.g., metastatic colorectal cancer, metastatic bladder cancer, metastatic breast cancer, metastatic kidney cancer, metastatic lung cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic prostate cancer, metastatic gastric cancer, metastatic thyroid cancer, metastatic uterine cancer, metastatic kidney cancer, metastatic cervical cancer, metastatic recurrent glioblastoma, or a combination thereof). In some embodiments, Log2(C1 / C0) < MAF threshold indicates a decrease in ctDNA MAF. C0 is the ctDNA MAF of the first sample, and C1 is the ctDNA MAF of one of the additional samples. In some embodiments, the MAF threshold is -0.05.

[0089] This method may further include determining the mutant allele frequency in one or more of the subject's ctDNA, PBMC, and BMMC. The mutant allele frequency in ctDNA can be detected, for example, using polymerase chain reaction (PCR), next-generation sequencing (NGS). In some embodiments, the mutant allele frequency in ctDNA is detected using droplet digital PCR (ddPCR). At least one of the first sample, one or more additional samples, and the second sample may be derived from the subject's whole blood, the subject's plasma, the subject's serum, or a combination thereof. In some embodiments, ctDNA is derived from the subject's whole blood, the subject's plasma, the subject's serum, or a combination thereof. In some embodiments, the subject whose ctDNA is analyzed is undergoing or will undergo treatment for cancer. The method can include analyzing the subject's ctDNA before treatment. The treatment can include one or more cycles, and the ctDNA is analyzed before, during, and after one or more cycles of treatment. For example, the ctDNA can be analyzed for two or more cycles of treatment, three or more cycles of treatment, or before, during, and after each cycle of treatment. Each treatment cycle can be at least 21 days, e.g., 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more days, or a range between any two of these values. In some embodiments, each treatment cycle is about 21 to about 28 days. In some embodiments, each treatment cycle is 21 to 28 days. In some embodiments, the subject is human.

[0090] Compositions and Kits Disclosed herein are compositions and kits for treating cancer. In some embodiments, the kit comprises a polo-like kinase 1 (PLK1) inhibitor; and a manual providing instructions for administering the PLK1 inhibitor together with an anti-angiogenic agent to a subject to treat metastatic cancer, wherein the subject has not previously been treated for cancer, or the subject has not previously been treated with angiogenesis inhibitors. In some embodiments, the kit comprises an anti-angiogenic agent. The cancer may be, for example, metastatic colorectal cancer, metastatic bladder cancer, metastatic breast cancer, metastatic kidney cancer, metastatic lung cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic prostate cancer, metastatic gastric cancer, metastatic thyroid cancer, metastatic uterine cancer, metastatic kidney cancer, metastatic cervical cancer, metastatic recurrent glioblastoma, or a combination thereof.

[0091] In some embodiments, the instructions include instructions regarding co-administration of a PLK1 inhibitor and an anti-angiogenic agent. In some embodiments, the instructions include instructions regarding sequential administration of a PLK1 inhibitor and an anti-angiogenic agent. In some embodiments, the instructions include instructions regarding orally administering a PLK1 inhibitor. In some embodiments, the instructions include instructions regarding orally administering an anti-angiogenic agent. In some embodiments, the instructions include instructions regarding intravenously administering an anti-angiogenic agent. The instructions may include that the subject has not had prior cancer treatment, for example, has not had prior cancer treatment that includes administration of an angiogenesis inhibitor (e.g., bev). The instructions may include a statement that the subject has undergone at least one prior treatment for the cancer. In some embodiments, the prior treatment does not include use of an anti-angiogenic agent, a PLK1 inhibitor, or both. In some embodiments, the instructions include a statement that the subject is in remission from the cancer. In some embodiments, a subject in remission from cancer is in complete remission (CR) or partial remission (PR).

[0092] In some embodiments, the instructions include administering to the subject each of the antiangiogenic agent and the PLK1 inhibitor at least twice per week in one cycle. In some embodiments, the instructions include administering to the subject each of the antiangiogenic agent and the PLK1 inhibitor at least five times per week in one cycle. In some embodiments, the instructions include administering the antiangiogenic agent, the PLK1 inhibitor, or both in at least 7-day cycles. In some embodiments, each treatment cycle is at least about 21 days. In some embodiments, each treatment cycle is about 21 days to about 28 days, e.g., 28 days. In some embodiments, the instructions include administering the PLK1 inhibitor for at least 4 days of the cycle. In some embodiments, the instructions include not administering the PLK1 inhibitor for at least one day of the cycle. In some embodiments, the instructions include administering the antiangiogenic agent daily. In some embodiments, the instructions include administering the antiangiogenic agent and the PLK1 inhibitor for at least two cycles. The instructions may include instructions for administering the anti-angiogenic agent daily, weekly, biweekly, every three weeks, every four weeks, or monthly.

[0093] The antiangiogenic agent can inhibit VEGF-A, VEGFR-1, VEGFR-2, VEGFR-3, EGFR, HER2, PDGFR family proteins, RAF, Kit (or c-Kit), FLT3, CSF-1R, RET, Abl, Itk, LcK, c-FMS, FGFR family proteins, c-Met, PlGF, TNF-α, IFNs, ILs, bFGF, mTOR, or any combination thereof. Antiangiogenic agents (e.g., angiogenesis inhibitors) include afatinib (Giotrif®), axitinib (Inlyta®), bevacizumab (Avastin®), cabozantinib (Cometriq®), cetuximab (Erbitux®), erlotinib (Tarceva®), everolimus (Afinitor®), gefitinib (Iressa®), imatinib (Gleevec®), lapatinib (Tykerb®), lenalidomide (Revlimid®), lenvatinib mesylate (Lenvima®), necitumumab (Polymerase Chain Reagents (PORC)), and rituximab (Tykerb®). The anti-cancer drug may be cyclophosphamide (Diabetes mellitus), cyclosporine ...

[0094] The PLK1 inhibitor may be selective and / or specific for PLK1. In some embodiments, the PLK1 inhibitor is a dihydropteridinone, a pyridopyrimidine, an aminopyrimidine, a substituted thiazolidinone, a pteridine derivative, a dihydroimidazo[1,5-f]pteridine, a meta-substituted thiazolidinone, a benzylstyryl sulfone analog, a stilbene derivative, or any combination thereof. In some embodiments, the PLK1 inhibitor is onvansertib, BI2536, volasertib (BI6727), GSK461364, AZD1775, CYC140, HMN-176, HMN-214, rigosertib (ON-01910), MLN0905, TKM-080301, TAK-960, or Ro3280. In some embodiments, the PLK1 inhibitor is onvansertib. In some embodiments, the antiangiogenic agent is bevacizumab and the PLK1 inhibitor is onvansertib. 2 ~90mg / m 2 In some embodiments, the instructions include administering the anti-angiogenic agent at a dose between 20 mg and 1200 mg. [Example]

[0095] Certain aspects of the above-described embodiments are disclosed in further detail in the following examples, which are not intended to limit the scope of the disclosure in any way.

[0096] Example 1 Clinical trial of onvansertib in combination with bevacizumab and FOLFIRI This example describes the results (as of July 25, 2022) of a Phase 1b / 2 clinical trial (NCT03829410) to determine the safety and efficacy of onvansertib in combination with FOLFIRI and bevacizumab (the current standard of care for second-line treatment of patients whose disease has relapsed or progressed on first-line treatment with oxaliplatin and a fluoropyrimidine). Patients received FOLFIRI and bevacizumab ("bev") at a standard dose and schedule on days 1 and 5 of a 28-day treatment cycle and orally administered onvansertib once daily on days 1-5 and 15-19 of a 28-day cycle. Patients were examined for disease response assessment at baseline and every 8 weeks during treatment. This trial was conducted to demonstrate that onvansertib safely complements and improves the efficacy of standard of care. The three efficacy endpoints in the phase 2 portion of the study were: (1) primary endpoint: objective response rate (or ORR); (2) secondary endpoints: progression-free survival and other disease response measures, including duration of response; and (3) one exploratory endpoint was the evaluation of the correlation between changes in KRAS mutation burden in circulating tumor DNA and radiographic disease response.

[0097] Table 1 shows study enrollment and patient characteristics. [Table 1] [Table 2] [Table 3]

[0098] Initial partial response rates (PRs) were observed up to 8 months into treatment. The combination of onvansertib and FOLFIR-bev was found to be well tolerated. Neutropenia was the most commonly reported TEAE, with more than half of the observed events being grade 3 or 4. Although 5FU bolus is part of standard FOLFIRI, it appears to increase the toxicity of the regimen, not its efficacy. Early in Part 1b of the study, based on investigator feedback, the protocol was amended to allow for discontinuation of 5FU bolus from the treatment regimen if grade 2 or higher neutropenia was observed. Discontinuation of 5FU bolus was subsequently shown to improve the severity of observed neutropenia, regardless of the use of growth factors.

[0099] The impact of first-line use of BEV on the efficacy of second-line use in prior clinical trials was minimal. In those previous studies (e.g., Hansen et al., Cancers 2021, 13, 1031; Tabernaro et al. Eur J Cancer, 2014, 50, 320-332; Bennouna et al., Lancet Oncol. 2013, 14, 29-37; Van Cutsem et al., J. Clin. Oncol. 2012, 30,3499-3506; Tabenaro et al., Lancet Oncol 2015; 16: 499-508; Beretta et al., Med Oncol (2013) 30:486; Moriwakij et al., Med Oncol (2012) 29:2842-2848), patients who had been previously treated with BEV ("prior BEV") and those who had not been previously treated with BEV ("BEV"). In the "bevacizumab-naive" group, mPFS (mo) was 6.7 months and 6.9 months, and mOS (mo) was 12.5 months and 13.9 months, respectively. The conclusion from previous studies is that in second-line treatment, mPFS and mOS are similar regardless of whether patients were bevacizumab-naive or had been previously treated with bevacizumab in first-line therapy. Regarding ORR, there was a 12-20% increase. Surprisingly, in this study (NCT03829410 "Onvansertib in Combination With FOLFIRI and Bevacizumab for Second-Line Treatment of Metastatic Colorectal Cancer Patients With a Kras Mutation"), we found that the "bevacizumab-naive" subset exhibited much greater ORR and mPFS than expected (see Figures 1 and 2). Regarding ORR among 33 patients previously treated with bevacizumab, 8 / 35 (22.9%) responded (CI: 10.4%-40.1%). The ORR of 15 bevacizumab-naive patients was 9 / 13 responders (69.2%) (CI: 38.6%-90.9%).Thus, the ORR in antiangiogenic-naïve patients was twice that in antiangiogenic-naïve patients, with an odds ratio of 0.14 (0.03-0.56, p=0.0049). PFS in bevacizumab-naïve patients also showed a striking improvement in bevacizumab-naïve patients. In this study, mPFS was 7.8 months in bevacizumab-naïve patients and 13.5 months in bevacizumab-naïve patients (p=0.14) (Figure 1). [Table 4]

[0100] ORR was consistently higher in bevacizumab-naive patients across all characteristics. No single patient characteristic explained the observed ORR differences. As shown in Figure 3 and Table 5, the lowest ORR in the bevacizumab-naive group was 50% and the highest was 80% at RP2D. The ORR range for bevacizumab-experienced patients was 14% to 31.3%, depending on the variable. [Table 5] The results presented here demonstrate striking improvements in multiple outcomes (e.g., ORR, PFS) in patients with metastatic cancer treated with the combination of onvansertib and bevacizumab in bevacizumab-naive patients. Without being bound by any particular theory, it is believed that there is a synergistic effect between onvansertib and bevacizumab in the bevacizumab-naive setting.

[0101] Example 2 Treatment benefits for Bev-naive patients Figure 4A shows the best radiographic response and duration of response for 66 evaluable patients (as of June 16, 2023). Figures 4A-4B show that the combination of FOLFIRI, bevacizumab, and ombansertib, the current standard of care (SoC) for patients with metastatic colorectal cancer, achieved higher response rates in bevacizumab-naive patients. In Figure 4A, **Historical controls were obtained from Bennouna et al., Lancet Oncol 2013;14:29-37; Giessen et al., Acta Oncologica 2015;54:187-193; Cremolini et al., Lancet Oncol 2020;21:497-507; Antoniotti et al., Correspondence Lancet Oncol June 2020. Giantonio et al., 2007, J Clin Oncol 25:1539-1544; Moriwaki et al., Med Oncol 2012;29:2842-2848; Beretta et al., Med Oncol 2013;30:486. Furthermore, Bev-naive patients demonstrated more durable responses (Figures 5A-5B). The PFS of treated patients was found to be superior to that of SoC historical controls, especially in BEV-naive patients. Progression-free survival from 66 evaluable patients (as of June 16, 2023) is shown in Figure 6 (Figure 6). ** Historical controls were Bennouna et al., Lancet Oncol 2013; 14: 29-37; Giessen et al., Acta Oncologica, 2015, 54: 187-193; Cremolini et al., Lancet Oncol 2020, 21: 497-507; Antoniotti et al., Correspondence Lancet Oncol June 2020. Giantonio et al., 2007, J Clin Oncol 25:1539-1544; Moriwaki et al., Med Oncol, 2012, 29:2842-2848; Beretta et al, Med Oncol 2013, 30:486). It was also observed that Bev-naive patients showed deeper tumor regression. Figure 7 shows baseline data across all doses. * The change in tumor size from the initial stage (as of June 16, 2023) is shown.

[0102] Example 3 Onvansertib plus bevacizumab is an effective combination in a KRAS-mutant CRC xenograft model Three KRAS mutant xenograft models (HCT116, LoVo, and SW620) were treated with vehicle, onvansertib, bevacizumab, or the combination of onvansertib and bevacizumab. As shown in Figure 8, the combination of onvansertib and bevacizumab demonstrated antitumor activity in the three models, including: (1) tumor stabilization was observed upon combination treatment; and (2) the combination demonstrated significantly superior antitumor activity compared to the single agents. Figure 9 demonstrates that the combination of onvansertib and bevacizumab reduced tumor vascularization, as indicated by the observation that tumors in mice treated with the combination of onvansertib and bevacizumab were smaller and appeared paler (less vascularized). Without being bound by any particular theory, it is believed that prior BEV treatment modulates genetic pathways that may confer resistance to BEV and onvansertib. The non-limiting method depicted in Figure 10 can be used to identify potential mechanisms of treatment resistance in patients with KRAS mutant mCRC exposed to BEV.

[0103] In at least some of the previously described embodiments, one or more elements used in one embodiment may be used interchangeably in another embodiment unless such substitution is technically infeasible. Those skilled in the art will appreciate that various other omissions, additions, and modifications may be made to the methods and structures described above without departing from the scope of the claimed subject matter. All such modifications and variations are intended to fall within the scope of the subject matter defined by the appended claims. With respect to the use of substantially any plural and / or singular term herein, those of ordinary skill in the art can interpret the plural to the singular and / or the singular to the plural as appropriate to the context and / or usage. The various singular / plural permutations may be explicitly indicated herein for the sake of brevity. As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. As used herein, "or" is intended to include "and / or" unless the context clearly indicates otherwise.

[0104] Those skilled in the art will generally understand that the terms used in this specification, and particularly in the appended claims (e.g., the body of the appended claims), are generally intended as "open" terms (e.g., the term "comprising" should be interpreted as "including, but not limited to," the term "having" should be interpreted as "having at least," the term "including" should be interpreted as "including, but not limited to," etc.). Those skilled in the art will further understand that where a specific number is intended in the introductory claim recitation, such intention will be expressly set forth in the claim; and, absent such recitation, no such intention exists. For example, as an aid to understanding, the following appended claims may include the use of the introductory phrases "at least one" and "one or more" to introduce claim recitations. However, the use of such phrases should not be construed as meaning that introducing a claim recitation with the indefinite article "a" or "an" means limiting a particular claim containing such an introduced claim recitation to embodiments containing only one such recitation, even if the same claim includes the introductory phrase "one or more" or "at least one" and an indefinite article such as "a" or "an" (e.g., "a" and / or "an" should be interpreted to mean "at least one" or "one or more"), nor should the use of a definite article to introduce a claim recitation be construed as meaning that a particular claim containing such an introduced claim recitation is limited to embodiments containing only one such recitation. Additionally, even if a specific number is explicitly stated in an introduced claim recitation, those skilled in the art will recognize that such a recitation should be interpreted to mean at least the recited number (e.g., a recitation simply stating "two recitations" without any other modifiers generally means at least two recitations, or more than two recitations).Furthermore, when notation similar to "at least one of A, B, and C, etc." is used, such syntax is generally intended in the sense that one of ordinary skill in the art would understand the notation (e.g., "a system having at least one of A, B, and C" includes, but is not limited to, systems having A alone, B alone, C alone, both A and B, both A and C, both B and C, and / or all of A, B, and C, etc.). When notation similar to "at least one of A, B, or C, etc." is used, such syntax is generally intended in the sense that one of ordinary skill in the art would understand the notation (e.g., "a system having at least one of A, B, or C" includes, but is not limited to, systems having A alone, B alone, C alone, both A and B, both A and C, both B and C, and / or all of A, B, and C, etc.). Furthermore, those skilled in the art will understand that virtually any alternative word and / or phrase expressing two or more alternative terms, whether in the specification, claims, or drawings, should be understood to contemplate the possibility of including one of those terms, either of those terms, or both of those terms.

[0105] Additionally, where features or aspects of the disclosure are described in terms of a Markush group, those skilled in the art will recognize that the disclosure is thereby also described in terms of any individual member or subgroup of members of the Markush group. As will be understood by those skilled in the art, all ranges disclosed herein encompass all possible subranges and combinations of subranges for all purposes, including in terms of providing a written description. Any recited range can be readily recognized as fully descriptive and allowing for the same range to be divided into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range set forth herein can be readily divided into a lower third, middle third, and upper third, etc. As will also be understood by those skilled in the art, all terms such as "up to," "at least," "greater than," "less than," etc., are inclusive of the recited number and refer to ranges that can be subsequently divided into subranges as described above. Finally, as will be understood by those skilled in the art, ranges include individual members. Thus, for example, a group having 1 to 3 items refers to groups having 1, 2, or 3 items. Similarly, a group having 1 to 5 items refers to groups having 1, 2, 3, 4, or 5 items. While various aspects and embodiments have been disclosed herein, other aspects and embodiments will be apparent to those skilled in the art. The various aspects and embodiments disclosed herein are for purposes of illustration and not limitation, with the true scope and spirit being indicated by the following claims.

Claims

1. A method for treating metastatic cancer in a subject, comprising administering to the subject a PLK1 inhibitor and an anti-angiogenic agent, thereby reducing or inhibiting the progression of the metastatic cancer, wherein the subject has no history of cancer treatment or the subject has no history of treatment involving the inhibition of angiogenesis.

2. A method for treating metastatic cancer in a subject, comprising administering to the subject a PLK1 inhibitor and an anti-angiogenic agent, thereby reducing or inhibiting the progression of the metastatic cancer, wherein the subject is known to have no history of cancer treatment or the subject is known to have no history of treatment involving the inhibition of angiogenesis.

3. 1. A method of treating metastatic cancer in a subject, comprising: Identifying subjects with metastatic cancer who have not received prior cancer treatment; and administering to said subject a PLK1 inhibitor and an anti-angiogenic agent, thereby reducing or inhibiting the progression of said metastatic cancer. A method comprising:

4. 1. A method of treating metastatic cancer in a subject, comprising: Identifying a subject with metastatic cancer who has not received prior treatment involving the inhibition of angiogenesis; and administering to said subject a PLK1 inhibitor and an anti-angiogenic agent, thereby reducing or inhibiting the progression of said metastatic cancer. A method comprising:

5. The method of any one of claims 1 to 4, comprising administering to the subject chemotherapy, the PLK1 inhibitor, and the anti-angiogenic agent.

6. The method of any one of claims 1 to 4, wherein the subject is chemotherapy naive.

7. The method of any one of claims 1 to 4, wherein the subject has not received chemotherapy for the metastatic cancer.

8. 8. The method of any one of claims 5 to 7, wherein the chemotherapy comprises treatment with FOLFIRI, abiraterone, FOLFOX, an anti-EGFR agent, a KRAS-directed inhibitor, gemcitabine, Abraxane, nanoliposomal irinotecan, 5-FU, FOLFIRINOX, FOLFOXIRI, or a combination thereof.

9. The method of any one of claims 1 to 8, wherein administration of the PLK1 inhibitor and the anti-angiogenic agent synergistically reduces or inhibits progression of the metastatic cancer compared to the additive effects of PLK1 inhibitor treatment alone, anti-angiogenic agent treatment alone, and / or PLK1 inhibitor treatment alone and anti-angiogenic agent treatment alone.

10. The method of any one of claims 1 to 9, wherein administration of the PLK1 inhibitor and the anti-angiogenic agent improves one or more therapeutic effects in the subject compared to a control or baseline.

11. 11. The method of claim 10, wherein administration of the PLK1 inhibitor and the anti-angiogenic agent reduces the oncogenic allele load in the subject compared to a subject who has previously undergone treatment involving inhibition of angiogenesis.

12. A method for improving objective response rate (ORR), progression-free survival (PFS), or both in the treatment of metastatic cancer, comprising administering a PLK1 inhibitor and an anti-angiogenic agent to a subject suffering from metastatic cancer, thereby improving the ORR and / or PFS of the subject, wherein the subject has no prior cancer treatment or the subject has no prior treatment involving inhibition of angiogenesis.

13. A method for improving objective response rate (ORR), progression-free survival (PFS), or both in the treatment of metastatic cancer, comprising administering a PLK1 inhibitor and an anti-angiogenic agent to a subject suffering from metastatic cancer, thereby improving the ORR and / or PFS of the subject, wherein the subject is known to have no prior cancer treatment or the subject is known to have no prior treatment involving the inhibition of angiogenesis.

14. The method of any one of claims 12-13, comprising identifying the subject as having metastatic cancer and having no prior cancer treatment.

15. The method of any one of claims 12-13, comprising identifying the subject as having metastatic cancer and having no prior treatment involving inhibition of angiogenesis.

16. 16. The method of any one of claims 12 to 15, wherein administration of the PLK1 inhibitor and the anti-angiogenic agent synergistically improves ORR and / or PFS in the subject compared to PLK1 inhibitor treatment alone, anti-angiogenic agent treatment alone, and / or the additive effects of PLK1 inhibitor treatment alone and anti-angiogenic agent treatment alone.

17. The method of any one of claims 12 to 15, wherein administration of the PLK1 inhibitor and the anti-angiogenic agent improves one or more therapeutic effects in the subject compared to a control or baseline, and optionally the one or more therapeutic effects include tumor size derived from metastatic cancer, objective response rate (ORR), duration of response, time to response, progression-free survival (PFS), overall survival (OS), disease control rate (DCR), oncogenic allele burden, or a combination thereof.

18. 16. The method of any one of claims 12-15, wherein administration of the PLK1 inhibitor and the anti-angiogenic agent improves ORR in the subject, improves PFS in the subject, improves OS in the subject, improves DCR in the subject, reduces oncogenic allele burden in the subject, or a combination thereof, compared to a subject that has received a previous treatment comprising inhibition of angiogenesis.

19. 19. The method of any one of claims 12-18, wherein administration of the PLK1 inhibitor and the anti-angiogenic agent improves ORR, PFS, or both in the subject by at least 50% compared to a subject that has received a previous treatment comprising inhibition of angiogenesis.

20. 20. The method of any one of claims 1 to 19, wherein the metastatic cancer is metastatic colorectal cancer, metastatic bladder cancer, metastatic breast cancer, metastatic kidney cancer, metastatic lung cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic prostate cancer, metastatic gastric cancer, metastatic thyroid cancer, metastatic uterine cancer, metastatic kidney cancer, metastatic cervical cancer, metastatic recurrent glioblastoma, or a combination thereof.

21. The method of any one of claims 1 to 20, wherein the PLK1 inhibitor is selective and / or specific for PLK1.

22. 22. The method of any one of claims 1 to 21, wherein the PLK1 inhibitor is onvansertib, BI2536, volasertib (BI 6727), GSK461364, AZD1775, CYC140, HMN-176, HMN-214, rigosertib (ON-01910), MLN0905, TKM-080301, TAK-960, or Ro3280.

23. 23. The method of any one of claims 1 to 22, wherein the subject has not previously undergone treatment including administration of an angiogenesis inhibitor, and optionally, the angiogenesis inhibitor is the same as the anti-angiogenic agent.

24. The method of any one of claims 1 to 23, wherein the anti-angiogenic agent is bevacizumab.

25. The method of any one of claims 23 to 24, wherein the angiogenesis inhibitor is bevacizumab.

26. The method of any one of claims 1 to 25, wherein the PLK1 inhibitor and the anti-angiogenic agent are administered simultaneously.

27. The method of any one of claims 1 to 25, wherein the PLK1 inhibitor and the anti-angiogenic agent are administered sequentially.

28. The method of claim 27, wherein the PLK1 inhibitor is administered before administration of the anti-angiogenic agent, and optionally, the PLK1 inhibitor is administered before administration of the anti-angiogenic agent on all days that the PLK1 inhibitor and the anti-angiogenic agent are administered to the subject.

29. 29. The method of claim 28, wherein the PLK1 inhibitor is administered about 30 minutes to about 5 hours before administration of the anti-angiogenic agent on a given day.

30. The method according to any one of claims 1 to 29, wherein the administration of the PLK1 inhibitor is oral administration, and the administration of the anti-angiogenic agent is intravenous or oral administration.

31. The method of any one of claims 1 to 30, wherein the antiangiogenic agent and the PLK1 inhibitor are each administered to the subject at least twice or at least five times per week in one cycle.

32. 32. The method of any one of claims 1-31, wherein the antiangiogenic agent, the PLK1 inhibitor, or both are administered in cycles of at least 7 days; optionally, each treatment cycle is at least about 21 days; and further optionally, each treatment cycle is from about 21 days to about 28 days.

33. The method of any one of claims 31 to 32, wherein the PLK1 inhibitor is administered on at least 4 days of the cycle.

34. The method of any one of claims 32-33, wherein the PLK1 inhibitor is not administered on at least one day of the cycle.

35. 35. The method of any one of claims 1 to 34, wherein the anti-angiogenic agent is administered daily, weekly, biweekly, every three weeks, every four weeks, or monthly.

36. The method of any one of claims 31 to 35, wherein the subject receives at least two cycles of the antiangiogenic agent and the PLK1 inhibitor.

37. 37. The method of any one of claims 1 to 36, wherein the antiangiogenic agent is bevacizumab and the PLK1 inhibitor is onvansertib.

38. Onvansertib 12 mg / m 2 ~90 mg / m 2 38. The method of claim 37, wherein the

39. 39. The method of any one of claims 37-38, wherein bevacizumab is administered at about 1 mg / kg to 20 mg / kg, optionally wherein bevacizumab is administered at about 5 mg / kg, about 7.5 mg / kg, about 10 mg / kg, or about 15 mg / kg.

40. 40. The method of any one of claims 1-39, wherein the subject has had at least one prior cancer treatment, optionally wherein the prior treatment did not include use of an anti-angiogenic agent, a PLK1 inhibitor, or both.

41. The method of any one of claims 1 to 40, wherein the one or more subjects have a history of cancer remission.

42. 42. The method of claim 41, wherein the remission history is complete remission (CR).

43. 42. The method of claim 41, wherein the remission history is partial remission (PR).

44. The method of any one of claims 1 to 43, further comprising one or more of: (1) determining the cancer status of the one or more subjects; (2) determining the responsiveness of the one or more subjects to PLK1 inhibitor treatment; and (3) administering one or more cancer therapeutic agents or therapies to the one or more subjects.

45. 45. The method of any one of claims 1 to 44, wherein the one or more subjects are human.

46. 45. The method of any one of claims 1-44, wherein the reduction or inhibition of cancer progression comprises an inhibition of the growth of one or more tumors in the one or more subjects and / or a reduction in the number of cancer cells detected in the one or more subjects by at least about 25%, 30%, 40%, 50%, 60%, or 70% compared to an untreated subject.

47. The method of any one of claims 1 to 44, wherein the reduction or inhibition of cancer progression comprises an inhibition of the growth of one or more tumors in the one or more subjects and / or a reduction in the number of cancer cells detected in the one or more subjects by at least about 25%, 30%, 40%, 50%, 60%, or 70% compared to the one or more subjects prior to administration of the PLK1 inhibitor and the anti-angiogenic agent.

48. 48. The method of claim 46 or 47, wherein the growth of at least one of the one or more tumors in the one or more subjects is reduced by at least about 25%, 30%, 40%, 50%, 60%, or 70% after one or more cycles of treatment.

49. 48. The method of claim 46 or 47, wherein the size / volume of at least one of the one or more tumors in the one or more subjects is reduced by at least about 25%, 30%, 40%, 50%, 60%, or 70% after one or more cycles of treatment.

50. 50. The method of any one of claims 44-49, wherein the one or more cancer therapeutic agents or therapies comprise FOLFIRI, abiraterone, FOLFOX, an anti-EGFR agent, a KRAS-directed inhibitor, gemcitabine, abraxane, nanoliposomal irinotecan, 5-FU, or a combination thereof, wherein the anti-EGFR agent is optionally cetuximab, and the KRAS-directed inhibitor is optionally a G12C inhibitor, a G12D inhibitor, or a combination thereof.

51. 51. The method of any one of claims 44-50, wherein determining the responsiveness of the one or more subjects comprises determining whether the subject is a responder to the treatment, whether the one or more subjects is in or heading towards complete recovery (CR), or whether the one or more subjects is in or heading towards partial remission (PR).

52. 52. The method of any one of claims 44-51, wherein determining the subject's responsiveness comprises determining the subject's objective response rate (ORR), duration of response, time to response, progression-free survival (PFS), overall survival (OS), disease control rate (DCR), oncogenic allele burden, or a combination thereof.

53. 53. The method of any one of claims 44-52, wherein determining the responsiveness of the one or more subjects comprises determining whether the one or more subjects have a partial response to the treatment, whether the subject has a complete response to the treatment, whether the subject has stable disease (SD), or whether the subject has progressive disease (PD).

54. Polo-like kinase 1 (PLK1) inhibitors, and a manual providing instructions for administering the PLK1 inhibitor in combination with an anti-angiogenic agent to a subject with metastatic cancer, wherein the subject has not received prior cancer treatment or the subject has not received prior treatment involving the inhibition of angiogenesis; Includes a kit.

55. Polo-like kinase 1 (PLK1) inhibitors, and a manual providing instructions for administering the PLK1 inhibitor in conjunction with an anti-angiogenic agent to a subject with metastatic cancer, wherein the subject is known to have no prior cancer treatment, or the subject is known to have no prior treatment involving the inhibition of angiogenesis; Includes a kit.

56. 55. The kit of claim 54, wherein the metastatic cancer is metastatic colorectal cancer, metastatic bladder cancer, metastatic breast cancer, metastatic kidney cancer, metastatic lung cancer, metastatic ovarian cancer, metastatic pancreatic cancer, metastatic prostate cancer, metastatic gastric cancer, metastatic thyroid cancer, metastatic uterine cancer, metastatic kidney cancer, metastatic cervical cancer, metastatic recurrent glioblastoma, or a combination thereof.

57. 57. The kit of any one of claims 54 to 56, wherein the instructions include instructions for co-administering the PLK1 inhibitor and the anti-angiogenic agent.

58. The kit of any one of claims 54 to 56, wherein the instructions include instructions for administering the PLK1 inhibitor and the anti-angiogenic agent sequentially.

59. The kit of any one of claims 54 to 58, wherein the instructions include (1) instructions for administering the PLK1 inhibitor orally, (2) instructions for administering the anti-angiogenic agent orally, (3) instructions for administering the anti-angiogenic agent intravenously, or any combination thereof.

60. 60. The kit of any one of claims 54-59, wherein the instructions include a statement that the subject has not previously received treatment including administration of an angiogenesis inhibitor, and optionally, the angiogenesis inhibitor is the same as the anti-angiogenic agent.

61. The kit of any one of claims 54 to 60, wherein the instructions include instructions for administering each of the antiangiogenic agent and the PLK1 inhibitor to the subject at least twice or at least five times per week in one cycle.

62. 62. The kit of any one of claims 54-61, wherein the instructions include instructions for administering the antiangiogenic agent, the PLK1 inhibitor, or both, in cycles of at least 7 days, optionally with each treatment cycle being at least about 21 days, and further optionally with each treatment cycle being from about 21 days to about 28 days.

63. 63. The kit of claim 62, wherein the instructions include instructions for administering the PLKl inhibitor on at least 4 days of the cycle.

64. The kit of any one of claims 62-63, wherein the instructions include instructions regarding not administering the PLK1 inhibitor on at least one day of the cycle.

65. 65. The kit of any one of claims 54-64, wherein the instructions include instructions for administering the anti-angiogenic agent daily, weekly, biweekly, every three weeks, every four weeks, or monthly.

66. The kit of any one of claims 61 to 65, wherein the instructions include instructions for administering the antiangiogenic agent and the PLK1 inhibitor for at least two cycles.

67. The kit of any one of claims 54 to 66, wherein the anti-angiogenic agent is bevacizumab.

68. The kit of any one of claims 54 to 67, wherein the PLK1 inhibitor is selective and / or specific for PLK1.

69. The kit of any one of claims 54 to 68, wherein the PLK1 inhibitor is onvansertib.

70. The above description is based on the use of onvansertib at 12 mg / m 2 ~90 mg / m 2 70. The kit of claim 69, comprising instructions for administering the

71. The kit of any one of claims 54 to 70, further comprising an anti-angiogenic agent.

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