Cancer treatment using MTDP inhibitors and PLK1 inhibitors

Abstract

Provided are methods, compositions, and kits for treating cancer in a subject, which can include administering to the subject an MTDP inhibitor (e.g., paclitaxel) and a PLK1 inhibitor (e.g., onvansertib) in amounts sufficient to inhibit or reduce the progression of the cancer.

Description

[Technical Field]

[0001] This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Patent Application No. 63 / 405,308, filed September 9, 2022, the contents of which are incorporated herein by reference in their entirety for all purposes.

[0002] Field This application relates generally to the treatment of cancer. More specifically, a combination therapy for treating cancer using a microtubule depolymerizing (MTDP) inhibitor in combination with a polo-like kinase 1 (PLK1) inhibitor is provided. [Background technology]

[0003] Related Field Description PLK1 is a serine / threonine kinase and the best-characterized member of a family of five closely related regulatory proteins. PLK-1 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.

[0004] Tubulin-targeting drugs are a diverse class of compounds that can bind to tubulin dimers. These compounds include natural or synthetic compounds that can bind to the taxane, colchicine, vinca, or other domains of tubulin dimers. These compounds can affect microtubule dynamics and / or polymerization. Some potent antimitotic agents can induce apoptosis, making them potential cancer therapeutics. Examples of tubulin-targeting drugs include MTDP inhibitors such as paclitaxel, docetaxel, epothilones, ixabepilone, and discodermolide. Summary of the Invention

[0005] There is a need to develop effective treatments for cancer patients, including those who are resistant to MTDP inhibitor treatment.

[0006] overview Included are methods, compositions, and kits for treating cancer. Some embodiments provide a method of treating cancer, the method comprising administering a microtubule depolymerizing (MTDP) inhibitor and a polo-like kinase 1 (PLK1) inhibitor to a subject with cancer, thereby inhibiting or reducing the progression of the cancer in the subject. In some embodiments, the subject has a hematological cancer or a solid cancer. The cancer may be breast cancer, which may be invasive breast cancer, unresectable breast cancer, locally advanced breast cancer, metastatic breast cancer, inflammatory breast cancer, triple-negative breast cancer (TNBC), hormone receptor / growth factor receptor-negative breast cancer, HER2-negative breast cancer, and / or hormone receptor-negative breast cancer. In some embodiments, the subject has TNBC. In some embodiments, the method includes administering to a subject with cancer a microtubule depolymerizing (MTDP) inhibitor and a polo-like kinase 1 (PLK1) inhibitor ... 3 Approximately 1000+ neutrophils per 1mm 3 In some embodiments, the subject is identified as having greater than about 100,000 platelets per mm, a total bilirubin of about 1.5 times the upper limit of institutional reference, and a serum creatinine glomerular filtration rate of less than about 1.5 times the upper limit of institutional reference or 60 mL / min, or a combination thereof. 3 Approximately 1000+ neutrophils per 1mm 3have more than about 100,000 platelets per minute, a total bilirubin level of about 1.5 times the upper limit of the facility's reference range, and a serum creatinine glomerular filtration rate of less than about 1.5 times the upper limit of the facility's reference range or 60 mL / min.

[0007] The PLK1 inhibitor and the MTDP inhibitor can be administered simultaneously. The PLK1 inhibitor and the MTDP inhibitor can also be administered sequentially. In some embodiments, the PLK1 inhibitor is administered orally, and the subject fasts for more than about 30 minutes before administration and fasts for about 4 hours after administration. In some embodiments, the MTDP inhibitor is administered intravenously, optionally through an in-line filter with a microporous membrane of 0.22 microns or less, and the subject is administered: (i) about 20 mg of dexamethasone orally about 12 hours and about 6 hours before administration or about 12 mg of dexamethasone intravenously about 60 minutes before administration; (ii) about 50 mg of diphenhydramine intravenously about 30 to about 60 minutes before administration; and / or (iii) about 300 mg of cimetidine intravenously about 30 to about 60 minutes before administration or about 20 mg of famotidine intravenously about 30 to about 60 minutes before administration.

[0008] The inhibition of cancer progression may be greater than the combined inhibition of progression caused by the MTDP inhibitor and the PLK1 inhibitor alone. In some embodiments, the subject achieves a complete response. In some embodiments, the subject has previously been treated with an MTDP inhibitor or a PLK1 inhibitor. In some embodiments, the subject has failed to respond to treatment with an MTDP inhibitor or a PLK1 inhibitor alone. In some embodiments, the subject is known to be resistant to MTDP inhibitor or PLK1 inhibitor therapy. In some embodiments, the MTDP inhibitor and the PLK1 inhibitor are each administered to the subject in 28-day cycles. In some embodiments, the MTDP inhibitor is administered to the subject in a 28-day cycle, and the PLK1 inhibitor is administered to the subject in a 28-day cycle, comprising about 14-28 days of administration and about 0-14 days of non-administration, preferably about 15-27 days of administration and about 1-13 days of non-administration, preferably about 16-26 days of administration and about 2-12 days of non-administration, preferably about 17-25 days of administration and about 3-11 days of non-administration, preferably about 18-24 days of administration and about 4-10 days of non-administration, preferably about 19-23 days of administration and about 5-9 days of non-administration, preferably about 20-22 days of administration and about 6-8 days of non-administration, preferably about 21 days of administration and about 7 days of non-administration, preferably the administration days are consecutive. In some embodiments, the MTDP inhibitor, the PLK1 inhibitor, or both are administered in a 28-day cycle.

[0009] Each treatment cycle can be at least about 28 days, or is at least about 28 days. In some embodiments, each treatment cycle is from about 14 days to about 28 days. The PLK1 inhibitor can be administered on at least 4 days of the cycle. In some embodiments, the PLK1 inhibitor is not administered on at least 1 day of the cycle. In some embodiments, the MTDP inhibitor is administered once daily or twice daily. In some embodiments, the MTDP inhibitor is administered once daily. In some embodiments, the subject receives at least two cycles of the MTDP inhibitor and the PLK1 inhibitor. MTDP inhibitors include, for example, paclitaxel, docetaxel, acetyltaxol, paclitaxel; lutetium Lu 177 Bipibotide tetraxetan; 7-hexanoyltaxol; Cabazitaxel; Larotaxel; Mirataxel; Ortataxel; Tesetaxel; Taxoplexin; Opaxil; Taxoplexin (DHA-paclitaxel); Poly(L-glutamic acid)-paclitaxel; Abraxane; SB-T-1214; SB-T1216; SB-T121602; SB-T-12854; DHA-SB-T 1214; abeotaxane, the abeotaxane optionally being abeotaxane 15a.2; docetaxel-d9-t-Boc; docetaxel-f3-t-Boc; cabazitaxel-7,10-d6; poly(glutamyl-glutamic acid)-taxane conjugate; or a pharmaceutically acceptable salt, hydrate, solvate, or prodrug thereof; or any combination thereof. In some embodiments, the MTDP inhibitor is paclitaxel. In some embodiments, the MTDP inhibitor is about 48 mg / m 2 Body surface area ~ approx. 80mg / m 2 A body surface area of ​​paclitaxel is administered to the subject.

[0010] The PLK1 inhibitor can be, for example, onvansertib (NMS-P937), BI2536, volasertib (BI 6727), GSK461364, adavosertib (AZD1775), CYC140, HMN-176, HMN-214, rigosertib (ON-01910), MLN0905, TKM-080301, TAK-960, GTPL10072, Ro3280; or a pharmaceutically acceptable salt, hydrate, solvate, or prodrug thereof; and any combination thereof. In some embodiments, the PLK1 inhibitor is onvansertib. In some embodiments, onvansertib is administered at a dose of about 6 mg / m 2 Body surface area ~ approx. 24mg / m 2 In some embodiments, onvansertib is administered at a dose of 9 mg / m 2 Body surface area, 12mg / m 2 body surface area, and 24 mg / m 2 It is administered in body surface area doses.

[0011] In some embodiments, the subject has received at least one prior cancer treatment. In some embodiments, the prior treatment does not include the use of an MTDP inhibitor, a PLK1 inhibitor, or both; optionally, the PLK1 inhibitor is onvansertib. In some embodiments, the subject has been in remission of the cancer, e.g., complete remission (CR) or partial remission (PR). In some embodiments, the method further comprises determining the subject's cancer status. The method can, for example, further comprise determining the subject's responsiveness to treatment with an MTDP inhibitor and a PLK1 inhibitor. In some embodiments, the method further comprises administering one or more cancer therapeutic agents or therapies for the cancer. In some embodiments, the subject is human.

[0012] Also disclosed herein is a method of sensitizing cancer cells to a microtubule depolymerization (MTDP) inhibitor, the method comprising contacting the cancer cells with a composition comprising a polo-like kinase 1 (PLK1) inhibitor, thereby sensitizing the cancer cells to the MTDP inhibitor. In some embodiments, the PLK1 inhibitor is onvansertib and / or the MTDP inhibitor is paclitaxel. The method can include contacting the cancer cells with the composition in vitro, ex vivo, and / or in vivo. In some embodiments, contacting the cancer cells with the composition is in a subject. In some embodiments, the subject is known to have failed to respond to or be resistant to an MTDP inhibitor. In some embodiments, the subject has previously been treated with an MTDP inhibitor. The subject can be a mammal, e.g., a human. The method can include determining sensitization of the cancer cells to the MTDP inhibitor after contacting with the composition.

[0013] In some embodiments, the method includes contacting the cancer cells with an MTDP inhibitor. The contacting of the cancer cells with the MTDP inhibitor can be performed in a subject. The method can include determining the subject's response to the MTDP inhibitor. In some embodiments, the contacting of the cancer cells with the MTDP inhibitor is simultaneous with or after contacting the cancer cells with the composition. In some embodiments, the subject is treated with 1 mm 3 Approximately 1000+ neutrophils per 1mm 3 have more than about 100,000 platelets per minute, a total bilirubin level of about 1.5 times the upper limit of the facility's reference range, and a serum creatinine glomerular filtration rate of less than about 1.5 times the upper limit of the facility's reference range or 60 mL / min.

[0014] Also disclosed are kits comprising a polo-like kinase 1 (PLK1) inhibitor; and a manual providing instructions for administering the PLK1 inhibitor in combination with a microtubule depolymerizing (MTDP) inhibitor to a subject in need thereof to treat cancer. The cancer can be, for example, a hematological cancer or a solid cancer, and optionally the cancer is breast cancer, which is optionally invasive breast cancer, unresectable breast cancer, locally advanced breast cancer, metastatic breast cancer, inflammatory breast cancer, triple-negative breast cancer (TNBC), hormone receptor / growth factor receptor-negative breast cancer, HER2-negative breast cancer, and / or hormone receptor-negative breast cancer. In some embodiments, the cancer is TNBC. In some embodiments, the subject is treated with a 1mm 3 Approximately 1000+ neutrophils per 1mm 3 have platelets greater than approximately 100,000 per minute, total bilirubin approximately 1.5 times the upper limit of the facility's reference range, and a serum creatinine glomerular filtration rate less than approximately 1.5 times the upper limit of the facility's reference range or 60 mL / min.

[0015] In some embodiments, the PLK1 inhibitor is onvansertib and / or the MTDP inhibitor is paclitaxel. The instructions may include instructions for co-administering the PLK1 inhibitor and the MTDP inhibitor simultaneously. In some embodiments, the instructions include instructions for co-administering the PLK1 inhibitor and the MTDP inhibitor sequentially. In some embodiments, the instructions include instructions for orally administering the PLK1 inhibitor and the MTDP inhibitor, wherein the subject fasts for more than about 30 minutes prior to administration and the subject fasts for about 4 hours after administration. In some embodiments, the instructions include instructions for administering the MTDP inhibitor intravenously, preferably through an in-line filter with a microporous membrane of 0.22 microns or less, and prior to administration, the subject is administered: (i) about 20 mg of dexamethasone orally about 12 hours and about 6 hours prior or about 12 mg of dexamethasone intravenously about 60 minutes prior; (ii) about 50 mg of diphenhydramine intravenously about 30 minutes to about 60 minutes prior; and (iii) about 300 mg of cimetidine intravenously about 30 minutes to about 60 minutes prior or about 20 mg of famotidine intravenously about 30 minutes to about 60 minutes prior.

[0016] In some embodiments, the instructions include a description that the subject has been previously treated with an MTDP inhibitor or a PLK1 inhibitor. In some embodiments, the instructions include a description that the subject did not respond to treatment with a single MTDP inhibitor or PLK1 inhibitor. In some embodiments, the instructions include a description that the subject is known to be resistant to MTDP inhibitor or PLK1 inhibitor therapy. In some embodiments, the instructions include a description of administering each of paclitaxel and onvansertib to the subject in a 28-day cycle. In some embodiments, the instructions include a description of administering paclitaxel and onvansertib to the subject in a cycle of at least five times per week. In some embodiments, the instructions include a description of administering the MTDP inhibitor, onvansertib, or both in a cycle of at least 7 days. In some embodiments, each treatment cycle is at least about 21 days. In some embodiments, each treatment cycle is from about 14 days to about 28 days. In some embodiments, the instructions include administering onvansertib for 28 days, including about 14-28 days of administration and about 0-14 days of non-administration, preferably about 15-27 days of administration and about 1-13 days of non-administration, preferably about 16-26 days of administration and about 2-12 days of non-administration, preferably about 17-25 days of administration and about 3-11 days of non-administration, preferably about 18-24 days of administration and about 4-10 days of non-administration, preferably about 19-23 days of administration and about 5-9 days of non-administration, preferably about 20-22 days of administration and about 6-8 days of non-administration, preferably about 21 days of administration and about 7 days of non-administration, preferably consecutive days of administration. In some embodiments, the instructions include not administering onvansertib on at least one day of the cycle. In some embodiments, the instructions include administering an MTDP inhibitor for 28 days. The MTDP inhibitor may be a reversible MTDP inhibitor. In some embodiments, the MTDP inhibitor is paclitaxel or a pharmaceutically acceptable salt, hydrate, solvate, or prodrug thereof.

[0017] In some embodiments, the description provides about 3 mg / m of onvansertib. 2 Body surface area ~ approx. 24mg / m 2 and / or administering paclitaxel at a dose of about 48 mg / m 2 Body surface area ~ approx. 80mg / m 2 In some embodiments, onvansertib is administered at a dose of 9 mg / m 2 Body surface area, 12mg / m 2 body surface area, and 24 mg / m 2 The kit is administered in a dose of 100 mg / kg body surface area. In some embodiments, the subject has previously been treated for cancer. In some embodiments, the prior treatment does not include the use of an MTDP inhibitor, onvansertib, or both. In some embodiments, the subject has been in remission of the cancer, e.g., complete remission (CR) or partial remission (PR). In some embodiments, the kit further comprises an MTDP inhibitor, preferably paclitaxel; and / or a PLK1 inhibitor, preferably onvansertib. [Brief explanation of the drawings]

[0018] [Figure 1] FIG. 1 shows non-limiting exemplary embodiments and data relating to a schematic diagram of a dose escalation decision map for a Phase 1b study according to the BOIN design. [Figure 2] FIG. 2 shows non-limiting exemplary embodiments and data relating to a Phase 2 trial schematic. [Figure 3] FIG. 3 shows non-limiting exemplary embodiments and data regarding the combination index (CI) of the paclitaxel-onvansertib combination in cell lines with mutant p53 (left) and wild-type p53 (right). [Figure 4] FIG. 4 shows non-limiting exemplary embodiments and data regarding the in vivo efficacy of onvansertib (O) alone or in combination with paclitaxel (P) against SUM159 xenografts. DETAILED DESCRIPTION OF THE INVENTION

[0019] 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.

[0020] 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.

[0021] 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 (i.e., an allopathic or osteopathic 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. In some embodiments, the patient is a human or an animal. In some embodiments, the patient is a mammal.

[0022] 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 active ingredients (e.g., taxane-derived diterpenoids, including paclitaxel, and onvansertib). 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.

[0023] 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. 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.

[0024] 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.

[0025] 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.

[0026] 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," "prevention," and grammatical variations thereof as used herein" refers 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.

[0027] As used herein, the terms "partial response," "partial remission," and "PR" 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 may 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 tumors can be detected by means well known to those skilled in the art, such as ELISA or other antibody-based tests. A partial response of target lesions may refer to at least a 30% decrease in the sum of the diameters of target lesions, based on the sum of the diameters at baseline.

[0028] As used herein, the terms "complete response," "complete remission," 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 pathological lymph nodes (whether target or non-target) with a short axis reduced to less than 10 mm. A complete response of non-target lesions includes the disappearance of all non-target lesions and normalization of tumor marker levels (all lymph nodes are free of pathological size (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.

[0029] 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, relative to the smallest sum during the study (including if the baseline sum was the smallest 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.

[0030] 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.

[0031] As used herein, the term "MTD" means maximum tolerated dose. As used herein, the term "DLT rate" means the dose-limiting toxicity rate. As used herein, "IC 50 The term "inhibitor concentration" refers to the inhibitor concentration that produces 50% of the maximum effect. As used herein, the term "SEM" means standard error of the mean. As used herein, the term "AUC(xy)" means area under the curve, where "x" is the start time (in hours) and "y" is the end time (in hours). As used herein, "C avg " means the average concentration. max The term "" refers to the maximum concentration. As used herein, the term "QD" means once daily. As used herein, the term "ANC" means absolute neutrophil count. As used herein, the term "EOT" means end of treatment. As used herein, the term "ALT" means alanine aminotransferase. As used herein, the term "AST" means aspartate aminotransferase.

[0032] As used herein, the term "CBC" means complete blood count. As used herein, the term "CT" means computed tomography. As used herein, the term "ctDNA" means circulating tumor DNA. As used herein, the term "ECOG" means Eastern Cooperative Oncology Group (Performance Score). As used herein, the term "ECG" means an electrocardiogram. As used herein, the term "MRI" means magnetic resonance imaging. As used herein, the term "PK" means pharmacokinetics. As used herein, the term "TNM" means tumor node metastasis. As used herein, the term "PBMC" means peripheral blood mononuclear cells. As used herein, the term "CNS" means the central nervous system.

[0033] As used herein, the term "concomitant medication" means that a medication (or treatment) other than the drug used in the study is taken or received by the patient during the study period (after administration of the first dose of study drug and before the final clinic evaluation visit). As used herein, the term "tolerable" means a dose level at which 1 / 6 or less participants experienced a DLT or a dose level that was declared an RP2D. As used herein, the term "adverse event" or "AE" means an unexpected medical occurrence in a subject administered a medicinal product that is not necessarily causally related to that treatment. An AE can be any untoward and unintended sign (including abnormal clinical laboratory findings), symptom, or disease temporarily associated with the use of an investigational product, whether or not related to the investigational medicinal product. Adverse events can include a worsening or progression of the disease being studied, a worsening or progression of a pre-existing condition or event, a concomitant illness, or a drug interaction. An expected variation in a pre-existing condition that does not amount to a clinically significant worsening or exacerbation is not considered an AE. Surgery is not an adverse event, but a treatment for a condition requiring surgery. A condition requiring surgery is an AE if the condition develops or is a worsening of a pre-existing condition. Disease progression is an efficacy endpoint, not an AE. A clinical event in the setting of disease progression is considered an AE if it cannot be clearly attributed to or is inconsistent with expected disease progression.

[0034] As used herein, the term "anticipated adverse events" means adverse events listed or characterized in the current adverse event listing, package insert (PI), investigator brochure (IB), or included in the informed consent document as a potential risk. As used herein, the term "unexpected adverse event" refers to an adverse event not listed or identified in a PI or current IB. This includes adverse events whose specificity or severity does not match the description in a PI or IB. For example, hepatic necrosis would not be expected by this definition. As used herein, "serious adverse event" or "SAE" means an AE that (1) results in death (i.e., the AE actually causes or leads to death), (2) is life-threatening (i.e., in the opinion of the investigator, the AE places the subject at risk of death, but does not include an AE that, if manifested in a more severe form, may have resulted in death), (3) requires or is prolonged hospitalization, (4) results in persistent or significant disability / incapacity (i.e., the AE results in substantial impairment of the subject's ability to perform normal life functions), or (5) results in a congenital anomaly / birth defect in a newborn / infant born to a mother exposed to an IMP.

[0035] As used herein, the term "definite AE" means that the AE is clearly related to the study treatment. As used herein, the term "likely AE" means that the AE is likely to be related to the study treatment. As used herein, the term "possible AE" means that the AE is possibly related to the study treatment. As used herein, the term "unlikely AE" means that the AE is suspected to be related to the study treatment. As used herein, the term "unrelated AE" means that the AE is not clearly related to the study treatment. As used herein, the term "expected disease progression" refers to events that are clearly associated with disease progression and whose clinical course is consistent with that expected for the patient's disease.

[0036] As used herein, the term "measurable lesion" means a lesion that can be accurately measured in at least one dimension (longest recorded diameter) as 20 mm or greater on a chest x-ray or 10 mm or greater by caliper on a CT scan, MRI, or clinical examination. Tumor lesions present in previously irradiated areas may or may not be considered measurable. Cystic lesions considered to represent cystic metastases are measurable lesions if they meet the definition of measurability above. However, if non-cystic lesions are also present in the same participant, they are target lesions. Clinical lesions are measurable if they are superficial (e.g., skin nodules and palpable lymph nodes) and 10 mm or greater in diameter as assessed with a caliper (e.g., skin nodules). As used herein, the term "malignant lymph node" means a pathologically enlarged, measurable lymph node measuring 15 mm or greater in short axis when assessed by CT scan. As used herein, the term "non-measurable disease" refers to small lesions (or lesion sites) less than 10 mm in their longest dimension or pathological lymph nodes with a short axis greater than or equal to 10 mm but less than 15 mm. Examples of non-measurable disease include bone lesions, leptomeningeal disease, ascites, pleural / pericardial effusions, cutaneous lymphangitis / pneumonia, inflammatory breast disease, abdominal masses (not trackable by CT or MRI), and cystic lesions. Cystic lesions that meet the radiographically defined criteria for a simple cyst are not malignant lesions (neither measurable nor non-measurable) but are simple cysts.

[0037] As used herein, the term "target lesion" refers to a fully measurable lesion, up to a maximum of two lesions per organ, up to a total of five lesions, representative of all involved organs. Target lesions are selected based on their size (the lesion with the largest diameter) that is representative of all involved organs, but must also be suitable for reproducible repeated measurements. If the largest lesion is not suitable for reproducible measurement, the next largest lesion that allows reproducible measurement is selected as the target lesion. As used herein, the term "non-target lesion" means any lesion (or site of disease) that is not a target lesion. Non-target lesions include any measurable lesion beyond the five target lesions.

[0038] 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 "time to progression" or "TTP" means the time from randomization (or enrollment) to progression. Participants with no reported progression are censored at the date of their last disease activity assessment. As used herein, the term "CXD1" means day 1 of cycle X. For example, C1D1 means day 1 of cycle 1, before the subject receives the intended treatment. C2D1 means day 1 of cycle 2.

[0039] cancer The methods, compositions, and kits disclosed herein can be used to treat cancer and / or tumors. In some embodiments, the method for treating cancer and / or tumors comprises administering an MTDP inhibitor (e.g., paclitaxel), or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, and a PLK1 inhibitor (e.g., onvansertib), or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, to a subject (e.g., a patient) in need thereof.

[0040] The methods, compositions, and kits disclosed herein can be used to treat various types of cancers and / or tumors. The cancers and / or tumors can be solid tumors, liquid tumors, or a combination thereof. In some embodiments, the cancers and / or tumors are hematological cancers or solid tumors. The cancers can be breast cancer, pancreatic cancer, gastric cancer, gastroesophageal cancer, esophageal cancer, lung cancer, prostate cancer, cervical cancer, colorectal cancer, thyroid cancer, bladder cancer, head and neck cancer, brain and central nervous system cancer, liver cancer, gallbladder cancer, bile duct cancer, ovarian cancer, vaginal cancer, kidney cancer, endometrial cancer, skin cancer, testicular cancer, thymic cancer, non-specific cancer, adenocarcinoma, leukemia, lymphoma, sarcoma, other neoplastic malignancies, and / or combinations thereof. The breast cancer may be invasive breast cancer, unresectable breast cancer, locally advanced breast cancer, metastatic breast cancer, inflammatory breast cancer, triple-negative breast cancer (TNBC), hormone receptor / growth factor receptor-negative breast cancer, HER2-negative breast cancer, and / or hormone receptor-negative breast cancer. The pancreatic cancer may be metastatic pancreatic ductal adenocarcinoma, pancreatic ductal adenocarcinoma, and / or borderline resectable pancreatic adenocarcinoma. The lung cancer may be lung adenocarcinoma and / or mesothelioma. The prostate cancer may be metastatic castration-resistant prostate cancer and / or prostate adenocarcinoma. The colorectal cancer may be metastatic colorectal cancer and / or colon cancer. The bladder cancer may be urothelial carcinoma. Head and neck cancers may be squamous cell carcinoma of the head and neck, locally advanced or metastatic cancer of the head and neck, oral cavity cancer, pharyngeal cancer, laryngeal cancer, recurrent squamous cell carcinoma of the larynx, salivary gland cancer, hypopharyngeal cancer, squamous cell carcinoma of the lip and oral cavity, tongue cancer, nasopharyngeal cancer, and / or oropharyngeal cancer. Brain and central nervous system cancers may be glioblastoma, glioma, and / or neuroblastoma. Liver cancer may be intrahepatic cholangiocarcinoma (iCCA) and / or hepatocellular carcinoma. Ovarian cancer may be fallopian tube cancer or peritoneal cancer. Kidney cancer may be renal cell carcinoma, advanced or metastatic renal cell carcinoma, and / or metastatic transitional cell carcinoma of the renal pelvis and ureter. Skin cancer may be melanoma, squamous cell carcinoma, and / or metastatic Merkel cell carcinoma. Leukemia may be acute myeloid leukemia and / or multiple myeloma. The lymphoma can be B-cell lymphoma, T-cell lymphoma, non-Hodgkin's lymphoma, and / or follicular lymphoma. The sarcoma can be rhabdomyosarcoma. In some embodiments, the cancer and / or tumor is a blood cancer or solid cancer. The blood cancer or solid cancer is not gastric cancer or lung cancer.Hematological or solid cancers include breast cancer (e.g., invasive breast cancer, unresectable breast cancer, locally advanced breast cancer, metastatic breast cancer, inflammatory breast cancer, TNBC, hormone receptor / growth factor receptor negative breast cancer, HER2 negative breast cancer, and / or hormone receptor negative breast cancer); pancreatic cancer (e.g., metastatic pancreatic ductal adenocarcinoma, pancreatic ductal adenocarcinoma, and / or borderline resectable pancreatic adenocarcinoma); gastroesophageal cancer; esophageal cancer; prostate cancer (e.g., metastatic castration-resistant prostate cancer and / or prostate adenocarcinoma); cervical cancer; colorectal cancer (e.g., metastatic colorectal cancer and / or colon cancer); thyroid cancer; bladder cancer (e.g., urothelial carcinoma); head and neck cancer (e.g., squamous cell carcinoma of the head and neck, locally advanced or metastatic carcinoma of the head and neck, oral cancer, pharyngeal cancer, laryngeal cancer, recurrent squamous cell carcinoma of the larynx, cancer of the salivary gland, hypopharyngeal cancer, squamous cell carcinoma of the lip and oral cavity, tongue cancer, nasopharyngeal cancer, and / or oropharyngeal cancer; brain and central nervous system cancers (e.g., glioblastoma, glioma, and / or neuroblastoma); liver cancer (e.g., intrahepatic cholangiocarcinoma (iCCA) and / or hepatocellular carcinoma); gallbladder cancer; bile duct cancer; ovarian cancer; vaginal cancer; kidney cancer (e.g., renal cell carcinoma, advanced or metastatic renal cell carcinoma, and / or metastatic transitional cell carcinoma of the renal pelvis and ureter); endometrial cancer; skin cancer (e.g., melanoma, squamous cell carcinoma, and / or metastatic Merkel cell carcinoma); testicular cancer; thymic carcinoma; non-specified carcinoma; adenocarcinoma; leukemia (e.g., acute myeloid leukemia); lymphoma; sarcoma (e.g., rhabdomyosarcoma); other neoplastic malignancies; and / or combinations thereof. In some embodiments, the cancer and / or tumor 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 T-cell lymphoma, B-cell lymphoma, non-Hodgkin's lymphoma ("NHL"), follicular lymphoma ("FL"), acute myeloid leukemia ("AML"), and multiple myeloma ("MM"). In some embodiments, the diseases or conditions provided herein include refractory or recurrent malignancies whose growth can be inhibited using the methods and compositions disclosed herein.

[0041] In some embodiments, the cancer and / or tumor is leukemia (e.g., AML); NHL; metastatic CRC; metastatic castration-resistant prostate cancer (mCRPC); pancreatic cancer (e.g., metastatic castration-resistant prostate cancer); adrenocortical carcinoma (ACC); breast cancer (e.g., TNBC); colorectal cancer (e.g., metastatic colorectal cancer with a KRAS mutation); small cell lung cancer (SCLC); ovarian cancer; and / or combinations thereof. In some embodiments, the cancer is an invasive cancer, unresectable locally advanced or metastatic disease, e.g., invasive breast cancer. In some embodiments, the cancer is an inflammatory cancer, e.g., breast cancer, including TNBC. In some embodiments, the cancer has a histological or cytological profile with ER≦10%, PR≦10%, and Her-2-neu negative (0-1+ by immunohistochemistry (IHC) or fluorescent in situ hybridization (FISH) negative) per ASCO / CAP 2018 guidelines.

[0042] The cancer and / or tumor may be a cancer and / or tumor having an abnormal alteration in the PLK1 gene or protein. For example, the abnormal alteration may include one or more PLK1 alterations and / or abnormal activation of PLK1, such as copy number alterations (CNAs), single nucleotide variants (SNVs), and gene rearrangements or fusions. Non-limiting exemplary cancers and / or tumors having PLK1 alterations include cancers having PLK1 gene or protein amplification, PLK1 gene or protein alterations, PLK1 gene deletions, PLK1 gene or protein overexpression, elevated PLK1 gene or protein expression, and / or combinations thereof. In some embodiments, the cancer and / or tumor may be a PLK1-amplified cancer, in which the PLK1 gene and / or protein is amplified, for example, as a result of gene doubling and / or aberrant gene transcription regulation. For example, a cancer with PLK1 amplification may be a cancer with elevated PLK1 mRNA and / or protein levels compared to healthy tissue. In some embodiments, kidney, lung, breast, colon, skin, and / or head and neck cancers may exhibit elevated PLK1 gene or protein amplification, and the resulting cancers and / or tumors are suitable for treatment with the methods and compositions disclosed herein. In heterogeneous cancer types, cancers / tumors may include subtypes with abnormally elevated expression of the PLK1 gene and / or protein. Non-limiting examples of cancers and / or tumors, including subtypes, with high PLK1 gene and / or protein expression include lymphomas (e.g., B-cell lymphoma, diffuse large B-cell lymphoma); testicular cancers (e.g., testicular germ cell carcinoma); cervical cancer; head and neck cancers (e.g., uveal melanoma, adenoid cystic carcinoma); ovarian cancer, uterine cancer (e.g., uterine carcinosarcoma); colorectal cancer; thyroid cancer (e.g., thymoma); bladder cancer; lung cancer (squamous cell lung cancer, adenocarcinoma, mesothelioma); uterine cancer; skin cancer (e.g., melanoma); sarcoma; brain tumors (e.g., glioblastoma, glioma); leukemia (e.g., acute myeloid leukemia); breast cancer; pancreatic cancer; bile duct cancer liver cancer; kidney cancer (e.g., renal cell carcinoma, clear cell renal cell carcinoma, chromophobe renal cell carcinoma, papillary renal cell carcinoma); neuroendocrine tumors (e.g., pheochromocytoma / paraganglioma).Cancers and / or tumors with amplified PLK1 may be lymph node-positive tumors, aggressive tumors, and / or invasive tumors. Cancers and / or tumors with amplified PLK1 may have a shorter disease-free survival period compared to cancers and / or tumors with normal levels of PLK1. Such cancers and / or tumors may have a high recurrence rate and / or be resistant to conventional therapies and / or monotherapy, such as chemotherapy and / or radiotherapy. Prostate cancer may be resistant to single-agent treatment with an MTDP inhibitor or to an MTDP inhibitor.

[0043] The cancer and / or tumor may be a breast cancer and / or breast tumor. In some embodiments, the breast cancer and / or breast tumor is unresectable. In some embodiments, the breast cancer and / or breast tumor is advanced and / or metastatic. In some embodiments, the breast cancer and / or breast tumor has cells with an unstable genome. In some embodiments, the breast cancer and / or breast tumor is of the basal-like subtype, characterized by epidermal growth factor receptor (EGFR) overexpression, phosphatase and tensin homolog (PTEN) deficiency, and TP53 gene (mutp53) mutations, and exhibits overall high genomic instability among subtypes. In some embodiments, the breast cancer and / or breast tumor has a low pathological complete response rate after neoadjuvant chemotherapy. In some embodiments, the breast cancer and / or breast tumor is inherently resistant to chemotherapy and / or has a low overall survival rate. In some embodiments, the breast cancer and / or tumor is invasive breast cancer, unresectable breast cancer, locally advanced breast cancer, metastatic breast cancer, inflammatory breast cancer, TNBC, hormone receptor / growth factor receptor-negative breast cancer, HER2-negative breast cancer, and / or hormone receptor-negative breast cancer. In some embodiments, the breast cancer is invasive breast cancer, unresectable breast cancer, locally advanced breast cancer, metastatic breast cancer, inflammatory breast cancer, TNBC, hormone receptor / growth factor receptor-negative breast cancer, HER2-negative breast cancer, hormone receptor-negative breast cancer, and / or combinations thereof. In some embodiments, the breast cancer and / or breast tumor is a biologically aggressive form of breast cancer. In some embodiments, the breast cancer and / or breast tumor lacks estrogen receptor (ER), lacks progesterone receptor, and lacks human epidermal growth factor receptor 2 (HER2) gene amplification. In some embodiments, the breast cancer and / or breast tumor is associated with a high mortality rate, with a median survival time of less than two years from the time of metastasis. In some embodiments, the breast cancer and / or breast tumor is TNBC.

[0044] Given the poor prognosis and short overall survival associated with TNBC, novel targeted and targeted combination therapies are needed. For example, targeted therapies for TNBC may include immune checkpoint inhibitors (CPIs). Given that over 60% of metastatic TNBC patients are PD-L1 negative (combined positive score (CPS) <10), in some cases, therapies can target programmed death 1 (PD-1) / PD-L1. In some cases, single-agent paclitaxel is the standard first-line treatment. In the KEYNOTE-355 trial, a chemotherapy (taxane or carboplatin plus gemcitabine) plus pembrolizumab (a monoclonal antibody against the PD-1 receptor) regimen significantly reduced the risk of death (CPS >10) by 27% in patients with metastatic TNBC whose tumors were strongly PD-L1 positive compared with chemotherapy versus placebo. Recently, the FDA approved sacituzumab govitecan, an antibody-drug conjugate directed against tumor-associated calcium signal transducer 2 (TROP2), as a second- and third-line treatment for metastatic TNBC. However, existing therapies are not ideal. Basal-like breast cancer is inherently resistant to chemotherapy, and patients with this type of breast cancer have a low overall survival rate, necessitating the development of novel targeted and targeted combination therapies. Among the heterogeneous TNBC subtypes, the basal-like subtype, identified by gene expression analysis, is characterized by overexpression of EGFR and PTEN and mutations in the TP53 gene, and exhibits the greatest overall genomic instability among the subtypes. Patients with basal-like breast cancer have a low rate of pathological complete response after neoadjuvant chemotherapy. PLK1 has been identified as a key gene for the proliferation and survival of genomically unstable breast cancer cells, including TNBC.

[0045] Paclitaxel is an effective agent for the treatment of metastatic breast cancer. Weekly paclitaxel has been shown to exhibit superior activity and cause less myelosuppression compared with an every-3-week schedule. Neuropathy, if present, is usually mild or moderate in severity and is generally reversible. A randomized controlled trial of paclitaxel 80 mg / m2, including 212 patients with metastatic breast cancer, was conducted. 2In a large phase 2 trial in which rituximab was administered weekly for 4 weeks in a 4-week cycle, the therapy was generally well tolerated. Grade 3 or 4 neutropenia occurred in 31 patients (15%). All but two patients who developed grade 3 or 4 neutropenia had received prior chemotherapy, five of whom had received high-dose chemotherapy. Grade 3 anemia occurred in 18 patients (9%). Grade 3 and 4 thrombocytopenia occurred in one patient each. Thirty patients (14%) had preexisting grade 1 neuropathy. Two of these patients eventually developed grade 3 neuropathy after five and 11 cycles of treatment. Overall, the incidence of neuropathy of any grade was 69%. However, only 20 patients (9%) developed grade 3 neuropathy, and no patients developed grade 4 neuropathy. The median number of treatment cycles until the onset of grade 2 or 3 neuropathy was five cycles (20 weeks) in all patients, including those with preexisting grade 1 toxicity, ranging from one to 13 cycles. Among 177 evaluable patients, the overall response rate was 21.5% (95% confidence interval, 15.4% to 27.5%). Responses occurred in 23 (17.6%) of 131 evaluable patients previously treated with anthracyclines and in 7 (15.6%) of 45 evaluable patients previously treated with taxanes. The median time to progression for evaluable patients was 142 days (4.7 months). The median time to progression for patients with no prior chemotherapy, those previously treated with one prior therapy, and those previously treated with two prior therapies for metastatic disease was 174 days (5.7 months), 140 days (4.6 months), and 85 days (2.7 months), respectively.

[0046] MTDP inhibitors and PLK inhibitors The methods, compositions, and kits disclosed herein can be used to treat cancers and / or tumors, such as breast cancer; pancreatic cancer; gastric cancer; gastroesophageal cancer; esophageal cancer; lung cancer; prostate cancer; cervical cancer; colorectal cancer; thyroid cancer; bladder cancer; head and neck cancer; brain and central nervous system cancer; liver cancer; gallbladder cancer; bile duct cancer; ovarian cancer; vaginal cancer; colorectal cancer; kidney cancer; endometrial cancer; skin cancer; testicular cancer; thymic cancer; non-specific cancer; adenocarcinoma; leukemia; lymphoma; sarcoma; other neoplastic malignancies; and / or combinations thereof. In some embodiments, a method for treating cancer and / or tumors includes administering a tubulin-targeting drug, such as an MTDP inhibitor (e.g., paclitaxel), or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, and a PLK1 inhibitor (e.g., onvansertib), or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, to a subject (e.g., patient) in need thereof. The method may include administering a pharmaceutically effective amount of the MTDP inhibitor (e.g., paclitaxel) and a pharmaceutically effective amount of the PLK1 inhibitor (e.g., onvansertib).

[0047] MTDP inhibitors Microtubules are highly dynamic polymers of tubulin that constitute the cytoskeleton and are crucial for cell shape, intracellular transport, cell division, and cancer. Microtubule-targeting drugs, such as compounds that inhibit microtubule dynamics (including spindle-microtubule dynamics), microtubule depolymerization inhibitors, and microtubule polymerization inhibitors, are important classes of antimitotic compounds with anticancer properties and can be used as first-line cancer therapies. Inhibition of microtubule dynamics, particularly spindle-microtubule dynamics, can prevent cells from completing mitosis. Cells blocked in a prometaphase / metaphase-like state can eventually undergo apoptosis. Tubulin-targeting drugs can be used as first-line cancer therapies. Generally, tubulin-targeting drugs interfere with microtubule dynamics by stabilizing or destabilizing microtubules, making them suitable as first-line cancer therapies. However, tubulin-targeting drugs also have drawbacks such as tissue specificity, inherent and / or acquired drug resistance, and systemic toxicity. Furthermore, the changes in microtubule dynamics induced by some tubulin-targeting drugs can be harmful to cancer therapy. For example, tubulin-targeting drugs can alter microtubule chromosomal attachment and chromosomal nondisjunction, causing chromosomal instability and aneuploidy. In some cases, tubulin-targeting drugs may promote tumorigenesis, cancer development, drug resistance, treatment failure, metastasis, poor prognosis, polyploid cell formation (including polyploid giant cells), migration, and invasive phenotypes.

[0048] Binding to the taxane site, located on the beta-tubulin monomers in the microtubule lumen, can stabilize the microtubule lattice. For example, paclitaxel or epothilones can stabilize the microtubule lattice by binding to the taxane site, although by different mechanisms. Other examples of drugs that bind to the taxane site include PM060184 (procabulin) and the covalent tubulin inhibitor zampanolide. Binding to the laulimalide or peloruside site, located in the outer-facing beta-tubulin pocket of microtubules, inhibits microtubule disassembly. For example, binding of laulimalide and peloruside to this site clamps the fibril and inhibits microtubule disassembly. Binding to the vinca domain, located at the interdimer interface between two tandemly aligned tubulin dimers, can inhibit tubulin polymerization. For example, binding of vincristine to the vinca domain can inhibit tubulin assembly by sequestrating tubulin into paracrystalline aggregates. Binding to the colchicine site, located in a deep pocket between tubulin dimers, inhibits microtubule assembly. For example, binding of colchicine, benzimidazoles (e.g., nocodazole), or combretastatins to the colchicine site inhibits microtubule assembly by preventing the conformational changes in tubulin required for assembly. Binding to the maytansine domain, located in the exposed β-tubulin pocket proximal to the vinca site, inhibits assembly at the plus end. For example, binding of maytansine and spongistatins to the maytansine domain inhibits the addition of new tubulin to the plus end. Binding to the pironetin site located on α-tubulin destabilizes microtubules. For example, binding of pironetin to this site disrupts longitudinal tubulin interactions and prevents heterodimer formation. Binding to the gatorubrin site located on α-tubulin proximal to the colchicine site inhibits tubulin assembly. For example, binding of sevipabulin to the gatorubin site allows the formation of a wedge at the tip of a microtubule via two tandemly aligned tubulin dimers.

[0049] MTDP inhibitors include, but are not limited to, diterpenoids, taxanes, taxane-derived diterpenoids, taxoids, and derivatives, analogs, precursors, and complexes thereof. In some embodiments, the MTDP inhibitor is paclitaxel, docetaxel, acetyltaxol, paclitaxel, lutetium Lu 177 bipibotide tetraxetan, 7-hexanoyltaxol, cabazitaxel, larotaxel, mirataxel, ortataxel, tesetaxel, taxoplexin, opaxiolide, taxoplexin (DHA-paclitaxel), poly(L-glutamic acid)-paclitaxel, abraxane, SB-T-1214, SB-T1216, SB-T121602, SB-T-12854, DHA-SB-T1214, or abeotaxane. The abeotaxane can be abeotaxane 15a.2; docetaxel-d9-t-Boc; docetaxel-f3-t-Boc; cabazitaxel-7,10-d6; poly(glutamyl-glutamic acid)-taxane and / or derivatives, analogs, precursors, and conjugates thereof. Non-limiting examples of microtubule-targeting drugs include compounds that target the taxane site of microtubules (e.g., paclitaxel); compounds that target the vinca domain of microtubules (e.g., vinflunine); compounds that target the colchicine domain (e.g., cyclohexanediones with distal 2-substituted benzofurans, heterocyclic nitrogen compounds such as plinabulin, verubulin, and ABT-751, combretastatins such as ombrabulin and phosbretabulin); and compounds that target other microtubule-binding sites (e.g., estramustine). Microtubule-targeting drugs also include other microtubule-destabilizing drugs, such as the antitussive drug noscapine; maytansine and / or maytansine / auristatin conjugates (e.g., TDM1, brentuximab vedotin, and SAR33419); rhizoxin; spongistatin; podophyllotoxin; steganacin; curacin; antimitotic herbicides that inhibit microtubule polymerization; antifungal and antihelminthic drugs; and certain psychoactive drugs (e.g., dilantin, vinblastine, chlorpromazine). Microtubule-targeting drugs also include other microtubule-stabilizing drugs, such as eleutherobin, sarcodictyin, laulimalide, radinaram, steroids, and polyisoprenylbenzophenones.

[0050] Many microtubule-targeting compounds are tissue-specific. For example, paclitaxel has been shown to be highly effective against ovarian, breast, and lung tumors, but to have little effect on many other solid tumors, such as kidney cancer, colon cancer, and some sarcomas. As another example, vinca alkaloids have often been found to be most effective against hematological cancers, but are often ineffective against many solid tumors. Drug resistance to microtubule-targeting compounds is a complex and largely unsolved problem. In some cases, drug resistance is correlated with the overexpression of ATP-dependent drug efflux pumps or ATP-binding cassettes, which are a type of membrane transport protein. Membrane pumps remove drugs at the intracellular level and may result in resistance to drugs with different chemical structures (e.g., both paclitaxel and vinca alkaloids). Tissue-specific differences in regulatory factors (e.g., differential expression of regulatory proteins, post-translational modifications of tubulin, and expression of different tubulin isotypes) may contribute to tissue-specific sensitivity and / or resistance to microtubule-targeting compounds.

[0051] In some embodiments, compounds that target microtubule taxane sites include paclitaxel, TL00139, and paclitaxel analogs, docetaxel (taxotere), epothilones (e.g., BMS-247550, epothilones B and D), ixabepilone, discodermolide, and other similar depolymerization inhibitors. In some embodiments, compounds that target microtubules near taxane sites include sarcodictyin and eleutherobin. Unlike vinca alkaloids, MTDP inhibitors, which bind to the taxane site of tubulin, stimulate microtubule polymerization and are an important class of compounds for the treatment of breast cancer, ovarian cancer, non-small cell lung cancer, and Kaposi's sarcoma. However, side effects include neurotoxicity and myelosuppression. MTDP inhibitors targeting the taxane site of the β subunit bind to the surface of polymerized microtubules. This stabilizes microtubules, increases microtubule polymerization, and enhances their affinity for neighboring tubulin molecules. Furthermore, low concentrations of paclitaxel can completely stabilize microtubule dynamics without increasing polymerization. For example, binding of a small number of paclitaxel molecules (e.g., one paclitaxel per several hundred tubulin molecules) can stabilize microtubule dynamics (e.g., reduce the rate or extent of microtubule shortening by approximately 50%). In HeLa cells, approximately 8 nM to approximately 10 nM paclitaxel can block mitosis by 50% without increasing microtubule-polymer mass. In general, inhibitor-mediated suppression of microtubule dynamics ultimately blocks mitosis and leads to apoptosis.

[0052] In some embodiments, the MTDP inhibitor is paclitaxel, a paclitaxel derivative or analog (e.g., docetaxel), or a pharmaceutically acceptable salt thereof. Paclitaxel is also known as 4,10-diacetic acid 2-benzoic acid 13-ester of 5β,20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytax-11-en-9-one with (2R,3S)-N-benzoyl-3-phenylisoserine, and has the empirical formula C 47 H 51 NO 14 Paclitaxel has a molecular weight of 853.9 and is a tricyclic diterpenoid containing a taxane ring (Formula 1). The structure of paclitaxel essential for antitumor activity includes the taxane ring, the C13 side chain, the oxetane ring, the hydroxyl group at the 2'-position, and the homochiral ester chain. The structure of paclitaxel not essential for antitumor activity includes the hydroxyl group at C7 and acetylation of the C10 hydroxyl group. [ka] (Formula 1)

[0053] Paclitaxel is generally obtained by semi-synthetic processes from the European yew (Taxus baccata). It is highly lipophilic, insoluble in water, and melts at approximately 216°C to 217°C. In some embodiments, paclitaxel is administered intravenously. In some embodiments, paclitaxel is administered orally. In some embodiments, paclitaxel is combined with an excipient. For example, paclitaxel can be formulated with surfactants, including heterogeneous nonionic surfactants such as Cremophor EL (polyoxyethylated castor oil), and / or dehydrated alcohol. Paclitaxel can be formulated as a nanomedicine. In some embodiments, paclitaxel is conjugated with albumin to form a 130 nm nanomedicine (e.g., nab-PTX). In some embodiments, paclitaxel is formulated in polymeric micelles using mPEG-PDLLA to form 25 nm nanomedicines, PVP-bPNIPAAM to form 80 nm to 100 nm nanomedicines, or N-tr-Lc methyl ester and N.13cr-Lc methyl ester to form 20 nm to 60 nm nanomedicines. In some embodiments, paclitaxel is formulated in liposomes using lecithin and cholesterol to form 400 nm nanomedicines. In some embodiments, paclitaxel is formulated in polymeric lipid nanoparticles using polyvinylpyrrolidone, cholesteryl sulfate, and caprylic acid to form 100 nm nanomedicines. In some embodiments, paclitaxel is formulated in an emulsion with monoolein, tricaprylin, and Tween 80. Paclitaxel can be formulated as a dimer. Paclitaxel can be administered orally, for example, by conjugating it to chitosan, lipid derivatives, nanococreates, hyaluronic acid-octadecylamine micelles, or oil-based nanocarriers. As another example, paclitaxel can be loaded into milk-derived exosomes.

[0054] Paclitaxel has been approved for use in, for example, ovarian cancer, breast cancer, non-small cell lung cancer, pancreatic cancer, bladder cancer, AIDS-related Kaposi's sarcoma, and gastric cancer. Paclitaxel is an effective drug for the treatment of metastatic breast cancer. Weekly paclitaxel has been demonstrated to exhibit superior activity and less myelosuppression compared with the every 3 weeks schedule. Neuropathy, if present, is usually mild or moderate in severity and generally reversible. A study of 212 patients with metastatic breast cancer receiving paclitaxel at 80 mg / m 2In a large phase 2 trial in which rituximab was administered weekly for 4 weeks in a 4-week cycle, the therapy was generally well tolerated. Grade 3 or 4 neutropenia occurred in 31 patients (15%). All but two patients who developed grade 3 or 4 neutropenia had received prior chemotherapy, five of whom had received high-dose chemotherapy. Grade 3 anemia occurred in 18 patients (9%). Grade 3 and 4 thrombocytopenia occurred in one patient each. Thirty patients (14%) had preexisting grade 1 neuropathy. Two of these patients eventually developed grade 3 neuropathy after five and 11 cycles of treatment. Overall, the incidence of neuropathy of any grade was 69%. However, only 20 patients (9%) developed grade 3 neuropathy, and no patients developed grade 4 neuropathy. The median number of treatment cycles until the onset of grade 2 or 3 neuropathy was five cycles (20 weeks) in all patients, including those with preexisting grade 1 toxicity, ranging from one to 13 cycles. Among 177 evaluable patients, the overall response rate was 21.5% (95% confidence interval, 15.4%–27.5%). Responses were observed in 23 (17.6%) of 131 evaluable patients previously treated with anthracyclines and 7 (15.6%) of 45 evaluable patients previously treated with taxanes. The median time to progression for evaluable patients was 142 days (4.7 months). The median time to progression for patients previously treated with chemotherapy for metastatic disease was 174 days (5.7 months), 140 days (4.6 months), and 85 days (2.7 months), respectively, for patients previously naïve, previously treated with one prior therapy, and previously treated with two prior therapies for metastatic disease. In some embodiments, paclitaxel is administered to the patient. Docetaxel has been used to treat breast cancer, lung cancer, and prostate cancer. MTDP inhibitors can be administered by any suitable route, including, but not limited to, oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal, epidural, and intranasal administration. Parenteral administration (e.g., injection) can include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

[0055] 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. PLK is also important for mitotic entry and progression, regulating cell entry into the G2 phase of the cell cycle by phosphorylating the forkhead box protein M1 (FOXM1), which then regulates the expression of cyclins and other genes required for cell progression through the cell cycle. PLK1 has been shown to be overexpressed in solid tumors and hematological malignancies, including breast cancer. Patients with breast cancers with high PLK1 expression have lower overall survival rates than those with low PLK1 expression. PLK1 expression levels were higher in TNBC compared with luminal A, luminal B, and HER-2-overexpressing breast cancers. Inhibition of PLK1 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 early preclinical development of PLK1-targeted drugs, cancer cells with mutated TP53 (mutp53) were more responsive than wild-type (wtp53) cell lines, demonstrating a higher IC 50The expression of PLK1 is low, consistent with the lack of mutp53-associated checkpoint control and genomic instability. These findings confirm the importance of PLK1 function for progression through the G2 and M phases of the cell cycle. Pyruvate dehydrogenase kinase 1 (PDK1), PLK1, and MYC have also been implicated in driving the expression of a set of genes associated with cancer stem cell self-renewal. Therefore, blocking PLK1 function and affecting the ability of cancer cells with unstable genomes to progress through mitosis can increase the overall sensitivity of cells to taxanes such as paclitaxel. PLK1 has been identified as a therapeutic target for TNBC through siRNA-mediated screening, and inhibition of PLK1 by siRNA-mediated knockdown or chemical inhibitors promoted cell cycle arrest and apoptosis in multiple TNBC lines. The lack of a druggable target contributes to the poor prognosis of TNBC. The relatively specific expression of PLK1 in TNBC tissues, as well as the efficacy and unique action profile of PLK1 inhibition, suggest that PLK1 is a promising molecular target for TNBC.

[0056] In a randomized phase II study of previously untreated AML patients who were unsuitable for induction therapy, intravenous administration of the pan-PLK inhibitor volasertib (BI6727) in combination with low-dose Ara-C (cytarabine) (LDAC) significantly improved overall survival compared with LDAC alone. A subsequent randomized phase III study failed to confirm any benefit of the combination and reported an increased risk of serious infections. PLK1 promotes homologous recombination (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. Onvansertib (also known as PCM-075, NMS-1286937, NMS-937, "the compound of formula (I)" in U.S. Pat. No. 8,927,530; 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), or a pharmaceutically acceptable salt, is a selective, ATP-competitive PLK1 inhibitor. Onvansertib can be formulated with excipients such as the free base, lactose monohydrate, pregelatinized starch, and glyceryl benzoate. In some embodiments, onvansertib is formulated for oral administration, such as in a hard gelatin capsule.

[0057] Biochemical assays demonstrated high specificity for PLK1 among a panel of 296 kinases, including other PLK members. Onvansertib 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 demonstrated a promising safety profile in a single-agent Phase I clinical trial. Further clinical trials of onvansertib include combination studies with abiraterone and prednisone in adult patients with metastatic castration-resistant prostate cancer, FOLFIRI and bevacizumab in adult patients with KRAS-mutated metastatic colorectal cancer, and nanoliposomal irinotecan and 5-FU in patients with metastatic pancreatic cancer. As described herein, in the presence of genomic instability, onvansertib synergizes with paclitaxel, subsequently achieving good antitumor activity at lower doses compared to the single agents and without drug-specific toxicity. Onvansertib also inhibited cell proliferation at nanomolar concentrations in AML cell lines and tumor growth in xenograft models of AML. Furthermore, onvansertib significantly augmented the antitumor activity of cytarabine in a disseminated AML model. [ka] Onvansertib

[0058] Onvansertib exhibits high potency in proliferation assays with low nanomolar activity against multiple cell lines derived from both solid and hematologic tumors. It has a relatively short half-life of 24 hours and is highly potent against the PLK1 enzyme (IC50 = 2 nM). In contrast, it exhibits low to no activity (IC50 > 10 μM) against a panel of 63 kinases other than PLK1, including PLK2 and PLK3 (IC50 > 500 nM). Onvansertib potently induces mitotic cell cycle arrest and subsequent apoptosis in cancer cell lines and inhibits xenograft tumor growth in mice at well-tolerated doses following oral administration through a distinct mechanism of action linked to PLK1. Furthermore, onvansertib has demonstrated activity in combination with approved cytotoxic agents, such as irinotecan, resulting in enhanced tumor regression compared with each agent alone in HT29 human colon adenocarcinoma xenografts, and in combination with cytarabine, it has been shown to extend survival in disseminated AML animal models. Onvansertib has favorable pharmacological parameters and good oral bioavailability in rodents and non-rodents. Its antitumor activity has also been demonstrated in various preclinical models using various dosing regimens, allowing for greater flexibility in dosing schedules, making it worthy of clinical consideration. Onvansertib offers several advantages over volasertib (BI6727, another PLK1 inhibitor), including higher potency and specificity for the PLK1 isoenzyme, as well as oral bioavailability. Furthermore, onvansertib has demonstrated antitumor activity in various preclinical models using various dosing regimens, allowing for greater flexibility in dosing schedules, making it worthy of clinical consideration.

[0059] 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 PCT Application No. PCT / US2021 / 013287, the contents of which are incorporated herein by reference in their entirety. The main metabolic pathways observed in various animal species were N-oxidation of the N-methyl-piperazine ring to give N-oxide M2 ​​and hydroxylation of the aliphatic carbon atom of the methylene bridge of the pyrazoloquinazoline moiety to give metabolite M1.Qualitatively, no significant differences in the metabolism of onvansertib were observed between species, but quantitatively, some differences were observed between species.

[0060] Using human liver microsomes, we investigated the potential inhibitory activity of onvansertib against the major human cytochrome P450 (CYP) isoforms (CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4) responsible for drug metabolism in the human liver. Onvansertib was able to inhibit the metabolic activity of CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 isoforms to various degrees, with 50% inhibitory concentration (IC50) values ​​ranging from 20 μM to 66 μM (Table 1). No significant inhibitory effect on CYP1A2 was detected. Considering that the appropriate concentration to achieve significant antitumor activity of this compound in mice is around 1 μM, onvansertib is unlikely to exhibit clinically relevant metabolic drug-drug interactions. Table 1 shows the IC50 values.50 The values ​​are shown as mean values±SEM (standard error of the mean). [Table 1]

[0061] To date, one Phase 1 safety study of onvansertib has been completed in adult patients with advanced / metastatic solid tumors at a single US trial site. First-cycle dose-limiting toxicity (DLT) and maximum tolerated dose (MTD) of onvansertib were administered orally for 5 consecutive days every 3 weeks (i.e., a 21-day treatment cycle). The safety profile of onvansertib was determined, the pharmacokinetics (PK) of onvansertib in plasma (at MTD) was determined, and antitumor activity was documented. A total of 21 patients were enrolled in one study, and 19 patients were treated. The first three dose levels (6 mg / m 2 / day, 12mg / m 2 / day, and 24 mg / m 2 No DLTs occurred at subsequent dose levels (48 mg / m 2 / day), DLT occurred in 2 of 3 patients. 2 An intermediate dose level of 24 mg / m was investigated. At this intermediate dose level, 4 patients were treated and 2 DLTs were observed. After further cohort expansion, the MTD was 24 mg / m . 2 The best treatment response observed was stable disease (SD), observed in 5 of 16 evaluable patients. Thrombocytopenia and neutropenia were the primary toxicities observed in this study, consistent with onvansertib's expected mechanism of action and preclinical results. These hematologic toxicities were reversible and typically resolved within 3 weeks. No other clinically relevant safety findings were observed with onvansertib as a single agent. No other mechanism-related, possibly expected, events, such as gastrointestinal disorders, mucositis, or alopecia, were observed, confirming that the bone marrow is the most sensitive target of onvansertib in humans on this schedule.

[0062] As disclosed herein, combination therapy using an MTDP inhibitor (including paclitaxel) and a PLK1 inhibitor (including onvansertib) is expected to result in significantly enhanced efficacy against cancer (e.g., prostate cancer, head and neck cancer, non-small cell lung cancer, intrahepatic cholangiocarcinoma, gastric cancer, urothelial cancer, small cell lung cancer, breast cancer, endometrial cancer, cervical cancer, rhabdomyosarcoma, cholangiocarcinoma, ovarian cancer, or a combination thereof), leading to predicted tumor regression and improved cancer survival. The tumor regression and cancer survival rates / lengths achieved by this combination may surprisingly be synergistic (i.e., greater than additive and superior to the combined antitumor efficacy achieved by the MTDP inhibitor and PLK1 inhibitor, respectively). For example, as described herein, the combination of onvansertib and paclitaxel demonstrated synergistic effects in in vitro and in vivo models of chemotherapy-resistant ovarian cancer. Treatment with onvansertib and paclitaxel demonstrated surprising synergistic effects in a triple-negative breast cancer model. Moreover, when used in combination, the concentration of PLK1 inhibitor required to achieve complete cell inhibition was surprisingly significantly lower compared to the single agents used to achieve the same inhibition.

[0063] Provided herein are methods, compositions, and kits for treating cancer in a subject (e.g., a human patient suffering from cancer). The methods include administering an MTDP inhibitor and a PLK1 inhibitor to the patient in a manner sufficient to inhibit or reduce the progression of the cancer. For example, the MTDP inhibitor and the PLK1 inhibitor can be administered simultaneously, separately, or sequentially to a subject with cancer. Combination treatment with onvansertib and an MTDP inhibitor is expected to be significantly more effective than combination treatment with another PLK inhibitor, BI2536, and MTDP for treating various cancers, including the treatment of prostate cancer and lung cancer (e.g., neuroendocrine prostate cancer). In some embodiments, the inhibition or reduction of cancer progression is potentiated or synergistic rather than merely additive (i.e., the inhibition is greater than the combined inhibition of progression caused by an MTDP inhibitor alone plus a PLK1 inhibitor alone). The potentiated or synergistic efficacy or inhibition of any combination of an MTDP inhibitor and a PLK1 inhibitor of the present disclosure may vary in different embodiments. In some embodiments, the potentiated or synergistic efficacy or inhibition of any combination of an MTDP inhibitor and a PLK1 inhibitor of the present disclosure is 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%, 7 0%, 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.

[0064] The molar ratio of the PLK1 inhibitor (e.g., onvansertib) to the MTDP inhibitor (e.g., paclitaxel) 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 MTDP inhibitor (e.g., paclitaxel) and a PLK1 inhibitor (e.g., onvansertib) is greater than the combined inhibition of progression caused by the MTDP inhibitor (e.g., paclitaxel) and the PLK1 inhibitor (e.g., onvansertib) alone, or is greater than, or is at least about, or is at most about, 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, or is greater than, or is at least about, or is at most about, 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, or is at most about, such a percentage greater. For example, the combination of an MTDP inhibitor 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 a single MTDP inhibitor (e.g., paclitaxel) and a single PLK1 inhibitor 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 MTDP inhibitor (e.g., paclitaxel) and a PLK1 inhibitor (e.g., onvansertib) is, for example, 50%, 60%, 70%, 80%, 90%, 100% or more greater than the combined inhibition of progression caused by the MTDP inhibitor (e.g., paclitaxel) and the PLK1 inhibitor alone.In some embodiments, the MTDP inhibitor is paclitaxel and the PLK1 inhibitor is onvansertib.

[0065] The methods disclosed herein may be effective against a variety of cancers, such as breast cancer; pancreatic cancer; gastric cancer; gastroesophageal cancer; esophageal cancer; lung cancer; prostate cancer; cervical cancer; colorectal cancer; thyroid cancer; bladder cancer; head and neck cancer; brain and central nervous system cancer; liver cancer; gallbladder cancer; bile duct cancer; ovarian cancer; vaginal cancer; colorectal cancer; kidney cancer; endometrial cancer; skin cancer; testicular cancer; thymic cancer; non-specific cancer; adenocarcinoma; leukemia; lymphoma; sarcoma; other neoplastic malignancies, or combinations thereof. As described herein, a patient may achieve a complete or partial response following treatment with an MTDP inhibitor and a PLK1 inhibitor. In some embodiments, the patient achieves a complete response. In some embodiments, the patient achieves a partial response. In some embodiments, the patient has not responded to treatment with an MTDP inhibitor (without a PLK1 inhibitor). In some embodiments, the patient has not responded to treatment with an MTDP inhibitor alone. The MTDP inhibitor and the PLK1 inhibitor can be administered to a patient in any manner believed to be effective for treating cancer. The MTDP inhibitor can be administered together with the PLK1 inhibitor or separately. When administered separately, the MTDP inhibitor can be administered before or after the PLK1 inhibitor, or in different administration cycles.

[0066] The MTDP inhibitor and the PLK1 inhibitor can each be administered on any schedule, such as once or more times per day or week; once, twice, three times, four times, five times, six times, or seven times per week (daily); or once or more weeks. The PLK1 inhibitor (e.g., onvansertib) can be administered, for example, orally. The MTDP inhibitor (e.g., paclitaxel) can be administered, for example, by intravenous infusion (e.g., over about 30 minutes). In some embodiments, the PLK1 inhibitor (e.g., onvansertib) is administered to the patient daily for 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 consecutive days in a cycle, e.g., the first 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 consecutive days of the cycle; the MTDP inhibitor (e.g., paclitaxel) is administered to the patient once each week that onvansertib is administered. The cycle can be, for example, 21 to 28 days long. In some embodiments, the PLK1 inhibitor is administered to the patient daily for the first 21 consecutive days of a 28-day cycle, and the patient is administered the MTDP inhibitor once a week for the first 3 weeks of the 28-day cycle. In some embodiments, neither the PLK1 inhibitor nor the MTDP inhibitor is administered on the last 7 days of the 28 cycles. The patient may receive one or more cycles of treatment / administration, for example, at least two cycles of treatment / administration. The administration schedules for the MTDP inhibitor and the PLK1 inhibitor may be the same or different for each treatment / administration cycle.

[0067] The MTDP inhibitor may be administered at any suitable dose, for example, at about, at least, or at most 5 mg / m 2 , 10 mg / m 2 , 15 mg / m 2 , 20 mg / m 2 , 25 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 / m2 、60mg / m 2 、65mg / m 2 、70mg / m 2 、75mg / m 2 、80mg / m 2 、85mg / m 2 、90mg / m 2 、95mg / m 2 、100mg / m 2 、105mg / m 2 、110mg / m 2 、115mg / m 2 、120mg / m 2 、125mg / m 2 、130mg / m 2 、135mg / m 2 、140mg / m 2 、145mg / m 2 、150mg / m 2 、155mg / m 2 、160mg / m 2 、165mg / m 2 、170mg / m 2 、175mg / m 2 、180mg / m 2 、185mg / m 2 、190mg / m 2 、195mg / m 2 、200mg / m 2 、205mg / m 2 、 210mg / m 2 、215mg / m 2 、220mg / m 2 、225mg / m 2 、230mg / m 2 、235mg / m 2 、240mg / m 2 、245mg / m 2 、250mg / m 2 、255mg / m 2 、260mg / m 2 、265mg / m 2 、270mg / m 2 、275mg / m 2 、280mg / m 2 、285mg / m 2The patient can be administered a dose of any of the following values: 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 21 mg / kg, 22 mg / kg, 23 mg / kg, 24 mg / kg, 25 mg / kg, 26 mg / kg, 27 mg / kg, 28 mg / kg, 29 mg / kg, 30 mg / kg, 31 mg / kg, 32 mg / kg, 33 mg / kg, 34 mg / kg, 35 mg / kg, 36 mg / kg, 37 mg / kg, 38 mg / kg, 39 mg / kg, 40 mg / kg, 41 mg / kg, 42 mg / kg, 43 mg / kg, 44 mg / kg, 45 mg / kg, 46 mg / kg, 47 mg / kg, 48 mg / kg, 49 mg / kg, 50 mg / kg, 51 mg / kg, 52 mg / kg, 53 mg / kg, 54 mg / kg, 55 mg / kg, 56 mg / kg, 57 mg / kg, 58 mg / kg, 59 mg / kg, 60 mg / kg, 61 mg / kg, 62 mg / kg, 63 mg / kg, 64 mg / kg, 65 mg / kg, 66 mg / kg, 67 mg / kg, 68 mg / kg, 69 mg / kg, 70 mg / kg, 71 mg / kg, 72 mg / kg, 7 2 In some embodiments, the MTDP inhibitor is paclitaxel, which is about, at least, or at most 38 mg / m 2 , 39 mg / m 2 , 40 mg / m 2 , 41 mg / m 2 , 42 mg / m 2 , 43 mg / m 2 、 44 mg / m 2 , 45 mg / m 2 、 46 mg / m 2 , 47 mg / m 2 、 48 mg / m 2 , 49 mg / m 2 、 50 mg / m 2 , 51 mg / m 2 , 52 mg / m 2 , 53 mg / m 2 , 54 mg / m 2 , 55 mg / m 2 , 56 mg / m 2 , 57 mg / m 2 , 58 mg / m 2 , 59 mg / m 2 , 60 mg / m 2 , 61 mg / m 2 , 62 mg / m 2 , 63 mg / m 2 , 64 mg / m 2 , 65 mg / m 2 , 66 mg / m 2 , 67 mg / m 2 , 68 mg / m 2 , 69 mg / m 2 , 70 mg / m 2 , 71 mg / m 2 , 72 mg / m 2 , 73 mg / m 2 , 74 mg / m 2 , 75 mg / m 2 , 76 mg / m2 , 77 mg / m 2 、 78 mg / m 2 , 79 mg / m 2 , 80 mg / m 2 , 81 mg / m 2 、 82 mg / m 2 , 83 mg / m 2 , 84 mg / m 2 , 88 mg / m 2 , 86 mg / m 2 , 87 mg / m 2 , 88 mg / m 2 , 89 mg / m 2 , 90 mg / m 2 , or a numerical dose between any two of these values.

[0068] The MTDP inhibitor can be administered to a patient once a week or twice a week. In some embodiments, the MTDP inhibitor is administered in a 14- to 28-day daily cycle. In some embodiments, the MTDP inhibitor is administered in a 14-, 15-, 16-, 17-, 18-, 19-, 20-, 21-, 22-, 23-, 24-, 25-, 26-, 27-, 28-, 29-, or 30-day cycle. In some embodiments, the MTDP inhibitor 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, or 30 of a cycle. In some embodiments, the MTDP inhibitor is administered on day 1, day 2, day 3, day 4, day 5, day 6, day 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, and / or day 30. In some embodiments, the MTDP inhibitor 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, and / or 30. For example, paclitaxel can be administered in a 5-, 6-, 7-, 8-, 9-, or 10-day cycle. Paclitaxel can be administered weekly in each week or selected weeks of the administration cycle. In some embodiments, paclitaxel can be administered in a 28-day cycle, administered weekly for 3 weeks (e.g., days 1, 8, and 15) and not administered on the remaining days of that cycle, including days 16 through 28.

[0069] 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. In some embodiments, the PLK1 inhibitor is 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 a patient at a dose exceeding 100 mg / kg / day. In some embodiments, onvansertib is administered to a patient daily. In some embodiments, onvansertib is administered in cycles of 5-14 days of daily onvansertib administration and 2-16 days without onvansertib administration. For example, in some embodiments, onvansertib is administered daily for 21 consecutive days of one cycle, followed by 7 days without onvansertib administration. In some embodiments, combination treatment with onvansertib and an MTDP inhibitor can be administered at the same dose as single-agent treatment with onvansertib or an MTDP inhibitor.

[0070] As will be understood by those skilled in the art, the amount and timing of the combined administration of the MTDP inhibitor and the 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 MTDP inhibitor and the PLK1 inhibitor can be formulated into a single pharmaceutical composition or two separate pharmaceutical compositions. The active ingredient can also be encapsulated in microcapsules prepared, for example, by coacervation techniques or interspecific polymerization, such as hydroxymethylcellulose or gelatin microcapsules and poly(methylmethacylate) microcapsules, colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or macroemulsions. The methods, compositions, kits, and systems disclosed herein can be applied to various types of subjects. For example, the subject may be undergoing cancer treatment, in remission from cancer, have undergone one or more cancer treatments, or be suspected of having cancer. The subject may have stage I cancer, stage II cancer, stage III cancer, and / or stage IV cancer. The cancer may be head and neck cancer, non-small cell lung cancer, intrahepatic cholangiocarcinoma, gastric cancer, urothelial cancer, small cell lung cancer, endometrial cancer, cervical cancer, rhabdomyosarcoma, cholangiocarcinoma, liver cancer, ovarian cancer, breast cancer, prostate cancer, colorectal cancer, pancreatic cancer, or a combination thereof. The cancer may be unresectable locally advanced or metastatic disease. These methods may further include administering an additional therapeutic intervention to the subject. The additional therapeutic intervention may include various therapeutic interventions other than administering a PLK1 inhibitor and an MTDP inhibitor, such as antibodies, adoptive T cell therapy, chimeric antigen receptor (CAR) T cell therapy, antibody-drug conjugates, cytokine therapy, cancer vaccines, checkpoint inhibitors, radiation therapy, surgery, chemotherapy drugs, or any combination thereof. The therapeutic intervention can be administered at any time during treatment, for example, when a subject has early-stage cancer. The therapeutic intervention may be more effective than when the therapeutic intervention is administered to a subject at a later time. Without being bound by a particular theory, it is believed that a PLK1 inhibitor (e.g., onvansertib) can sensitize cells (e.g., cancer cells) to MTDP inhibitor treatment to achieve effective cancer treatment.

[0071] Administration and Pharmacokinetics The treatment of the present disclosure may include administering a PLK1 inhibitor (e.g., onvansertib) for a desired duration in one or more cycles of treatment, and administering an MTDP inhibitor. Daily or weekly administration (e.g., intravenous administration) of an MTDP inhibitor can be, or can be about, 0.01 mg, 0.05 mg, 0.1 mg, 0.15 mg, 0.2 mg, 0.25 mg, 0.3 mg, 0.35 mg, 0.4 mg, 0.45 mg, 0.5 mg, 0.55 mg, 0.6 mg, 0.65 mg, 0.7 mg, 0.75 mg, 0.8 mg, 0.85 mg, 0.9 mg, 0.95 mg, 1 mg, 5 mg, 10 mg, 20 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1000 mg, 1100 mg, 1200 mg, or a value or range between any two of these values. During treatment of a subject, the daily or weekly dose of the MTDP inhibitor can be adjusted (e.g., increased or decreased within a range). Daily or weekly administration of the MTDP inhibitor can be in different amounts depending on the day or week. For example, treatment can include daily or weekly administration of the MTDP inhibitor at 0.1 mg to 20 mg in week 1, 0.25 mg to 50 mg in week 2, 0.5 mg to 100 mg in week 3, 1 mg to 200 mg in week 4, and 2 mg to 400 mg from week 5 onward. For example, treatment can include daily or weekly administration of the MTDP inhibitor at 0.1 mg to 100 mg on day 1, 0.2 mg to 200 mg on day 2, 0.4 mg to 400 mg on day 3, and 0.4 mg to 400 mg or 0.6 mg to 600 mg from day 4 onward. For example, the MTDP inhibitor is paclitaxel and is administered in a daily or weekly dose of about 0.01 mg, 0.05 mg, 0.1 mg, 0.15 mg, 0.2 mg, 0.25 mg, 0.3 mg, 0.35 mg, 0.4 mg, 0.45 mg, 0.5 mg, 0.55 mg, 0.6 mg, 0.65 mg, 0.7 mg, 0.75 mg, 0.8 mg, 0.85 mg, 0.9 mg, 0.95 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, or any value or range between any two of these values.

[0072] In some embodiments, the MTDP inhibitor is administered daily or weekly at a dose of about 15 mg / m 2 ~about 275mg / m 2For example, an MTDP inhibitor (e.g., paclitaxel) can be administered at a dose of about 5 mg / m 2 , 10 mg / m 2 , 15 mg / m 2 , 20 mg / m 2 , 25 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 , 75 mg / m 2 , 80 mg / m 2 , 85 mg / m 2 , 90 mg / m 2 , 95 mg / m 2 , 100 mg / m 2 , 105 mg / m 2 , 110 mg / m 2 , 115 mg / m 2 , 120 mg / m 2 , 125 mg / m 2 , 130 mg / m 2 , 135 mg / m 2 , 140 mg / m 2 , 145 mg / m 2 , 150 mg / m 2 , 155 mg / m 2 , 160 mg / m 2 , 165 mg / m 2 , 170 mg / m 2 , 175 mg / m 2 , 180 mg / m 2 , 185 mg / m 2 , 190 mg / m 2 , 195 mg / m 2 , 200 mg / m 2 , 205 mg / m 2 , 210 mg / m 2 , 215 mg / m 2 , 220 mg / m 2 , 225 mg / m 2 , 230 mg / m 2, 235 mg / m 2 , 240 mg / m 2 , 245 mg / m 2 , 250 mg / m 2 , 255 mg / m 2 , 260 mg / m 2 , 265 mg / m 2 , 270 mg / m 2 , 275 mg / m 2 , 280 mg / m 2 , 285 mg / m 2 , or a value or range between or about any two of these values. In some embodiments, the MTDP inhibitor is administered daily or weekly at a unit dose of 38 mg / m 2 , 39 mg / m 2 , 40 mg / m 2 , 41 mg / m 2 , 42 mg / m 2 , 43 mg / m 2 , 44 mg / m 2 , 45 mg / m 2 , 46 mg / m 2 , 47 mg / m 2 , 48 mg / m 2 , 49 mg / m 2 , 50 mg / m 2 , 51 mg / m 2 , 52 mg / m 2 , 53 mg / m 2 , 54 mg / m 2 , 55 mg / m 2 , 56 mg / m 2 , 57 mg / m 2 , 58 mg / m 2 , 59 mg / m 2 , 60 mg / m 2 , 61 mg / m 2 , 62 mg / m 2 , 63 mg / m 2 、 64 mg / m 2 , 65 mg / m 2 , 66 mg / m 2 , 67 mg / m 2 , 68 mg / m 2 , 69 mg / m 2 , 70 mg / m 2 , 71 mg / m 2, 72 mg / m 2 , 73 mg / m 2 , 74 mg / m 2 , 75 mg / m 2 , 76 mg / m 2 , 77 mg / m 2 , 78 mg / m 2 , 79 mg / m 2 , 80 mg / m 2 , 81 mg / m 2 , 82 mg / m 2 , 83 mg / m 2 , 84 mg / m 2 , 88 mg / m 2 , 86 mg / m 2 , 87 mg / m 2 , 88 mg / m 2 , 89 mg / m 2 , 90 mg / m 2 Or at or about a drug / body surface area unit dose of any value or range between any two of these values.

[0073] Each treatment / administration cycle can be of various lengths, e.g., at least 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or more days. In some embodiments, the MTDP inhibitor is given daily, semi-weekly, or weekly for 3 weeks in a 28-day cycle. In exemplary embodiments, the MTDP inhibitor is administered for 1 to 10 cycles, e.g., 1 to 9 cycles, 1 to 8 cycles, 1 to 7 cycles, 1 to 6 cycles, 1 to 5 cycles, 1 to 4 cycles, 1 to 3 cycles, 1 to 2 cycles, or 1 cycle. Administration of the MTDP inhibitor (and / or one or more chemotherapeutic agents) can be daily or weekly, and / or with rest periods between doses. The drug holiday can 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. In some embodiments, the drug holiday can be 6 days and / or 13 days. In some embodiments, the daily or weekly dose of the MTDP inhibitor can be adjusted (e.g., increased or decreased) during treatment of a subject. The daily or weekly administration of the MTDP inhibitor can be in amounts that vary from day to day or week to week. For example, treatment can begin with 80 mg / m on day 1 every week. 2 , 64 mg / m on day 8 2 , and 48 mg / m on day 15. 2For example, treatment may involve administration of an MTDP inhibitor daily or weekly at 0.1 mg to 20 mg in week 1, 0.25 mg to 50 mg in week 2, 0.5 mg to 100 mg in week 3, 1 mg to 200 mg in week 4, and 2 mg to 400 mg from week 2 onwards. For example, treatment may involve administration of an MTDP inhibitor daily or weekly at 0.1 mg to 100 mg on day 1, 0.2 mg to 200 mg on day 2, 0.4 mg to 400 mg on day 3, and 0.4 mg to 400 mg or 0.6 mg to 600 mg from day 4 onwards. For example, the MTDP inhibitor can be administered in a daily or weekly dose of about 0.01 mg, 0.05 mg, 0.1 mg, 0.15 mg, 0.2 mg, 0.25 mg, 0.3 mg, 0.35 mg, 0.4 mg, 0.45 mg, 0.5 mg, 0.55 mg, 0.6 mg, 0.65 mg, 0.7 mg, 0.75 mg, 0.8 mg, 0.85 mg, 0.9 mg, 0.95 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, or any value or range between any two of these values. In some embodiments, the daily or weekly dose of the MTDP inhibitor is 0.005 mg / m 2 , 0.01 mg / m 2 , 0.05 mg / m 2 , 0.1 mg / m 2 , 0.15 mg / m 2 , 0.2 mg / m 2 , 0.25 mg / m 2 , 0.3 mg / m 2 , 0.35 mg / m 2 , 0.4 mg / m 2 , 0.45 mg / m 2 , 0.5 mg / m 2 , 0.55 mg / m 2 , 0.6 mg / m 2 , 0.65 mg / m 2 , 0.7 mg / m 2 , 0.75 mg / m 2 , 0.8 mg / m 2 , 0.85 mg / m 2 , 0.9 mg / m 2 , 0.95 mg / m 2 , 1 mg / m 2 , 2 mg / m 2, 3 mg / m 2 , 4 mg / m 2 , 5 mg / m 2 , 6 mg / m 2 , 7 mg / m 2 , 8 mg / m 2 , 9 mg / m 2 , 10 mg / m 2 or a value or range between any two of these values, or about such a dose. In some embodiments, the patient receives an effective dose of a corticosteroid (e.g., dexamethasone), diphenhydramine, and / or an H2 antagonist (e.g., cimetidine or famotidine) prior to administration of the MTDP inhibitor.

[0074] The maximum concentration (C) of the MTDP inhibitor in the blood of a subject when the MTDP inhibitor is administered alone or in combination with a PLK1 inhibitor. max ) (during or after treatment) can be about 1 pg / mL (picogram / mL) to about 10 μg / mL (microgram / mL). For example, when an MTDP inhibitor is administered alone or in combination with a PLK1 inhibitor, the C of the MTDP inhibitor in the blood of the subject can be maxは、1pg / mL、5pg / mL、10pg / mL、20pg / mL、30pg / mL、40pg / mL、50pg / mL、60pg / mL、70pg / mL、80pg / mL、90pg / mL、100pg / mL、150pg / mL、200pg / mL、250pg / mL、 300pg / mL、350pg / mL、400pg / mL、450pg / mL、500pg / mL、1000pg / mL、5000pg / mL、10000pg / mL、50000pg / mL、100000pg / mL(0.1μg / mL)、0.2μg / mL、0.3μg / mL mL、0.4μg / mL、0.5μg / mL、0.6μg / mL、0.7μg / mL、0.8μg / mL、0.9μg / mL、1μg / mL、1.1μg / mL、1.2μg / mL、1.3μg / mL、1.4μg / mL、1.5μg / mL、1.6μg / mL、1.7μg / mL mL、1.8μg / mL、1.9μg / mL、2μg / mL、2.1μg / mL、2.2μg / mL、2.3μg / mL、2.4μg / mL、2.5μg / mL、2.6μg / mL、2.7μg / mL、2.8μg / mL、2.9μg / mL、3μg / mL、3.1μg / mL 、3.2μg / mL、3.3μg / mL、3.4μg / mL、3.5μg / mL、3.6μg / mL、3.7μg / mL、3.8μg / mL、3.9μg / mL、4μg / mL、4.1μg / mL、4.2μg / mL、4.3μg / mL、4.4μg / mL、4.5μg / mL 、4.6μg / mL、4.7μg / mL、4.8μg / mL、4.9μg / mL、5μg / mL、5.1μg / mL、5.2μg / mL、5.3μg / mL、5.4μg / mL、5.5μg / mL、5.6μg / mL、5.7μg / mL、5.8μg / mL、5.9μg / mL 、6μg / mL、6.1μg / mL、6.2μg / mL、6.3μg / mL、6.4μg / mL、6.5μg / mL、6.6μg / mL、6.7μg / mL、6.8μg / mL、6.9μg / mL、7μg / mL、7.1μg / mL、7.2μg / mL、7.3μg / mL .4μg / mL、7.5μg / mL、7.6μg / mL、7.7μg / mL、7.8μg / mL、7.9μg / mL、8μg / mL、8.1μg / mL、8.2μg / mL、8.3μg / mL、8.4μg / mL、8.5μg / mL、8.6μg / mL、8.7μg / mL.It can be at or about 8 μg / mL, 8.9 μg / mL, 9 μg / mL, 9.1 μg / mL, 9.2 μg / mL, 9.3 μg / mL, 9.4 μg / mL, 9.5 μg / mL, 9.6 μg / mL, 9.7 μg / mL, 9.8 μg / mL, 9.9 μg / mL, 10 μg / mL, a range between any two of these values, or any value between 1 pg / mL and 10 μg / mL.

[0075] Treatments of the present disclosure can include administration of a PLK1 inhibitor (e.g., onvansertib) for a desired duration in one or more cycles. In some embodiments, the PLK1 inhibitor (e.g., onvansertib) is administered for 1 to 10 cycles, e.g., 1 to 9 cycles, 1 to 8 cycles, 1 to 7 cycles, 1 to 6 cycles, 1 to 5 cycles, 1 to 4 cycles, 1 to 3 cycles, 1 to 2 cycles, or 1 cycle. Each treatment cycle can be of various lengths, e.g., at least 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or more days.

[0076] The PLK inhibitor (and / or one or more chemotherapeutic agents) may be administered daily or with a drug holiday between administration days. The drug holiday may be, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or more days. Administration may be once, twice, three times, four times, or more times daily when the PLK inhibitor (and / or one or more chemotherapeutic agents) is administered to a patient. Administration may be, for example, once every 2, 3, 4, 5, 6, or 7 days. The desired length of time can vary, for example, 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, or more days. Each treatment cycle can be of various lengths, for example, at least 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or more days. For example, one cycle of treatment can include administration of a PLK1 inhibitor (e.g., onvansertib) and / or one or more chemotherapeutic agents 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 one or more chemotherapeutic agents 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)). The administration of the PLK1 inhibitor (e.g., onvansertib) and / or one or more chemotherapeutic agents in one cycle of treatment may be continuous or separated by one or more periods (e.g., a 1- or 2-day break). In some embodiments, treatment involves administration of a PLK1 inhibitor (e.g., onvansertib) for 5 days in one 21- to 28-day cycle.In some embodiments, the PLK1 inhibitor (e.g., onvansertib) is administered daily for 21 days, followed by a 7-day rest period. In some embodiments, the PLK1 inhibitor (e.g., onvansertib) is administered orally. In some embodiments, the PLK1 inhibitor (e.g., onvansertib) is administered without a catch-up dose.

[0077] 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 15-24) in a 28-day cycle. The 20 days can be, for example, 10 days (e.g., days 1-10) of continuous daily administration and another 10 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, a PLK1 inhibitor (e.g., onvansertib) is administered to a subject in need thereof for 21 days (e.g., days 1-21) in a 28-day cycle. In some embodiments, for example, if a patient is identified as having low resistance to a PLK1 inhibitor (e.g., onvansertib), 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, consecutive daily administrations for 10 days (e.g., days 1-10) or two daily administrations for each of 5 days (e.g., days 1-5 and days 15-19). In some embodiments, the PLK1 inhibitor (e.g., onvansertib) 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 treatment cycles. In the case of combination treatment, the administration cycles, administration schedules, and / or dosages of the MTDP inhibitor and the PLK1 inhibitor can be the same or different. In the case of combination treatment, the administration cycle, administration schedule, and / or dosage of the MTDP inhibitor can be adjusted according to the administration cycle, administration schedule, and / or dosage of the PLK1 inhibitor. For example, the MTDP inhibitor (e.g., paclitaxel) can be administered three times in a 28-day cycle (e.g., daily administration on days 1, 8, and 15) corresponding to the 28-day cycle of administration of the PLK1 inhibitor (e.g., onvansertib).

[0078] Treatment can be, for example, at a daily dose of 6 mg / m 2~90mg / m 2 For example, treatment can include administration of a PLK1 inhibitor (e.g., onvansertib) at or about 6 mg / m of drug / body surface area. 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 number or range between any two of these values, or 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 7 or 21 mg / m 8 or 22 mg / m 9 or 23 mg / m 10 or 24 mg / m 2 or 24 mg / m 3 or 24 mg / m 4 or 24 mg / m 5 or 24 mg / m 6 or 24 mg / m 7 or 24 mg / m 8 or 24 mg / m 9 or 24 mg / m 10 or 24 mg / m 10 or 24 mg / m 10 or 24 mg / m 2 or 24 mg / m 3 ... 2 In some embodiments, the PLK 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. 2In some embodiments, the PLK inhibitor (e.g., onvansertib) is administered at 8 mg / m every day (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 single cycle of treatment (e.g., the first, second, third, and subsequent cycles).

[0079] 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 MTDP inhibitor. max ) (during or after treatment) can be about 100 nmol / L to about 1500 nmol / L. For example, when a PLK1 inhibitor (e.g., onvansertib) is administered alone or in combination with an MTDP inhibitor, 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 a PLK1 inhibitor (e.g., onvansertib) in the blood of a subject versus time when the PLK1 inhibitor (e.g., onvansertib) is administered as a single agent or in combination with an MTDP inhibitor (e.g., AUC during the first 24 hours after administration). 0-24 ) can be about 1,000 nmol / L-h to about 400,000 nmol / L-h. For example, when a PLK1 inhibitor (e.g., onvansertib) is administered alone or in combination with an MTDP inhibitor, the AUC (e.g., AUC during the first 24 hours after administration) of a plot of the concentration of the PLK1 inhibitor (e.g., onvansertib) in the subject's blood versus time can be calculated.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.

[0080] 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 MTDP inhibitor max ) can be about 1 hour to about 5 hours. For example, when a PLK1 inhibitor (e.g., onvansertib) is administered alone or in combination with an MTDP inhibitor, 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 MTDP inhibitor 1 / 2 ) can be about 10 hours to about 60 hours. For example, when a PLK1 inhibitor (e.g., onvansertib) is administered alone or in combination with an MTDP inhibitor, 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. Patients receiving one or more cycles of an MTDP inhibitor in combination with one or more cycles of a PLK1 inhibitor may experience well-tolerated AEs, including potentially undetectable overt AEs or overt SAEs. A less likely but noteworthy outcome is that the patient likely or even unlikely to experience an AE or SAE. In some embodiments, treatment with the MTDP inhibitor and PLK1 inhibitor combination therapy may result in a significant therapeutic effect. A therapeutic effect greater than predicted from in vitro or in silico analysis indicates a surprising result. A therapeutic dose lower than predicted from in vitro or in silico analysis indicates a surprising result. The combination therapy is expected to mitigate disease progression in patients. A highly positive outcome is that the combination therapy may result in stable disease. Less likely but noteworthy outcomes are a complete response or remission of the cancer, a progression-free survival or overall survival rate greater than predicted from in vitro or in silico analysis, or the absence of measurable disease, target lesions, or malignant lymph nodes.

[0081] Additional cancer medications or therapies The methods, compositions, and kits disclosed herein can be used to treat cancer, e.g., prostate cancer. In some embodiments, a method for treating cancer includes administering an MTDP inhibitor and a PLK1 inhibitor (e.g., onvansertib) to a subject (e.g., a patient) in need thereof. The method may include administering a therapeutically effective amount of an MTDP inhibitor and a therapeutically effective amount of a PLK1 inhibitor. The treatment may include administration of at least one additional cancer therapeutic agent or cancer therapy. The treatment may include administration of a therapeutically effective amount of at least one additional cancer therapeutic agent or cancer therapy. The MTDP inhibitor and the cancer therapeutic agent or cancer therapy can be co-administered, for example, simultaneously or sequentially. The PLK1 inhibitor (e.g., onvansertib) and the cancer therapeutic agent or cancer therapy can be co-administered, for example, simultaneously or sequentially.

[0082] Methods for predicting / determining treatment efficacy and cancer status Also disclosed herein are methods, compositions, kits, and systems for predicting / determining the clinical outcome of a cancer combination therapy of the present disclosure, monitoring the combination therapy, predicting / determining a subject's responsiveness to the combination therapy, determining the cancer status in a subject, and improving the outcome of the combination therapy. These methods, compositions, kits, and systems can be used to predict / determine the clinical outcome of a cancer therapy using a combination of an MTDP inhibitor and a PLK1 inhibitor of the present disclosure, monitor the combination therapy, predict / determine a subject's responsiveness to the combination therapy, determine the cancer status in a subject, improve the outcome of the combination therapy, guide the combination therapy, provide combination therapy recommendations, and / or reduce or avoid ineffective combination therapy. ctDNA can be analyzed to predict / determine the clinical outcome of a cancer therapy, monitor the cancer therapy, predict / determine a subject's responsiveness to the cancer therapy, determine the cancer status in a subject, improve the outcome of the cancer therapy, guide the cancer therapy, provide treatment recommendations, and / or reduce or avoid ineffective cancer therapy. Such analysis of ctDNA is described in PCT Application No. PCT / US2021 / 013287, the contents of which are incorporated herein by reference in their entirety.

[0083] A method of determining a subject's responsiveness to a combination therapy comprising an MTDP inhibitor and a PLK1 inhibitor of the present disclosure can include, for example, analyzing circulating tumor DNA (ctDNA) of a subject with cancer who is undergoing treatment and / or who has received a combination therapy, 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 for a CR, or whether the subject is in or heading for 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. 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 end 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 end of a second cycle of the combination treatment.

[0084] 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. Disclosed herein is a method for determining the cancer status of a subject, comprising analyzing the subject's circulating tumor DNA (ctDNA), thereby determining the cancer status of the subject. The subject may be currently receiving a combination treatment comprising an MTDP inhibitor 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).

[0085] 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. 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. In some embodiments, this 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 a 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.

[0086] 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 as or different from the current or prior concomitant 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, 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., ovarian cancer, breast cancer, prostate cancer, colorectal cancer, pancreatic cancer, or a combination thereof). In some embodiments, the mutant allele frequency is the MAF of one or more driver mutations of cancer (e.g., ovarian cancer, breast cancer, prostate cancer, colorectal cancer, pancreatic cancer, 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 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.

[0087] In some embodiments, the first sample contains ctDNA from the subject before treatment, and one of the additional samples contains ctDNA from the subject after treatment. In some embodiments, the driver mutations are 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, and JAK3. , KDM6A, KIT, KMT2A, KRAS, LUC7L2, MAP2K1, MPL, MYC, MYD88, NF1, NOTCH1, NPM1, NRAS, PDGFRA, PHF6, PPM1D, PTEN, PTPN11, RAD21, RBBP6, RPS14, RUNX1, SETBP1, SF3B1, SH2B3, SLC29A1, SMC1A, SMC3, SRSF2, STAG2, STAT3, TET2, TP53, U2AF1, U2AF2, WT1, XPO1, and ZRSR2. In some embodiments, at least one of the one or more driver mutations is a mutation in said 75 genes. In some embodiments, the one or more driver mutations is a mutation in said 75 genes.

[0088] The driver mutation or at least one of the one or more driver mutations can be in a gene selected from the group consisting of 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.

[0089] 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.

[0090] Disclosed herein is a method for improving cancer treatment outcomes. The method may include detecting a mutant allele frequency in circulating tumor DNA (ctDNA) obtained from the subject at a first time point in a first sample before the subject receives a combination treatment comprising an MTDP inhibitor and a PLK1 inhibitor of the present disclosure; 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; 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, discontinuing the combination treatment on the subject and / or initiating a different combination treatment on the subject. A decrease in the 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.

[0091] Also included herein are methods of treating cancer. The methods can include administering a combination treatment comprising an MTDP inhibitor and a PLK1 inhibitor of the present disclosure to a subject in need thereof; determining a decrease in the 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 mutant allele frequency in 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.

[0092] The first time point can be, for example, before or immediately before the combination therapy. At least one of the one or more additional time points can be, for example, at or after the end of at least one cycle of the combination therapy. In some embodiments, the cycle of the combination therapy is the first cycle of the combination therapy. In some embodiments, the first time point is before or immediately before the first cycle of the combination therapy, and the one or more additional time points are at or after the end of the second cycle of the combination therapy. In some embodiments, the first cycle of the combination therapy is immediately before the second cycle of the combination therapy. 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 the one or more additional samples, and / or the average mutant allele frequency in each of the first sample and the one or more additional samples. In some embodiments, the mutant allele frequency is the mutant allele frequency (MAF) of a driver mutation of cancer (such as ovarian cancer, breast cancer, prostate cancer, colorectal cancer, pancreatic cancer, or a combination thereof). In some embodiments, the mutant allele frequency is the mutant allele frequency (MAF) of one or more driver mutations of cancer (such as ovarian cancer, breast cancer, prostate cancer, colorectal cancer, pancreatic cancer, 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.

[0093] Driver mutations include those 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, KIT, KMT2A, KRAS, LUC7L2, MAP2K1, MPL, and MYC. , MYD88, NF1, NOTCH1, NPM1, NRAS, PDGFRA, PHF6, PPM1D, PTEN, PTPN11, RAD21, RBBP6, RPS14, RUNX1, SETBP1, SF3B1, SH2B3, SLC29A1, SMC1A, SMC3, SRSF2, STAG2, STAT3, TET2, TP53, U2AF1, U2AF2, WT1, XPO1, and ZRSR2, or at least one of the one or more driver mutations is a mutation in one of said 75 genes, and / or the one or more driver mutations is a mutation in said 75 genes. 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 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.

[0094] In some embodiments, the method further comprises determining the mutant allele frequency in one or more of the subject's ctDNA, PBMCs, and BMMCs. The mutant allele frequency in ctDNA can be detected using, for example, polymerase chain reaction (PCR) or 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, the one or more additional samples, and the second sample can be derived from the subject's whole blood, the subject's plasma, the subject's serum, or a combination thereof. In some embodiments, the ctDNA is derived from the subject's whole blood, the subject's plasma, the subject's serum, or a combination thereof.

[0095] 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.

[0096] Compositions and Kits Disclosed herein are compositions and kits for treating cancer. The kit may include a PLK1 inhibitor and a manual providing instructions for administering the PLK1 inhibitor in combination with an MTDP inhibitor to a subject to treat cancer. The kit may include an MTDP inhibitor. The cancer may be, for example, ovarian cancer, breast cancer, prostate cancer, colorectal cancer, pancreatic cancer, or a combination thereof. In some embodiments, the subject has cancer (e.g., head and neck cancer, non-small cell lung cancer, small cell lung cancer, intrahepatic cholangiocarcinoma, gastric cancer, urothelial carcinoma, breast cancer, endometrial cancer, cervical cancer, rhabdomyosarcoma, cholangiocarcinoma, glioblastoma, low-grade glioma, thyroid carcinoma, gallbladder cancer, ovarian cancer, prostate cancer, or a combination thereof). In some embodiments, the instructions include instructions for administering a PLK inhibitor and an MTDP inhibitor simultaneously. In some embodiments, the instructions include instructions for co-administering a PLK inhibitor and an MTDP inhibitor sequentially. In some embodiments, the instructions include instructions for orally administering a PLK1 inhibitor. In some embodiments, the instructions include instructions for orally administering an MTDP inhibitor.

[0097] In some embodiments, the description includes a description that the subject has had a history of treatment with an MTDP inhibitor. In some embodiments, the description includes a description that the subject did not respond to treatment with an MTDP inhibitor alone. In some embodiments, the description includes a description that the subject is known to be resistant to MTDP inhibitor therapy. In some embodiments, the instructions 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 MTDP inhibitor, a PLK 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 has been in complete remission (CR) or partial remission (PR).

[0098] The instructions may include administering each of the MTDP inhibitor and the PLK1 inhibitor to the subject for one cycle at least twice per week. In some embodiments, the instructions include administering each of the MTDP inhibitor and the PLK1 inhibitor to the subject for one cycle at least five times per week. In some embodiments, the instructions include administering the MTDP inhibitor, the PLK1 inhibitor, or both for one cycle of at least 7 days. In some embodiments, each treatment cycle is at least about 21 days. In some embodiments, each treatment cycle is from about 21 days 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 one day of the cycle. In some embodiments, the instructions include administering the MTDP inhibitor daily. In some embodiments, the instructions include administering the MTDP inhibitor and the PLK1 inhibitor for at least two cycles.

[0099] The MTDP inhibitor can be paclitaxel or a pharmaceutically acceptable salt thereof. In some embodiments, the PLK1 inhibitor is selective and / or specific to 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 (BI 6727), 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 description provides a dose of the PLK1 inhibitor at 8 mg / m 2 ~90mg / m2 In some embodiments, the instructions include instructions for administering the MTDP inhibitor at 0.01 mg to 1200 mg (e.g., a daily dose of 0.01 mg to 10 mg administered orally).

[0100] The methods, compositions, and kits disclosed herein can also be used to sensitize cancer cells to one or more MTDP inhibitors. The method may include contacting cancer cells with a composition comprising a PLK1 inhibitor (e.g., onvansertib), or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, thereby sensitizing the cancer cells to the one or more MTDP inhibitors. Contacting the cancer cells with the composition can be performed in vitro, ex vivo, in vivo, or any combination. In some embodiments, contacting the cancer cells with the composition occurs within a subject. In some embodiments, the cancer cells are contacted with the composition in cell culture. The subject may be a mammal, such as a human. Sensitizing cancer cells can increase the responsiveness of cancer cells to one or more MTDP inhibitors by 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or a range between any two of these values, or by about such a percentage. Sensitizing cancer cells can increase the responsiveness of cancer cells to one or more MTDP inhibitors by at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or a range between any two of these values, or by at least about such a percentage. In some embodiments, the increased responsiveness of cancer cells is compared to untreated cancer cells. Sensitization of cancer cells can increase the responsiveness of a subject having cancer cells to one or more MTDP inhibitors by 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or a range between any two of these values, or by about such a percentage.Sensitization of cancer cells can increase the responsiveness of a subject having cancer cells to one or more MTDP inhibitors by at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or a range between any two of these values, or by at least about such a percentage. The increased responsiveness of a subject having cancer cells is, in some embodiments, compared to a subject not treated with the composition.

[0101] The method may include determining sensitization of the cancer cells to one or more MTDP inhibitors after contact with the PLK1 inhibitor. The method may include contacting the cancer cells with one or more MTDP inhibitors simultaneously with and / or after contact with the PLK1 inhibitor. In some embodiments, contacting the cancer cells with one or more MTDP inhibitors occurs in a subject. The subject may be a mammal, e.g., a human. The subject may, for example, be a subject who has not responded to or is known to be resistant to a single MTDP inhibitor. The subject may, for example, be a subject who has previously been treated with one or more MTDP inhibitors. In some embodiments, the method includes determining the subject's response to the one or more MTDP inhibitors. [Example]

[0102] 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.

[0103] Example 1 Onvansertib synergizes with paclitaxel in SCLC cell lines Five small cell lung cancer (SCLC) cell lines were treated with various doses of onvansertib and paclitaxel for 6 days. Cell viability was measured using the CellTiterGlo® assay. Synergy of drug combinations was calculated using the SynergyFinder application. Bliss-independent synergy scores were assessed for each of the SHP77, DMS53, DMS114, H1417, and H69 cell lines. A positive Bliss-independent synergy score indicated synergy between onvansertib and paclitaxel at a given concentration. In these cell lines, strong synergy (indicated by a high positive synergy score) was observed at an onvansertib concentration of approximately 25 nM and over a wide range of paclitaxel concentrations (approximately 0.03 to 500 nM). Additionally, we examined cell viability in SHP77, DMS53, DMS114, H1417, and H69 cell lines treated with various doses of paclitaxel in the absence or presence of onvansertib. Expected viability was calculated using the Bliss independence model of drug additive. The difference between predicted and observed viability in the presence of onvansertib (approximately 25 nM) suggested synergistic effects between onvansertib and paclitaxel. For example, in DMS53 cells, the predicted and observed viability of onvansertib in combination with paclitaxel was approximately 75% and 50%, respectively (i.e., 25% predicted tumor cell inhibition vs. 50% observed tumor cell inhibition), representing an approximately 100% increase in efficacy of the drug combination compared to the combined inhibition caused by paclitaxel and onvansertib alone. Taken together, these results demonstrate that the combination of paclitaxel and ombansertib has an enhanced or synergistic effect in inhibiting cancer progression.

[0104] Example 2 Onvansertib synergizes with paclitaxel in TNCB cell lines We evaluated the activity of the PLK1 inhibitor GSK461364 alone and in combination with the MTDP inhibitor docetaxel in several TNBC cell models. Surprisingly, the antitumor activity of the PLK1 inhibitor GSK461364 showed significant synergy when combined with the MTDP inhibitor docetaxel at concentrations significantly lower than those required for single-agent activity. Surprisingly, the combination of the PLK1 inhibitor GSK461364 and the MTDP inhibitor docetaxel significantly reduced the clonogenic potential and stem cell fraction of two well-studied TNBC cell lines, SUM149 and SUM159. Without being bound by theory, it is believed that PLK1 is required for entry into mitosis upon recovery from DNA damage-induced G2 arrest. Preliminary data suggest that PLK1 is a key functional gene in the basal-like cell line SUM149. The synergistic combination of PLK1 inhibition and chemotherapy, for example with taxanes, can specifically block the G2-M transition, induce aberrant mitotic exit and apoptosis, and eliminate stem cell-like resistant tumor clones.

[0105] As shown in Figure 3, we calculated the combination index (CI) for the combination of paclitaxel and onvansertib in cell lines with p53 mutations (TNBC: SUM149 and SUM159; Luminal: SUM52 and T47D) and wild-type p53 (TNBC: SUM1315; Luminal: MCF7; Normal: MCF10A). The observed synergy (CI < 1) was independent of TNBC vs. ER+ / Luminal classification but dependent on p53 mutation status. These data suggest that patients with mutant p53 breast cancer may be more sensitive to the PLK1 inhibitor onvansertib. We investigated the effects of PLK1 inhibition with onvansertib alone or in combination with paclitaxel in a mouse xenograft model of mesenchymal breast cancer, SUM159. A total of 35 mice were randomized to receive oral (PO) vehicle + intraperitoneal (IP) vehicle (N = 8), PO onvansertib 120 mg / kg on days 1 and 2 weekly + IP vehicle (N = 9), IP paclitaxel 10 mg / kg on day 1 weekly + PO vehicle (N = 8), or PO onvansertib 120 mg / kg on days 1 and 2 weekly + IP paclitaxel 10 mg / kg on day 1 weekly (N = 10) (Figure 4). Onvansertib and paclitaxel demonstrated similar tumor growth inhibition compared to control (difference at day 21 = -0.406 and -0.337, p = 0.262 and 0.340, respectively). The combination of onvasertib and paclitaxel was significantly superior to single-agent treatment with differences of -1.346 compared to onvasertib alone and -1.414 compared to paclitaxel alone (p<0.0001, p<0.0001, respectively).

[0106] 9 mg / m on days 1-21 of each 28-day treatment cycle to increase exposure to onvansertib and optimize efficacy 2 Based on the PK data from a completed phase 1 onvansertib monotherapy study, onvansertib concentrations reached their effective level threshold (IC , taking into account the effect of plasma protein binding) by day 7 (48 hours after the last dose of onvansertib). 50 The tumors were then treated with onvansertib for 14 days during the 21-day cycle, reducing the tumors' response to onvansertib to less than 10-fold (Figure 1). Longer treatment interruptions could allow tumors to recover and continue to grow. A more continuous dosing regimen of onvansertib in patients with mCRPC was under clinical trial (NCT03414034). This trial included patients receiving onvansertib 12 mg / m 2Treatment was performed in combination with abiraterone QD for 14 days (days 1-14) in 21-day cycles (starting on day 1 and continuing without interruption throughout each cycle) (14+7 regimen). As of December 4, 2020, nine patients had been treated with this regimen, with no reported grade 2 or higher hematologic toxicities. A completed phase 1 study of onvansertib alone showed that neutropenia and thrombocytopenia were the primary on-target toxicities of onvansertib. Onvansertib 12 mg / m on a 14+7 regimen was associated with a significant improvement in overall survival. 2 Neutrophil and platelet depletion was not observed in patients with mCRPC treated with 12 mg / m 2 Long-term exposure to onvansertib treatment in mCRPC was shown to be safe and well tolerated. Of nine patients treated with onvansertib in a 14+7 regimen, five were evaluable for efficacy, and three (60%) met the primary efficacy endpoint. Another dosing regimen tested in mCRPC trials (18 mg / m in 14-day cycles) 2 With a 5-day regimen of onvansertib (5+9 regimen), 3 of 10 evaluable patients (30%) met the primary efficacy endpoint. Although preliminary, these data suggest that a more continuous dosing regimen of onvansertib may enhance efficacy.

[0107] Pharmacokinetic data from a completed phase 1 study of onvansertib monotherapy were used to simulate drug exposure to onvansertib under different dosing regimens. The effective level threshold for onvansertib was calculated using the IC500-IC5000, taking into account the effects of plasma protein binding. 50 To assess drug exposure, the maximum concentration (C max ), area under the curve from 0 to 336 hours (AUC (0-336) ), average concentration (C avg ;AUC (0-336) / 336 hours), and 10 × IC 50The time to exceed the IC50 threshold was calculated (Table 2). In Table 2, the dosing schedule is shown along with the number of days on which QD onvansertib is administered and the number of days on which QD onvansertib is not administered in any given treatment cycle. For example, a 5+9 dosing schedule represents administration of QD onvansertib on days 1-5 of a 14-day treatment schedule. 10×IC50 of onvansertib 50 was calculated as 32.5 mg / mL to account for the effect of plasma protein binding. 50 This simulation was performed using a more continuous dosing regimen (12 mg / m 2 10+4 regimen (12 mg / m in a 14-day cycle) 2 Onvansertib for 10 days) or 14+7 plan) avg increased, but C max is 15 mg / m 2 It was shown that the 5+9 plan was lower than the 10×IC plan during the 14-day period. 50 For times exceeding 12 mg / m 2 In patients treated with 15 mg / m for 10 days 2 The mean mean mean 12 mg / m2 was 263 h and 148 h, respectively, compared with patients treated with 12 mg / m2 for 5 days. 2 The 10+4 regimen may result in more pronounced and durable responses in patients. [Table 2] The dose and dosing schedule of onvansertib for this study were selected based on an analysis of pharmacokinetic (PK), pharmacodynamic, safety, and efficacy data from a Phase 1 dose-escalation study. Altering the dose and schedule of onvansertib, as well as coadministration with paclitaxel, may alter the toxicity of the dosing regimen. Therefore, the starting dose selected for this study was 9 mg / m for 21 consecutive days. 2 / day, followed by 7 days off (4-week cycle). 2 The starting dose of 0.001 / day allowed for continuous exposure to onvansertib during weekly paclitaxel infusions.

[0108] Example 3 Background of correlation studies This example describes the use of PLK1 inhibition with onvansertib in combination with a taxane for the treatment of patients with TNBC, and the use of TP53 mutations as a biomarker of response. Based on the TCGA bioportal, TP53 is one of the most commonly mutated genes in breast cancer. Mutation frequency varies depending on the histological and biochemical characteristics of the breast cancer, with higher rates in ductal than lobular, lymph node-positive than lymph node-negative, estrogen receptor (ER)-negative than ER-positive, and HER2-positive than HER2-negative breast cancers. As mentioned above, mutation prevalence also depends on the molecular subtype of the cancer, being most prevalent in triple-negative breast cancer (TNBC) and least prevalent in luminal A subtypes. Furthermore, there is some evidence that TP53 mutations are associated with shorter OS in patients with luminal B and TNBC. Currently, there are no targeted therapies for the treatment of patients with TP53-mutated breast cancer, and patients receive standard chemotherapy. Because TP53-mutated breast cancers are genomically unstable and have difficulty progressing through mitosis, PLK1 is particularly important in these cells and therefore a potential drug target. Consequently, blocking PLK1 function can affect the ability of mutp53-harboring cancer cells to progress through mitosis, thereby increasing their overall sensitivity to chemotherapy drugs such as paclitaxel. Several other genes, including RB, CCND1, BIRC5, BUB1, and PTEN, are also involved in the genomic instability of basal-like breast cancer. With recent advances in immuno-oncology, PLK1 is being explored as an immune modulator in cancer treatment. Cancers with high PLK1 expression tend to exhibit reduced immune activity, including decreased HLA expression and reduced infiltration of B cells, NK cells, and tumor-infiltrating lymphocytes. On the other hand, increased tumor immunity increases the sensitivity of cancer cells to PLK1 inhibitors. The primary mechanism underlying the association between PLK1 and tumor immunity may be aberrant cell cycle and p53 pathways in cancer. Therefore, the combination of PLK1 inhibition and immunotherapy can achieve synergistic antitumor effects.

[0109] In both study phases, baseline biopsies were required if the disease was safe for biopsy. Samples underwent whole-exome sequencing (WES) to capture (1) TP53 mutations and the mutational status of most genes involved in genomic instability, (2) cell cycle proteins (Rb, CCND1, BIRC5, BUB1), and (3) global mutational signatures. Fresh baseline biopsy samples were also subjected to RNA sequencing to assess gene expression and breast molecular subtypes. If subjects did not undergo a baseline biopsy or if the tissue obtained from this biopsy was deemed insufficient, archival tissue was examined for TP53 status. Blood samples were collected at baseline, C3D1, and EOT for assessment of circulating tumor DNA (ctDNA). In addition to evaluating genomic alterations in DNA extracted from blood, the findings were compared with parallel sequencing analysis performed on tissue biopsies from the same subjects to identify similarities and potential differences. Because tissue biopsies are invasive and costly, this analysis aimed to determine whether ctDNA sequencing could serve as an alternative to potentially important tissue-based sequencing. Quantitative changes in ctDNA were assessed throughout the treatment course to evaluate whether there were genomic alterations that could be associated with improved clinical outcomes with onvansertib and paclitaxel and to assess biomarkers of response. To evaluate the role of onvansertib in improving antitumor immunity, peripheral blood mononuclear cells (PBMCs) were collected and analyzed at C1D1, C1D15, and EOT.

[0110] Example 4 Clinical trial of onvansertib and paclitaxel combination treatment in TNBC patients with unresectable locally advanced or metastatic disease Patient selection Patients aged 18 years or older with histologically confirmed invasive breast cancer, unresectable locally advanced or metastatic disease, Eastern Cooperative Oncology Group (ECOG) performance status 0 or 1, and no prior cancer therapy were enrolled, regardless of ethnicity, race, or sex. Unresectable locally advanced or metastatic invasive breast cancer includes inflammatory breast cancer and TNBC. Histologically or cytologically confirmed TNBC is defined as estrogen receptor ≤10%, progesterone receptor ≤10%, and Her-2-neu negative according to the ASCO / CAP 2018 guidelines (IHC 0-1+ or FISH negative). Patients with treated brain metastases who were radiographically stable and had discontinued steroid therapy for at least 4 weeks prior to enrollment were also eligible. Patients with asymptomatic small incidental brain metastases not requiring immediate steroid-containing treatment were also eligible. However, patients with estrogen receptor / progesterone receptor ≥1% were not permitted to receive concomitant endocrine therapy. In addition, the patient's organ function must be as follows: [Table 3]

[0111] Patients were excluded if: (1) they received anticancer chemotherapy or biologic therapy within 21 days or 5 half-lives (whichever was shorter) before the first dose of study drug; (2) they received palliative radiotherapy within 2 weeks of enrollment; (3) they received more than 3 lines of chemotherapy for metastatic disease in phase 2 (no restriction on prior lines in the dose-escalation cohort); (4) they had disease recurrence or progression within 6 months after exposure to the most recent taxane-based therapy in neoadjuvant, field, or metastatic disease; or (5) they underwent major surgery within 6 weeks before starting treatment. (6) pregnant or breastfeeding; (7) gastrointestinal (GI) disorders that, in the investigator's judgment, may significantly impair oral drug absorption (e.g., ileus, active Crohn's disease, ulcerative colitis, extensive gastric or small bowel resection); (8) unable or unwilling to swallow the study drug; (9) including, but not limited to, ongoing or active infection, clinically significant non-healing or healing wound, symptomatic congestive heart failure (CHF) of New York Heart Association (NYHA) functional class II or higher, or unstable angina. (10) Patients with known active infection with COVID-19 or human immunodeficiency virus (HIV) with measurable viral titers and / or active infection with hepatitis B or C (patients vaccinated against hepatitis B virus (HBV) are eligible; patients with HIV and CD4+ T cell (CD4+) counts of 350 cells / μL or more are not eligible). Subjects were eligible if they had been on established ART for at least 4 weeks and had an HIV viral load of less than 400 copies / mL before enrollment; (11) had clinically significant ascites or pleural effusion; (12) had known hypersensitivity to paclitaxel; (13) had grade 2 or greater peripheral neuropathy; (14) had a history of other malignancies, excluding adequately treated nonmelanoma skin cancer, curatively treated noninvasive cervical cancer, or other solid tumors that had been curatively treated and had been symptom-free for at least 2 years;(15) Active medical conditions that make protocol treatment unsafe or impair the patient's ability to tolerate the study drug; (16) Fridericia-corrected QT interval (QTcF) > 480 milliseconds. QTcF is calculated as the arithmetic mean of QTcF on triplicate ECGs. Triplicate ECGs may be repeated once during screening if there is a correctable cause of QT prolongation (e.g., drugs, hypokalemia). The results can be used to determine eligibility; (17) if concomitant use of drugs known to prolong the QT / QTc interval is planned; (18) if risk factors for torsade de pointes, including a family history of long QT syndrome or uncorrected hypokalemia, are present; or (19) if strong inhibitors of CYP3A4, such as atazanavir, ceritinib, clarithromycin, cobicistat and cobicistat-containing combination drugs, darunavir, idelalisib, indinavir, itraconazole, ketoconazole, lonafarnib, lopinavir, mifepristone, nefazodone, If you have used or will use: nelfinavir, ombitasvir-paritaprevir-ritonavir, ombitasvir-paritaprevir-ritonavir + dasabuvir, posaconazole, ritonavir and ritonavir-containing combinations, saquinavir, telithromycin, tucatinib, and voriconazole; and / or strong inducers of CYP3A4 such as apalutamide, carbamazepine, enzalutamide, fosphenytoin, lumacaftor, lumacaftor-ivacaftor, mitotane, phenobarbital, phenytoin, primidone, and rifampin (rifampicin).

[0112] Pre-treatment criteria Patients with laboratory values ​​meeting the following criteria will be treated: (1) absolute neutrophil count >1000 / mm3; (2) platelet count >100,000 / mm3; (3) total bilirubin 1.5 × ULN (institutional standard); and (4) serum creatinine <1.5 × ULN (institutional standard) or calculated growth factor receptor regulating flow (GFR) 60 mL / min.

[0113] Onvansertib and paclitaxel combination therapy Patients who met the following pretreatment criteria within 72 hours of Day 1 of Cycle 1 were treated with the onvansertib-paclitaxel combination in a Phase 1b clinical trial managed using a BOIN design and a Phase 2 clinical trial managed using a Simon two-stage design with the recommended Phase 2 dose (RP2D) for onvansertib administration. Treatment and study visits occurred within + / - 3 days. Patients were divided into three cohorts, with up to nine patients treated at each dose level over 28 consecutive days, according to the dose escalation / deescalation decision map in Figure 1, the dose escalation / deescalation schedule in Table 4, the dosing schedule description in Table 5, and the escalation / deescalation rules in Table 6. Patients began at dose level 0 and received oral onvansertib daily for the first 21 days of a 28-day cycle. Patients also received paclitaxel at 80 mg / m on days 1, 8, and 15 of each 28-day cycle. 2 The dose was administered intravenously at a dose of 0.001 mg / kg / day, which could be reduced at the investigator's discretion. Acceptable forms of paclitaxel included nab-paclitaxel (albumin-bound paclitaxel). In Table 4, onvansertib was administered orally once daily for 21 consecutive days, followed by a 7-day rest period, for a 28-day cycle. In Table 6, "exclusion" means that doses above the current dose were excluded due to excessive toxicity at those doses, and no further patients would be treated at those doses. When a dose was excluded, the dose was automatically tapered to the next lower level. When the lowest dose was excluded, the study was stopped for safety reasons, and no dose was selected as the MTD. If the current dose was the lowest and the protocol indicated dose reduction, the next patient cohort was treated with the lowest dose unless the number of DLTs reached the exclusion boundary, at which point the study for safety purposes was terminated. If the current dose was the highest and the protocol indicated dose escalation, the next patient cohort was treated with the highest dose. If no action (ie, escalation, tapering, or elimination) was invoked, new patients were treated at the current dose. [Table 4] [Table 5] [Table 6]

[0114] Onvansartib was administered orally to patients in approximately 8 ounces (approximately 240 mL) of ice-free water once every 24 hours for 21 days of a 28-day cycle, following a fast of at least 30 minutes, according to Table 5 (once-daily for 21 days followed by 7 days off). If a dose was delayed, it could be administered up to 4 hours after the scheduled time. Food intake could be resumed after 4 hours, and no "make-up" doses were given in the event of vomiting. During Phase 1b, onvansartib was administered in the clinic before paclitaxel on Days C1D1 (Cycle 1 Day 1), C1D8, C1D15, and C2D1. On all other days (including all days of Phase 2), onvansartib was administered at home or in the clinic at approximately the same time each day. All patients were pretreated with corticosteroids, diphenhydramine, and H2 antagonists. Patients were pretreated with dexamethasone, diphenhydramine, and cimetidine or famotidine before paclitaxel administration, as described in Table 5. Paclitaxel pretreatment could be initiated after onvansertib administration. Paclitaxel was administered intravenously once weekly for 3 weeks every 4 weeks according to the institution's package insert. An in-line filter with a microporous membrane of 0.22 microns or less was used. Body surface area (BSA) for the dosing regimen was calculated using the DuBois formula in accordance with DFCI policy. For paclitaxel and onvansertib, BSA was determined on day 1 of each cycle. The dose of onvansertib was rounded to the nearest 5 mg. No investigational or commercially available drugs or therapies other than those listed below may be administered to treat participants' malignancies.

[0115] Phase 1b trial The test was 12 mg / m 2Patients were started with 100 mg of onvansertib. If the observed dose-limiting toxicity (DLT) rate was ≤0.236, the next patient cohort was treated at the next higher dose level. If the observed DLT rate was ≥0.359, the next patient cohort was treated at the next lower dose level. The target DLT rate was 0.3 or 30%. For overdose control, dose levels j and above were excluded from further consideration if Pr(pj > 0.3 | data) > 0.95 (where pj = true DLT rate for dose level j, j = 1,…,3). If the lowest dose was excluded, the study was stopped for safety reasons. The study was terminated when the maximum sample size of 24 patients was reached or when 9 patients continued to receive the same dose according to the escalation / de-escalation rules in Table 6.

[0116] DLTs with 95% confidence intervals were determined according to CTACE version 5.0 after the first cycle of treatment. The maximum tolerated dose (MTL) of onvansertib was 9 mg / m 2 ~18mg / m 2 demonstrated that the combination of onvansertib and paclitaxel was safe and well-tolerated, and this dose can be used as the recommended dose for phase 2. Serious side effects, including grade 3 and grade 4 toxicities, were recorded separately. The proportion of patients experiencing serious side effects, including grade 3 and grade 4 toxicities, did not exceed 16%, indicating that the combination of onvansertib and paclitaxel was safe and well-tolerated. The RP2D was determined using the shiny app "BOIN" by a pre-specified isotonic regression. The isotonic estimate of the toxicity rate closest to the target toxicity rate was selected as the RP2D. In the case of ties, a higher dose level was selected if the isotonic estimate was lower than the target toxicity rate, and a lower dose level was selected if the isotonic monotonic estimate was equal to or greater than the target toxicity rate.

[0117] C max The pharmacokinetics of onvansertib and paclitaxel combination treatment, including AUC, plasma half-life, and plasma clearance, were also determined. The patient sample size was determined by running 10,000 simulations (using the shiny app "BOIN," available at www.trialdesign.org) under various scenarios. Table 7 shows that approximately 14–16 subjects were required. The trial operational characteristics in Table 7 indicate that in Phase 1b trials, the true MTD (if present) was selected with a high probability, and more patients were assigned to the dose level with the DLT rate closest to the target value of 0.3. [Table 7]

[0118] Phase 2 trial This phase 2 study was a single-arm, two-stage Simon design to determine the objective response rate (ORR) of the onvansertib-paclitaxel combination treatment using RECIST 1.1 criteria. Patients in the first, second, and third lines of treatment were enrolled in the phase 2 study. The objective response rate for paclitaxel alone in the third line was set at 10%, with a 10% objective response rate considered clinically unremarkable and expected for paclitaxel alone. A 15% objective response rate was considered clinically noteworthy, with a target objective response rate of 25%. A Simon two-stage design was used with a one-sided type I error of 10% and 80% power to detect the difference between the null ORR of 10% and the target objective response rate of 25%. The first phase of this design began with 13 patients. Two or more responses indicated no benefit, and accumulation continued into the second phase, enrolling 21 additional patients. Six or more responses among 34 patients indicated the combination had clinical benefit meriting further testing (precisely α = 0.095). If the true response rate were 10% or 25%, the probability of stopping the trial after the first phase would be 62% or 13%, respectively. The safety and tolerability of the combination of onvansertib and paclitaxel were also evaluated. Treatment-related toxicities were summarized by maximum grade and terminology using CTCAE v4.0 and reported with 90% binomial exact confidence intervals. Progression-free survival (PFS) was defined as the time from randomization (or enrollment) to progression (per RECIST 1.1) or death from any cause, whichever occurred first, and was assessed using RECIST 1.1 criteria for combination therapy. Patients surviving without progression were censored at the date of the last disease assessment. PFS and overall survival (OS) were reported using the Kaplan-Meier method and are presented with 95% confidence intervals.

[0119] Determination of dose-limiting toxicities (DLTs) DLTs were measured after the first cycle of 28-day therapy in Phase 1b. DLTs were defined as toxicity potentially related to a clinical trial and met one or more of the following criteria: (1) death not clearly attributable to the underlying disease or external causes, (2) grade 3 or higher nonhematologic toxicity with the following exceptions: (3) liver function changes meeting Hy's rule, (4) grade 3 or higher febrile neutropenia, (5) grade 4 or higher neutropenia or thrombocytopenia lasting more than 7 days, and (6) grade 3 or higher thrombocytopenia accompanied by bleeding. Exceptions to grade 3 nonhematologic toxicity included: (1) grade 3 or higher electrolyte abnormalities associated with clinical symptoms, regardless of duration, (2) grade 3 or higher nausea, vomiting, or diarrhea lasting less than 72 hours and for which antiemetics or other supportive care was appropriate, or (3) grade 3 or higher fatigue lasting less than 7 days, or (4) grade 3 or higher electrolyte abnormalities lasting less than 72 hours without clinical symptoms. A DLT may lead to drug withholding until the adverse event (AE) resolved to Grade 1 or less. If the DLT did not resolve within 2 weeks, the patient was withdrawn from the study. Patients were unevaluable and eligible for replacement if (1) they discontinued for reasons other than a DLT, or (2) they received less than 75% (16 of 21 days) of the planned onvansertib dose in Cycle 1 for reasons other than a DLT-related discontinuation. Patients who experienced a DLT and subsequently resolved could continue on an appropriately reduced onvansertib dose if the investigator determined there would be clinical benefit.

[0120] Adverse events, dose delays and modifications All adverse events (AEs), including expected, unexpected, and severe adverse events (SAEs), were monitored and reported in a timely manner according to the clinical trial protocol (Table 8). The severity of AEs was assessed using the latest version of the NCI-CTCAE (Version 5.0). For each SAE, the highest severity grade was reported. Onvansertib-related AEs were generally reversible and generally resolved within 3 weeks. Examples of onvansertib-related AEs include myelosuppression (e.g., anemia, leukopenia, thrombocytopenia, and neutropenia, including febrile neutropenia) and fatigue. Examples of paclitaxel-related AEs include anaphylaxis, a severe hypersensitivity reaction characterized by dyspnea and hypotension, angioedema, and generalized urticaria requiring treatment, myelosuppression (primarily neutropenia) and nadir neutrophils, and severe conduction abnormalities. In Table 8, # indicates that if an event was expected in the protocol and did not require prompt reporting, the event did not need to be reported. * means that AEs occurring within 30 days of a participant enrolled and actively participating in the study or the last intervention must be reported within 1 business day of learning of the event, and & means that events occurring with a severity greater than that listed as expected in the protocol and / or current consent form are considered unexpected, reportable events. [Table 8]

[0121] Patients were provided with appropriate monitoring and supportive care per the clinical trial protocol. Toxicity-related dose delays could be maintained per the clinical trial protocol for up to 28 days. Toxicity-related dose modifications could be maintained up to 18 mg / m per Table 9. 2 of onvansertib, 48 mg / m 2In Table 9, onvansertib was administered orally once daily for 21 consecutive days in every 28-day cycle, and paclitaxel was administered intravenously on days 1, 8, and 15 of every 28-day cycle. Dose escalation within patients was permitted according to the clinical trial protocol. Potential clinical trial-related toxicities were managed according to the clinical trial protocol. For example, hematologic toxicity, nonhematologic toxicity, hepatotoxicity, and neuropathy were managed according to Table 10. Patients could also be withdrawn from treatment and / or the study according to the clinical trial protocol. Regarding the management of hematologic toxicities in Table 7, a superscript "1" means that study treatment should be discontinued if blood counts do not recover to Grade 2 or higher within 4 weeks of treatment interruption, and a superscript "2" means that study treatment should be discontinued if the absolute neutrophil count (ANC) is at baseline grade or below Grade 2 (1000 / mm3). 3 This means that a new cycle may be initiated for patients who have recovered within 24 hours (or more). Regarding the management of non-hematologic toxicities in Table 7, participants who experience intolerable grade 2 events may have their study drug dose withheld and / or reduced at the investigator's discretion. The exceptions, grade 3 nausea and vomiting, did not require routine prophylactic antiemetic therapy at the start of study treatment. However, patients should receive appropriate antiemetic therapy at the first episode of nausea or vomiting and as needed thereafter, according to local practice guidelines. [Table 9] [Table 10]

[0122] Pharmacological assessment The Phase 1b and Phase 2 clinical trials were conducted according to the study calendar (Table 11). Evaluations were performed according to the clinical trial protocol. Pharmacokinetic evaluations were collected for Phase 1b patients at the following time points according to the clinical trial protocol: C1D1 (pre-paclitaxel administration, 1, 2, 3, 4, and 8 hours post-paclitaxel administration), C1D8 (2 hours post-paclitaxel administration), C1D15 (2 hours post-paclitaxel administration), and C2D1 (pre-paclitaxel administration). Abbreviations used in Table 11 include AE ​​(adverse event), ALT (alanine aminotransferase), AST (aspartate aminotransferase), CBC (complete blood count), CT (computed tomography), ctDNA (circulating tumor DNA), ECOG (Eastern Cooperative Oncology Group (performance score)), ECG (electrocardiogram), EOT (end of treatment), MRI (magnetic resonance imaging), PK (pharmacokinetics), and TNM (tumor lymph node metastasis), PBMC (peripheral blood mononuclear cells).In Table 11, medical history includes relevant medical history and recording of TNM stage at diagnosis, recording the start and end dates of previous treatment (recording the reason if not tolerated), physical examination includes height (at screening only), weight, vital signs, and general physical examination; blood test panel (including blood tests and CBC with differential tests) includes sodium, potassium, chloride, bicarbonate, calcium, blood urea nitrogen, creatinine, glucose, albumin, alkaline phosphatase, total bilirubin, AST, and ALT (CBC and blood Testing was permitted up to 48 hours prior to Days 1, 8, and 15, with all visits within a ±3-day window; blood samples for PK analysis were collected on Days 1, 8, and 15 of Cycle 1 and on Day 1 of Cycle 2 only for the Phase 1b part (C1D1 samples were collected pre-dose and 1, 2, 3, 4, and 8 hours post-dose; C1D8 and C1D15 samples were collected 2 hours post-dose; C2D1 samples were collected pre-dose; this applies only to Phase 1b); and for Phases 1b and 2 of this study, blood samples were collected pre-dose if readily available. A baseline biopsy is required after initiation (if available, the facility should confirm access to archival tissue; biopsy at EOT is optional); blood samples for ctDNA assessment (collected in two 10 mL archival tubes) will be collected at C1D1, C3D1, and EOT; chest / abdominal / pelvic imaging, preferably CT scan, must be obtained from subjects within 4 weeks of initiation for disease assessment, and brain MRI (or if subject is able to undergo MRI) is required for subjects with symptoms suggestive of possible central nervous system (CNS) metastatic disease. If not possible, a brain contrast CT scan must be obtained; CT or MRI repeats were performed every 2 cycles (± 1 week) until EOT (e.g., C2D15), with EOT assessment occurring within 28 days (± 5 days) after the last onvansertib dose; subjects received oral onvansertib for 21 consecutive days followed by a 7-day break (onvansertib was administered in the clinic on days 1, 8, and 15 of cycle 1 and day 1 of cycle 2, but was otherwise taken at home); blood for PBMCs was collected in four 8 mL CPT tubes on C1D1, C1D15, and EOT. [Table 11]

[0123] Disease response and progression were assessed using the new international criteria proposed by the Response Evaluation Criteria in Solid Tumors (RECIST) guidelines (version 1.1), which used the change in the longest diameter of the tumor lesion (unidimensional measurement) and the shortest diameter in the case of malignant lymph nodes. Target disease response was assessed in patients with measurable disease at baseline, who had received at least one cycle of therapy, and whose disease had been reassessed. Non-target disease was assessed in patients with measurable disease at baseline but who did not meet the definition of measurable disease, who had received at least one cycle of therapy, and whose disease had been reassessed.

[0124] Assessment of response was based on the presence, absence, or obvious progression of disease. Exemplary responses for patients with measurable disease can be found in Table 12. In Table 12, measurable disease can be target disease. In Table 12, * indicates that response confirmation was only in non-randomized trials with response as the primary endpoint, ** means that in exceptional circumstances, overt progression in non-target disease may be recognized as disease progression. Furthermore, in Table 12, participants who demonstrate an overall deterioration in health status requiring treatment discontinuation without objective evidence of disease progression at that time are reported as "disease worsening." Every effort should be made to document objective progression, even after treatment discontinuation. Typical responses for patients with non-measurable disease can be found in Table 13. In Table 13, non-measurable disease may be non-target disease. In Table 13, "non-CR / non-PD" is preferred over "stable disease" for non-target disease because SD is increasingly being used as an efficacy endpoint in some trials. Therefore, it is not recommended to classify patients with non-measurable disease in this category. [Table 12] [Table 13]

[0125] 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.

[0126] 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 intended as "open" terms that 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 clearly stated 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.

[0127] 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 pharmaceutical composition for use in a method of treating cancer, comprising a polo-like kinase 1 (PLK1) inhibitor, wherein the method comprises administering a microtubule depolymerization (MTDP) inhibitor and a polo-like kinase 1 (PLK1) inhibitor to a subject having cancer, thereby inhibiting or reducing the progression of cancer in the subject.

2. The subject has a blood cancer or solid tumor, Optionally, the cancer may be breast cancer, and further optionally, the breast cancer may be invasive breast cancer, unresectable breast cancer, locally advanced breast cancer, metastatic breast cancer, inflammatory breast cancer, triple-negative breast cancer (TNBC), hormone receptor / growth factor receptor-negative breast cancer, HER2-negative breast cancer, hormone receptor-negative breast cancer, or a combination thereof. The pharmaceutical composition according to claim 1.

3. (a) The object is 1 mm 3 Approximately 1000 or more neutrophils per unit area, or 1 mm 3 Each has more than 100,000 platelets. or (b) The subject has a total bilirubin level approximately 1.5 times the upper limit of the institutional reference value, and / or a serum creatinine glomerular filtration rate lower than approximately 1.5 times the upper limit of the institutional reference value, or a serum creatinine glomerular filtration rate lower than 60 mL / min. Here, optionally, the method may include identifying subjects with cancer as those having more than approximately 1,000 neutrophils per 1 mm³, more than approximately 100,000 platelets per 1 mm³, total bilirubin approximately 1.5 times the upper limit of the institutional reference value, and / or a serum creatinine glomerular filtration rate lower than approximately 1.5 times the upper limit of the institutional reference value or a serum creatinine glomerular filtration rate lower than 60 mL / min. The pharmaceutical composition according to claim 1.

4. The pharmaceutical composition according to claim 1, wherein the PLK1 inhibitor and the MTDP inhibitor are administered simultaneously or sequentially.

5. (a) The PLK1 inhibitor is administered orally, and optionally the subject may fast for more than 30 minutes before administration and / or the subject may fast for about 4 hours after administration. and / or (b) The administration of the MTDP inhibitor is intravenous, and optionally via an in-line filter having a microporous membrane of 0.22 microns or less, and optionally before administration, the subject may be given (i) an oral dose of about 20 mg of dexamethasone about 12 hours and about 6 hours prior, or an intravenous dose of about 12 mg of dexamethasone about 60 minutes prior; (ii) an intravenous dose of about 50 mg of diphenhydramine about 30 minutes to about 60 minutes prior; and / or (iii) an intravenous dose of about 300 mg of cimetidine about 30 minutes to about 60 minutes prior, or an intravenous dose of about 20 mg of famotidine about 30 minutes to about 60 minutes prior. The pharmaceutical composition according to claim 4.

6. (a) The inhibition of cancer progression is greater than the combined inhibition of progression caused by the MTDP inhibitor monotherapy and the PLK1 inhibitor monotherapy, (b) The subject achieves complete success, (c) The subject has a history of treatment with an MTDP inhibitor or a PLK1 inhibitor. (d) The subject did not respond to treatment with the MTDP inhibitor monotherapy or the PLK1 inhibitor monotherapy, and / or (e) The subject is known to be resistant to MTDP inhibitor or PLK1 inhibitor therapy, The pharmaceutical composition according to claim 1.

7. (a) The MTDP inhibitor and the PLK1 inhibitor are administered to the subjects in 28-day cycles. (b) The MTDP inhibitor is administered to the subject in a 28-day cycle, and the PLK1 inhibitor is administered to the subject in a 28-day cycle, comprising approximately 14 to 28 days of administration and approximately 0 to 14 days of non-administration, and optionally comprising approximately 21 days of administration of the PLK1 inhibitor and approximately 7 days of non-administration of the PLK1 inhibitor. and / or (c) The MTDP inhibitor, the PLK1 inhibitor, or both are administered in a 28-day cycle. The pharmaceutical composition according to claim 1.

8. (a) Each treatment cycle is at least about 28 days, or each treatment cycle is about 14 to about 28 days. (b) The PLK1 inhibitor is administered for at least 7 days, at least 14 days, or at least 21 days of a cycle, or the PLK1 inhibitor is not administered for at least 1 day, at least 3 days, or at least 7 days of a cycle. (c) The MTDP inhibitor is administered once or twice a week, or the MTDP inhibitor is administered once a week for two or three consecutive weeks in one cycle. and / or (d) The subject receives at least two cycles of administration of the MTDP inhibitor and the PLK1 inhibitor. The pharmaceutical composition according to claim 1.

9. (a) The MTDP inhibitor is paclitaxel, docetaxel, acetyltaxol, paclitaxel; lutetium Lu 177 bipivotidotetraxetan; 7-hexanoyltaxol; cabazitaxel; larotaxel; mirataxel; ortataxel; tesetaxel; taxoplexin; opaxio; taxoplexin (DHA-paclitaxel); poly(L-glutamic acid)-paclitaxel; abraxane; SB-T-1214; SB-T1216; SB-T121602; SB-T-12854; DHA-SB-T1214; aveotaxane, the aveotaxane may optionally be aveotaxane 15a.2; docetaxel-d 9 -t-Boc; docetaxel-f3-t-Boc; cabazitaxel-7,10-d 6 ; poly(glutamyl-glutamic acid)-taxane complexes; or pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof; or any combination thereof, Optionally, the MTDP inhibitor may be paclitaxel. Furthermore, the subject may optionally be administered paclitaxel at a dose of approximately 48 mg / m² to approximately 80 mg / m² of body surface area. and / or (b) The PLK1 inhibitor is onvancertib (NMS-P937), BI2536, voracertib (BI 6727), GSK461364, adavocertib (AZD1775), CYC140, HMN-176, HMN-214, rigosertib (ON-01910), MLN0905, TKM-080301, TAK-960, GTPL10072, Ro3280; or pharmaceutically acceptable salts, hydrates, solvates or prodrugs thereof; and any combination thereof. Optionally, the PLK1 inhibitor may be onbancertib. Furthermore, onbansertib may be administered at doses of approximately 6 mg / m² to approximately 24 mg / m², and further optionally, at doses of 9 mg / m², 12 mg / m², or 18 mg / m². The pharmaceutical composition according to claim 1.

10. (a) The subject has a history of at least one cancer treatment, Optionally, the treatment history does not have to include the use of MTDP inhibitors, PLK1 inhibitors, or both; further optionally, the PLK1 inhibitor may be onvancertib. and / or (b) The subject was in a state of cancer remission, Optionally, the subject may be one who was in a state of complete remission (CR) or partial remission (PR). The pharmaceutical composition according to claim 1.

11. (a) The method further comprises determining the state of the cancer of the subject, (b) The method further includes determining the responsiveness of the subject to treatment with the MTDP inhibitor and the PLK1 inhibitor, (c) The method further comprises administering one or more cancer drugs or therapies to the cancer, and / or (d) The subject is a human, A pharmaceutical composition according to any one of claims 1 to 10.

12. A composition for use in an in vivo method for sensitizing cancer cells to a microtubule depolymerization (MTDP) inhibitor, comprising a polo-like kinase 1 (PLK1) inhibitor, wherein the method comprises contacting the cancer cells with the composition within a subject to thereby sensitize the cancer cells to the MTDP inhibitor, optionally, (a) The PLK1 inhibitor may be onban sertib, and / or the MTDP inhibitor may be paclitaxel. (b) The subject may be known to have not responded to or to be resistant to the MTDP inhibitor or another MTDP inhibitor. (c) The subject may have a history of treatment with the MTDP inhibitor or another MTDP inhibitor. (d) The subject may be a mammal, and optionally the mammal may be a human. (e) The method may include determining the subject's response to the MTDP inhibitor. and / or (f) The object is 1 mm 3 Approximately 1000 or more neutrophils per unit area, or 1 mm 3 The above composition may also include subjects having more than approximately 100,000 platelets per unit.

13. An in vitro or ex vivo method for sensitizing cancer cells to a microtubule depolymerization (MTDP) inhibitor, comprising contacting cancer cells with a composition comprising a polo-like kinase 1 (PLK1) inhibitor, thereby sensitizing the cancer cells to the MTDP inhibitor.

14. (a) The method comprises determining the sensitization of the cancer cells to the MTDP inhibitor after contact with the composition, and / or (b) The method comprises contacting the cancer cells with the MTDP inhibitor, optionally, the contact of the cancer cells with the MTDP inhibitor may occur simultaneously with or after contacting the cancer cells with the composition. The composition according to claim 12 or the method according to claim 13.

15. Polo-like kinase 1 (PLK1) inhibitors; and A manual providing instructions for administering the PLK1 inhibitor in combination with a microtubule depolymerization (MTDP) inhibitor to patients who require it for cancer treatment. This kit includes, optionally (a) The PLK1 inhibitor may be onbansertib, and / or the MTDP inhibitor may be paclitaxel, and further optionally: (i) The instructions may include instructions to administer the paclitaxel and onbansertib to the subject in 28-day cycles. (ii) The instructions may include instructions to administer paclitaxel and onbansertib to the subject in a cycle of at least five times per week. (iii) The instructions may include instructions that the administration of the MTDP inhibitor, onvansertive, or both shall constitute one cycle of at least 7 days, Furthermore, each treatment cycle may optionally be at least approximately 21 days, or approximately 14 to approximately 28 days. (iv) The instructions may include instructions to administer onvansertive daily for 14 to 21 consecutive days in a 28-day cycle. (v) The instructions may include instructions not to administer onbanserutib for three, five, or seven consecutive days in the cycle, and / or (vi) said instruction comprises administering omburtamib at a dose of about 3 mg / m 2 body surface area to about 24 mg / m 2 body surface area, and / or administering paclitaxel at about 48 mg / m 2 body surface area to about 80 mg / m 2 body surface area, and optionally, said instruction may comprise administering omburtamib at 9 mg / m 2 body surface area, 12 mg / m 2 body surface area or 18 mg / m 2 body surface area. and / or (b) (i) The instructions may include instructions to administer the PLK1 inhibitor and the MTDP inhibitor simultaneously or sequentially. (ii) The instructions may include instructions to administer the PLK1 inhibitor orally, to fast for more than 30 minutes prior to administration, and to fast for approximately 4 hours after administration. (iii) The instructions may include instructions for intravenous administration of the MTDP inhibitor, which may optionally be administered intravenously via an in-line filter having a microporous membrane of 0.22 microns or less, and optionally, before administration, the subject may be given (i) an oral dose of about 20 mg of dexamethasone about 12 hours and about 6 hours prior, or an intravenous dose of about 12 mg of dexamethasone about 60 minutes prior; (ii) an intravenous dose of about 50 mg of diphenhydramine about 30 minutes to about 60 minutes prior; and / or (iii) an intravenous dose of about 300 mg of cimetidine about 30 minutes to about 60 minutes prior, or an intravenous dose of about 20 mg of famotidine about 30 minutes to about 60 minutes prior. (iv) The instructions may include instructions to administer to subjects with a history of treatment with MTDP inhibitors or PLK1 inhibitors. (v) The instructions may include instructions to administer to subjects who have not responded to treatment with the MTDP inhibitor or PLK1 inhibitor monotherapy, (vi) The instructions may include instructions to administer to subjects known to be resistant to MTDP inhibitors or PLK1 inhibitor therapy, and / or (vii) A kit in which the instructions include instructions to administer the MTDP inhibitor weekly for three consecutive weeks in one cycle.

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