Cancer treatment using MTDP inhibitors and PLK1 inhibitors

Abstract

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

Description

[0001] Related Applications

[0002] This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Patent Application Serial No. 63 / 405,308, filed on September 9, 2022, the contents of which are incorporated herein by reference in their entirety for all purposes.

[0003] background

[0004] field

[0005] The present application generally relates to the treatment of cancer. More specifically, provided is a combination therapy for treating cancer using a microtubule depolymerization (MTDP) inhibitor in combination with a polo-like kinase 1 (PLK1) inhibitor.

[0006] Description of Related Technology

[0007] PLK1 is a serine / threonine kinase and is the best characterized member of the family of five closely related regulatory proteins. PLK-1 is a master regulator of mitosis, controlling cell entry and progression into and through mitosis. PLK1 performs several important functions during the mitotic (M) phase of the cell cycle, including regulation of centrosome maturation and spindle assembly, removal of cohesin from chromosome arms, inactivation of anaphase-promoting complex / cyclosome (APC / C) inhibitors, and regulation of mitotic exit and cytokinesis. PLK1 plays a key role in centrosome function and the assembly of the bipolar spindle. PLK1 controls the interaction of the kinetochore with the spindle microtubules, which is required for successful separation and segregation of chromatids into appropriate mother and daughter cells. PLK1 also acts as a negative regulator of p53 family members, leading to the ubiquitination and subsequent degradation of p53 / TP53, inhibiting the pro-apoptotic function mediated by p73 / TP73 and the phosphorylation / degradation of the cofactor bora of Aurora kinase A. During different stages of mitosis, PLK1 localizes to the centrosome, kinetochore, and central spindle. PLK1 is aberrantly overexpressed in a variety of human cancers and is associated with cell proliferation and poor prognosis.

[0008] Tubulin targeting agents are compounds of various types that can be combined with tubulin dimers. The compound includes natural or synthetic compounds that can be combined with the taxane site, colchicine domain, vinca domain or other domains of tubulin dimers. The compound can affect microtubule dynamics and / or polymerization. Some compounds are strong anti-mitotic agents that cause apoptosis, and are candidates for cancer therapeutic agents. The example of tubulin targeting agents includes MTDP inhibitors, such as paclitaxel, docetaxel, epothilone, ixabepilone and discodermolide.

[0009] There is a need to develop effective treatments for cancer patients, including patients who are resistant to treatment with MTDP inhibitors.

[0010] Overview

[0011] Provided are methods, compositions and kits for treating cancer. Some embodiments provide a method for treating cancer, wherein the method comprises: administering a microtubule depolymerization (MTDP) inhibitor and a Polo-like kinase 1 (PLK1) inhibitor to a subject suffering from cancer, thereby inhibiting or reducing the progression of cancer in the subject. In some embodiments, the subject suffers from a blood cancer or a solid cancer. The cancer can be breast cancer, wherein the breast cancer can 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 suffers from TNBC. In some embodiments, the method comprises identifying a subject suffering from cancer as having 1% or more of leukemia per mm 3 More than about 1000 neutrophils per mm 3 In some embodiments, the subject has a total bilirubin of about 1.5 times the upper limit of the institutional normal value, and a glomerular filtration rate of less than about 1.5 times the upper limit of the institutional normal value or 60 mL / min of serum creatinine, or a combination thereof. 3 More than about 1000 neutrophils per mm 3 Greater than about 100,000 platelets, total bilirubin about 1.5 times the upper limit of institutional normal, and a glomerular filtration rate less than about 1.5 times the upper limit of institutional normal or a serum creatinine of 60 mL / min.

[0012] PLK1 inhibitors and MTDP inhibitors can be co-administered simultaneously. PLK1 inhibitors and MTDP inhibitors can also be administered sequentially. In some embodiments, the administration of the PLK1 inhibitor is oral, wherein before administration, the subject fasts for more than about 30 minutes, and wherein after administration, the subject fasts for about 4 hours. In some embodiments, the administration of the MTDP inhibitor is intravenous, optionally through an in-line filter with a microporous membrane of no more than 0.22 microns, wherein before administration, the subject is administered ((i) about 20 mg of dexamethasone orally administered about 12 mg of dexamethasone or about 60 minutes before administration; (ii) about 50 mg of diphenhydramine administered intravenously before administration of about 30 minutes to about 60 minutes; and / or (iii) about 300 mg of cimetidine administered intravenously before administration of about 30 minutes to about 60 minutes or about 20 mg of famotidine administered intravenously before administration of about 30 minutes to about 60 minutes.

[0013] The inhibition of cancer progression can be greater than the combined inhibition of progression caused by a single MTDP inhibitor plus a single PLK1 inhibitor. In some embodiments, the subject achieves a complete response. In some embodiments, the subject has been treated with a prior MTDP inhibitor or PLK1 inhibitor. In some embodiments, the subject has no response to treatment with a single MTDP inhibitor or PLK1 inhibitor. 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 a 28-day cycle. 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, 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, and preferably the days of administration are consecutive. In some embodiments, the MTDP inhibitor, the PLK1 inhibitor, or both are administered in a 28-day cycle.

[0014] Each cycle of treatment can be, or is at least about 28 days. In some embodiments, each cycle of treatment is about 14 days to about 28 days. The PLK1 inhibitor can be administered on at least four days of the cycle. In some embodiments, the PLK1 inhibitor is not administered on at least one day of the cycle. In some embodiments, the MTDP inhibitor is administered once a day or twice a day. In some embodiments, the MTDP inhibitor is administered once a day. In some embodiments, the subject undergoes at least two cycles of administration of the MTDP inhibitor and the PLK1 inhibitor. The MTDP inhibitor can be, for example, paclitaxel, docetaxel, acetyltaxol, paclitaxel; Lu 177vipivotide tetraxetan; 7-hexanoyltaxol; cabazitaxel; larotaxel; milataxel; ortataxel; tesetaxel; taxoprexin; opaxio; taxoprexin (DHA-paclitaxel); poly (L-glutamic acid) -paclitaxel; abraxane; SB-T-1214; SB-T1216; SB-T121602; SB-T-12854; DHA-SB-T1214; abeo-taxane, wherein the abeo-taxane is optionally abeo-taxane 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, about 48 mg / m 2 Body surface area: about 80 mg / m 2 Paclitaxel by body surface area.

[0015] The PLK1 inhibitor can be, for example, onvansertib (NMS-P937), BI2536, volasertib (BI6727), 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, the dosage is about 6 mg / m 2 Body surface area: about 24 mg / m 2In some embodiments, onvansertib is administered at a dose of 9 mg / m 2 Body surface area, 12 mg / m 2 Body surface area and 24 mg / m 2 Dosing of onvansertib was based on body surface area.

[0016] 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; and optionally, the PLK1 inhibitor is onvansertib. In some embodiments, the subject is in cancer remission, such as in complete remission (CR) or in partial remission (PR). In some embodiments, the method further includes determining the subject's cancer state. The method may, for example, also include determining the subject's responsiveness to treatment with an MTDP inhibitor and a PLK1 inhibitor. In some embodiments, the method further includes administering one or more cancer therapeutic agents or therapies for cancer. In some embodiments, the subject is human.

[0017] The present disclosure also includes a method for sensitizing cancer cells to microtubule depolymerization (MTDP) inhibitors, the method comprising: contacting 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 may 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 the subject's body. In some embodiments, the subject has no response to the MTDP inhibitor or is known to be resistant to the MTDP inhibitor. In some embodiments, the subject has a prior treatment with an MTDP inhibitor. The subject may be a mammal, such as a human. The method may include determining the sensitization of the cancer cells to the MTDP inhibitor after contact with the composition.

[0018] In some embodiments, the method includes contacting the cancer cells with an MTDP inhibitor. Contacting the cancer cells with the MTDP inhibitor can occur in a subject. The method can include determining the subject's response to the MTDP inhibitor. In some embodiments, contacting the cancer cells with the MTDP inhibitor is simultaneous with contacting the cancer cells with the composition, or after contacting the cancer cells with the composition. In some embodiments, the subject has 100 μg / mm 3 More than about 1000 neutrophils per mm 3Greater than about 100,000 platelets, total bilirubin about 1.5 times the upper limit of institutional normal, and a glomerular filtration rate less than about 1.5 times the upper limit of institutional normal or a serum creatinine of 60 mL / min.

[0019] The present disclosure also includes a kit comprising: a Polo-like kinase 1 (PLK1) inhibitor; and a manual providing instructions for co-administering the PLK1 inhibitor with a microtubule depolymerization (MTDP) inhibitor to a subject in need thereof to treat cancer. The cancer can be, for example, a blood cancer or a solid cancer, and optionally the cancer is breast cancer, wherein the breast cancer 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 has 100% or more of the breast cancer per mm 3 More than about 1000 neutrophils per mm 3 Greater than about 100,000 platelets, total bilirubin about 1.5 times the upper limit of institutional normal, and a glomerular filtration rate less than about 1.5 times the upper limit of institutional normal or a serum creatinine of 60 mL / min.

[0020] 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 administering the PLK1 inhibitor orally, wherein prior to administration, the subject fasts for more than about 30 minutes, and wherein after administration, the subject fasts for about 4 hours.

[0021] In some embodiments, the instructions include instructions for administering the MTDP inhibitor intravenously, preferably through an in-line filter having a microporous membrane no larger than 0.22 microns, wherein prior to administration, the subject (i) is orally administered about 20 mg of dexamethasone between about 12 hours and about 6 hours prior or is administered about 12 mg of dexamethasone intravenously about 60 minutes prior; (ii) is administered about 50 mg of diphenhydramine intravenously about 30 minutes to about 60 minutes prior; and / or (iii) is administered about 300 mg of cimetidine intravenously about 30 minutes to about 60 minutes prior or is administered about 20 mg of famotidine intravenously about 30 minutes to about 60 minutes prior.

[0022] In some embodiments, the instructions include instructions that the subject has received prior MTDP inhibitor or PLK1 inhibitor treatment. In some embodiments, the instructions include instructions that the subject has not responded to treatment with a single MTDP inhibitor or PLK1 inhibitor. In some embodiments, the instructions include instructions that the subject is known to be resistant to MTDP inhibitor or PLK1 inhibitor therapy. In some embodiments, the instructions include instructions for administering each of paclitaxel and onvansertib to the subject in a cycle of 28 days. In some embodiments, the instructions include instructions for administering each of paclitaxel and onvansertib to the subject in a cycle of at least five times in a week. In some embodiments, the instructions include instructions for administering MTDP inhibitors, onvansertib, or both in a cycle of at least 7 days. In some embodiments, each cycle of treatment is at least about 21 days. In some embodiments, each cycle of treatment is about 14 days to about 28 days. In some embodiments, the instructions include instructions for administering onvansertib for 28 days, including administration of about 14-28 days and non-administration of about 0-14 days, preferably administration of about 15-27 days and non-administration of about 1-13 days, preferably administration of about 16-26 days and non-administration of about 2-12 days, preferably administration of about 17-25 days and non-administration of about 3-11 days, preferably administration of about 18-24 days and non-administration of about 4-10 days, preferably administration of about 19-23 days and non-administration of about 5-9 days, preferably administration of about 20-22 days and non-administration of about 6-8 days, preferably administration of about 21 days and non-administration of about 7 days, and preferably the days of administration are continuous. In some embodiments, the instructions include instructions for not administering onvansertib for at least one day in the cycle. In some embodiments, the instructions include instructions for administering MTDP inhibitors for 28 days. The MTDP inhibitor can be a reversible MTDP inhibitor. In some embodiments, the MTDP inhibitor is paclitaxel or a pharmaceutically acceptable salt, hydrate, solvate, or prodrug thereof.

[0023] In some embodiments, the instructions include administering at about 3 mg / m 2 Body surface area: about 24 mg / m 2 Onvansertib is administered at a dose of approximately 48 mg / m 2 Body surface area: about 80 mg / m 2 In some embodiments, paclitaxel is administered at a dose of 9 mg / m 2 Body surface area, 12 mg / m 2 Body surface area and 24 mg / m 2Onvansertib is administered at a dose of body surface area. In some embodiments, the subject has received at least one prior treatment 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 is in cancer remission, for example, in complete remission (CR) or in partial remission (PR). In some embodiments, the kit also includes an MTDP inhibitor, preferably paclitaxel; and / or a PLK1 inhibitor, preferably onvansertib BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Depicted are non-limiting exemplary embodiments and data relating to a schematic diagram of a dose escalation decision diagram for a Phase 1b trial with a BOIN design.

[0026] Figure 2 Depicted are non-limiting exemplary embodiments of schematic diagrams and data relating to a Phase 2 trial.

[0027] Figure 3 Depicted are non-limiting exemplary embodiments and data relating to the combination index (CI) of the paclitaxel-onvansertib combination in cell lines with mutant p53 (left) and wild-type p53 (right).

[0028] Figure 4 Depicted are non-limiting exemplary embodiments and data relating to the in vivo efficacy of onvansertib (O) alone or in combination with paclitaxel (P) against SUM159 xenografts. DETAILED DESCRIPTION

[0029] In the following detailed description, reference is made to the accompanying drawings, which form a part of the detailed description. In the drawings, similar symbols generally identify similar components unless the 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 accompanying drawings, may be arranged, substituted, combined, separated, and designed in many different configurations, all of which are expressly contemplated herein and constitute a part of the disclosure herein.

[0030] All patents, published patent applications, other publications, and sequences from GenBank and other databases mentioned herein are incorporated by reference in their entirety for the relevant art.

[0031] definition

[0032] Unless otherwise defined, the technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art to which the present disclosure belongs. See, for example, Singleton et al., Dictionary of Microbiology and Molecular Biology 2nd Edition, 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 the purpose of the present disclosure, the following terms are defined below.

[0033] 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, crustaceans, reptiles and particularly 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 and particularly humans.

[0034] As used herein, "patient" refers to a subject who is treated by a medical professional such as a physician (i.e., an allopathic medicine physician or an osteopathic medicine physician) or a veterinary physician in an 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.

[0035] As used herein, "administration" or "administering" refers to a method of giving a dose of a pharmaceutically active ingredient to a vertebrate.

[0036] As used herein, "dose" refers to the amount of the combination of active ingredients (eg, taxane-derived diterpenoids (including paclitaxel) and onvansertib).

[0037] As used herein, "unit dose" refers to the amount of therapeutic agent administered to a patient in a single dose.

[0038] As used herein, the term "daily dose" or "daily dosage" refers to the total amount of a pharmaceutical composition or therapeutic agent to be taken within 24 hours.

[0039] As used herein, the term "delivery" refers to methods, formulations, techniques and systems for delivering pharmaceutical compositions or therapeutic agents to a patient as needed to safely achieve their desired therapeutic effects. In some embodiments, an effective amount of a composition or agent is formulated for delivery to a patient's bloodstream.

[0040] As used herein, the term "formulated" or "formulation" refers to the process of combining different chemical substances including one or more pharmaceutically active ingredients to produce a dosage form. In some embodiments, two or more pharmaceutically active ingredients can be co-formulated into a single dosage form or combined dosage unit, or formulated separately and then combined into a combined dosage unit. Sustained release formulations are formulations designed to slowly release the therapeutic agent in vivo over an extended period of time, while immediate release formulations are formulations designed to quickly release the therapeutic agent in vivo over a shortened period of time.

[0041] As used herein, the term "pharmaceutically acceptable" indicates that the indicated material does not have the properties that would cause a reasonably prudent medical practitioner to avoid administering the material to a patient in view of the disease or condition to be treated and the corresponding route of administration. For example, such materials are generally required to be substantially sterile.

[0042] 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 encapsulating material, which is involved in carrying or transporting any supplement or composition or its components from one organ or part of the body to another organ or part of the body, or delivering the agent to a diseased tissue or a tissue adjacent to a diseased tissue. A carrier or excipient can be used to produce a composition. A carrier or excipient can be selected to facilitate the administration of a drug or prodrug. Examples of carriers include calcium carbonate, calcium phosphate, a variety of sugars such as lactose, glucose or sucrose, or various types of starch, cellulose derivatives, gelatin, vegetable oils, polyethylene glycols and physiologically compatible solvents. Examples of physiologically compatible solvents include sterile solutions of water for injection (WFI), saline solutions and dextrose.

[0043] As used herein, the term "pharmaceutically acceptable salt" refers to any acid addition salt or base addition salt whose counterions are non-toxic to the patient at the pharmaceutical dose of the salt. Many pharmaceutically acceptable salts are well known in the pharmaceutical field. If pharmaceutically acceptable salts of the compounds of the present disclosure are used in these compositions, these salts are preferably derived from inorganic or organic acids and bases. Such acid salts include the following: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate (ethanesulfonate), fumarate, lucoheptanoate (lucoheptanoate), glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate , oxalate, pamoate, pectinate, persulfate, 3-phenyl-propionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, toluenesulfonate, undecanoate, hydrohalide (e.g., hydrochloride and hydrobromide), sulfate, phosphate, nitrate, sulfamate, malonate, salicylate, methylene-bis-b-hydroxynaphthoate, gentisate, isethionate, di-p-toluoyl tartrate, ethanesulphonate, cyclohexylsulfamate, quinate, and the like. Pharmaceutically acceptable base addition salts include, but are not limited to, those derived from alkali metal bases or alkaline earth metal bases or conventional organic bases (such as triethylamine, pyridine, piperidine, morpholine, N-methylmorpholine), ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts of organic bases such as dicyclohexylamine salts, N-methyl-D-glucamine salts, and salts of amino acids such as arginine, lysine, and the like.

[0044] As used herein, the term "hydrate" refers to a complex formed by the combination of water molecules and molecules or ions of a solute. As used herein, the term "solvate" refers to a complex formed by the combination of solvent molecules and molecules or ions of a solute. The solvent can be an organic compound, an inorganic compound, or a mixture of the two. Solvates are intended to include hydrates, hemihydrates, channel hydrates, and the like. Some examples of solvents include, but are not limited to, methanol, N,N-dimethylformamide, tetrahydrofuran, dimethyl sulfoxide, and water.

[0045] As used herein, "therapeutically effective amount" or "pharmaceutically effective amount" refers to the amount of a therapeutic agent that has a therapeutic effect. The dosage of a pharmaceutically active ingredient useful in treatment when administered alone or in combination with one or more additional therapeutic agents is a therapeutically effective amount. Therefore, as used herein, a therapeutically effective amount refers to the amount of a therapeutic agent that produces the desired therapeutic effect as judged based on clinical trial results and / or model animal studies. The therapeutically effective amount will vary depending on the compound, the disease, disorder or condition and its severity, and the age, weight, etc. of the mammal to be treated. The dosage can be conveniently administered, for example, in divided doses up to four times a day or in a sustained release form.

[0046] As used herein, the terms "treat," "treatment," or "treating" refer to the administration of a therapeutic agent or pharmaceutical composition to a subject for preventive and / or therapeutic purposes. The term "prophylactic treatment" refers to the treatment of a subject who has not yet shown symptoms of a disease or condition, but is susceptible to or otherwise 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 the administration of a treatment to a subject who already has a disease or condition. As used herein, a "therapeutic effect" alleviates one or more symptoms of a disease or disorder to some extent. For example, a therapeutic effect can be observed by a reduction in subjective discomfort conveyed by the subject (e.g., reduced discomfort recorded in a self-administered patient questionnaire).

[0047] As used herein, the terms "prophylaxis," "prevent," "preventing," "prevention," and grammatical variations thereof refer to the prophylactic treatment of a subclinical disease state in a subject (e.g., a mammal (including a human)) to reduce the probability of the clinical disease state occurring. The method can partially or completely delay or eliminate the onset or recurrence of one or more of the disorder or condition and / or its attendant symptoms, or prevent the subject from acquiring or reacquiring the disorder or condition, or reduce the risk of the subject acquiring or reacquiring the disorder or condition or its attendant symptoms. Subjects are selected for prophylactic therapy based on factors known to increase the risk of developing a clinical disease state compared to the general population. "Preventive" therapy can be divided into (a) primary prevention and (b) secondary prevention. Primary prevention is defined as treatment of a subject who has not yet presented with a clinical disease state, while secondary prevention is defined as preventing a second occurrence of the same or similar clinical disease state.

[0048] As used herein, each of the terms "partial response" and "partial remission" and "PR" refers to an improvement in the cancer state as measured by, for example, tumor size and / or cancer marker levels in response to treatment. In some embodiments, a "partial response" means that a tumor or a blood marker indicating a tumor is reduced by about 50% in size or level in response to treatment. The treatment can be any treatment directly directed at the cancer, including but not limited to chemotherapy, radiotherapy, hormone therapy, surgery, cell or bone marrow transplantation, and immunotherapy. The size of the tumor can be detected by clinical or radiological means. Markers indicating tumors can be detected by means familiar to technicians, for example, ELISA or other antibody-based tests. A partial response of a target lesion can refer to a reduction of at least 30% in the sum of the diameters of the target lesions, with reference to the sum of the baseline diameters.

[0049] As used herein, each of the terms "complete response" or "complete remission" or "CR" means that the cancerous state as measured by, for example, tumor size and / or cancer marker levels has disappeared after treatment, including but not limited to chemotherapy, radiotherapy, hormone therapy, surgery, cell or bone marrow transplantation, and immunotherapy. The presence of a tumor can be detected by clinical or radiological means. Markers indicating a tumor can be detected by means well known to technicians, for example, ELISA or other antibody-based tests. However, a "complete response" does not necessarily indicate that the cancer has been cured. Relapse may occur after a complete response. A complete response of a target lesion includes the disappearance of all target lesions and any pathological lymph nodes (whether target or non-target), with the short axis reduced to <10 mm. A complete response of a non-target lesion includes the disappearance of all non-target lesions and the normalization of tumor marker levels (the size of all lymph nodes must be non-pathological (short axis <10 mm)). If the tumor markers are initially above the upper limit of normal, they need to be normalized so that the patient is considered a complete clinical response of the non-target lesion. Duration of overall CR was measured from the time the first CR measurement criteria were met until the first day of objectively documented disease progression or death from any cause. Participants without reported events were censored at the time of the last disease assessment.

[0050] As used herein, the term "stable disease" or "SD" means neither sufficient shrinkage to qualify as PR nor sufficient increase to qualify as progressive disease (PD), with reference to the smallest sum of diameters on study. The duration of stable disease is from the start of treatment until the criteria for progression are met, with reference to the smallest measurement recorded since the start of treatment (including the baseline measurement).

[0051] As used herein, the term "progressive disease" or "PD" when referring to target lesions means an increase of at least 20% in the sum of the diameters of the target lesions, with the minimum sum in the study as a reference (if the baseline sum is the minimum sum in the study, this includes the baseline sum). In addition to a relative increase of 20%, the sum must also show an absolute increase of at least 5 mm. (Note: The appearance of one or more new lesions is also considered progression). When progressive disease or PD refers to non-target lesions, it means the appearance of one or more new lesions and / or clear progression of existing non-target lesions. Clear progression should not generally take precedence over target lesion status. It must represent a change in the overall disease state, not an increase in a single lesion.

[0052] As used herein, the term "best overall response" means the best response recorded from the beginning of treatment until disease progression / recurrence (using the minimum measurement recorded since the start of treatment as a reference for progressive disease). The patient's best response distribution depends on the realization of both the measurement value and the confirmation criteria. From the time of meeting the measurement criteria of CR or PR (based on the first record) until objectively recording recurrent or progressive disease (using the minimum measurement recorded since the start of treatment as a reference for progressive disease), or the first day of death due to any reason. Participants with no reported events were censored at the last disease assessment.

[0053] As used herein, the term "MTD" means maximum tolerated dose.

[0054] As used herein, the term "DLT rate" means dose limiting toxicity rate.

[0055] As used herein, the term “IC 50 ” refers to the inhibitory drug concentration that produces 50% of the maximal effect.

[0056] As used herein, the term "SEM" means standard error of the mean.

[0057] As used herein, the term "AUC(xy)" means the area under the curve, where "x" is the start time in hours and "y" is the end time in hours.

[0058] As used herein, the term “C avg " means the average concentration. As used herein, the term "C max ” means maximum concentration.

[0059] As used herein, the term "QD" means once daily.

[0060] As used herein, the term "ANC" means absolute neutrophil count.

[0061] As used herein, the term "EOT" means end of treatment.

[0062] As used herein, the term "ALT" means alanine aminotransferase.

[0063] As used herein, the term "AST" means aspartate aminotransferase.

[0064] As used herein, the term "CBC" means complete blood count.

[0065] As used herein, the term "CT" means computed tomography.

[0066] As used herein, the term "ctDNA" means circulating tumor DNA.

[0067] As used herein, the term "ECOG" means Eastern Cooperative Oncology Group (performance score).

[0068] As used herein, the term "ECG" means electrocardiogram.

[0069] As used herein, the term "MRI" means magnetic resonance imaging.

[0070] As used herein, the term "PK" means pharmacokinetics.

[0071] As used herein, the term "TNM" means tumor, node, metastasis.

[0072] As used herein, the term "PBMC" refers to peripheral blood mononuclear cells.

[0073] As used herein, the term "CNS" means central nervous system.

[0074] As used herein, the term "concomitant medication" means a medication (or treatment) taken or received by the patient during the study (after administration of the first dose of study drug and prior to the final study visit assessment) in addition to the medication used in the study.

[0075] As used herein, the term "tolerable" means a dose level where ≤ 1 / 6 of participants experienced a DLT, or a dose level declared as the RP2D.

[0076] As used herein, the term "adverse event" or "AE" means an unfavorable medical occurrence in a subject to which a drug product is administered that does not necessarily have a causal relationship with the treatment. An AE may be an adverse and unexpected sign (including abnormal laboratory findings), symptom, or disease that is temporarily associated with the use of an investigational product, whether or not related to the investigational drug product. Adverse events may include worsening or aggravation of the disease being studied; worsening or aggravation of a pre-existing condition or event; a concurrent disease; or a drug interaction. Expected fluctuations in pre-existing conditions that do not represent a clinically significant worsening or aggravation are not considered AEs. Surgeries are not adverse events; they are treatment measures for conditions that require surgery. If a condition progresses to the point of requiring surgery or a pre-existing condition worsens to the point of requiring surgery, the condition is an AE. Disease progression is an efficacy endpoint and is not an AE. If a clinical event in a disease progression situation cannot be clearly attributed to the expected disease progression or is consistent with the expected disease progression, the clinical event is considered an AE.

[0077] As used herein, the term "anticipated adverse event" refers to an adverse event that is listed or characterized in the current Adverse Event Listing, Package Insert (PI), Investigator's Brochure (IB), or included as a potential risk in the informed consent document.

[0078] As used herein, the term "unexpected adverse event" refers to an adverse event that is not listed in the PI or the current IB or is unidentified. This includes adverse events that are inconsistent in specificity or severity with the description in the PI or IB. For example, according to this definition, hepatic necrosis would be unexpected.

[0079] As used herein, the term "serious adverse event" or "SAE" means an AE that: (1) results in death (i.e., the AE actually caused or contributed to death); (2) is life-threatening (i.e., the investigator believes the AE placed the subject at immediate risk of death, but does not include AEs that could have caused death if they occurred in a more severe form); (3) requires or prolongs hospitalization; (4) results in persistent or significant disability / incapacity (i.e., the AE caused a serious interruption in the subject's ability to perform normal life functions); or (5) results in congenital anomalies / birth defects in newborns / infants born to mothers exposed to the IMP.

[0080] As used herein, the term "clear AE" means that the AE is clearly related to the study treatment.

[0081] As used herein, the term "probable AE" means that the AE is very likely related to the study treatment.

[0082] As used herein, the term "possible AE" means that the AE may be related to the study treatment.

[0083] As used herein, the term "unlikely AE" means that the AE is doubtfully related to the study treatment.

[0084] As used herein, the term "unrelated AE" means an AE that is clearly not related to study treatment.

[0085] As used herein, the term "expected disease progression" means events that are clearly related to disease progression, and the clinical course is consistent with what should be expected for the patient's disease.

[0086] As used herein, the term "measurable lesions" means lesions that can be accurately measured as ≥20 mm in at least one dimension (the longest diameter to be recorded) by chest x-ray or ≥10 mm by CT scan, MRI, or by clinical examination caliper. Tumor lesions located in previously irradiated areas may or may not be considered measurable. Cystic lesions that are considered to represent cystic metastases are measurable lesions if they meet the definition of measurability described above. However, if non-cystic lesions are also present in the same participant, they are target lesions. Clinical lesions are measurable when they are superficial (e.g., skin nodules and palpable lymph nodes) and the diameter assessed using calipers is ≥10 mm (e.g., skin nodules).

[0087] As used herein, the term "malignant lymph node" means a pathologically enlarged and measurable lymph node with a short axis of ≥ 15 mm when assessed by CT scan.

[0088] As used herein, the term "non-measurable disease" means a small lesion (or disease site) with a longest diameter <10 mm or a pathological lymph node with a short axis of ≥10 to <15 mm. Bone lesions, leptomeningeal disease, ascites, pleural / pericardial effusion, cutaneous lymphangitis / pneumonia, inflammatory breast disease, abdominal masses (no CT or MRI follow-up) and cystic lesions are examples of non-measurable disease. Cystic lesions that meet the criteria for simple cysts defined by radiology are not malignant lesions (neither measurable nor non-measurable), but are simple cysts.

[0089] As used herein, the term "target lesion" means all measurable lesions, up to a maximum of 2 lesions per organ, and a total of 5 lesions, which represent all involved organs. Target lesions are selected based on their size (lesions with the longest diameter), representing all involved organs, but in addition should also be those lesions suitable for repeatable repeated measurements. When the largest lesion itself is not suitable for repeatable measurement, the next largest lesion that can be repeatedly measured is the target lesion.

[0090] As used herein, the term "non-target lesions" means all lesions (or sites of disease) that are not target lesions. Non-target lesions include any measurable lesions in excess of 5 target lesions.

[0091] As used herein, the term "overall survival" or "OS" means the time from randomization (or enrollment) to death due to any cause. Surviving participants were censored at the last known alive date.

[0092] As used herein, the term "progression-free survival" or "PFS" refers to the time from randomization (or enrollment) to the earlier of progression or death from any cause. Participants who survived without disease progression were censored at the date of the last disease assessment.

[0093] As used herein, the term "time to progression" or "TTP" means the time from randomization (or enrollment) to progression. Participants without reported progression were censored at the date of the last disease assessment.

[0094] As used herein, the term "CXD1" means Day 1 of Cycle X. For example, C1D1 means Day 1 of Cycle 1, which is before the subject receives the intended treatment. C2D1 means Day 1 of Cycle 2.

[0095] cancer

[0096] 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, stereoisomer thereof, and a PLK1 inhibitor (e.g., onvansertib) or a pharmaceutically acceptable salt, solvate, stereoisomer thereof to a subject (e.g., patient) in need thereof.

[0097] Methods, compositions and kits disclosed herein can be used to treat various types of cancer and / or tumors.Cancer and / or tumors can be solid tumors, liquid tumors or combinations thereof.In some embodiments, cancer and / or tumors are blood cancers or solid cancers.Cancer 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, unspecified cancer, adenocarcinoma, leukemia, lymphoma, sarcoma, other neoplastic malignancies and / or combinations thereof.Breast cancer can 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. Pancreatic cancer can be metastatic pancreatic ductal adenocarcinoma, pancreatic ductal adenocarcinoma and / or borderline resectable pancreatic adenocarcinoma. Lung cancer can be lung adenocarcinoma and / or mesothelioma. Prostate cancer can be metastatic castration-resistant prostate cancer and / or prostate adenocarcinoma. Colorectal cancer can be metastatic colorectal cancer and / or colon cancer. Bladder cancer can be urothelial carcinoma. Head and neck cancer can be head and neck squamous cell carcinoma, locally advanced or metastatic head and neck cancer, oral cancer, pharyngeal cancer, laryngeal cancer, recurrent laryngeal squamous cell carcinoma, salivary gland cancer, hypopharyngeal cancer, lip and oral squamous cell carcinoma, tongue cancer, nasopharyngeal carcinoma and / or oropharyngeal cancer. Brain and central nervous system cancer can be glioblastoma, glioma and / or neuroblastoma. Liver cancer can be intrahepatic bile duct carcinoma (iCCA) and / or hepatocellular carcinoma. Ovarian cancer can be fallopian tube cancer, peritoneal cancer. Kidney cancer can be renal cell carcinoma, advanced or metastatic renal cell carcinoma and / or metastatic transitional cell carcinoma of the renal pelvis and ureter. Skin cancer can be melanoma, squamous cell carcinoma and / or metastatic Merkel cell carcinoma. Leukemia can be acute myeloid leukemia and / or multiple myeloma. Lymphoma can be B cell lymphoma, T cell lymphoma, non-Hodgkin lymphoma and / or follicular lymphoma. Sarcoma can be rhabdomyosarcoma. In some embodiments, the cancer and / or tumor is a blood cancer or a solid cancer. The blood cancer or solid cancer is not gastric cancer or lung cancer.Hematologic cancers 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 cancer); 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., head and neck squamous cell carcinoma, locally advanced or metastatic head and neck cancer, oral cancer, pharyngeal cancer, laryngeal cancer, recurrent laryngeal squamous cell carcinoma); cancer of the ovary (e.g., pancreatic cancer, pancreatic cancer, ovarian ... In some embodiments, 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, cancer is a liquid tumor. In some embodiments, cancer is a blood cancer. In some embodiments, cancer includes blood cancer. Non-limiting examples of blood cancer include T cell lymphoma, B cell lymphoma, non-Hodgkin 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, the growth of which can be inhibited using the methods and compositions disclosed herein.

[0098] In some embodiments, the cancer and / or tumor is a 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.

[0099] In some embodiments, the cancer is an invasive cancer of unresectable locally advanced or metastatic disease, such as invasive breast cancer. In some embodiments, the cancer is an inflammatory cancer, such as breast cancer, including TNBC. In some embodiments, according to the ASCO / CAP 2018 guidelines, the cancer has a histological or cytological profile of ER≤10%, PR≤10%, Her-2-neu negative (0-1+ by immunohistochemistry (IHC) or fluorescent in situ hybridization (FISH)-negative).

[0100] Cancer and / or tumor can be a cancer and / or tumor with abnormal changes to PLK1 gene or protein. For example, abnormal changes can include one or more PLK1 changes and / or abnormal activation of PLK1, such as copy number changes (CNA), single nucleotide variations (SNV) and gene rearrangements or fusions. Non-limiting exemplary cancers and / or tumors with PLK1 changes include cancers with PLK1 gene or protein amplification, PLK1 gene or protein modification, PLK1 gene deletion, PLK1 gene or protein overexpression, PLK1 gene or protein expression increase and / or its combination. In some embodiments, cancer and / or tumor can be a cancer with PLK1 amplification, wherein the PLK1 gene and / or protein is amplified, for example, due to gene duplication and / or abnormal gene transcription control. For example, compared with healthy tissue, a cancer with PLK1 amplification can be a cancer with higher PLK1 mRNA and / or protein levels. In some embodiments, kidney, lung, breast, colon, skin and / or head and neck may exhibit high PLK1 gene or protein amplification, and the resulting cancer and / or tumor is suitable for treatment by the methods and compositions disclosed herein. Among heterogeneous cancer types, cancer / or tumors may include subtypes with abnormally high expression of PLK1 gene and / or protein. Non-limiting examples of cancers and / or tumors having a subtype containing high PLK1 gene and / or protein expression include lymphoma (e.g., B-cell lymphoma, diffuse large B-cell lymphoma); testicular cancer (e.g., testicular germ cell carcinoma); cervical cancer; head and neck cancer (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, adenocarcinoma, mesothelioma); uterine cancer; skin cancer (e.g., melanoma); sarcoma; brain tumor (e.g., glioblastoma, glioma); leukemia (e.g., acute myeloid leukemia); breast cancer; pancreatic cancer; bile duct cancer (e.g., cholangiocarcinoma); 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, invasive tumors and / or infiltrating tumors. Cancers and / or tumors with amplified PLK1 may have a shorter disease-free survival period than cancers and / or tumors with normal levels of PLK1. Cancers and / or tumors may show high recurrence and / or resistance to traditional and / or monotherapy (such as chemotherapy and / or radiotherapy). Prostate cancer may be resistant to MTDP inhibitors or monotherapy with MTDP inhibitors.

[0101] Cancer and / or tumor can be breast cancer and / or breast tumor.In some embodiments, breast cancer and / or breast tumor are locally unresectable.In some embodiments, breast cancer and / or breast tumor are advanced and / or metastatic.In some embodiments, breast cancer and / or breast tumor have cells containing unstable genome.In some embodiments, breast cancer and / or breast tumor are basal-like subtypes, characterized in that the overexpression of epidermal growth factor receptor (EGFR), the deletion of phosphatase and tensin homologue (PTEN), mutation in TP53 gene (mutp53), and show high overall genome instability in subtype.In some embodiments, breast cancer and / or breast tumor have low pathological complete response rate after neoadjuvant chemotherapy.In some embodiments, breast cancer and / or breast tumor are essentially resistant to chemotherapy and / or have poor overall survival rate.In some embodiments, breast cancer and / or tumor are 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, 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 its combination. In some embodiments, breast cancer and / or breast tumor is a biological invasive form of breast cancer. In some embodiments, breast cancer and / or breast tumor lack estrogen receptor (ER), lack progesterone receptor, and lack human epidermal growth factor receptor 2 (HER2) gene amplification. In some embodiments, breast cancer and / or breast tumor are associated with high mortality, wherein the median survival period is less than two years from the time of transfer. In some embodiments, breast cancer and / or breast tumor is TNBC.

[0102] New targeted therapies and targeted combination therapies are needed for such TNBC with poor prognosis and short overall survival. For example, targeted therapy for TNBC can involve immune checkpoint inhibitors (CPIs). More than 60% of metastatic TNBCs have negative PD-L1 expression (combined positive score (CPS) <10), and in some cases, the therapy can target programmed death 1 (PD-1) / PD-L1. In some cases, paclitaxel alone is the standard first-line therapy. In the KEYNOTE-355 study, chemotherapy (taxane or carboplatin plus gemcitabine) and pembrolizumab (a monoclonal antibody directed against the PD-1 receptor) significantly (27%) reduced the risk of death in patients with metastatic TNBC whose tumors were strongly PD-L1 positive (CPS>10) compared with chemotherapy versus placebo. Recently, the FDA approved sacituzumab govitecan, an antibody-drug conjugate directed against tumor-associated calcium signaling protein 2 (TROP2), as a second- and third-line therapy for metastatic TNBC. However, existing therapies are suboptimal.

[0103] New targeted therapies and targeted combination therapies are needed for basal-like breast cancer, which is intrinsically resistant to chemotherapy, and patients with this type of breast cancer have poor overall survival. Among heterogeneous TNBC, the basal-like subtype, identified by gene expression analysis, is characterized by overexpression of EGFR and PTEN and mutations in the TP53 gene, exhibiting 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 an important gene for the growth and survival of genomically unstable breast cancer cells, including TNBC.

[0104] Paclitaxel is an effective drug for the treatment of metastatic breast cancer. Once-weekly paclitaxel has been shown to have better activity and less myelosuppression than a once-every-3-week schedule. When present, neuropathy is usually mild or moderate and is usually reversible. In a study of 212 patients with metastatic breast cancer, paclitaxel 80 mg / m weekly was given. 2In a large phase 2 trial lasting 4 weeks, with each 4-week cycle, the therapy was generally well tolerated. Grade 3 or 4 neutropenia occurred in 31 patients (15%). Except for 2 patients who developed grade 3 or 4 neutropenia, all patients had received prior chemotherapy, and five of these patients had received prior high-dose chemotherapy. Grade 3 anemia occurred in 18 patients (9%). One patient experienced grade 3 and 4 thrombocytopenia, respectively. 30 patients (14%) with pre-existing grade 1 neuropathy were included. Two of these patients eventually developed grade 3 neuropathy after 5 and 11 courses of treatment. Overall, the incidence of neuropathy of any grade was 69%. However, grade 3 neuropathy occurred in only 20 patients (9%), and no patient experienced grade 4 neuropathy. The median number of cycles of treatment before the development of grade 2 or 3 neuropathy (including patients with preexisting grade 1 toxicity) was 5 cycles (20 weeks), with a range of 1 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 of 131 evaluable patients (17.6%) who had received prior anthracycline therapy, and in 7 of 45 evaluable patients (15.6%) who had received prior taxane therapy. The median time to progression in evaluable patients was 142 days (4.7 months). The median time to progression was 174 days (5.7 months) for patients who had not received prior chemotherapy for metastatic disease, 140 days (4.6 months), and 85 days (2.7 months) for patients who had received two prior regimens.

[0105] MTDP inhibitors and PLK inhibitors

[0106] The methods, compositions and kits disclosed herein can be used to treat cancer 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; unspecified cancer; adenocarcinoma; leukemia; lymphoma; sarcoma; other neoplastic malignancies; and / or combinations thereof. In some embodiments, the method for treating cancer and / or tumors comprises administering a microtubule protein targeting agent, such as an MTDP inhibitor (e.g., paclitaxel) or a pharmaceutically acceptable salt, solvate, stereoisomer thereof and a PLK1 inhibitor (e.g., onvansertib) or a pharmaceutically acceptable salt, solvate, stereoisomer thereof, to a subject (e.g., a patient) in need thereof. The method can include administering a pharmaceutically effective amount of an MTDP inhibitor (eg, paclitaxel) and a pharmaceutically effective amount of a PLK1 inhibitor (eg, onvansertib).

[0107] MTDP inhibitors

[0108] Microtubules are highly dynamic polymers of tubulin that constitute the cytoskeleton, which are essential for cell shape, intracellular transport, cell division and cancer. Microtubule targeting agents, such as compounds that inhibit microtubule dynamics (including spindle-microtubule dynamics), microtubule depolymerization inhibitors and microtubule polymerization inhibitors, are a group of important anti-mitotic compounds with anti-cancer properties, and can be used as first-line cancer therapeutic agents. The inhibition of microtubule dynamics, particularly the inhibition of spindle-microtubule dynamics, can prevent cells from completing mitosis. Cells blocked in the prometaphase / metaphase state can eventually undergo apoptosis.

[0109] Tubulin targeting agents can be used as first-line cancer therapeutic agents. Generally, tubulin targeting agents interfere with microtubule dynamics by stabilizing or destabilizing microtubules, making them suitable first-line cancer therapeutic agents. However, tubulin targeting agents also have tissue specificity, innate and / or acquired drug resistance, and systemic toxicity. In addition, changes in the microtubule dynamics induced by some tubulin targeting agents may be harmful to cancer treatment. For example, tubulin targeting agents can change microtubule chromosome attachment and chromosome missegregation, leading to chromosome instability and aneuploidy. In some cases, tubulin targeting agents can drive tumorigenesis, cancer development, drug resistance, treatment failure, metastasis, poor prognosis, polyploid cell formation (including polyploid giant cells), migration ability, and invasion phenotype.

[0110] Binding to the taxane site at the β-tubulin monomer located in the lumen of the microtubule can stabilize the microtubule lattice. For example, the binding of paclitaxel or epothilone to the taxane site can stabilize the microtubule lattice, although by different mechanisms. Other examples of agents that bind to the taxane site include PM060184 (plocabulin) and the covalent tubulin inhibitor zampanolide. Binding to the laulimalide or peloruside site located at the β-tubulin pocket facing the outside of the microtubule inhibits microtubule disassembly. For example, the binding of laulimalide and peloruside to this site clamps the protofilaments and prevents microtubule disassembly. Binding to the vinca domain located at the inter-dimer interface between two longitudinally arranged tubulin dimers can inhibit tubulin polymerization. For example, the binding of vincristine to the vinca domain can inhibit tubulin assembly by sequestering tubulin into paracrystalline aggregates. Binding to the colchicine site located in a deep pocket between tubulin dimers inhibits microtubule formation. For example, binding of colchicine, benzimidazoles (e.g., nocodazole), or combretastatin to the colchicine site can inhibit microtubule polymerization by preventing conformational changes in tubulin required for polymerization. Binding to the maytansine domain located on an exposed β-tubulin pocket near the vinca site inhibits polymerization at the plus end. For example, binding of maytansine and spongistatin to the maytansine domain can inhibit new tubulin from being added to the plus end. Binding to the pironetin site located on α-tubulin destabilizes microtubules. For example, binding of pironetin to this site can inhibit longitudinal tubulin interactions and prevent heterodimer formation. Binding to the gatorbulin site on α-tubulin located near the colchicine site inhibits tubulin polymerization. For example, binding of cevipabulin to the gatorbulin site can create a wedge by two longitudinally aligned tubulin dimers at the tip of a microtubule.

[0111] MTDP inhibitors include, but are not limited to, diterpenoids; taxanes; taxane-derived diterpenes; taxanes; and derivatives, analogs, precursors, and conjugates thereof. In some embodiments, the MTDP inhibitor is paclitaxel, docetaxel, acetyltaxol, paclitaxel; lutetium Lu 177vipivotide tetraxetan; 7-hexanoyltaxol; cabazitaxel; larotaxel; milataxel; ortataxel; tesetaxel; taxoprexin; opaxio; taxoprexin (DHA-paclitaxel); poly (L-glutamic acid)-paclitaxel; abraxane; SB-T-1214; SB-T1216; SB-T121602; SB-T-12854; DHA-SB-T1214; abeo-taxane. The abeo-taxane may be abeo-taxane 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.

[0112] Non-limiting examples of microtubule targeting agents 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., cyclohexanedione with a distal 2-substituted benzofuran, nitrogen heterocyclic compounds such as plinabulin, verubulin and ABT-751, combretastatins such as ombrabulin and fosbretabulin; and compounds that target other microtubule binding sites (e.g., estramustine. Microtubule targeting agents also include other microtubule destabilizing agents, such as the antitussive drug noscapine; maytansine and / or maytansine / auristatin conjugates (e.g., TDM1, brentuximab vedotin and SAR33419); rhizoxin; spongistatins; podophyllotoxin; steganacins; curacins; antimitotic herbicides that inhibit microtubule polymerization; antifungal and anthelmintic agents; and certain psychoactive drugs (e.g., Dilantin, Vinblastine, chlorpromazine). Microtubule targeting agents also include other microtubule stabilizers such as Eleutherobin, Sarcodictyin, Lorimamine, rhazinalam, steroids, and polyisoprenyl benzophenones.

[0113] Many microtubule targeting compounds are tissue specific. For example, paclitaxel has been proposed to be very effective against ovarian, breast, and lung tumors, but has little effect against many other solid tumors such as kidney cancer, colon cancer, and some sarcomas. As another example, vinca alkaloids have been found to be generally most effective against blood cancers, but generally ineffective against many solid tumors.

[0114] Drug resistance to microtubule targeting compounds is a complex and largely unsolved problem. In some cases, drug resistance is associated with overexpression of ATP-dependent drug efflux pumps or ATP-binding cassettes (a class of membrane transporters). Membrane pumps remove drugs at the intracellular level and can lead to drug resistance to drugs with different chemical structures (e.g., paclitaxel and vinca alkaloids). Tissue-specific differences in regulatory factors (e.g., differences in regulatory protein expression, post-translational modifications of tubulin, and the expression of different tubulin isotypes) can contribute to tissue-specific sensitivity and / or resistance to microtubule targeting compounds.

[0115] In some embodiments, compounds targeting the taxane site of microtubules include paclitaxel, TL00139 and paclitaxel analogs, docetaxel, epothilones (e.g., BMS-247550, epothilones B and D), ixabepilone, discolone and other similar depolymerization inhibitors. In some embodiments, compounds targeting the vicinity of the taxane site of microtubules include stoloniferol and acanthopanax.

[0116] Unlike vinca alkaloids, MTDP inhibitors that bind to the taxane site of tubulin stimulate microtubule polymerization and are an important group of compounds for treating, for example, breast cancer, ovarian cancer, non-small cell lung cancer and Kaposi's sarcoma. However, side effects include neurotoxicity and bone marrow suppression. MTDP inhibitors that target the taxane site in the β-subunit bind to the surface of polymerized microtubules. This stabilizes the microtubules and increases microtubule polymerization and increases its affinity for adjacent tubulin molecules. In addition, at low paclitaxel concentrations, polymerization does not increase but microtubule dynamics can be completely stabilized. For example, the binding of a small amount of paclitaxel molecules (e.g., one paclitaxel per several hundred tubulin molecules) can stabilize the dynamics of microtubules (e.g., reducing the rate or degree of microtubule shortening by about 50%). In HeLa cells, paclitaxel at about 8nM to about 10nM can block mitosis to a semi-maximal extent without increasing microtubule-polymer mass. Typically, inhibitor-mediated inhibition of microtubule dynamics ultimately blocks mitosis and leads to apoptosis.

[0117] In some embodiments, the MTDP inhibitor is paclitaxel, a paclitaxel derivative or analog (e.g., docetaxel), or a pharmaceutically acceptable salt thereof. Paclitaxel, also known as 5β,20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytax-11-ene-9-one 4,10-diacetate 2-benzoate 13-ester with (2R,3S)-N-benzoyl-3-phenylisoserine, has the empirical formula C 47 H 51 NO 14 , and the molecular weight is 853.9, and is a tricyclic diterpenoid compound with a taxane ring (Formula 1). The structure of paclitaxel necessary for antitumor activity includes a taxane ring, a C13 side chain, an oxetane ring, a hydroxyl group at the 2' position, and a homochiral ester chain. The structure of paclitaxel that is not necessary for antitumor activity includes a hydroxyl group at C7 and acetylation of the C10 hydroxyl group.

[0118]

[0119] Paclitaxel is usually obtained from Taxus baccata via a semi-synthetic process, is highly lipophilic, insoluble in water, and melts at about 216°C-217°C. In some embodiments, paclitaxel is administered by intravenous injection. In some embodiments, paclitaxel is administered orally. In some embodiments, paclitaxel is combined with an additive. For example, paclitaxel can be formulated with a surfactant including a heterogeneous nonionic surfactant such as Cremophor EL (polyoxyethylated castor oil) and / or dehydrated alcohol.

[0120] Paclitaxel can be formulated into nano drugs. In some embodiments, paclitaxel is combined with albumin to form 130nm nano drugs (e.g., nab-PTX). In some embodiments, paclitaxel is formulated in polymer micelles, such as forming 25nm nano drugs with mPEG-PDLLA, forming 80nm-100nm nano drugs with PVP-bPNIPAAM, and forming 20nm-60nm nano drugs with N-tr-Lc methyl ester and N.13cr-Lc methyl ester. In some embodiments, paclitaxel is formulated in liposomes such as with lecithin and cholesterol to form 400nm nano drugs. In some embodiments, paclitaxel is formulated in polymer lipid nanoparticles such as with polyvinyl pyrrolidone, cholesterol sulfate and caprylic acid to form 100nm nano drugs. In some embodiments, paclitaxel is formulated in emulsions with monoolein, tricaprylin and Tween80. Paclitaxel can be formulated as a dimer.

[0121] Paclitaxel can be administered orally, for example, paclitaxel can be conjugated to chitosan, lipid derivatives, nanocochlear, hyaluronic acid-octadecylamine micelles, or oil-based nanocarriers. As another example, paclitaxel can be loaded into milk-derived exosomes.

[0122] Paclitaxel has been approved for, 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. Once-weekly paclitaxel has been shown to have better activity and less myelosuppression than a once-every-3-week schedule. When present, neuropathy is usually mild or moderate and is usually reversible. In a study of 212 patients with metastatic breast cancer, paclitaxel 80 mg / m once weekly was given for 1 week. 2In a large phase 2 trial lasting 4 weeks, with each 4-week cycle, the therapy was generally well tolerated. Grade 3 or 4 neutropenia occurred in 31 patients (15%). Except for 2 patients who developed grade 3 or 4 neutropenia, all patients had received prior chemotherapy, and 5 of these patients had received prior high-dose chemotherapy. Grade 3 anemia occurred in 18 patients (9%). One patient experienced grade 3 and 4 thrombocytopenia, respectively. 30 patients (14%) with pre-existing grade 1 neuropathy were included. Two of these patients eventually developed grade 3 neuropathy after 5 and 11 courses of treatment. Overall, the incidence of neuropathy of any grade was 69%. However, grade 3 neuropathy occurred in only 20 patients (9%), and no patient experienced grade 4 neuropathy. The median number of courses of treatment before the development of grade 2 or 3 neuropathy (including patients with pre-existing grade 1 toxicity) was 5 courses (20 weeks), ranging from 1 to 13 courses. Among 177 evaluable patients, the overall response rate was 21.5% (95% confidence interval, 15.4% to 27.5%). Responses occurred in 23 of 131 evaluable patients who had received prior anthracycline therapy (17.6%), and in 7 of 45 evaluable patients who had received prior taxane therapy (15.6%). The median time to progression for evaluable patients was 142 days (4.7 months). The median time to progression for patients who had not received prior chemotherapy for metastatic disease, had received one prior regimen, and had received two prior regimens was 174 days (5.7 months), 140 days (4.6 months), and 85 days (2.7 months), respectively. In some embodiments, paclitaxel is administered to the patient. Docetaxel has been used to treat breast, lung, and prostate cancers.

[0123] 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.

[0124] PLK1 inhibitors

[0125] 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 steps of the cell cycle, including mitotic entry, centrosome maturation, bipolar spindle formation, chromosome segregation, and cytokinesis. It is also essential for entry and progression through mitosis, regulating cell passage through the G2 phase of the cell cycle by phosphorylating forkhead box protein M1 (FOXM1), which then regulates the expression of cell cycle proteins and other genes necessary for cell passage through the cell cycle. PLK1 has been shown to be overexpressed in solid tumors and hematological malignancies, including breast cancer. Breast cancer patients with high PLK1 expression have a lower overall survival than breast cancer patients with low PLK1 expression. PLK1 expression levels are higher in TNBC compared to luminal A, luminal B, and HER-2 overexpressing breast cancer. Inhibition of PLK1 induces G2-M arrest and subsequent apoptosis in cancer cells and has emerged as a promising targeted therapy. Several PLK inhibitors have been studied in clinical trials. In early preclinical development of PLK1-targeted drugs, cancer cells with TP53 mutations (mutp53) were more responsive and had lower IC s than cell lines with wild-type (wtp53) 50 , which is consistent with the lack of checkpoint control and genomic instability associated with mutp53. These observations 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 shown to be important 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 may increase the overall sensitivity of cells to taxanes such as paclitaxel. Through siRNA-mediated screening, PLK1 has been identified as a therapeutic target for TNBC, and inhibition of PLK1 by siRNA-mediated knockdown or chemical inhibitors promotes cell cycle arrest and apoptosis in multiple TNBC lines. The lack of a druggable target is the reason for the poor prognosis of TNBC. In addition to the relatively specific expression of PLK1 in TNBC tissues, the effectiveness and unique spectrum of action of PLK1 inhibition indicate that PLK1 is a promising molecular target for TNBC.

[0126] In a study of patients who had never received treatment In a randomized phase II study of patients with AML who were not suitable for induction therapy, intravenous administration of the pan-PLK inhibitor volasertib (BI6727) in combination with low-dose LDAC) showed a significant increase in OS when compared with Ara-C (cytarabine) (LDAC) alone. A subsequent randomized phase III study did not find a benefit from the combination and described 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 HR-mediated DNA repair.

[0127] Onvansertib (also known as PCM-075, NMS-1286937, NMS-937, "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, for example, with additives such as free base, lactose monohydrate, pregelatinized starch, and behenic acid glyceride. In some embodiments, onvansertib is formulated for oral administration, such as in a hard gelatin capsule.

[0128] Biochemical assays show that onvansertib has high specificity for PLK1 in a group of 296 kinases (including other PLK members). In models of both solid malignancies and hematological malignancies, onvansertib has strong in vitro and in vivo antitumor activity. Onvansertib is the first PLK1-specific ATP-competitive inhibitor administered by the oral route that has been entered into clinical trials with proven antitumor activity in different preclinical models. Onvansertib showed a promising safety profile as a single agent in a Phase I clinical trial. In addition, clinical studies of onvansertib include onvansertib combined with abiraterone and prednisone to treat adult patients with metastatic castration-resistant prostate cancer, onvansertib combined with FOLFIRI and bevacizumab to treat adult patients with KRAS-mutated metastatic colorectal cancer, and onvansertib combined with nanoliposome irinotecan and 5-FU to treat patients with metastatic pancreatic cancer. As described herein, onvansertib can synergize with paclitaxel in the presence of genomic instability and thereafter achieve good antitumor activity at lower doses compared to single agents and without drug-specific toxicity.

[0129] Onvansertib also inhibited cell proliferation of AML cell lines and tumor growth in xenograft models of AML at nanomolar concentrations. In addition, onvansertib significantly increased the antitumor activity of cytarabine in a diffuse model of AML.

[0130]

[0131] Onvansertib showed high potency in proliferation assays against a broad range of cell lines from both solid tumors and hematological malignancies, with low nanomolar activity. Onvansertib has a relatively short half-life of 24 hours and is highly potent against the PLK1 enzyme ([IC 50 ] = 2 nM). In contrast, low or no activity was observed for a panel of 63 kinases except PLK1 (IC 50 >500nM), including PLK members PLK2 and PLK3 (IC 50>10 μM). After oral administration at a well-tolerated dose in mice, onvansertib effectively causes mitotic cell cycle arrest and subsequent apoptosis in cancer cell lines, and inhibits xenograft tumor growth, with a clear PLK1-related mechanism of action. In addition, onvansertib shows activity in combination therapy with approved cytotoxic drugs (such as irinotecan), where there is enhanced tumor regression in HT29 human colon adenocarcinoma xenografts compared to each agent alone, and shows extended survival of animals in a diffusion model of AML in combination therapy with cytarabine. Onvansertib has favorable pharmacological parameters and good oral bioavailability in rodents and non-rodent species, as well as proven anti-tumor activity in different non-clinical models using a variety of dosing regimens, which can provide a high degree of flexibility for dosing schedules, ensuring research in clinical settings. Onvansertib has several advantages over volasertib (BI6727, another PLK1 inhibitor), including a higher degree of potency and specificity for PLK1 isozymes and a higher degree of oral bioavailability. In addition, onvansertib has demonstrated antitumor activity in different nonclinical models using multiple dosing regimens, which can provide flexibility in dosing schedules and therefore ensure studies in the clinical setting.

[0132] A phase I, first-in-human, dose-escalation study of onvansertib in patients with advanced / metastatic solid tumors identified neutropenia and thrombocytopenia as the main dose-limiting toxicities. These hematological toxicities are expected based on the drug's mechanism of action and are reversible, with recovery occurring within 3 weeks. The half-life of onvansertib was determined to be between 20 and 30 hours. The oral bioavailability of onvansertib, coupled with its short half-life, provides an opportunity for a convenient, controlled, and flexible dosing schedule, with the potential to minimize toxicity and improve the therapeutic window. Pharmacodynamic and biomarker studies have been conducted, including baseline genomic profiling, continuous monitoring of mutant allele fractions in plasma, and the extent of PLK1 inhibition in circulating blasts to identify biomarkers associated with clinical response, and are described in CT application No. PCT / US2021 / 013287, the contents of which are incorporated herein by reference in their entirety.

[0133] The major metabolic pathways found in different animal species were N-oxidation of the N-methyl-piperazine ring to produce the N-oxide M2, and hydroxylation on the aliphatic carbon atom of the methylene bridge of the pyrazoloquinazoline moiety to produce the metabolite M1. Qualitatively, no significant differences in the metabolism of onvansertib were observed between species, and quantitatively, some differences were observed across species.

[0134] The potential inhibitory ability of onvansertib against the major human cytochrome P450 (CYP) isoforms responsible for human hepatic drug metabolism (CYP1A2, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4) was investigated using human liver microsomes. Onvansertib was able to inhibit the metabolic activity of CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 isoforms to varying degrees, with 50% inhibitory concentrations (IC 50 ) values ​​ranged from 20 μM to 66 μM (Table 1). No significant inhibition of CYP1A2 was detected. Considering that the relevant concentration for significant antitumor activity of this compound in mice is approximately 1 μM, it is believed that onvansertib is less likely to exhibit clinically relevant metabolic drug-drug interactions. In Table 1, IC 50 Shown are mean ± SEM (standard error of the mean).

[0135] Table 1: Summary of mean inhibitory potency of ONVANSERTIB against human hepatic cytochrome P450S

[0136] P450 enzymes Enzyme response <![CDATA[IC 50 (μM)]]> CYP1A2 Tacrine 1-Hydroxylation >100 CYP2C8 Paclitaxel 6-hydroxylation 20.2±1.6 CYP2C9 Diclofenac 4-hydroxylation 20.4±3.2 CYP2C19 Mephenytoin 4-hydroxylation 36.9±15.7 CYP2D6 Bufuralol 1-hydroxylation 26.8±5.4 CYP3A4 Testosterone 6β-hydroxylation 52.7±9.8 CYP3A4 Midazolam 1'-Hydroxylation 66.2±4.0

[0137] To date, a Phase 1 safety study of onvansertib in adult patients with advanced / metastatic solid tumors has been completed at a single center in the United States. The first cycle of oral onvansertib for 5 consecutive days every 3 weeks (i.e., 21-day treatment cycle) dose-limiting toxicity (DLT) and maximum tolerated dose (MTD) were conducted. The safety profile of onvansertib has been determined to determine the pharmacokinetics (PK) of onvansertib in plasma (MTD) and to record any antitumor activity. In a study, a total of 21 patients were enrolled and 19 patients were treated. The first 3 dose levels (6 mg / m 2 / day, 12mg / m 2 / day and 24mg / m 2 No DLT occurred at subsequent dose levels (48 mg / m 2 / day), 2 of 3 patients experienced DLT. 2 At the intermediate dose level, 4 patients were treated and 2 DLTs were observed. After further cohort expansion, the MTD was determined to be 24 mg / m 2 / day. The best treatment response observed was stable disease (SD), which occurred in 5 of 16 evaluable patients. The study identified thrombocytopenia and neutropenia as the main toxicities. This is consistent with the expected mechanism of action of onvansertib and the results from preclinical studies. These hematological toxicities are reversible and usually recover within 3 weeks. No other clinically relevant safety findings were seen with onvansertib as a single-agent treatment. No other mechanism-related, potentially expected events, such as gastrointestinal disorders, mucositis, and alopecia, were observed, confirming that bone marrow is the most sensitive target of onvansertib in humans under this schedule.

[0138] As disclosed herein, combination therapy using MTDP inhibitors (including paclitaxel) and PLK1 inhibitors (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 bile duct cancer, gastric cancer, urothelial carcinoma, small cell lung cancer, breast cancer, endometrial cancer, cervical cancer, rhabdomyosarcoma, bile duct cancer, ovarian cancer, or a combination thereof), resulting in tumor regression and improved cancer survival. The tumor regression and cancer survival rate / duration produced by the combination can be surprisingly synergistic (i.e., beyond additivity, superior to the cumulative anti-tumor efficacy caused by MTDP inhibitors and PLK1 inhibitors alone). For example, as described herein, the combination of onvansertib and paclitaxel shows synergy in in vitro and in vivo models of chemotherapy-resistant ovarian cancer. In a triple-negative breast cancer model, treatment with onvansertib and paclitaxel is surprisingly synergistic. In addition, surprisingly, when combined together, the concentration of PLK1 inhibitors for obtaining complete cell inhibition is significantly lower than that of a single agent for obtaining the same inhibition.

[0139] Provided herein are methods, compositions and kits for treating cancer in subjects (e.g., human patients suffering from cancer). The method includes administering an MTDP inhibitor and a PLK1 inhibitor to a patient in a manner sufficient to inhibit or reduce the progression of cancer. For example, an MTDP inhibitor and a PLK1 inhibitor can be administered to a subject suffering from cancer simultaneously, individually or sequentially. It is expected that the combination therapy of onvansertib and an MTDP inhibitor is significantly more effective for various cancer treatments, including the treatment of prostate cancer and lung cancer (e.g., neuroendocrine prostate cancer), than the combination therapy of another PLK inhibitor BI2536 and MTDP.

[0140] In some embodiments, the inhibition or reduction of cancer progression is not merely additive, but is enhanced or synergistic (i.e., the inhibition is greater than the combined inhibition of progression caused by the MTDP inhibitor alone plus the PLK1 inhibitor alone). In different embodiments, the enhanced or synergistic potency or inhibition of any combination of the MTDP inhibitor and the PLK1 inhibitor of the present disclosure may be different. In some embodiments, the enhanced or synergistic potency or inhibition of any combination of the MTDP inhibitor and the PLK1 inhibitor of the present disclosure is less than, about less than, at least less than, at least about less than, up to or up to about less than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 60%, 70%, 75%, 80%, 85%, 90%, 90%, 95%, 10 ... %, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 110%, 120%, 130%, 140%, 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.

[0141] 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 less than, about less than, at least less than, at least about less than, up to about less than, or up to 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, as compared to the combined inhibition of progression caused by an MTDP inhibitor (e.g., paclitaxel) plus a PLK1 inhibitor (e.g., onvansertib) alone. For example, the combination of an MTDP inhibitor and a PLK1 inhibitor can cause 50%, 60%, 70%, 80%, 90% or more inhibition of cancer progression (cancer cell viability of 50%, 40%, 30%, 20%, 10% or less), while under the same conditions, the combined inhibition of a single MTDP inhibitor (e.g., paclitaxel) plus a single PLK1 inhibitor can be 10%, 20%, 25%, 30% or less inhibition of cancer progression (cancer cell viability of 90%, 80%, 75%, 70% or more). Thus, the enhanced or synergistic efficacy or inhibition of cancer progression caused by a 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 higher than the combined inhibition of progression caused by a single MTDP inhibitor (e.g., paclitaxel) plus a single PLK1 inhibitor. In some embodiments, the MTDP inhibitor is paclitaxel and the PLK1 inhibitor is onvansertib.

[0142] The methods disclosed herein can be effective against various cancers, for example, 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; unspecified cancer; adenocarcinoma; leukemia; lymphoma; sarcoma; other neoplastic malignancies or a combination thereof.

[0143] As described herein, patients can achieve a complete response or a partial response after 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 does not respond to treatment with one or more MTDP inhibitors (without a PLK1 inhibitor). In some embodiments, the patient does not respond to treatment with a single MTDP inhibitor.

[0144] The MTDP inhibitor and the PLK1 inhibitor can be administered to the patient in any manner that is considered effective for treating cancer. The MTDP inhibitor can be administered together with the PLK1 inhibitor, or administered alone with the PLK1 inhibitor. When administered alone, the MTDP inhibitor can be administered before or after the PLK1 inhibitor, or in different administration cycles.

[0145] The MTDP inhibitor and PLK1 inhibitor can each be administered on any schedule, such as once or more than once a day or a week; once, twice, three times, four times, five times, six times or seven times a week (daily); for one week or more than one week; etc. The PLK1 inhibitor (e.g., onvansertib) can, for example, be administered orally. The MTDP inhibitor (e.g., paclitaxel) can, for example, be administered by intravenous infusion (e.g., over about 30 minutes). In some embodiments, the PLK1 inhibitor (e.g., onvansertib) is or is only administered to the subject daily for 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 consecutive days during the cycle, for example, the first 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 consecutive days of the cycle; and the MTDP inhibitor (e.g., paclitaxel) is or is administered to the patient once in each week of onvansertib administration. The length of the cycle can be, for example, 21-28 days. In some embodiments, the PLK1 inhibitor is administered to the patient daily for the first 21 consecutive days during a 28-day cycle, and the MTDP inhibitor is administered to the patient once a week for the first 3 weeks of the 28-day cycle. In some embodiments, in the last 7 days of the 28-day cycle, neither the PLK1 inhibitor nor the MTDP inhibitor is administered to the patient. The patient may undergo one or more cycles of treatment / administration, such as at least two cycles of treatment / administration. The schedule of administration of the MTDP inhibitor and the PLK1 inhibitor may be the same or different in each cycle of treatment / administration.

[0146] The MTDP inhibitor can be administered to the patient at any suitable dose, for example, about, at least, or at most the following dose: 5 mg / m 2 , 10mg / m 2 , 15mg / m 2 , 20mg / m 2 , 25mg / m 2 , 30mg / m 2 , 35mg / m 2 , 40mg / m 2 45mg / m 2 , 50mg / m 2 , 55mg / m 2 , 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 2 , or a number between any two of these values. A dosage unit based on body weight (mg / kg) can be converted to another unit (e.g., mg / m 2 ), as will be understood by those skilled in the art. In some embodiments, the MTDP inhibitor is paclitaxel, which is administered at a dose of about, at least, or at most: 38 mg / m 2 , 39mg / m 2 , 40mg / m2 、41mg / m 2 、42mg / m 2 、43mg / m 2 、44mg / m 2 、45mg / m 2 、46mg / m 2 、47mg / m 2 、48mg / m 2 、49mg / m 2 、50mg / m 2 、51mg / m 2 、52mg / m 2 、53mg / m 2 、54mg / m 2 、55mg / m 2 、56mg / m 2 、57mg / m 2 、58mg / m 2 、59mg / m 2 、60mg / m 2 、61mg / m 2 、62mg / m 2 、63mg / m 2 、64mg / m 2 、65mg / m 2 、66mg / m 2 、67mg / m 2 、68mg / m 2 、69mg / m 2 、70mg / m 2 、71mg / m 2 、72mg / m 2 、73mg / m 2 、74mg / m 2 、75mg / m 2 、76mg / m 2 、77mg / m 2 、78mg / m 2 、79mg / m 2 、80mg / m 2 、81mg / m 2 、82mg / m 2 、83mg / m 2 、84mg / m 2 、88mg / m 2 、86mg / m 2 、87mg / m 2 、88mg / m 2 、89mg / m 2 、90mg / m2 , or a number between any two of these values.

[0147] MTDP inhibitors can be administered to patients once a week or twice a week. In some embodiments, MTDP inhibitors are administered in a cycle of 14-28 days of daily administration. In some embodiments, MTDP inhibitors are administered in a cycle of 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 days. In some embodiments, MTDP inhibitors are administered on the 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th day of the 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 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. For example, paclitaxel can be administered in a cycle of 5, 6, 7, 8, 9, or 10 days. Paclitaxel can be administered weekly in a selected week of a cycle of administration. In some embodiments, paclitaxel is administered in a cycle of 28 days, wherein weekly administration continues for three weeks (e.g., on days 1, 8, and 15), and is not administered on the remaining days of the cycle (including days 16-28).

[0148] Similarly, any PLK1 inhibitor now known or later discovered 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 thiazolinone, a pteridine derivative, a dihydroimidazo[1,5-f]pteridine, a meta-substituted thiazolinone, 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.

[0149] In some embodiments, the PLK1 inhibitor is onvansertib. In these embodiments, onvansertib is administered to the patient at any appropriate dose, for example, less than 12 mg / m 2 , less than or equal to 24 mg / m 2 , or greater than 24 mg / m 2 In some embodiments, onvansertib is administered to the patient daily. In some embodiments, onvansertib is administered in a cycle of 5-14 days of daily administration of onvansertib and 2-16 days of no onvansertib administration. For example, in some embodiments, onvansertib is administered daily for 21 consecutive days in the cycle, and then onvansertib is not administered for 7 consecutive days. In some embodiments, combination therapy with onvansertib and an MTDP inhibitor can be administered at the same dose as a monotherapy of onvansertib or an MTDP inhibitor.

[0150] As will be appreciated by those skilled in the art, the amount of MTDP inhibitor and PLK1 inhibitor co-administered and the time of co-administration may depend on the type (species, sex, age, weight, etc.) and condition of the subject being treated and the severity of the disease or condition being treated. The MTDP inhibitor and PLK1 inhibitor may be formulated as a single pharmaceutical composition or two separate pharmaceutical compositions. The active ingredients may also be embedded in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, such as, respectively, in hydroxymethylcellulose or gelatin microcapsules and poly (methyl methacrylate) microcapsules, embedded in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or macroemulsions.

[0151] The methods, compositions, kits and systems disclosed herein can be applied to different types of subjects. For example, the subject can be a subject receiving cancer treatment, a subject in cancer remission, a subject who has received one or more cancer treatments, or a subject suspected of having cancer. The subject can suffer from phase I cancer, phase II cancer, phase III cancer and / or phase IV cancer. Cancer can be head and neck cancer, non-small cell lung cancer, intrahepatic bile duct cancer, gastric cancer, urothelial carcinoma, small cell lung cancer, endometrial cancer, cervical cancer, rhabdomyosarcoma, bile duct cancer, liver cancer, ovarian cancer, breast cancer, prostate cancer, colorectal cancer, pancreatic cancer or a combination thereof. Cancer can be unresectable locally advanced or metastatic disease. The method can also include applying additional therapeutic interventions to the subject. Additional therapeutic interventions can include therapeutic interventions different from applying PLK1 inhibitors and MTDP inhibitors, such as antibodies, adoptive T cell therapy, chimeric antigen receptor (CAR) T cell therapy, antibody-drug conjugates, cytokine therapy, cancer vaccines, checkpoint inhibitors, radiotherapy, surgery, chemotherapeutic agents or any combination thereof. Therapeutic intervention can be applied at any time of treatment, such as when the subject suffers from early stage cancer. The therapeutic intervention can be more effective than applying the therapeutic intervention to the subject at a later time. Without being bound by any particular theory, it is believed that PLK1 inhibitors (e.g., onvansertib) can sensitize cells (e.g., cancer cells) to MTDP inhibitor treatment to achieve effective cancer treatment.

[0152] Dosage and Pharmacokinetics

[0153] Treatments of the present disclosure may include administration of a PLK1 inhibitor (eg, onvansertib) for the desired duration, and administration of an MTDP inhibitor in one or more treatment cycles.

[0154] The daily or weekly administration (e.g., intravenous administration) of the MTDP inhibitor can be at or 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 an amount or range between any two of these values. During the treatment of the subject, the daily dose or weekly dose of the MTDP inhibitor can be adjusted (for example, increased or decreased with the range). The daily or weekly administration of the MTDP inhibitor can be applied in different amounts on different days or during different weeks. For example, treatment can include applying the MTDP inhibitor daily or weekly at 0.1 mg to 20 mg during the 1st week, at 0.25 mg to 50 mg during the 2nd week, at 0.5 mg to 100 mg during the 3rd week, at 1 mg to 200 mg during the 4th week, and at the 5th week and thereafter at 2 mg to 400 mg. For example, treatment can include applying the MTDP inhibitor daily or weekly at 0.1 mg to 100 mg on the 1st day, at 0.2 mg to 200 mg on the 2nd day, at 0.4 mg to 400 mg on the 3rd day, and at the 4th day and thereafter at 0.4 mg to 400 mg or 0.6 mg to 600 mg. For example, the MTDP inhibitor is paclitaxel and is administered at 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 an amount or range between any two of these values.

[0155] In some embodiments, the MTDP inhibitor may be administered at about 15 mg / m 2 About 275 mg / m 2 The drug / body surface area unit dose is administered daily or weekly. For example, an MTDP inhibitor (e.g., paclitaxel) can be administered at or about: 5 mg / m 2 , 10mg / m 2 , 15mg / m 2、20mg / m 2 、25mg / m 2 、30mg / m 2 、35mg / m 2 、40mg / m 2 、45mg / m 2 、50mg / m 2 、55mg / m 2 、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 / m2 , 265mg / m 2 , 270mg / m 2 , 275mg / m 2 , 280mg / m 2 , 285mg / m 2 , or a number or range between any two of these values. In some embodiments, the MTDP inhibitor can be administered daily or weekly at, or about, the following drug / body surface area unit dosage: 38 mg / m 2 , 39mg / m 2 , 40mg / m 2 41mg / m 2 42mg / m 2 43mg / m 2 44mg / m 2 45mg / m 2 46mg / m 2 47mg / m 2 48mg / m 2 , 49mg / m 2 , 50mg / m 2 , 51mg / m 2 , 52mg / m 2 , 53mg / m 2 , 54mg / m 2 , 55mg / m 2 , 56mg / m 2 , 57mg / m 2 , 58mg / m 2 , 59mg / m 2 , 60mg / m 2 , 61mg / m 2 , 62mg / m 2 , 63mg / m 2 , 64mg / m 2 , 65mg / m 2 , 66mg / m 2 , 67mg / m 2 , 68mg / m 2 , 69mg / m 2 , 70mg / m 2 , 71mg / m 2 , 72mg / m 2 , 73mg / m 2 , 74mg / m 2 , 75mg / m 2 , 76mg / m 2 , 77mg / m 2 , 78mg / m 2, 79mg / m 2 , 80mg / m 2 , 81mg / m 2 , 82mg / m 2 , 83mg / m 2 , 84mg / m 2 , 88mg / m 2 , 86mg / m 2 , 87mg / m 2 , 88mg / m 2 , 89mg / m 2 , 90mg / m 2 Or a number or range between any two of these values.

[0156] Each cycle of treatment / administration can have various lengths, for example, at least 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days or more. In some embodiments, the MTDP inhibitor is administered daily, half a week or weekly for three weeks in a 28-day cycle. In an exemplary embodiment, the MTDP inhibitor is administered for 1 to 10 cycles, for example, 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. The administration of the MTDP inhibitor (and / or one or more chemotherapeutic agents) can be daily or weekly and / or with one or more interruptions between administrations. The interruption 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 interruption 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 over a range) during the treatment of the subject. The daily or weekly administration of the MTDP inhibitor can be administered in different amounts on different days or during different weeks. For example, treatment can include taking 80 mg / m 2 On the 8th day, 64 mg / m 2 and 48 mg / m on day 15 2Administer the MTDP inhibitor weekly. For example, treatment can include administering the MTDP inhibitor daily or weekly at 0.1 mg to 20 mg during week 1, 0.25 mg to 50 mg during week 2, 0.5 mg to 100 mg during week 3, 1 mg to 200 mg during week 4, and 2 mg to 400 mg in week 5 and beyond. For example, treatment can include administering the 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 on day 4 and beyond. For example, the MTDP inhibitor can be administered daily or weekly at a dosage 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 an amount or range between any two of these values. In some embodiments, the daily or weekly dosage of the MTDP inhibitor can be or is about 0.005 mg / m 2 , 0.01mg / m 2 , 0.05mg / m 2 , 0.1mg / m 2 , 0.15mg / m 2 , 0.2mg / m 2 , 0.25mg / m 2 , 0.3mg / m 2 , 0.35mg / m 2 , 0.4mg / m 2 , 0.45mg / m 2 , 0.5mg / m 2 , 0.55mg / m 2 , 0.6mg / m 2 , 0.65mg / m 2 , 0.7mg / m 2 , 0.75mg / m 2 , 0.8mg / m 2 , 0.85mg / m 2 , 0.9mg / m 2 , 0.95mg / m 2 , 1mg / m 2 , 2mg / m 2 , 3mg / m 2, 4mg / m 2 , 5mg / m 2 , 6mg / m 2 , 7mg / m 2 , 8mg / m 2 , 9mg / m 2 , 10mg / m 2 , or a number or range between any two of these values. In some embodiments, an effective dose of a corticosteroid (e.g., dexamethasone), diphenhydramine, and / or an H2 antagonist (e.g., cimetidine or famotidine) is administered to the patient prior to administration of the MTDP inhibitor.

[0157] When the MTDP inhibitor is administered alone or in combination with a PLK1 inhibitor, the maximum concentration (C max ) (during or after treatment) can be about 1 pg / mL (picogram / mL) to about 10 μg / mL (microgram / mL). For example, when the MTDP inhibitor is administered alone or in combination with a PLK1 inhibitor, the C of the MTDP inhibitor in the blood of the subject is about 1 pg / mL (picogram / mL) to about 10 μg / mL (microgram / mL). maxIt can be the following or approximately the following: 1 pg / mL, 5 pg / mL, 10 pg / mL, 20 pg / mL, 30 pg / mL, 40 pg / mL, 50 pg / mL, 60 pg / mL, 70 pg / mL, 80 pg / mL, 90 pg / mL, 100 pg / mL, 150 pg / mL, 200 pg / mL, 250 pg / mL, 300 pg / mL, 350 pg / mL, 400 pg / mL, 450 pg / mL, 500 pg / mL, 1000 pg / mL, 5000 pg / mL, 10000 pg / mL, 50000 pg / mL, 100000 pg / mL (0.1 μg / mL), 0.2 μg / mL, 0.3 μg / mL, 0.4 μg / mL, 0.5 μg / mL, 0.6 μg / mL, 0.7 μg / mL, 0.8 μg / mL, 0.9 μg / mL, 1 μg / mL, 1.1 μg / mL, 1.2 μg / mL, 1.3 μg / mL, 1.4 μg / mL, 1.5 μg / mL, 1.6 μg / mL, 1.7 μg / mL, 1.8 μg / mL, 1.9 μg / mL, 2 μg / mL, 2.1 μg / mL, 2.2 μg / mL, 2.3 μg / mL, 2.4 μg / mL, 2.5 μg / mL, 2.6 μg / mL, 2.7 μg / mL, 2.8 μg / mL, 2.9 μg / mL, 3 μg / mL, 3.1 μg / mL, 3.2 μg / mL, 3.3 μg / mL, 3.4 μg / mL, 3.5 μg / mL, 3.6 μg / mL, 3.7 μg / mL, 3.8 μg / mL, 3.9 μg / mL, 4 μg / mL, 4.1 μg / mL, 4.2 μg / mL, 4.3 μg / mL, 4.4 μg / mL, 4.5 μg / mL, 4.6 μg / mL, 4.7 μg / mL, 4.8 μg / mL, 4.9 μg / mL, 5 μg / mL, 5.1 μg / mL, 5.2 μg / mL, 5.3 μg / mL, 5.4 μg / mL, 5.5 μg / mL, 5.6 μg / mL, 5.7 μg / mL, 5.8 μg / mL, 5.9 μg / mL, 6 μg / mL, 6.1 μg / mL, 6.2 μg / mL, 6.3 μg / mL, 6.4 μg / mL, 6.5 μg / mL, 6.6 μg / mL, 6.7 μg / mL, 6.8 μg / mL, 6.9 μg / mL, 7 μg / mL, 7.1 μg / mL, 7.2 μg / mL, 7.3 μg / mL, 7.4 μg / mL, 7.5 μg / mL, 7.6 μg / mL, 7.7 μg / mL, 7.8 μg / mL, 7.9 μg / mL, 8 μg / mL, 8.1 μg / mL, 8.2 μg / mL, 8.3 μg / mL, 8.4 μg / mL, 8.5 μg / mL, 8.6 μg / mL, 8.7μg / mL, 8.8μg / mL, 8.9μg / mL, 9μg / mL, 9.1μg / mL, 9.2μg / mL, 9.3μg / mL, 9.4μg / mL, 9.5μg / mL, 9.6μg / mL, 9.7μg / mL, 9.8μg / mL, 9.9μg / mL, 10μg / mL, a range between any two of these values, or any value between 1pg / mL and 10μg / mL. .

[0158] The treatment of the present disclosure can include administering a PLK1 inhibitor (onvansertib) for a desired duration in one or more cycles. In some embodiments, a 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 have various lengths, e.g., at least 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, or more.

[0159] Administration of the PLK inhibitor (and / or one or more chemotherapeutic agents) can be once daily or with one or more interruptions between days of administration. Interruptions 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. On the day that the PLK1 inhibitor (and / or one or more chemotherapeutic agents) is administered to the patient, administration can be once, twice, three times, four times or more. Administration can be, for example, once every two days, every three days, every four days, every five days, every six days or every seven days. The length of the desired duration 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 have a variety of 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, a single 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 a cycle (e.g., in a cycle of at least 21 days (e.g., 21 to 28)). In some embodiments, treatment may include administration of a PLK1 inhibitor (e.g., onvansertib) and / or one or more chemotherapeutic agents for at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or a range between any two of these values ​​in a cycle (e.g., a cycle of at least 21 days (e.g., 21 to 28 days)). Administration of a PLK1 inhibitor (e.g., onvansertib) and / or one or more chemotherapeutic agents in a single cycle of treatment may be continuous or have one or more intervals (e.g., interrupted for one or two days). In some embodiments, treatment includes administration of a PLK1 inhibitor (e.g., onvansertib) for 5 days in a cycle of 21 to 28 days. In some embodiments, a PLK1 inhibitor (e.g., onvansertib) is administered daily for 21 days, followed by an interruption of 7 days. In some embodiments, the PLK1 inhibitor (eg, onvansertib) is administered orally.In some embodiments, the PLK1 inhibitor (eg, onvansertib) is administered without any catch-up dose.

[0160] In some embodiments, the PLK1 inhibitor (e.g., onvansertib) is administered to a subject in need of the treatment on day 20 (e.g., day 1-10 and day 15-24) of a 28-day cycle. The 20 days can be, for example, continuous daily administration for 10 days (e.g., day 1-10) and another continuous daily administration for 10 days (e.g., day 15-24), or continuous daily administration for 4 groups of 5 days each (e.g., day 1-5, day 8-12, day 15-19, and day 22-26). In some embodiments, the PLK1 inhibitor (e.g., onvansertib) is administered to a subject in need of the treatment on day 21 (e.g., day 1-21) of a 28-day cycle. In some embodiments, for example, when a patient is identified as having low tolerance to a PLK1 inhibitor (e.g., onvansertib), a PLK1 inhibitor is administered to a subject in need of the treatment for 10 days (e.g., days 1-5 and days 15-19) during a 28-day cycle. These 10 days can be, for example, continuous daily administration for 10 days (e.g., days 1-10) or two continuous daily administrations each for five days (e.g., days 1-5 and days 15-19). In some embodiments, a PLK1 inhibitor (e.g., onvansertib) is administered to a subject in need of the treatment every day throughout the cycle (e.g., daily administration for 28 days in a 28-day cycle). Depending on the need for cancer progression inhibition / reversal in the subject, the subject can receive one, two, three, four, five, six or more cycles of treatment. For combination therapy, the administration cycle, dosing schedule and / or dosage amount of the MTDP inhibitor and the PLK1 inhibitor can be the same or different. For combination therapy, the administration cycle, dosing regimen and / or dosage amount of the MTDP inhibitor can be adjusted according to the administration cycle, dosing schedule and / or dosage amount of the PLK1 inhibitor. For example, an MTDP inhibitor (e.g., paclitaxel) can be administered three times in a 28-day cycle (e.g., at daily doses on days 1, 8, and 15), which corresponds to a 28-day cycle for administering a PLK1 inhibitor (e.g., onvansertib).

[0161] Treatment may include 6 mg / m 2 -90 mg / m 2 Drug / body surface area or about 6 mg / m 2 -90 mg / m 2Drug / body surface area PLK1 inhibitor (e.g., onvansertib) is administered, for example, as a daily dose. For example, treatment may include daily administration of the following, or about the following, of a PLK1 inhibitor (e.g., onvansertib): 6 mg / m 2 , 8mg / m 2 , 10mg / m 2 , 12mg / m 2 , 14mg / m 2 , 16mg / m 2 , 18mg / m 2 , 20mg / m 2 , 23mg / m 2 , 27mg / m 2 , 30mg / m 2 , 35mg / m 2 , 40mg / m 2 45mg / m 2 , 50mg / m 2 , 55mg / m 2 , 60mg / m 2 , 65mg / m 2 , 70mg / m 2 , 80mg / m 2 , 85mg / m 2 , 90mg / m 2 , a number or range between any two of these values, or 8 mg / m 2 -90 mg / m 2 In some embodiments, for a subject, the daily dose of a PLK1 inhibitor (e.g., onvansertib) can be adjusted (e.g., increased or decreased from a range) during treatment or during a single cycle of treatment (e.g., a first cycle, a second cycle, a third cycle, and subsequent cycles). In some embodiments, a PLK inhibitor (e.g., onvansertib) is administered at 12 mg / m 2 Administered on 20 days (e.g., days 1-10 and days 15-24) during a 28-day cycle. In some embodiments, a PLK inhibitor (e.g., onvansertib) is administered at 15 mg / m 2 Administered on 10 days (e.g., days 1-5 and 15-19) during a 28-day cycle. In some embodiments, a PLK inhibitor (e.g., onvansertib) is administered at 8 mg / m 2 or 10 mg / m 2Administration daily (e.g., days 11-28) during a 28-day cycle. In some embodiments, for a subject, the daily dose of a PLK1 inhibitor (e.g., onvansertib) can be adjusted (e.g., increased or decreased within a range) during treatment or during a single cycle of treatment (e.g., first cycle, second cycle, third cycle, and subsequent cycles).

[0162] When the PLK1 inhibitor is administered alone or in combination with the MTDP inhibitor, the maximum concentration (C max ) (during or after treatment) can be about 100 nmol / L to about 1500 nmol / L. For example, when the PLK1 inhibitor is administered alone or in combination with the MTDP inhibitor, the C of the PLK1 inhibitor (e.g., onvansertib) in the blood of the subject is about 100 nmol / L to about 1500 nmol / L. max It can be, or is about 100nmol / L, 200nmol / L, 300nmol / L, 400nmol / L, 500nmol / L, 600nmol / L, 700nmol / L, 800nmol / L, 900nmol / L, 1000nmol / L, 1100nmol / L, 1200nmol / L, 1300nmol / L, 1400nmol / L, 1500nmol / L, a range between any two of these values, or any value between 200nmol / L and 1500nmol / L.

[0163] When the PLK1 inhibitor is administered alone or in combination with the MTDP inhibitor, the area under the curve (AUC) of a plot of the concentration of the PLK1 inhibitor (e.g., onvansertib) in the blood of a subject over time (e.g., AUC in the first 24 hours after administration) is 0-24 ) can be about 1000 nmol / L·h to about 400000 nmol / L·h. For example, when the PLK1 inhibitor is administered alone or in combination with the MTDP inhibitor, the AUC of a plot of the concentration of the PLK1 inhibitor (e.g., onvansertib) in the blood of a subject over time (e.g., the AUC in the first 24 hours after administration) 0-24) can be or be about 1000 nmol / L·h, 5000 nmol / L·h, 10000 nmol / L·h, 15000 nmol / L·h, 20000 nmol / L·h, 25000 nmol / L·h, 30000 nmol / L·h, 35000 nmol / L·h, 40000 nmol / L·h, a range between any two of these values, or any value between 1000 nmol / L·h and 400000 nmol / L·h.

[0164] When the PLK1 inhibitor is administered alone or in combination with the MTDP inhibitor, the time to reach the maximum concentration of the PLK1 inhibitor (eg, onvansertib) in the blood of the subject (T max ) can be about 1 hour to about 5 hours. For example, when the PLK1 inhibitor is administered alone or in combination with the MTDP inhibitor, the time to reach the maximum concentration of the PLK1 inhibitor (e.g., onvansertib) in the blood of the subject (T max ) can be or be approximately 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 between 1 hour and 5 hours.

[0165] When the PLK1 inhibitor is administered alone or in combination with the MTDP inhibitor, the elimination half-life (T 1 / 2 ) can be about 10 hours to about 60 hours. For example, when the PLK1 inhibitor is administered alone or in combination with the MTDP inhibitor, the elimination half-life (T) of the PLK1 inhibitor (e.g., onvansertib) in the blood of the subject is about 10 hours to about 60 hours. 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 between 10 hours and 60 hours.

[0166] Patients administered one or more dose cycles of an MTDP inhibitor in combination with one or more dose cycles of a PLK1 inhibitor may exhibit very tolerable AEs, including undetectable clear AEs or clear SAEs in some cases. A significant but unlikely result is the finding that the patient has no possible or even possible AEs or SAEs. In some embodiments, treatment with a combination therapy of an MTDP inhibitor and a PLK1 inhibitor may result in a significant therapeutic effect. A therapeutic effect higher than the therapeutic effect predicted in vitro or by computer indicates a surprising result. A therapeutic dose lower than the therapeutic dose predicted in vitro or by computer indicates a surprising result. It is expected that the combined therapy may reduce the progression of the patient's disease. A highly positive result is the finding that the combined therapy may result in stable disease. A significant but unlikely result is the finding of a complete response or complete remission of cancer, progression-free survival or overall survival exceeding the predicted value of in vitro or computer analysis, or the absence of any measurable lesions, any target lesions, or any malignant lymph nodes.

[0167] Additional cancer therapeutic agents or therapies

[0168] The methods, compositions and kits disclosed herein can be used to treat cancer, such as prostate cancer. In some embodiments, the 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 administering at least one additional cancer therapeutic agent or cancer therapy. The treatment may include administering 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 may be co-administered, for example, simultaneously or sequentially. The PLK1 inhibitor (e.g., onvansertib) and the cancer therapeutic agent or cancer therapy may be co-administered, for example, simultaneously or sequentially.

[0169] Methods for predicting / determining cancer treatment efficacy and status

[0170] Also disclosed herein are methods, compositions, kits, and systems for predicting / determining the clinical outcomes of cancer combination therapies of the present disclosure, monitoring combination therapies, predicting / determining the responsiveness of subjects to combination therapies, determining the state of cancer in subjects, and improving the results of combination therapies. Methods, compositions, kits, and systems can be used to guide combination therapies, provide combination therapy suggestions, and reduce or avoid unnecessary ineffective combination therapies for patients. ctDNA can be analyzed to predict / determine the clinical outcomes of cancer treatment using a combination of MTDP inhibitors and PLK1 inhibitors of the present disclosure, monitor combination therapy, predict / determine the responsiveness of subjects to combination therapy, determine the state of cancer in subjects, improve combination therapy outcomes, guide combination therapy, provide combination therapy suggestions, and / or reduce or avoid ineffective combination therapy. ctDNA can be analyzed to predict / determine the clinical outcomes of cancer treatment, monitor cancer treatment, predict / determine the responsiveness of subjects to cancer treatment, determine the state of cancer in subjects, improve cancer treatment outcomes, guide cancer treatment, provide treatment suggestions, and / or reduce or avoid ineffective cancer treatment. Analysis of such ctDNA has been described in PCT Application No. PCT / US2021 / 013287, the contents of which are incorporated herein by reference in their entirety.

[0171] The method for determining the responsiveness of a subject to a combination therapy including an MTDP inhibitor and a PLK1 inhibitor of the present disclosure may include, for example, analyzing circulating tumor DNA (ctDNA) of a subject with cancer, the subject being treated and / or having received the combination therapy, thereby determining the responsiveness of the subject to the combination therapy. In some embodiments, determining the responsiveness of the subject includes determining whether the subject is a responder to the treatment, whether the subject is in or will be in CR, or whether the subject is in or will be in partial remission (PR). For example, analyzing ctDNA may include: detecting the variant allele frequency in ctDNA in a first sample obtained from the subject at a first time point, detecting the variant allele frequency in ctDNA obtained from the subject in one or more other samples at one or more other time points, and determining the difference in the variant allele frequency in ctDNA between at least one of the first sample and one or more other samples, and the reduction in the variant allele frequency in at least one of the other samples relative to the first sample indicates that the subject is responsive to cancer treatment.

[0172] In some embodiments, the first time point is before or immediately before the combination therapy, 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 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 is at or after the end of the second cycle of the combination therapy.

[0173] In some embodiments, the first cycle of the combination therapy is immediately prior to the second cycle of the combination therapy. In some embodiments, the method includes continuing the combination therapy for the subject if the subject is indicated as responding to the combination therapy. In some embodiments, the method includes stopping the combination therapy for the subject and / or starting a different combination therapy for the subject if the subject is not indicated as responding to the combination therapy.

[0174] Disclosed herein are methods for determining the cancer status of a subject, including analyzing the subject's circulating tumor DNA (ctDNA), thereby determining the cancer status of the subject. The subject may be a subject who is undergoing a current combination therapy including an MTDP inhibitor and a PLK1 inhibitor of the present disclosure, a subject who has received a prior combination therapy of the present disclosure, and / or a subject in cancer remission. A subject in cancer remission may be in complete remission (CR) or in partial remission (PR).

[0175] In some embodiments, analyzing ctDNA includes detecting variant allele frequencies in ctDNA. In some embodiments, analyzing ctDNA includes: detecting variant allele frequencies in ctDNA obtained from a subject in a first sample at a first time point, detecting variant allele frequencies in ctDNA obtained from a subject in one or more additional samples at one or more additional time points, and determining the difference in variant allele frequencies in ctDNA between at least one of the first sample and one or more additional samples, an increase in variant allele frequencies in one or more additional samples relative to the first sample indicating that the subject is at risk of cancer recurrence or is at cancer recurrence.

[0176] In some embodiments, the first time point is before or immediately before the combination therapy, and the one or more additional time points are 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.

[0177] In some embodiments, the method includes initiating additional treatment of the subject if the subject is indicated as being at risk of cancer recurrence. The additional treatment can be the same as or different from the current combination therapy or a prior combination therapy.

[0178] The variant allele frequency in ctDNA can be determined, for example, by the total mutation count in the ctDNA of each of the first sample and the one or more additional samples, or by the average variant allele frequency in the ctDNA of each of the first sample and the one or more additional samples. In some embodiments, the variant allele frequency is the mutant allele frequency (MAF) of a driver mutation of a cancer (e.g., ovarian cancer, breast cancer, prostate cancer, colorectal cancer, pancreatic cancer, or a combination thereof). In some embodiments, the variant allele frequency is the MAF of one or more driver mutations of a 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 in the first sample, and C1 is the ctDNA MAF in one of the additional samples. In some embodiments, the MAF threshold is from 0.01 to -0.10 or about 0.01 to -0.10. In some embodiments, the MAF threshold is 0.06 or about 0.05. In some embodiments, the MAF threshold is 0.05 or about 0.05.

[0179] In some embodiments, the first sample comprises ctDNA from a subject before treatment, and one of the additional samples comprises ctDNA from a subject after treatment. In some embodiments, the driver mutation is a mutation in one of 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, J AK2, 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 these 75 genes. In some embodiments, one or more driver mutations are mutations in these 75 genes.

[0180] The driver mutation or at least one of the one or more driver mutations may be located 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 located 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 the variant allele frequency in one or more of the subject's ctDNA, PBMCs, and BMMCs.

[0181] ctDNA can be analyzed using, for example, polymerase chain reaction (PCR), next generation sequencing (NGS), and / or droplet digital PCR (ddPCR). Samples disclosed herein can be derived from, for example, whole blood of a subject, plasma of a subject, serum of a subject, or a combination thereof. In some embodiments, ctDNA is from whole blood of a subject, plasma of a subject, serum of a subject, or a combination thereof.

[0182] In some embodiments, the method includes analyzing the ctDNA of the subject before treatment. In some embodiments, treatment includes one or more cycles, and ctDNA is analyzed before, during, and after each cycle of treatment. Each cycle of treatment can be at least 21 days. In some embodiments, each cycle of treatment is about 21 days to about 28 days. In some embodiments, the subject is human.

[0183] Disclosed herein are methods for improving cancer treatment outcomes. The method may include: detecting a variant allele frequency in circulating tumor DNA (ctDNA) obtained from a subject in a first sample at a first time point before the subject undergoes a combination therapy comprising an MTDP inhibitor and a PLK1 inhibitor of the present disclosure; detecting a variant allele frequency in ctDNA obtained from the subject in one or more additional samples at one or more additional time points after the subject undergoes the combination therapy; determining the difference in variant allele frequency in ctDNA between at least one of the first and one or more additional samples; and continuing the combination therapy for the subject if the subject is indicated to respond to the combination therapy, or stopping the combination therapy for the subject and / or starting a different cancer therapy for the subject if the subject is not indicated to respond to the combination therapy. 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 combination therapy.

[0184] Also disclosed herein are methods for treating cancer. The method may include: administering a combination therapy including an MTDP inhibitor and a PLK1 inhibitor of the present disclosure to a subject in need thereof; determining a decrease in the variant allele frequency in a second sample of the subject obtained at a second time point after the subject receives the combination therapy relative to the variant allele frequency in the first sample of the subject obtained at a first time point before the subject receives the combination therapy; and continuing the combination therapy. In some embodiments, the subject is a subject newly diagnosed with cancer, such as a subject who has not received any prior cancer treatment before the combination therapy. In some embodiments, the subject has received prior cancer treatment and is in cancer remission, such as a subject who is in complete remission (CR) or in partial remission (PR) after receiving prior combination therapy.

[0185] 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 a 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.

[0186] The variant allele frequency in ctDNA can be determined, for example, by the total mutation count in the ctDNA of each of the first sample and the one or more additional samples, and / or by the average variant allele frequency in the ctDNA of each of the first sample and the one or more additional samples. In some embodiments, the variant allele frequency is the mutant allele frequency (MAF) of a driver mutation of a cancer (e.g., ovarian cancer, breast cancer, prostate cancer, colorectal cancer, pancreatic cancer, or a combination thereof). In some embodiments, the variant allele frequency is the mutant allele frequency (MAF) of one or more driver mutations of a 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 in the first sample, and C1 is the ctDNA MAF in one of the additional samples. In some embodiments, the MAF threshold is -0.05.

[0187] The driver mutation can be, for example, one of the one or more driver mutations, or a mutation in at least one of the one or more driver mutations, wherein the one or more driver mutations are mutations in one of 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, 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, and / or one or more driver mutations are mutations in these 75 genes. In some embodiments, the driver mutation or at least one of the one or more driver mutations is located 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 located in a gene selected from the group consisting of DNMT3A, TET2, NPM1, SRSF2, NRAS, CDKN2A, SF3B1, FLT3, ASXL1, SRSF2, IDH2, NRAS, and SF3B1.

[0188] In some embodiments, the method also includes determining the variant allele frequency in one or more of the subject's ctDNA, PBMC and BMMC. For example, the variant allele frequency in ctDNA can be detected using polymerase chain reaction (PCR) or next generation sequencing (NGS). In some embodiments, the variant allele frequency in ctDNA is detected using droplet digital PCR (ddPCR).

[0189] At least one of the first sample, one or more additional samples, and the second sample can be derived from whole blood of the subject, plasma of the subject, serum of the subject, or a combination thereof. In some embodiments, the ctDNA is from whole blood of the subject, plasma of the subject, serum of the subject, or a combination thereof.

[0190] In some embodiments, the subject whose ctDNA is analyzed is experiencing or will experience cancer treatment. The method may include analyzing the ctDNA of the subject before treatment. The treatment may include one or more cycles, and the ctDNA is analyzed before, during, and after one or more cycles of treatment. For example, ctDNA can be analyzed before, during, and after two or more cycles of treatment, three or more cycles of treatment, or each cycle of treatment. Each cycle of treatment can be at least 21 days, for example, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days or more or any two of these values. In some embodiments, each cycle of treatment is about 21 days to about 28 days. In some embodiments, each cycle of treatment is 21 to 28 days. In some embodiments, the subject is human.

[0191] Compositions and kits

[0192] Disclosed herein are compositions and kits for treating cancer. The kit may include: a PLK1 inhibitor; and a manual providing instructions for co-administering the PLK1 inhibitor and the MTDP inhibitor to a subject to treat cancer. The kit may include the MTDP inhibitor. The cancer may be, for example, ovarian cancer, breast cancer, prostate cancer, colorectal cancer, pancreatic cancer, or a combination thereof.

[0193] In some embodiments, the subject has cancer (e.g., head and neck cancer, non-small cell lung cancer, small cell lung cancer, intrahepatic bile duct cancer, gastric cancer, urothelial carcinoma, breast cancer, endometrial cancer, cervical cancer, rhabdomyosarcoma, bile duct cancer, glioblastoma, low-grade glioma, thyroid cancer, gallbladder cancer, ovarian cancer, prostate cancer, or a combination thereof). In some embodiments, the instructions include instructions for co-administering a PLK inhibitor and an MTDP inhibitor simultaneously. In some embodiments, the instructions include instructions for sequentially co-administering a PLK inhibitor and an MTDP inhibitor. 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.

[0194] In some embodiments, the instructions include a statement that the subject has received prior treatment with an MTDP inhibitor. In some embodiments, the instructions include a statement that the subject has not responded to treatment with an MTDP inhibitor alone. In some embodiments, the instructions include a statement that the subject is known to be resistant to MTDP inhibitor therapy.

[0195] In some embodiments, the instructions include an instruction that the subject has received at least one prior treatment for the cancer. In some embodiments, the prior treatment does not include the use of an MTDP inhibitor, a PLK inhibitor, or both. In some embodiments, the instructions include an instruction that the subject is in remission of the cancer. In some embodiments, the subject in remission of the cancer is in complete remission (CR) or in partial remission (PR).

[0196] The instructions may include instructions for administering each of the MTDP inhibitor and the PLK1 inhibitor to the subject in a cycle of at least twice a week. In some embodiments, the instructions include instructions for administering each of the MTDP inhibitor and the PLK1 inhibitor to the subject in a cycle of at least five times a week. In some embodiments, the instructions include instructions for administering the MTDP inhibitor, the PLK1 inhibitor, or both in a cycle of at least 7 days. In some embodiments, each cycle of treatment is at least about 21 days. In some embodiments, each cycle of treatment is about 21 days to about 28 days. In some embodiments, the instructions include instructions for administering the PLK1 inhibitor on at least four days of the cycle. In some embodiments, the instructions include instructions for not administering the PLK1 inhibitor on at least one day of the cycle. In some embodiments, the instructions include instructions for administering the MTDP inhibitor daily. In some embodiments, the instructions include instructions for administering the MTDP inhibitor and the PLK1 inhibitor for at least two cycles.

[0197] The MTDP inhibitor can be paclitaxel or a pharmaceutically acceptable salt thereof. In some embodiments, the PLK1 inhibitor is selective and / or specific for PLK1. In some embodiments, the PLK1 inhibitor is a dihydropteridinone, a pyridopyrimidine, an aminopyrimidine, a substituted thiazolinone, a pteridine derivative, a dihydroimidazo[1,5-f]pteridine, a meta-substituted thiazolinone, 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.

[0198] In some embodiments, the instructions include for use at 8 mg / m 2 -90 mg / m 2 Instructions for administering the PLK1 inhibitor. In some embodiments, the instructions include instructions for administering the MTDP inhibitor at 0.01 mg-1200 mg (eg, a daily dose of 0.01 mg-10 mg administered orally).

[0199] 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, stereoisomer thereof, thereby sensitizing cancer cells to one or more MTDP inhibitors. Contacting cancer cells with the composition may occur in vitro, ex vivo, in vivo, or any combination. In some embodiments, contacting cancer cells with the composition is in the body of a subject. In some embodiments, contacting cancer cells with the composition in cell culture. The subject may be a mammal, such as a human. Sensitization of cancer cells may increase the responsiveness of cancer cells to one or more MTDP inhibitors by or about 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. Sensitization of cancer cells can increase the responsiveness of cancer cells to one or more MTDP inhibitors by at least or about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or any range between two of these values. In some embodiments, the increase in responsiveness of cancer cells is relative 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 at least or about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or any range between two of these values. Sensitization of cancer cells can increase the responsiveness of a subject having cancer cells to one or more MTDP inhibitors by at least or about 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. In some embodiments, the increase in responsiveness of a subject having cancer cells is relative to a subject not treated with the composition.

[0200] The method may include determining the sensitization of cancer cells to one or more MTDP inhibitors after contacting the cancer cells with the PLK1 inhibitor. The method may include contacting the cancer cells with the one or more MTDP inhibitors while contacting the cancer cells with the PLK1 inhibitor and / or after contacting the cancer cells with the PLK1 inhibitor. In some embodiments, contacting the cancer cells with the one or more MTDP inhibitors occurs in the subject. The subject may be a mammal, such as a human. The subject may be, for example, a subject that does not respond to a single MTDP inhibitor or is known to be resistant to a single PARP inhibitor. The subject may be, for example, a subject that has been previously treated with one of the one or more MTDP inhibitors. In some embodiments, the method includes determining the response of the subject to the one or more MTDP inhibitors.

[0201] Example

[0202] Some aspects of the embodiments discussed above are disclosed in further detail in the following examples, which are not intended in any way to limit the scope of the present disclosure.

[0203] Example 1

[0204] Onvansertib synergizes with paclitaxel in SCLC cell lines

[0205] Five small cell lung cancer (SCLC) cell lines were treated with different doses of onvansertib and paclitaxel for 6 days. The assay measures cell viability. The SynergyFinder application is used to calculate the synergy of drug combination. The Bliss independence synergy score for each cell line was tested in SHP77, DMS53, DMS114, H1417 and H69 cell lines, respectively. The positive Bliss independence synergy score indicates the synergy of onvansertib and paclitaxel at a given concentration. In these cell lines, at the ovansertib concentration of about 25nM, a strong synergistic effect (as indicated by the high positive synergy score) was observed in the paclitaxel concentration (about 0.03nM-500nM) of a wide range. In addition, the viability of the cell treated with paclitaxel of various doses in the absence or presence of onvansertib was also tested in SHP77, DMS53, DMS114, H1417 and H69 cell lines, respectively. The Bliss independence model using drug additive effect (additivity) is used to calculate the expected viability. In the presence of onvansertib (about 25nM), the difference between the expected viability and the observed viability indicates the synergy between onvansertib and paclitaxel. For example, in DMS53 cells, for the combination of onvansertib and paclitaxel, the expected and observed viabilities are about 75% and about 50%, respectively (i.e., 25% cancer cell inhibition is expected, while 50% cancer cell inhibition is observed), which means that the efficacy of the drug combination is increased by about 100% compared to the combined inhibition caused by paclitaxel alone plus onvansertib alone. In short, these results demonstrate the enhanced or synergistic efficacy of the combination of paclitaxel and onvansertib in inhibiting cancer progression.

[0206] Example 2

[0207] Onvansertib synergizes with paclitaxel in TNBC cell lines

[0208] The activity of PLK1 inhibitor GSK461364 alone and together with MTDP inhibitor docetaxel in several TNBC cell models was evaluated. Surprisingly, when PLK1 inhibitor GSK461364 was combined with MTDP inhibitor docetaxel at a concentration significantly lower than that required for single-agent activity, the anti-tumor activity was significantly synergistic. In addition, it was surprisingly found that PLK1 inhibitor GSK461364 plus MTDP inhibitor docetaxel could significantly reduce the clonal potential and stem cell fraction of SUM149 and SUM159 cells (two well-studied TNBC cell lines). Without being bound by a particular theory, it is believed that PLK1 is a fact necessary for mitosis entry during recovery from G2 arrest induced by DNA damage. Preliminary data show that PLK1 is a functional key gene in basal-like cell line SUM149. The synergistic combination of PLK1 inhibition and chemotherapy using, for example, taxanes can specifically inhibit G2-M transition, induce abnormal mitosis exit and apoptosis, and eliminate stem cell-like resistant tumor clones.

[0209] like Figure 3 As shown in Figure 2, the combination index (CI) of the combination of paclitaxel and onvansertib was calculated 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 was dependent on p53 mutation status. These data suggest that patients with mutant p53 breast cancer may be more sensitive to the PLK1 inhibitor onvansertib.

[0210] The effect of PLK1 inhibition via treatment with onvansertib alone or in combination with paclitaxel on a mouse xenograft model of mesenchymal breast cancer SUM159 was examined. A total of 35 mice were randomized to receive oral (PO) vehicle plus intraperitoneal (IP) vehicle (N=8), oral onvansertib 120 mg / Kg plus intraperitoneal vehicle on days 1 and 2 of each week (N=9), intraperitoneal paclitaxel 10 mg / Kg plus oral vehicle on day 1 of each week (N=8), and oral onvansertib 120 mg / Kg plus intraperitoneal paclitaxel 10 mg / Kg on day 1 of each week (N=10) ( Figure 4). Onvansertib and paclitaxel exhibited similar tumor growth inhibition when compared to control (difference = -0.406 and -0.337 at day 21, p = 0.262 and 0.340, respectively). The combination of onvansertib and paclitaxel was significantly superior to single-agent treatment, with a difference of -1.346 compared to onvansertib alone and a difference of -1.414 compared to paclitaxel alone (p < 0.0001 and p < 0.0001, respectively).

[0211] 9 mg / m2 is recommended on days 1 to 21 of each 28-day treatment cycle. 2 The dose and schedule of onvansertib were adjusted to increase exposure to onvansertib and optimize efficacy. Based on PK data from the completed Phase 1 single-agent study of onvansertib, onvansertib concentrations decreased to their effective threshold level (10×IC 50 The following table shows that the tumor was not exposed to onvansertib for a period of 14 days in a 21-day cycle (to account for the effects of plasma protein binding). Prolonged treatment interruptions may allow tumors to recover and continue to grow.

[0212] A more continuous onvansertib regimen is being studied in patients with mCRPC (NCT03414034). In this study, patients were treated with onvansertib 12 mg / m in 21-day cycles. 2 QD combined with abiraterone QD treatment lasts for 14 days (Day 1 to Day 14) (starting on Day 1 and continuing uninterrupted in each cycle) (14+7 regimen). As of December 4, 2020, 9 patients have been treated with this regimen and no grade 2 or higher hematologic toxicity has been reported. Although the completed Phase 1 single-agent study of onvansertib showed that neutropenia and thrombocytopenia were the main on-target toxicities of onvansertib, the use of onvansertib 12mg / m 2 Patients with mCRPC treated with 12 mg / m 2 Treatment with onvansertib was safe and well tolerated. Of the 9 patients treated with onvansertib in the 14+7 regimen, 5 were evaluable for efficacy, and 3 (60%) achieved the primary efficacy endpoint. In the alternative dosing regimen tested in the mCRPC study (18 mg / m 2With 5 days of onvansertib treatment (5+9 regimen), 3 of 10 evaluable patients (30%) achieved the primary efficacy endpoint. Although preliminary, these data suggest that more continuous dosing of onvansertib may improve efficacy.

[0213] The pharmacokinetic data from the completed phase 1 single-dose study of onvansertib were used to simulate onvansertib drug exposure in different dosing regimens. The onvansertib effective level threshold was calculated as 10×IC 50 To induce the effect of plasma protein binding. For evaluating drug exposure, the maximum concentration (C max ), area under the curve (AUC (0-336) ), average concentration (C avg ; Calculated as AUC (0-336) / 336 hours) and above 10×IC 50 In Table 2, the dosing schedule represents the number of days on which onvansertib was received QD and the number of days on which onvansertib was not received QD in any given treatment cycle. For example, the 5+9 dosing schedule represents QD dosing of onvansertib on days 1 to 5 of a 14-day treatment schedule. 10×IC 50 The onvansertib dose was calculated to be 32.5 mg / mL to account for the effects of plasma protein binding. Calculation of a 14-day cycle above 10×IC 50 The simulation showed that although the more continuous dosing regimen (12 mg / m 2 10+4 regimen (12 mg / m in a 14-day cycle) 2 C of onvansertib in the onvansertib treatment regimen (10 days) or 14+7 regimen) avg Increase, but C max Less than 15 mg / m 2 5+9 regimen. In addition, 2 Compared with patients treated for 5 days, patients treated with 12 mg / m 2 Patients treated for 10 days had a 10×IC 50 Therefore, 12 mg / m 2 The 10+4 regimen produced deeper and more durable responses in patients.

[0214] Table 2: Simulated ONVANSERTIB pharmacokinetic results using different doses and dosing schedules

[0215]

[0216] The dose and schedule of onvansertib used in this study were selected based on analysis of pharmacokinetic (PK), pharmacodynamic, safety, and efficacy data from the Phase 1 dose-escalation study. Changes in the dose and schedule of onvansertib and in combination with paclitaxel may alter the toxicity of the regimen. Therefore, the starting dose selected for this study was 9 mg / m 2 / day for 21 consecutive days, then stop taking (off) for 7 days (4-week cycle). 2 A starting dose of 1:1 / day allowed continuous exposure to onvansertib during weekly paclitaxel infusions.

[0217] Example 3

[0218] Related research background

[0219] This embodiment describes the use of onvansertib to inhibit PLK1 in combination with taxanes for the treatment of TNBC patients and TP53 mutations as a biomarker of response. According to TCGAbioportal, TP53 is one of the most mutated genes in breast cancer. The frequency of mutations varies with the histological and biochemical characteristics of breast cancer, more common in ductal cases than in lobular cases, more common in lymph node-positive cases than in lymph node-negative cases, more common in estrogen receptor (ER) negative cases than in ER positive cases, and more common in HER2 positive cases than in HER2 negative cases. As described above, the incidence of mutations also depends on the molecular subtype of cancer, most common in triple negative breast cancer (TNBC), and least common in luminal A subtype. In addition, there are several evidences that TP53 mutations are associated with shorter OS in patients with luminal B and TNBC. At present, there is no targeted therapy for treating breast cancer patients with TP53 mutations, and patients receive standard chemotherapy. Since breast cancer with TP53 mutations is genomically unstable and difficult to pass through mitosis, PLK1 is particularly important in these cells and is therefore a potential drug target. Therefore, blocking PLK1 function can affect the ability of cancer cells with mutp53 to pass through mitosis, and thus can increase the overall sensitivity of cells to chemotherapeutic agents such as paclitaxel. Several other genes are associated with genomic instability in basal-like breast cancer, including RB, CCND1, BIRC5, BUB1 and PTEN. With the progress of immuno-oncology in recent years, PLK1 has been explored as an immunomodulator in cancer treatment. Cancers with higher PLK1 expression levels tend to have lower immune activity, such as lower HLA expression and reduced infiltration of B cells, NK cells and tumor-infiltrating lymphocytes. On the other hand, elevated tumor immunity increases the sensitivity of cancer cells to PLK1 inhibitors. The main mechanism of the potential association between PLK1 and tumor immunity may lie in the abnormal cell cycle and p53 pathways in cancer. Therefore, PLK1 inhibition and immunotherapy combination can achieve synergistic anti-tumor efficacy.

[0220] During both phases of the study, a baseline biopsy was performed if the disease could be safely biopsied. Whole exome sequencing (WES) was performed on the samples 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) the overall mutational signature.

[0221] RNA sequencing was also performed on fresh baseline biopsy samples to assess gene expression and breast molecular subtypes. If subjects did not undergo a baseline biopsy, or tissue collected at that biopsy was considered inappropriate, archived tissue was examined for TP53 status.

[0222] Blood will be collected at baseline, C3D1, and EOT for circulating tumor DNA (ctDNA) assessment. In addition to evaluating genomic alterations in DNA extracted from the blood, study findings will be compared to parallel sequencing analyses performed on tissue biopsies from the same subjects to compare similarities and any potential differences. This analysis aims to determine whether ctDNA sequencing can be a possible alternative to tissue-based sequencing, which will be critical as tissue biopsies are both invasive and expensive. Quantitative changes in ctDNA will be assessed throughout treatment to evaluate if there are genomic alterations that may be associated with improved clinical outcomes with onvansertib and paclitaxel and to assess biomarkers of response. Peripheral blood mononuclear cells (PBMCs) will be collected and analyzed at C1D1, C1D15, and EOT to evaluate the effect of onvansertib in improving anti-tumor immunity.

[0223] Example 4

[0224] Onvansertib and Taxane in Patients with TNBC with Unresectable Locally Advanced or Metastatic Disease Clinical trials of combination therapy with alcohol

[0225] Patient selection

[0226] Patients aged ≥18 years with histologically confirmed invasive breast cancer with unresectable locally advanced or metastatic disease, an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1, and not receiving any other cancer therapy were enrolled, regardless of race, ethnicity, and sex. Invasive breast cancer with unresectable locally advanced or metastatic disease can include inflammatory breast cancer and TNBC. According to the ASCO / CAP 2018 guidelines, histologically or cytologically confirmed TNBC is defined as estrogen receptor ≤10%, progesterone receptor ≤10%, Her-2-neu negative (IHC 0-1+ or FISH negative). Patients who have received treatment for brain metastases, have stable imaging for at least four weeks before enrollment, and have stopped steroid therapy are eligible. Patients with small, asymptomatic incidental brain metastases that do not require immediate treatment (including steroids) are also eligible. However, patients with estrogen receptor / progesterone receptor ≥1% are not allowed to receive concomitant endocrine therapy. In addition, patients must have the following organ function.

[0227] Table 3: Organ function of patients

[0228] Function value leukocyte ≥ 3,000 / mcL Absolute neutrophil count ≥1,500 / mcL platelets ≥ 100,000 / mcL (≥ 100,000 / mcL) Hemoglobin ≥ 9.0 g / dL (14.0 g / dL) Total bilirubin ≤1.5× institutional upper limit of normal (ULN) AST(SGOT) / ALT(SGPT) ≤3×Institutional ULN Creatinine ≤1.5ULN or creatinine clearance >50 mL / min, calculated by Cockcroft-Gault equation

[0229] Patients were excluded if they: (1) had received anticancer chemotherapy or biologic therapy within 21 days or 5 half-lives (whichever was shorter) before the first dose of study drug; (2) had received palliative radiation therapy ≤ 2 weeks from enrollment; (3) had > 3 lines of chemotherapy for metastatic disease in the Phase 2 portion (with no restriction on prior lines in the dose-escalation cohort); (4) had recurrent or progressive disease less than 6 months after the last exposure to any taxane-based therapy in the neoadjuvant, adjuvant, or metastatic setting; (5) had undergone major surgery within 6 weeks before the start of treatment; (6) were pregnant or breastfeeding; (7) had a disease that the investigator considered to be significant (i) gastrointestinal (GI) disorders that significantly impair absorption of oral doses (e.g., ileus, active Crohn's disease, ulcerative colitis, extensive gastric and small bowel resection); (ii) inability or unwillingness to swallow study medication; (iii) uncontrolled concurrent illness, including but not limited to ongoing or active infection, clinically significant nonhealing or healing wounds, symptomatic congestive heart failure (CHF) according to the New York Heart Association (NYHA) functional class II or higher, unstable angina, clinically significant arrhythmias, significant pulmonary disease (shortness of breath at rest or with mild exertion), uncontrolled infection, or psychiatric illness that would limit compliance with study requirements. Medical / social status; (10) known to have active COVID-19 or human immunodeficiency virus (HIV) infection with measurable viral titers, and / or active hepatitis B or C infection (patients receiving hepatitis B virus (HBV) immunization are eligible; subjects living with HIV and CD4+ T cell (CD4+) count ≥350 cells / μL are eligible; subjects who have received established ART for at least four weeks before enrollment and HIV viral load less than 400 copies / mL are eligible; (11) have clinically significant ascites or pleural effusion; (12) are known to be hypersensitive to paclitaxel; (13) have grade 2 or higher peripheral Neuropathy; (14) History of other malignancies, except: adequately treated non-melanoma skin cancer, curatively treated cervical carcinoma in situ, or other solid tumors that have been curatively treated and have no evidence of disease for >2 years; (15) Active disease that would make protocol treatment hazardous or compromise the patient's ability to receive study medication; (16) Fridericia-corrected QT interval (QTcF) >480 milliseconds. QTcF is calculated as the arithmetic mean of QTcF on triplicate ECGs. In the case of potential correctable causes of QT prolongation (e.g., medication, hypokalemia), triplicate ECGs may be repeated once during the screening period.Results may be used to determine eligibility; (17) planned concomitant use of medications known to prolong the QT / QTc interval; (18) risk factors for torsades de pointes, including a family history of long QT syndrome or uncorrected hypokalemia; or (19) use or plan to use strong inhibitors of CYP3A4, such as atazanavir, ceritinib, clarithromycin, cobicistat and coformulations containing cobicistat, darunavir, idelalisib, indinavir, itraconazole, ketoconazole, lonafarnib, lopinavir, mifepristone, nefazodone, nelfinavi, ombitasvir-paritoprevir, or sedative agents. taprevir-ritonavir, ombitasvir-paritrimvir-ritonavir plus dasabuvir, posaconazole, ritonavir and ritonavir-containing co-formulations, 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).

[0230] Pre-treatment criteria

[0231] Patients who meet the following laboratory test criteria are treated: (1) absolute neutrophil count >1000 / mm 3 ; (2) Platelet count>100,000 / mm 3 ; (3) total bilirubin 1.5×ULN (institution); and (4) serum creatinine <1.5×ULN (institution) or calculated glomerular filtration rate (GFR) 60 mL / min.

[0232] Onvansertib and paclitaxel combination therapy

[0233] In the Phase 1b clinical trial managed using the BOIN design and the Phase 2 clinical trial managed using the Simon two-stage design, patients who met the above pre-treatment criteria were treated with the onvansertib-paclitaxel combination within 72 hours of Day 1 of Cycle 1, using the recommended Phase 2 dose (RP2D) of onvansertib. Treatment and study visits were performed within a window of + / - 3 days. Figure 1 The patients were divided into 3 cohorts, with up to 9 patients in each cohort treated with a 28-day cycle per dose. Starting at dose level 0, patients were administered an oral dose of onvansertib daily for the first 21 days of a 28-day cycle. Patients were also treated with 80 mg / m2 of onvansertib on days 1, 8, and 15 of a 28-day cycle. 2 The dose of paclitaxel is administered intravenously, and the principal investigator may reduce the dose as appropriate. Acceptable forms of paclitaxel include nab-paclitaxel (albumin-bound paclitaxel). In Table 4, oral onvansertib is administered once a day for 21 consecutive days, followed by 7 days of drug withdrawal to include a complete cycle of 28 days. In Table 6, "elimination" means eliminating the current and higher doses and preventing any future patients from being treated with these doses because these doses are too toxic. When the dose is eliminated, the dose is automatically reduced to the next lower level. When the lowest dose is eliminated, the trial is stopped for safety reasons, and no dose is selected as MTD. If the current dose is the lowest dose and the rule indicates a dose reduction, the next patient cohort is treated with the lowest dose, unless the number of DLTs reaches the elimination boundary, at which point the trial is terminated for safety reasons. If the current dose is the highest dose and the rule indicates a dose increase, the next patient cohort is treated with the highest dose. If no operation (i.e., increasing, decreasing, or eliminating) is triggered, new patients are treated with the current dose.

[0234] Table 4: Dose escalation schedule

[0235] Dosage Levels Onvansertib Dosage (QD) Paclitaxel dosage -1 <![CDATA[9mg / m 2 ]]> <![CDATA[80mg / m 2 ]]> 0 (starting dose level) <![CDATA[12mg / m 2 ]]> <![CDATA[80mg / m 2 ]]> 1 <![CDATA[18mg / m 2 ]]> <![CDATA[80mg / m 2 ]]>

[0236] Table 5: Description of the protocol

[0237]

[0238] Table 6: Increasing / decreasing rules for BOIN design when the target DLT rate is 0.3

[0239]

[0240] According to Table 5, for 21 days in a 28-day cycle, once every 24 hours, Onvansertib is administered orally to the patient with approximately 8 ounces (approximately 240 mL) of ice-free water after fasting for more than 30 minutes (once a day, for 21 days, then 7 days of drug withdrawal). The missed dose can be administered up to 4 hours after the scheduled time. Eating can be resumed after four hours, and even after vomiting, the "supplementary" dose should not be taken. During Phase 1b, before the administration of paclitaxel in C1D1 (Day 1 of Cycle 1), C1D8, C1D15, and C2D1, a dose of onvansertib should be administered in the clinic. On all other days (including all Phase 2), onvansertib should be administered at home or in the clinic at approximately the same time every day.

[0241] All patients were pretreated with corticosteroids, diphenhydramine and H2 antagonists. Before applying paclitaxel, patients were premedicated with dexamethasone, diphenhydramine and cimetidine or famotidine, as described in Table 5. The premedication of paclitaxel can start after the application of onvansertib. Paclitaxel is administered intravenously according to the package insert of the institution, once a week, for 3 weeks in every 4 weeks. An online filter with a microporous membrane not more than 0.22 microns is used. According to DFCI policy, the body surface area (BSA) for the dosage regimen is calculated by the DuBois formula. For paclitaxel and onvansertib, BSA is determined on the 1st day of each cycle. The dose of onvansertib is rounded to the nearest 5mg. Except as described below, the research or commercial agent or therapy intended to treat the participant's malignant tumor shall not be used.

[0242] Phase 1b trial

[0243] The test was performed at 12 mg / m 2 onvansertib is started. If the observed dose-limiting toxicity (DLT) rate is ≤0.236, the next cohort of patients is treated at the next higher dose level. If the observed DLT rate is ≥0.359, the next cohort of patients is treated at the next lower dose level. The target DLT rate is 0.3% or 30%. For the purpose of overdose control, if Pr(pj>0.3|data)>0.95, dose j and higher levels are eliminated from further examination, where pj = the true DLT rate at dose level j, and j = 1,…,3. If the lowest dose is eliminated, the trial is stopped for safety reasons. The trial is completed when the maximum sample size of 24 is reached or 9 patients continue to receive the same dose according to the ascending / descending rules in Table 6.

[0244] After the first treatment cycle, DLTs with 95% confidence intervals were determined according to CTACE version 5.0. The maximum tolerated dose (MTL) of onvansertib was 9 mg / m 2 Up to 18 mg / m 2 , indicating that the combination of onvansertib and paclitaxel is safe and well tolerated, and the dose can be the recommended Phase 2 dose. Severe adverse reactions, including grade 3 and 4 toxicities, were recorded separately. The proportion of patients experiencing severe adverse reactions (such as grade 3 and 4 toxicities) did not exceed 16%, indicating that the combination of onvansertib and paclitaxel is safe and well tolerated.

[0245] The RP2D was determined by previously specified isotonic regression and using the shiny app "BOIN". The isotonic estimate of the toxicity rate that was closest to the target toxicity rate was selected as the RP2D. If there was a tie, a higher dose level was selected when the isotonic estimate was lower than the target toxicity rate, and a lower dose level was selected when the isotonic estimate was greater than or equal to the target toxicity rate.

[0246] The pharmacokinetics of onvansertib and paclitaxel combination therapy were also determined, including C max , AUC, plasma half-life and plasma clearance.

[0247] The patient sample size was determined by simulating 10,000 trials under different scenarios (using the shiny app "BOIN" available at www.trialdesign.org). Table 7 shows that approximately 14-16 subjects are needed. The trial operating characteristics in Table 7 show that the Phase 1b trial selected the true MTD (if any) with a high probability and assigned more patients to the dose level with the DLT rate closest to the target 0.3.

[0248] Table 7: Expected samples for different toxicity profiles and test handling characteristics The amount

[0249]

[0250]

[0251] Phase 2 trials

[0252] The Phase 2 trial was a single-arm, Simon two-stage design that used RECIST 1.1 criteria to determine the objective response rate of onvansertib-paclitaxel combination therapy. The Phase 2 study enrolled patients in the 1st-3rd line. The objective response rate of paclitaxel alone in the 3rd line setting was set to 10%, an objective response rate of 10% was not clinically significant, and was expected for paclitaxel alone. An objective response rate of 15% was clinically significant, and the target objective response rate was 25%. Using a Simon two-stage design, a one-sided type I error of 10% and a power of 80% were used to detect the difference between the null of 10% ORR and the target of 25% objective response rate.

[0253] The first stage of the design started with 13 patients. The results of 2 or more responses indicated non-ineffectiveness and continued to accumulate to the second stage, which included another 21 patients. 6 or more responses in 34 patients indicated that the combination treatment had a clinical benefit worthy of further study (precise α=0.095). A true response rate of 10% or 25% indicated that the probability of stopping the study after the first stage was 62% or 13%, respectively.

[0254] The safety and tolerability of the combination of onvansertib and paclitaxel were also evaluated. Treatment-related toxicities were summarized by highest grade and term using CTCAE v4.0 and reported with 90% binomial exact confidence intervals.

[0255] The progression-free survival (PFS) of the combined treatment was also evaluated using the RECIST 1.1 criteria, defined as the time from randomization (or registration) to progression (according to RECIST 1.1) or death due to any cause (whichever is earlier). Surviving patients without disease progression were censored on the date of the last disease assessment. PFS and overall survival (OS) were described using the Kaplan-Meier method and presented with 95% confidence intervals.

[0256] Determination of dose-limiting toxicity (DLT)

[0257] DLTs were measured from Phase 1b after the first cycle of 28 days of treatment. DLTs were defined as toxicities that were possibly related to the clinical trial and met one or more of the following criteria: (1) death not clearly due to the underlying disease or external causes; (2) Grade ≥3 non-hematologic toxicity, except as follows; (3) changes in liver function consistent with Hy's law; (4) Grade ≥3 febrile neutropenia; (5) Grade ≥4 neutropenia or thrombocytopenia lasting for >7 days; and (6) Grade ≥3 thrombocytopenia with bleeding. Exceptions to Grade 3 non-hematologic toxicity include: (1) Grade ≥3 electrolyte abnormalities with clinical symptoms, regardless of duration; (2) Grade ≥3 nausea or vomiting or diarrhea for <72 hours with adequate antiemetic or other supportive care; (3) Grade ≥3 fatigue lasting <7 days; or (4) Grade ≥3 electrolyte abnormalities lasting <72 hours without clinical symptoms.

[0258] DLTs may lead to discontinuation until the adverse event (AE) resolves to ≤ Grade 1. If a DLT does not resolve within 2 weeks, the patient is discontinued from the study. Patients are not evaluable and may be replaced if (1) discontinuation is due to reasons other than DLT, or (2) the patient receives less than 75% of the planned onvansertib dose (16 of 21 days) in Cycle 1 for any reason other than DLT-related interruption. Patients who experience a DLT that subsequently resolves may be allowed to continue onvansertib at an appropriate reduction in dose if the investigator determines that clinical benefit can be obtained.

[0259] Adverse Events, Dose Delays, and Dose Modifications

[0260] All adverse events (AEs) were monitored and reported in a timely manner according to the clinical trial protocol, including expected AEs, unexpected AEs and SAEs (Table 8). The severity of AEs was graded using the latest version of NCI-CTCAE 5.0. For each SAE, the highest severity grade was reported. Onvansertib-related AEs are generally reversible and typically recover within 3 weeks. Examples of onvansertib-related AEs may include bone marrow suppression (e.g., anemia, leukopenia, thrombocytopenia and neutropenia, including febrile neutropenia) and fatigue. Examples of paclitaxel-related AEs may include allergic reactions and severe hypersensitivity reactions, characterized by dyspnea and hypotension, angioedema and systemic urticaria requiring treatment; bone marrow suppression (mainly neutropenia) and neutrophil nadirs; and severe conduction abnormalities. In Table 8, # indicates that the event does not need to be reported if it is expected as listed in the protocol and does not require expedited reporting; * indicates that for participants enrolled and actively participating in the study or for AEs occurring within 30 days of the last intervention, the event must be reported within 1 working day of becoming aware of the event; and & indicates that the event exhibits a greater severity than expected as listed in the protocol and / or current informed consent and will be considered unexpected and reportable.

[0261] Table 8: Reportable adverse events (AEs) in DF / HCC

[0262]

[0263] Provide patients with appropriate monitoring and supportive care according to the clinical trial protocol. According to the clinical trial protocol, toxicity-related dose delays can last up to 28 days. Toxicity-related dose adjustments are made according to Table 9. The maximum dose of onvansertib is 18 mg / m 2 The maximum dose of paclitaxel was 48 mg / m 2 . In Table 9, onvansertib is administered orally once daily for 21 consecutive days of each 28-day cycle, while paclitaxel is administered intravenously (IV) once on days 1, 8, and 15 of each 28-day cycle. Intra-patient dose escalation may be allowed according to the clinical trial protocol. Toxicity potentially related to the clinical trial is managed according to the clinical trial protocol. For example, hematological toxicity, non-hematological toxicity, hepatotoxicity, and neuropathy are managed according to Table 10. Patients may also discontinue treatment and / or study according to the clinical trial protocol. For the management of hematological toxicity in Table 7, the superscript "1" indicates that study treatment should be discontinued if blood cell counts have not recovered to Grade 2 or better within 4 weeks of dosing interruption; and the superscript "2" indicates that for absolute neutrophil count (ANC) to resolve to baseline grade or ≤Grade 2 (≥1000 / mm 3) can start a new cycle. For the management of non-hematologic toxicity in Table 7, participants who experience intolerable grade 2 events may have their study drug suspended and / or the dose reduced at the discretion of the treating investigator. For grade 3 nausea and vomiting in exceptional circumstances, routine prophylactic antiemetic therapy is not required at the start of study treatment. However, patients should receive appropriate antiemetic therapy at the first onset of nausea or vomiting and thereafter as needed, according to local treatment practice guidelines.

[0264] Table 9: Dose reduction

[0265]

[0266] Table 10: Toxicity Management

[0267]

[0268]

[0269] Pharmacological determination

[0270] 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. According to the clinical trial protocol, pharmacokinetic evaluations of Phase 1b patients were collected at the following time points: C1D1 (before paclitaxel administration and 1, 2, 3, 4, 8 hours after paclitaxel administration), C1D8 (2 hours after paclitaxel administration), C1D15 (2 hours after paclitaxel administration) and C2D1 (before paclitaxel administration).

[0271] The 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), TNM (tumor, lymph node, metastasis), and PBMC (peripheral blood mononuclear cells). In Table 11, the medical history includes the record of relevant medical history and TNM stage at diagnosis; the start and end dates of previous treatment should be recorded (if intolerance, the reason should be recorded); the physical examination should include height (only at screening), weight, vital signs, and general physical examination; blood chemistry panel (including blood chemistry and complete blood count with differential (CBC with Differential) including sodium, potassium, chloride, bicarbonate, calcium, blood urea nitrogen, creatinine, glucose, albumin, alkaline phosphatase, total bilirubin, AST, ALT (CBC and blood chemistry tests can be performed up to 48 hours before Days 1, 8, and 15, and all visits have a ±3-day window); blood samples for PK analysis will be obtained only during Days 1, 8, and 15 of Cycle 1 and Day 1 of Cycle 2 in the Phase 1b portion (C1D1 samples should be collected before dosing and Collect 1, 2, 3, 4, and 8 hours after dosing; C1D8 and C1D15 samples should be collected 2 hours after dosing; C2D1 sample should be collected before dosing; this applies only to Phase 1b); In Phase 1b and Phase 2 studies, a baseline biopsy is required after consent is obtained when readily available (if available, sites should confirm that archival tissue is available; biopsy at EOT is optional); Blood samples for ctDNA assessment should be collected on C1D1, C3D1, and EOT (collected in two 10 mL Streck tube); for disease assessment, subjects must have chest / abdomen / pelvic imaging, preferably using CT scan, within 4 weeks before starting, and an MRI of the brain (or contrast CT scan of the brain if the subject cannot have MRI) must be obtained in subjects with symptoms suggestive of possible central nervous system (CNS) metastatic disease; CT or MRI restaging should be performed every 2 cycles (±1 week) (e.g., C2D15) until EOT, and EOT assessment should be performed within 28 days (±5 days) after the last dose of onvansertib; subjects were given onvansertib orally for 21 consecutive days, followed by 7 days of rest (onvansertib was administered in the clinic on Days 1, 8, and 15 of Cycle 1 and Day 1 of Cycle 2, but was taken at home otherwise); and blood for PBMCs was collected in four 8 mL CPT tubes at C1D1, C1D15, and EOT.

[0272] Table 11: Study Calendar for Phase 1B and Phase 2 Trials

[0273]

[0274]

[0275] Disease response and progression were evaluated using the new international criteria proposed in the Response Evaluation Criteria in Solid Tumors (RECIST) guidelines (version 1.1). The RECIST criteria use changes in the largest diameter of the tumor lesion (a one-dimensional measurement) and the shortest diameter in the case of malignant lymph nodes.

[0276] Patients who had measurable disease at baseline, had received at least one cycle of treatment, and had their disease reassessed were evaluated for target disease response. Patients who had evaluable lesions at baseline but did not meet the definition of measurable disease, had received at least one cycle of treatment, and had their disease reassessed were evaluated for non-target disease.

[0277] Response assessment is based on the presence, absence, or clear progression of lesions. Exemplary responses for patients with measurable disease can be found in Table 12. In Table 12, measurable disease can be a target disease. In Table 12, * indicates that response confirmation is only used for non-randomized trials with response as the primary endpoint; and ** indicates that in special circumstances, clear progression of non-target lesions can be accepted as disease progression. In addition, in Table 12, participants with an overall deterioration in health status, requiring discontinuation of treatment, and without objective evidence of disease progression at the time should be reported as "symptom worsening." Even after discontinuation of treatment, every effort should be made to record objective progression. Exemplary responses for patients with unmeasurable disease can be found in Table 13. In Table 13, unmeasurable disease can be a non-target disease. In Table 13, for non-target diseases, "non-CR / non-PD" takes precedence over "stable disease" because SD is increasingly used as an endpoint for efficacy assessment in some trials. Therefore, when there are no measurable lesions, it is not recommended to be included in this category.

[0278] Table 12: Treatment Findings for Participants with Measurable Disease

[0279]

[0280]

[0281] Table 13: Treatment Findings for Participants with Non-Measurable Disease

[0282] Non-target lesions New lesions Overall Response CR no CR Non-CR / Non-PD no Non-CR / Non-PD* Not all rated no Not rated Clear PD Yes or No PD any yes PD

[0283] In at least some previously described embodiments, one or more elements used in the embodiments may be used interchangeably in another embodiment, unless such replacement is technically infeasible. It will be appreciated by those skilled in the art 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 changes are intended to fall within the scope of the subject matter defined by the appended claims.

[0284] Regarding the use of any plural and / or singular terms in general herein, those skilled in the art can translate the plural into the singular and / or translate the singular into the plural according to the context and / or application. Various singular / plural arrangements can be explicitly set forth herein for clarity. As used in this specification and the appended claims, unless the context clearly indicates otherwise, the singular forms "a", "an", and "the" include plural indicators. Unless otherwise indicated, any reference to "or" herein is intended to include "and / or".

[0285] Those skilled in the art will understand that, in general, the terms used herein, and particularly the terms used in the appended claims (e.g., the bodies of the appended claims), are generally intended to be "open-ended" terms (e.g., the term "including" should be understood to mean "including but not limited to," the term "having" should be understood to mean "having at least," the term "include" should be understood to mean "including but not limited to," etc.). Those skilled in the art will also understand that if a specific number of claim recitations is intended to be introduced, such intent will be explicitly recited in the claim, and in the absence of such a recitation, such intent is absent. For example, as an aid to understanding, the appended claims may contain usage of the introductory phrases "at least one" and "one or more" to introduce claim recitations. However, the use of such phrases should not be interpreted as meaning that the introduction of a claim recitation by the indefinite article "a" or "an" limits any particular claim containing such introduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the introductory phrases "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 as meaning "at least one" or "one or more"); this also applies to the use of definite articles used to introduce claim recitations. In addition, even if a specific number of an introduced claim recitation is explicitly recited, one skilled in the art will recognize that such recitation should be understood to mean at least the recited number (e.g., the mere recitation of "two recitations" without other modifiers means at least two recitations, or means two or more recitations). In addition, in those cases where similar conventions to "at least one of A, B, and C, etc." are used, generally, such constructions are intended in a sense that a person skilled in the art will understand the meaning of the convention (e.g., "a system having at least one of A, B, and C" will include but is not limited to systems having only A, only B, only C, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). In those cases where similar conventions to "at least one of A, B, or C, etc." are used, generally, such constructions are intended in a sense that a person skilled in the art will understand the meaning of the convention (e.g., "a system having at least one of A, B, or C" will include but is not limited to systems having only A, only B, only C, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). It should also be understood by those skilled in the art that any transitional words and / or phrases that actually represent two or more optional terms, whether in the specification, claims, or drawings, should be understood to contemplate the possibility of including one of the terms, either of the terms, or both of the terms.

[0286] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0287] As will be understood by those skilled in the art, for any and all purposes, such as in providing written description, all scopes disclosed herein also encompass any and all possible sub-ranges and combinations of sub-ranges.Any listed scope can be easily identified as fully describing and enabling the same scope to be decomposed into at least equal half, one-third, one-quarter, one-fifth, one-tenth, etc. As non-limiting examples, each scope discussed herein can be easily decomposed into lower third, middle third, and upper third, etc. As will be understood by those skilled in the art, all languages, such as "up to", "at least", "greater than", "less than", etc., include stated numbers, and refer to the scope of the sub-ranges discussed above that can be subsequently decomposed. Finally, as will be understood by those skilled in the art, scope includes each individual member.Therefore, for example, a group with 1-3 articles refers to a group with 1, 2 or 3 articles.Similarly, a group with 1-5 items refers to a group with 1, 2, 3, 4 or 5 items, etc.

[0288] Although various aspects and embodiments are 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 are not intended to be limiting, with the true scope and spirit being indicated by the appended claims.

Claims

1. A method for treating cancer, comprising: A microtubule depolymerization (MTDP) inhibitor and a Polo-like kinase 1 (PLK1) inhibitor are administered to a subject having cancer, thereby inhibiting or reducing the progression of the cancer in the subject.

2. The method of claim 1, wherein the subject has a hematological cancer or a solid cancer.

3. The method of claim 1, wherein the cancer is breast cancer, wherein the breast cancer 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, hormone receptor negative breast cancer, or a combination thereof.

4. The method of any one of claims 1-3, wherein the subject has TNBC.

5. The method of any one of claims 1-4, comprising identifying the subject with cancer as having 3 More than about 1000 neutrophils per mm 3 A subject having greater than about 100,000 platelets, a total bilirubin of about 1.5 times the upper limit of the institutional normal value, and a glomerular filtration rate of less than about 1.5 times the upper limit of the institutional normal value or a serum creatinine of 60 mL / min, or a combination thereof.

6. The method according to any one of claims 1-5, wherein the subject has 3 More than about 1000 neutrophils, or 100 neutrophils per mm 3 More than about 100,000 platelets.

7. The method of any one of claims 1-5, wherein the subject has a total bilirubin of about 1.5 times the institutional upper limit of normal, and / or has a glomerular filtration rate of less than about 1.5 times the institutional upper limit of normal or a serum creatinine of 60 mL / min.

8. The method of any one of claims 1-7, wherein the PLK1 inhibitor and the MTDP inhibitor are co-administered simultaneously.

9. The method of any one of claims 1-7, wherein the PLK1 inhibitor and the MTDP inhibitor are administered sequentially.

10. The method of any one of claims 1-9, wherein said administration of said PLK1 inhibitor is oral, and optionally, said subject fasts for more than about 30 minutes prior to said administration and / or said subject fasts for about 4 hours after said administration.

11. The method of any one of claims 1-10, wherein the administration of the MTDP inhibitor is intravenous, optionally through an in-line filter having a microporous membrane of no greater than 0.22 microns, and further optionally wherein prior to administration, the subject is (i) orally administered about 20 mg of dexamethasone between about 12 hours and about 6 hours prior or intravenously administered about 12 mg of dexamethasone about 60 minutes prior; (ii) administered about 50 mg of diphenhydramine about 30 minutes to about 60 minutes prior intravenously; and / or (iii) administered about 300 mg of cimetidine about 30 minutes to about 60 minutes prior intravenously or about 20 mg of famotidine about 30 minutes to about 60 minutes prior intravenously.

12. The method of any one of claims 1-11, wherein the inhibition of cancer progression is greater than the combined inhibition of progression caused by the MTDP inhibitor alone plus the PLK1 inhibitor alone.

13. The method of any one of claims 1-12, wherein the subject achieves a complete response.

14. The method according to any one of claims 1-13, wherein the subject has received prior treatment with an MTDP inhibitor or a PLK1 inhibitor.

15. The method of any one of claims 1-14, wherein the subject is unresponsive to treatment with the MTDP inhibitor or PLK1 inhibitor alone.

16. The method of any one of claims 1-14, wherein the subject is known to be resistant to MTDP inhibitor or PLK1 inhibitor therapy.

17. The method of any one of claims 1-16, wherein the MTDP inhibitor and the PLK1 inhibitor are each administered to the subject in a 28-day cycle.

18. The method of any one of claims 1-17, wherein 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, the cycle comprising about 14-28 days of administration and about 0-14 days of no administration, and optionally about 21 days of administration of the PLK1 inhibitor and about 7 days of no administration of the PLK1 inhibitor.

19. The method of any one of claims 1-18, wherein the MTDP inhibitor, the PLK1 inhibitor, or both are administered in a 28-day cycle.

20. The method of any one of claims 1-16, wherein each cycle of treatment is at least about 28 days.

21. The method of any one of claims 1-16, wherein each cycle of treatment is from about 14 days to about 28 days.

22. The method of any one of claims 1-21, wherein the PLK1 inhibitor is administered on at least seven days, at least fourteen days, or at least twenty-one days of a cycle.

23. The method of any one of claims 1-21, wherein the PLK1 inhibitor is not administered on at least one day, at least three days, or at least seven days of a cycle.

24. The method of any one of claims 1-23, wherein the MTDP inhibitor is administered once or twice per week.

25. The method of any one of claims 1-23, wherein the MTDP inhibitor is administered once a week for two or three consecutive weeks in a cycle.

26. The method of any one of claims 1-25, wherein the subject undergoes at least two cycles of administration of the MTDP inhibitor and the PLK1 inhibitor.

27. The method according to any one of claims 1-26, wherein the MTDP inhibitor is paclitaxel, docetaxel, acetyltaxol, paclitaxel; Lu 177vipivotide tetraxetan; 7-hexanoyltaxol; cabazitaxel; larotaxel; milataxel; ortataxel; tesetaxel; taxoprexin; opaxio; taxoprexin (DHA-paclitaxel); poly(L-glutamic acid)-paclitaxel; abraxane; SB-T-1214; SB-T1216; SB-T121602; SB-T-12854; DHA-SB-T1214; abeo-taxane, wherein the abeo-taxane is optionally abeo-taxane 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; and any combination thereof.

28. The method of any one of claims 1-26, wherein the MTDP inhibitor is paclitaxel.

29. The method of any one of claims 1-28, wherein about 48 mg / m 2 Body surface area: about 80 mg / m 2 Paclitaxel by body surface area.

30. The method of any one of claims 1-29, wherein the PLK1 inhibitor is 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; optionally, wherein the PLK1 inhibitor is onvansertib.

31. The method of claim 30, wherein onvansertib is administered at about 6 mg / m 2 Body surface area: about 24 mg / m 2 The dosage is administered per body surface area, optionally at 9 mg / m 2 Body surface area, 12 mg / m 2 Body surface area or 18 mg / m 2 Dosage administration by body surface area.

32. The method of any one of claims 1-31, wherein the subject has received at least one prior cancer treatment.

33. The method of claim 32, wherein the prior treatment did not include use of an MTDP inhibitor, a PLK1 inhibitor, or both; and optionally, the PLK1 inhibitor is onvansertib.

34. The method of any one of claims 1-33, wherein the subject is in cancer remission.

35. The method of any one of claims 1-34, wherein the subject in cancer remission is in complete remission (CR) or in partial remission (PR).

36. The method of any one of claims 1-35, further comprising determining the cancer status of the subject.

37. The method of any one of claims 1-36, further comprising determining the subject's responsiveness to treatment with the MTDP inhibitor and the PLK1 inhibitor.

38. The method of any one of claims 1-37, further comprising administering one or more cancer therapeutic agents or therapies directed against the cancer.

39. The method of any one of claims 1-38, wherein the subject is a human.

40. A method of sensitizing cancer cells to a microtubule depolymerization (MTDP) inhibitor, the method comprising: Cancer cells are contacted with a composition comprising a Polo-like kinase 1 (PLK1) inhibitor, thereby sensitizing the cancer cells to the MTDP inhibitor; optionally, wherein the PLK1 inhibitor is onvansertib and / or the MTDP inhibitor is paclitaxel.

41. The method of claim 40, wherein contacting the cancer cells with the composition occurs in vitro, ex vivo, and / or in vivo.

42. The method of any one of claims 40-41, wherein contacting the cancer cells with the composition is in a subject; optionally, wherein the subject is unresponsive to, or is known to be resistant to, the MTDP inhibitor or another MTDP inhibitor.

43. The method of any one of claims 40-42, wherein the subject has prior treatment with the MTDP inhibitor or another MTDP inhibitor.

44. The method of any one of claims 40-43, wherein the subject is a mammal, and optionally the mammal is a human.

45. The method of any one of claims 40-44, comprising determining sensitization of the cancer cells to an MTDP inhibitor after contacting the cancer cells with the composition.

46. ​​The method of any one of claims 40-45, comprising contacting the cancer cell with the MTDP inhibitor, optionally wherein contacting the cancer cell with the MTDP inhibitor occurs in the subject.

47. The method of any one of claims 40-46, comprising determining the subject's response to the MTDP inhibitor.

48. The method of any one of claims 40-47, wherein contacting the cancer cell with the MTDP inhibitor is simultaneous with contacting the cancer cell with the composition, or after contacting the cancer cell with the composition.

49. The method of any one of claims 40-48, wherein the subject has 3 More than about 1000 neutrophils, or 100 neutrophils per mm 3 More than about 100,000 platelets.

50. A medicine kit comprising: polo-like kinase 1 (PLK1) inhibitors; and A manual is provided with instructions for co-administering the PLK1 inhibitor with a microtubule depolymerization (MTDP) inhibitor to a subject in need thereof for treating cancer.

51. The kit of claim 50, wherein the cancer is a hematological cancer or a solid cancer, and optionally the cancer is breast cancer, wherein the breast cancer 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, hormone receptor negative breast cancer, or a combination thereof; optionally, wherein the cancer is TNBC.

52. The kit of any one of claims 50-51, wherein the subject has 3 More than about 1000 neutrophils per mm 3 Greater than about 100,000 platelets, total bilirubin about 1.5 times the upper limit of institutional normal, and a glomerular filtration rate less than about 1.5 times the upper limit of institutional normal or a serum creatinine of 60 mL / min.

53. The kit of any one of claims 50-52, wherein the PLK1 inhibitor is onvansertib and / or the MTDP inhibitor is paclitaxel.

54. The kit of any one of claims 50-53, wherein the instructions include instructions for co-administering the PLK1 inhibitor and the MTDP inhibitor simultaneously.

55. The kit of any one of claims 50-54, wherein the instructions include instructions for sequential co-administration of the PLK1 inhibitor and the MTDP inhibitor.

56. The kit of any one of claims 50-55, wherein the instructions comprise instructions for administering the PLK1 inhibitor orally, wherein prior to the administering, the subject fasts for more than about 30 minutes, and wherein after the administering, the subject fasts for about 4 hours.

57. The kit of any one of claims 50-56, wherein the instructions include instructions for administering the MTDP inhibitor intravenously, optionally through an in-line filter having a microporous membrane of no greater than 0.22 microns, and optionally prior to administration, the subject (i) is orally administered about 20 mg of dexamethasone between about 12 hours and about 6 hours before or is intravenously administered about 12 mg of dexamethasone about 60 minutes before; (ii) is intravenously administered about 50 mg of diphenhydramine about 30 minutes to about 60 minutes before; and / or (iii) is intravenously administered about 300 mg of cimetidine about 30 minutes to about 60 minutes before or is intravenously administered about 20 mg of famotidine about 30 minutes to about 60 minutes before.

58. The kit of any one of claims 50-57, wherein the instructions include instructions for administration to a subject who has received prior treatment with an MTDP inhibitor or a PLK1 inhibitor.

59. The kit of any one of claims 50-58, wherein the instructions include instructions for administration to a subject who has not responded to treatment with the MTDP inhibitor or PLK1 inhibitor alone.

60. The kit of any one of claims 50-59, wherein the instructions include instructions for administration to a subject known to be resistant to MTDP inhibitor or PLK1 inhibitor therapy.

61. The kit of any one of claims 50-60, wherein the instructions comprise instructions for administering each of the paclitaxel and onvansertib to the subject in cycles of 28 days.

62. The kit of any one of claims 50-61, wherein the instructions comprise instructions for administering each of the paclitaxel and onvansertib to the subject in cycles of at least five times a week.

63. The kit of any one of claims 50-62, wherein the instructions comprise instructions for administering the MTDP inhibitor, onvansertib, or both in a cycle of at least 7 days.

64. The kit of claim 63, wherein each cycle of treatment is at least about 21 days, or from about 14 days to about 28 days.

65. The kit of any one of claims 50-64, wherein the instructions comprise instructions for administering onvansertib daily for 14-21 consecutive days in a 28-day cycle.

66. The kit of any one of claims 50-65, wherein the instructions include instructions for not administering onvansertib for three, five, or seven consecutive days in a cycle.

67. The kit of any one of claims 50-66, wherein the instructions include instructions for administering the MTDP inhibitor weekly for three consecutive weeks in a cycle.

68. The kit of any one of claims 50-67, wherein the MTDP inhibitor is a reversible MTDP inhibitor.

69. The kit of any one of claims 50-68, wherein the MTDP inhibitor is paclitaxel or a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof.

70. The kit of any one of claims 50-69, wherein the instructions include for use at about 3 mg / m 2 Body surface area: about 24 mg / m 2 Onvansertib is administered at a dose of approximately 48 mg / m 2 Body surface area: about 80 mg / m 2 Instructions for administering paclitaxel at a dose of 9 mg / m 2 Body surface area, 12 mg / m 2 Body surface area or 18 mg / m 2 Instructions for dosing of onvansertib by body surface area.

71. The kit of any one of claims 50-70, wherein the subject has received at least one prior treatment for the cancer.

72. The kit of claim 71, wherein the prior treatment did not include the use of an MTDP inhibitor, onvansertib, or both.

73. The kit of any one of claims 50-72, wherein the subject is in cancer remission.

74. The kit of claim 73, wherein the subject in cancer remission is in complete remission (CR) or in partial remission (PR).

75. The kit according to any one of claims 50-74, further comprising the MTDP inhibitor, preferably paclitaxel; and / or a PLK1 inhibitor, preferably onvansertib.

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