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WO2019165473A1 - Procédés de traitement du cancer comprenant des inhibiteurs de cdc7 - Google Patents

Procédés de traitement du cancer comprenant des inhibiteurs de cdc7 Download PDF

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Publication number
WO2019165473A1
WO2019165473A1 PCT/US2019/019676 US2019019676W WO2019165473A1 WO 2019165473 A1 WO2019165473 A1 WO 2019165473A1 US 2019019676 W US2019019676 W US 2019019676W WO 2019165473 A1 WO2019165473 A1 WO 2019165473A1
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Prior art keywords
cancer
day
inhibitor
tumor
sra141
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PCT/US2019/019676
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English (en)
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Christian Andrew HASSIG
Ryan James HANSEN
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Sierra Oncology Inc
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Sierra Oncology Inc
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Priority to JP2021516638A priority Critical patent/JP7579781B2/ja
Priority to AU2019350581A priority patent/AU2019350581B2/en
Priority to US17/275,732 priority patent/US20210393620A1/en
Priority to CN201980069832.XA priority patent/CN113348020A/zh
Priority to PCT/US2019/048657 priority patent/WO2020068347A1/fr
Priority to CA3113621A priority patent/CA3113621A1/fr
Priority to KR1020217010445A priority patent/KR20210064252A/ko
Priority to CN202511057693.2A priority patent/CN121081469A/zh
Priority to EP19866944.2A priority patent/EP3856352A4/fr
Publication of WO2019165473A1 publication Critical patent/WO2019165473A1/fr
Anticipated expiration legal-status Critical
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid 
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/451—Non condensed piperidines, e.g. piperocaine having a carbocyclic group directly attached to the heterocyclic ring, e.g. glutethimide, meperidine, loperamide, phencyclidine, piminodine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4965—Non-condensed pyrazines
    • A61K31/497—Non-condensed pyrazines containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A61K31/675—Phosphorus compounds having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • Cancer is a group of diseases caused by uncontrolled, unlimited growth of cells within a living body. Since cancer cells usually grow faster than normal cells, cancers are capable of being treated by controlling the replication of DNA during cell division, particularly during the division of chromosomes.
  • Cdc7 is a serine-threonine protein kinase enzyme which is essential for the initiation of DNA replication in the cell cycle. Specifically, Cdc7 forms a complex with cofactors such as Dbf4 (ASK), and phosphorylates its substrate, MCM (mini-chromosome maintenance) proteins. It is purported that this phosphorylation results in assembly of Cdc45 and a DNA polymerase on the DNA to form an MCM complex, thereby initiating the DNA replication.
  • ASK Dbf4
  • MCM mini-chromosome maintenance
  • Cdc7 As an anticancer target since the expression level of Cdc7 is frequently elevated in various cancer cell lines and human tumor tissues. It has been found that Cdc7 is overexpressed not only in commonly established cell lines derived from human tumors, but also in cells taken from live tissues.
  • Cdc7 inhibitors have been demonstrated to effect the growth of human tumor cells, such as HeLa and HCT116 cells, while exhibiting only limited effects on normal cells.
  • Described herein is a method of treating a cancer, the method comprising
  • a therapeutically effective amount of a Cdc7 inhibitor wherein the therapeutically effective amount is between an absolute dose of 10-400 mg/day or between 10-1000 mg/day.
  • the subject is a human.
  • the therapeutically effective amount is at least 10 mg/day, at least 20 mg/day, at least 40 mg/day, at least 80 mg/day, at least 160 mg/day, or at least 320 mg/day. In some embodiments, the therapeutically effective amount is at least 15 mg/day, at least 25 mg/day, at least 50 mg/day, at least 100 mg/day, at least 150 mg/day, at least 200 mg/day, at least 250 mg/day, at least 300 mg/day, or at least 350 mg/day.
  • the Cdc7 inhibitor is administered orally.
  • the Cdc7 inhibitor is administered daily. In some embodiments, the Cdc7 inhibitor is administered daily. In some embodiments, the Cdc7 inhibitor is administered daily.
  • the Cdc7 inhibitor is administered for at least 5 consecutive days, at least 7 consecutive days, or at least 14 consecutive days.
  • the Cdc7 inhibitor is administered following a dosing schedule selected from the group consisting of: 5 days of dosing followed by 2 days of non-dosing each week; 1 week of daily dosing followed by 1, 2, or 3 weeks of non-dosing; 2 or 3 weeks of daily dosing followed by 1, or 2 weeks of non dosing; and dosing on days 2 and 3 of a weekly cycle.
  • the Cdc7 inhibitor is administered for at least 5 consecutive days, at least 7 consecutive days, or at least 14 consecutive days.
  • the Cdc7 inhibitor is administered following a dosing schedule selected from the group consisting of: 5 days of dosing followed by 2 days of non-dosing each week; 1 week of daily dosing followed by 1, 2, or 3 weeks of non-dosing; 2 or 3 weeks of daily dosing followed by 1, or 2 weeks of non dosing; and dosing on
  • therapeutically effective amount is administered in a single dose once a day. In some embodiments, half of the therapeutically effective amount is administered twice a day.
  • the cancer is selected from the group consisting of: melanoma, uterine cancer, thyroid cancer, blood cancer, bladder cancer, breast cancer, cervical cancer, colorectal cancer (CRC), gastric cancer, endometrial cancer, hepatocellular cancer, leukemia, lymphoma, myeloma, non-small cell lung cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain cancer, sarcoma, small cell lung cancer, neuroblastoma, and head and neck cancer.
  • melanoma uterine cancer, thyroid cancer, blood cancer, bladder cancer, breast cancer, cervical cancer, colorectal cancer (CRC), gastric cancer, endometrial cancer, hepatocellular cancer, leukemia, lymphoma, myeloma, non-small cell lung cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain cancer, sarcoma, small cell lung cancer, neuroblastoma, and head and neck cancer.
  • the cancer is a blood cancer selected from the group consisting of: acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), chronic eosinophilic leukemia, and diffuse large B-cell lymphoma (DLBCL).
  • AML acute myeloid leukemia
  • CML chronic myelogenous leukemia
  • DLBCL diffuse large B-cell lymphoma
  • the cancer is AML.
  • the cancer is metastatic colorectal cancer (mCRC).
  • the mCRC is not categorized as having a high microsatellite instability (MSI- H) status.
  • MSI-H status is determined by detection of repetitive DNA sequences selected from the group consisting of: mononucleotide repeat markers, dinucleotide repeat markers, quasimonomorphic markers, and combinations thereof.
  • the detection if performed by a method selected from the group consisting of: PCR analysis, multiplexed PCR analysis, capillary electrophoresis, DNA sequencing, and combinations thereof.
  • a tumor associated with the cancer comprises a phenotype selected from the group consisting of: chromosome instability (CIN), a spindle checkpoint assembly defect, a mitosis defect, a Gl/S checkpoint defect, and combinations thereof.
  • a tumor associated with the cancer comprises a Wnt signaling pathway mutation.
  • the Wnt signaling pathway mutation is selected from the group consisting of: an Adenomatous polyposis coli (APC) gene mutation, a FAT1 mutation, a FAT4 mutation, and combinations thereof.
  • APC Adenomatous polyposis coli
  • the method further comprises screening the tumor associated using either archival or fresh tumor biopsy.
  • the screening comprises examining a pattern of chromosome separation by histochemical staining, examining pharmacodynamic markers by histochemical staining, or a combination thereof.
  • the screening further comprises determining whether the tumor exhibits aberrant mitosis.
  • the screening is performed before the administering of the Cdc7 inhibitor. In some embodiments, the screening is performed after the
  • the method results in a plasma Cmax greater than 600, greater than 1000, or greater than 1400 ng/mL of the Cdc7 inhibitor in the subject after
  • the method results in an AUCiast greater than 5800, greater than 11900, or greater than 16400 ng.h/mL of the SRA141 compound in the subject after administration.
  • the method results in an intra-tumoral concentration of greater than 500 ng/mL, greater than 600 ng/mL, greater than 900 ng/mL, or greater than 1300 ng/mL of the Cdc7 inhibitor in the subject after administration.
  • the intra-tumoral concentration is reached after multiple doses of the SRA141 compound.
  • the intra-tumoral concentration is reached after a single dose of the Cdc7 inhibitor.
  • the method results in in vivo inhibition of MCM2
  • the in vivo inhibition of MCM2 phosphorylation is at amino acid residues Ser40 or Ser53. In some embodiments, the in vivo inhibition of MCM2 phosphorylation is in a tumor associated with the cancer. In some embodiments, the in vivo inhibition of MCM2 phosphorylation in the tumor associated with the cancer is at least 50% relative to an untreated tumor sample. In some embodiments, the untreated tumor sample is a biopsy obtained prior to administration of the Cdc7 inhibitor to the subject. In some embodiments, the in vivo inhibition of MCM2 phosphorylation is after a single dose of the
  • the in vivo inhibition of MCM2 phosphorylation is in skin of the subject. In some embodiments, the in vivo inhibition of MCM2 phosphorylation is measured by Western blot analysis, immunohistochemistry (IHC), or liquid chromatography- mass spectrometry (LC/MS). In some embodiments, the in vivo inhibition of MCM2 phosphorylation is measured in a biopsy of the subject. In some embodiments, the in vivo inhibition of MCM2 phosphorylation is measured after multiple doses of the Cdc7 inhibitor. In some embodiments, the in vivo inhibition of MCM2 phosphorylation is measured after the Cdc7 inhibitor reaches a steady state plasma concentration.
  • the method results in growth inhibition of a tumor associated with the cancer.
  • the growth inhibition of the tumor is a minimum growth inhibition of at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% relative to an untreated tumor.
  • the growth inhibition of the tumor is a minimum growth inhibition of at least 47% relative to an untreated tumor.
  • the growth inhibition of the tumor is a minimum growth inhibition of at least 93% relative to an untreated tumor.
  • the method results in a regression of a tumor associated with the cancer. In some embodiments, the regression is a complete regression.
  • the method results in cytotoxicity of a tumor associated with the cancer. In some embodiments, the method results in at least a 30% decrease in the sum of diameters of tumors associated with the cancer. In some embodiments, the method results in a partial response, a complete response, or stable disease in the subject.
  • the method further comprises administering to the subject a second therapeutically effective amount of one or more additional treatments.
  • the one or more additional treatments comprises an anti-neoplastic agent.
  • the anti -neoplastic agent is selected from the group consisting of: a DNA polymerase inhibitor, a receptor tyrosine kinase inhibitor, a mammalian target of rapamycin
  • the anti -neoplastic agent comprises a mitogen activated protein kinase (MAPK) pathway inhibitor.
  • MAPK mitogen activated protein kinase
  • the MAPK inhibitor is Trametinib.
  • the anti -neoplastic agent comprises a retinoid pathway regulator.
  • the retinoid pathway regulator is the RXR agonist Bexarotene or the RAR agonist Tretinoin (all-trans retinoic acid, ATRA).
  • the wherein the anti -neoplastic agent comprises an apoptosis regulator.
  • the apoptosis regulator comprises an apoptosis inducer.
  • the apoptosis inducer comprises a BCL-2 inhibitor.
  • the retinoid pathway regulator is the RXR agonist Bexarotene or the RAR agonist Tretinoin (all-trans retinoic acid, ATRA).
  • the wherein the anti -neoplastic agent comprises an apoptosis regulator.
  • the apoptosis regulator comprises an apoptosis inducer.
  • the apoptosis inducer comprises a BCL-2 inhibitor.
  • the BCL-2 inhibitor is ABT-199.
  • the anti -neoplastic agent comprises a phosphatidylinositol-4,5-bisphosphate 3 kinase (PI3K) pathway inhibitor.
  • the PI3K pathway inhibitor is Copanlisib.
  • the anti- neoplastic agent comprises a PARP inhibitor.
  • the PARP inhibitor is BMN673.
  • the anti -neoplastic agent comprises an Aurora B kinase inhibitor.
  • the Aurora B kinase inhibitor is Barasertib.
  • the one or more additional treatments is administered daily.
  • the Cdc7 inhibitor and the one or more additional treatments demonstrate synergistic effects.
  • the method comprises a Cdc7 inhibitor that is a furanone derivative represented by the formula (I):
  • the A of formula (I) is -COOR1. In some embodiments, the A of formula (I) is a hydrogen atom. In some embodiments, the furanone derivative has the structure of the following compound (I-A), compound (I-B), compound (I-C), compound (I- D), or compound (I-E):
  • the Cdc7 inhibitor is any of the following compounds: formula (I-F): 2-(pyridin-4-yl)-l,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridinone
  • Figure 1A is a graph showing tumor volumes of animals treated with SRA141 monotherapy in a mouse xenograft model of acute myeloid leukemia.
  • Figure IB is a graph showing tumor weights of animals treated with SRA141 monotherapy in a mouse xenograft model of acute myeloid leukemia.
  • Figure 1C is a graph showing body weight change of animals treated with SRA141 monotherapy in a mouse xenograft model of acute myeloid leukemia.
  • Figure 2 A is a graph showing mean tumor volumes of animals treated with SRA141 monotherapy in a rat xenograft model of acute myeloid leukemia.
  • Figure 2B is a graph showing body weight change of animals treated with SRA141 monotherapy in a rat xenograft model of acute myeloid leukemia.
  • Figure 2C is a graph showing tumor volumes of individual subjects treated with SRA141 monotherapy in a rat xenograft model of acute myeloid leukemia.
  • Figure 3A is a graph showing the half maximal inhibitory concentration (ICso) of SRA141 in a panel of hematological cancer derived cell lines.
  • Figure 3B shows a summary of determined ICso values broken down by cancer type.
  • Figure 3C illustrates sensitivity to SRA141 between cancer and normal cells.
  • Figure 3D shows activity of Cdc7 inhibitors SRA141, TAK-931, and LY-3177833 in numerous cell lines.
  • Figure 3E shows ICso values using four orthogonal assays designed to measure ATP levels (CTG); metabolic activity (CellTiter-Blue (CTB)); DNA content of the cells
  • Figure 3F shows ICso values determined by CTG and CyQuant at l44h following differing exposure times to SRA141.
  • Figure 4 A is a graph showing mean tumor volumes of animals treated with SRA141 monotherapy in a patient derived xenograft (PDX) model of colorectal cancer (CRC).
  • PDX patient derived xenograft
  • CRC colorectal cancer
  • Figure 4B is a graph showing body weight change of animals treated with SRA141 monotherapy in a patient derived xenograft (PDX) model of colorectal cancer (CRC).
  • PDX patient derived xenograft
  • CRC colorectal cancer
  • Figure 5 is a graph showing tumor volumes of individual subjects of animals treated with SRA141 monotherapy in a patient derived xenograft (PDX) model of colorectal cancer (CRC).
  • PDX patient derived xenograft
  • CRC colorectal cancer
  • Figure 6 is a graph showing mean tumor volumes of animals treated with SRA141 monotherapy on Day 32 (D32) of treatment in a patient derived xenograft (PDX) model of colorectal cancer (CRC).
  • PDX patient derived xenograft
  • CRC colorectal cancer
  • Figure 7 A is a graph showing mean tumor volumes of animals treated with SRA141 in a rat xenograft model of colorectal cancer.
  • Figure 7B shows graphs of percent body weight change, tumor concentration of SRA141, and an immunoblot showing reduction of phosphorylated MCM2 12 hours post- treatment with SRA141 in a rat xenograft model of colorectal cancer.
  • Figure 8 are graphs showing inhibition of cell growth (cell viability compared to 0 hr and 72 hr untreated control) of cells lines sequentially treated (pre-treated) with SRA141 in combination with anti-neoplastic agents.
  • Figure 9 shows tables depicting synergy scores for growth inhibition of cell lines co- treated with SRA141 (“Sierra compound 1”) in combination with anti -neoplastic agents.
  • Figure 10A shows tables depicting synergy scores for growth inhibition of cell lines co-treated with SRA141 (“Sierra compound 1”) in combination with anti-neoplastic agents.
  • Figure 10B is a graph showing percent growth inhibition of HT-29 cells treated with gemcitabine alone or in combination with SRA141.
  • Figure 11A shows potent inhibition of Cdc7 by SRA141 in an in vitro biochemical assay.
  • Figure 11B shows residence time for SRA141 binding to Cdc7 and dissociation kinetics.
  • Figure 12 shows results of kinome screening assays of SRA141 and TAK931.
  • Figure 13 shows Colo-205 cells treated with SRA141 for 8 (Fig. 13A) to 24 (Fig. 13B) hours at concentrations between 0.033 and 3.3 mM, and for 3, 6, and 24 hours (Fig.
  • Figure 14 is a graph showing quantification of phosphorylation on Ser53 of MCM2 following treatment with SRA141 at 0.1 pM and 1 pM for 24 hours.
  • Figure 15 shows flow cytometry data from cells treated with SRA141 during S phase (Fig. 15A) and at the beginning of M phase (Fig. 15B); assessment of cell cycle and DNA markers in cells treated with SRA141 (Fig. 15C); and assessment of the percent of mitotic cells in a population of cells treated a Cdc7 inhibitor (Fig. 15D).
  • Figure 16 shows a summary of SRA141 sensitivity in cell lines with and without APC mutations.
  • Figure 17 is a graph showing mean tumor volumes of animals treated with SRA141 monotherapy in BALB/c mice bearing Colo-205 tumor xenografts.
  • Figure 18 is a graph showing mean tumor volumes of animals treated with SRA141 monotherapy in BALB/c mice bearing SW620 tumor xenografts.
  • Figure 19 is a graph showing mean tumor volumes of animals treated with SRA141 monotherapy in BALB/c mice bearing A20 tumor xenografts.
  • Figure 20 is a graph showing mean tumor volumes of animals treated with SRA141 monotherapy in CB-17 SCID mice bearing MDA-MB-486 breast tumor xenografts.
  • Figure 21 shows pMCM2 levels decreased following a single SRA141 administration in BALB/c mice bearing subcutaneous SW620 tumors (pMCM2 levels shown in Fig. 21A, % inhibition quantified and normalized to actin in Fig. 21B).
  • Figure 22 shows pMCM2 levels following SRA141 treatment in female Rowett nude rats bearing subcutaneous Colo-205 tumors.
  • Figure 23 shows SRA141 plasma concentrations determined by LC-MS/MS in non tumor bearing female nude rats.
  • Figure 24A shows immunohistochemistry assessments of total MCM2, phosphorylated MCM2, and gH2AX in tumor and surrogate tissue from a rat xenograft model of leukemia treated with and without SRA141.
  • Figure 24B shows immunohistochemistry assessments of phosphorylated MCM2 in tumor and surrogate tissue from a rat xenograft model of leukemia.
  • Figure 24C shows immunohistochemistry and H&E assessments of tumor from a rat xenograft model of leukemia treated with SRA141 and vehicle.
  • Figure 25A shows results of evaluation of pMCM2-S40 expression in xenograft tumors following treatment with SRA141 at various dosages.
  • Figure 25B shows results of evaluation of pMCM2-S40 expression in skin samples from rat xenografts following treatment with SRA141 at various dosages.
  • Figure 26 shows results of evaluation of total MCM2, pMCM2-S53, pMCM2-S40, and gH2AX in normal human skin.
  • Figure 27 shows results of cell viability assays performed with SRA141 in
  • Figure 28 shows results of treatment with SRA141 in cells following inhibition of anti-apoptotic genes by RNAi knockdown (Fig. 28A) or treatment with ABT-199 (Fig. 28B).
  • Disclosed herein are methods of inhibiting tumor growth in a subject, e.g., a human, by administration of an effective amount of a Cdc7 inhibitor. Also disclosed herein are methods of inhibiting tumor growth in a subject, e.g., a human, by administration of an effective amount of a Cdc7 inhibitor in a combination therapy.
  • the practice of the present invention includes the use of conventional techniques of organic chemistry, molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are within the skill of the art.
  • Compounds utilized in the present invention possess asymmetric carbon atoms (optical centers) or double bonds; the racemates, diastereomers, geometric isomers, regioisomers and individual isomers (e.g., separate enantiomers) are all intended to be encompassed within the scope of the present invention.
  • the compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds.
  • the compounds may be radiolabeled with radioactive isotopes, such as for example, and without limitation, tritium (3 ⁇ 4), iodine- 125 ( 125 I), or carbon-l4 ( 14 C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.
  • subject refers to any mammal including humans, and mammals such as those animals of veterinary and research interest that are including, but not limited to:
  • administering or“administration of’ a drug and/or therapy to a subject (and grammatical equivalents of this phrase) refers to both direct or indirect administration, which may be administration to a subject by a medical professional, may be self- administration, and/or indirect administration, which may be the act of prescribing or inducing one to prescribe a drug and/or therapy to a subject.
  • coadministration refers to two or more compounds administered in a manner to exert their pharmacological effect during the same period of time. Such coadministration can be achieved by either simultaneous, contemporaneous, or sequential administration of the two or more compounds.
  • the term“treating” or“treatment of” a disorder or disease refers to taking steps to alleviate the symptoms of the disorder or disease, e.g., tumor growth or cancer, or otherwise obtain some beneficial or desired results for a subject, including clinical results. Any beneficial or desired clinical results may include, but are not limited to, alleviation or amelioration of one or more symptoms of cancer or conditional survival and reduction of tumor load or tumor volume; diminishment of the extent of the disease; delay or slowing of the tumor progression or disease progression; amelioration, palliation, or stabilization of the tumor and/or the disease state; or other beneficial results.
  • the term“in situ” or“in vitro” refers to processes that occur in a living cell growing separate from a living organism, e.g. , growing in tissue culture.
  • in vivo refers to processes that occur in a living organism.
  • Cdc7 refers to a cell division cycle 7 serine/threonine-protein kinase that is encoded by a CDC7 gene.
  • the methods of the invention include administration of an effective amount a Cdc7 inhibitor.
  • the present disclosure provides for a method of treatment wherein the effective amount of a Cdc7 inhibitor is administered to a subject.
  • the term “effective amount” or“therapeutically effective amount” refers to an amount that is effective to ameliorate a symptom of a disease, e.g. an amount that is effective to inhibit the growth of a tumor.
  • the effective amount of a Cdc7 inhibitor is less than maximum tolerated dose (MTD).
  • the effective amount of a Cdc7 inhibitor is less than 1000 mg/day, less than 500 mg/day, or less than 400 mg/day.
  • the effective amount of a Cdc7 inhibitor is less than 300 mg/day, less than 200 mg/day, less than 150 mg/day, less than 100 mg/day, or less than 75 mg/day. In some aspects, the effective amount of a Cdc7 inhibitor is less than 324 mg/day. In some aspects, the effective amount of a Cdc7 inhibitor is 324 mg/day. In some aspects, the effective amount of a Cdc7 inhibitor is at least 10 mg/day. In some aspects, the effective amount of a Cdc7 inhibitor is between 10-400 mg/day. In some aspects, the effective amount of a Cdc7 inhibitor is between 10-324 mg/day. In some aspects, the effective amount of a Cdc7 inhibitor is between 40-400 mg/day.
  • the effective amount of a Cdc7 inhibitor is at least 10 mg/day, at least 20 mg/day, at least 40 mg/day, at least 80 mg/day, at least 160 mg/day, or at least 320 mg/day. In some aspects, the effective amount of a Cdc7 inhibitor is at least 15 mg/day, at least 25 mg/day, at least 50 mg/day, at least 75 mg/day, at least 100 mg/day, at least 150 mg/day, at least 200 mg/day, at least 250 mg/day, at least 300 mg/day, or at least 350 mg/day.
  • the effective amount of a Cdc7 inhibitor is 10 mg/day, 20 mg/day, 40 mg/day, 80 mg/day, 160 mg/day, 320 mg/day, 325 mg/day, 350 mg/day, or 400 mg/day. In some aspects, the effective amount of a Cdc7 inhibitor is 15 mg/day, 25 mg/day, 50 mg/day, 75 mg/day, 100 mg/day,
  • the effective amount of a Cdc7 inhibitor is administered as a monotherapy.
  • the effective amount of the Cdc7 inhibitor monotherapy is less than a maximum tolerated dose (MTD). In some aspects, the effective amount of the Cdc7 inhibitor monotherapy is less than 1000 mg/day, less than 500 mg/day, or less than 400 mg/day. In some aspects, the effective amount of the Cdc7 inhibitor monotherapy is less than 300 mg/day, less than 200 mg/day, less than 150 mg/day, less than 100 mg/day, or less than 75 mg/day. In some aspects, the effective amount of the Cdc7 inhibitor monotherapy is less than 324 mg/day. In some aspects, the effective amount of the SRA141 monotherapy is 324 mg/day.
  • MTD maximum tolerated dose
  • the effective amount of the Cdc7 inhibitor monotherapy is at least 10 mg/day. In some aspects, the effective amount of the Cdc7 inhibitor monotherapy is between 10-400 mg/day. In some aspects, the effective amount of the Cdc7 inhibitor monotherapy is between 10-324 mg/day. In some aspects, the effective amount of the Cdc7 inhibitor monotherapy is between 40-400 mg/day. In some aspects, the effective amount of the Cdc7 inhibitor monotherapy is at least 10 mg/day, at least 20 mg/day, at least 40 mg/day, at least 80 mg/day, at least 160 mg/day, or at least 320 mg/day.
  • the effective amount of the Cdc7 inhibitor monotherapy is at least 15 mg/day, at least 25 mg/day, at least 50 mg/day, at least 75 mg/day, at least 100 mg/day, at least 150 mg/day, at least 200 mg/day, at least 250 mg/day, at least 300 mg/day, or at least 350 mg/day.
  • the effective amount of the SRA141 monotherapy is 10 mg/day, 20 mg/day, 40 mg/day, 80 mg/day, 160 mg/day, 320 mg/day, 325 mg/day, 350 mg/day, or 400 mg/day.
  • the effective amount of the SRA141 monotherapy is 15 mg/day, 25 mg/day, 50 mg/day, 75 mg/day, 100 mg/day, 150 mg/day, 200 mg/day, 250 mg/day, 300 mg/day, or 350 mg/day.
  • the methods of the invention include a combination therapy administering an effective amount of a Cdc7 inhibitor and coadministering a second effective amount of one or more additional treatments.
  • Additional treatments include, but are not limited to, administering a
  • chemotherapeutic agent administering an antibody or antibody fragment (such as an immune checkpoint inhibitors), administering a radiation treatment, and administering a combination thereof.
  • Additional treatments also include, but are not limited to, administering an anti neoplastic agent, such as a DNA polymerase inhibitor, a receptor tyrosine kinase inhibitor, a mammalian target of rapamycin (mTOR) pathway inhibitor.
  • Anti -neoplastic agents can also include a mitogen activated protein kinase (MAPK) pathway inhibitor, such as Trametinib.
  • Anti -neoplastic agents can also include a retinoid pathway regulator, such as is the RXR agonist Bexarotene, and the RAR agonist Tretinoin.
  • Anti -neoplastic agents can also include an apoptosis regulator, such as comprises an apoptosis inducer, including, but not limited, to an anti-Bcl-2 agent (e.g, ABT-199).
  • Anti-neoplastic agents can also include
  • PI3K phosphatidylinositol-4,5-bisphosphate 3 kinase pathway inhibitors
  • Anti-neoplastic agents can also include PARP inhibitors, such as BMN673.
  • Anti -neoplastic agents can also include ATM kinase inhibitors, such as KU-60019.
  • Anti neoplastic agents can also include Aurora B kinase inhibitors, such as Barasertib.
  • Anti neoplastic agents can also include tyrosine threonine kinase (TTK) inhibitors, such as an inhibitor of monopolar spindle 1 kinase (Mpsl) (e.g, CFI-402257).
  • Anti-neoplastic agents can also include inhibitors of epidermal growth factor (EGF), such as Erlotinib.
  • the anti-neoplastic agent is gemcitabine. In some embodiments that anti neoplastic agent is not gemcitabine.
  • Additional treatments can also include a combination of additional treatments, such as a combination of anti -neoplastic agents.
  • the effective amount of a Cdc7 inhibitor is administered to a subject as a combination therapy with a second effective amount of an additional treatment.
  • the second effective amount is an amount from about
  • the second effective amount of the additional treatment is 0.001, 0.005, 0.010, 0.020, 0.050, 0.1, 0.2, 0.5, 1.0, 2.0, 5.0, 10.0 or
  • the second effective amount of the additional treatment is between 10-2000 mg/m 2 /day. In some embodiments the second effective amount of the additional treatment is between 50-1250 mg/m 2 /day. In some embodiments the second effective amount of the additional treatment is 50 mg/m 2 /day, 100 mg/m 2 /day, 150 mg/m 2 /day, 200 mg/m 2 /day, 300 mg/m 2 /day, 400 mg/m 2 /day, 500 mg/m 2 /day, 600 mg/m 2 /day, 700 mg/m 2 /day, 800 mg/m 2 /day, 900 mg/m 2 /day, 1000 mg/m 2 /day, 1100 mg/m 2 /day, or 1250 mg/m 2 /day.
  • Coadministered encompasses methods where a Cdc7 inhibitor and the additional treatment are given simultaneously, where a Cdc7 inhibitor and the additional treatment are given sequentially, and where either one of, or both of, a Cdc7 inhibitor and the additional treatment are given intermittently or continuously, or any combination of: simultaneously, sequentially, intermittently and/or continuously.
  • intermittent administration is not necessarily the same as sequential because intermittent also includes a first administration of an agent and then another administration later in time of that very same agent.
  • intermittent administration also encompasses sequential administration in some aspects because intermittent
  • administration does include interruption of the first administration of an agent with an administration of a different agent before the first agent is administered again. Further, the skilled artisan will also know that continuous administration can be accomplished by a number of routes including intravenous drip or feeding tubes, etc.
  • the term“coadministered” encompasses any and all methods where the individual administration of a Cdc7 inhibitor and the individual administration of the additional treatment to a subject overlap during any timeframe.
  • the present disclosure provides for methods where either one of, or both of, or any combination thereof, a Cdc7 inhibitor and/or an additional treatment are administered by a route selected from the group consisting of: intravenous, subcutaneous, cutaneous, oral, intramuscular, and intraperitoneal. In some aspects, the present disclosure provides for methods where either one of, or both of, or any combination thereof, a Cdc7 inhibitor and/or a additional treatment are administered intravenously. In some aspects, the present disclosure provides for methods where either one of, or both of, or any combination thereof, a Cdc7 inhibitor and/or an additional treatment are administered orally.
  • the unit dose forms of the present disclosure may be administered in the same or different physicals forms, i.e., orally via capsules or tablets and/or by liquid orally or via intravenous infusion, and so on.
  • the unit dose forms for each administration may differ by the particular route of administration.
  • Several various dosage forms may exist for either one of, or both of, a Cdc7 inhibitor and an additional treatment. Because different medical conditions can warrant different routes of administration, the same components of a combination of a Cdc7 inhibitor and an additional treatment described herein may be exactly alike in composition and physical form and yet may need to be given in differing ways and perhaps at differing times to alleviate the condition.
  • a condition such as persistent nausea, especially with vomiting, can make it difficult to use an oral dosage form, and in such a case, it may be necessary to administer another unit dose form, perhaps even one identical to other dosage forms used previously or afterward, with an inhalation, buccal, sublingual, or suppository route instead or as well.
  • the specific dosage form may be a requirement for certain combinations of a Cdc7 inhibitor and an additional treatment, as there may be issues with various factors like chemical stability or pharmacokinetics.
  • the frequency of administration of a Cdc7 inhibitor or the additional treatment to a subject includes, but is not limited to, Qld, Q2d, Q3d, Q4d, Q5d, Q6d, Q7d, Q8d, Q9d, QlOd, Ql4d, Q2ld, Q28d, Q30d, Q90d, Ql20d, Q240d, or Q365d.
  • QnD or qnd refers to drug administration once every“n” days.
  • QD refers to once every day or once daily dosing
  • Q2D refers to a dosing once every two days
  • Q7D refers to a dosing once every 7 days or once a week
  • Q5D refers to dosing once every 5 days
  • a Cdc7 inhibitor and the additional treatment are administered on different schedules.
  • the frequency of administration of a Cdc7 inhibitor or the additional treatment to a subject includes, but is not limited to: 5 days of dosing followed by 2 days of non-dosing each week; 1 week of daily dosing followed by 1, 2, or 3 weeks of non-dosing; 2 or 3 weeks of daily dosing followed by 1, or 2 weeks of non-dosing; twice daily dosing; or dosing on days 2 and 3 of a weekly cycle.
  • SRA141 and the additional treatment are administered on different schedules.
  • the present disclosure provides for methods where either one of or both of or any combination thereof a Cdc7 inhibitor and/or the additional treatment are
  • the present disclosure provides for methods comprising administering either one of, or both of, or any combinations thereof, a Cdc7 inhibitor or the additional treatment, to a subject with at least ten (10) minutes, fifteen (15) minutes, twenty (20) minutes, thirty (30) minutes, forty (40) minutes, sixty (60) minutes, two (2) hours, three (3) hour, four (4) hours, six (6) hours, eight (8) hours, ten (10) hours, twelve (12) hours, fourteen (14) hours, eighteen (18) hours, twenty -four (24) hours, thirty-six (36) hours, forty-eight (48) hours, three (3) days, four (4) days, five (5) days, six (6) days, seven (7) days, eight (8) days, nine (9) days, ten (10) days, eleven (11) days, twelve (12) days, thirteen (13) days, fourteen (14) days, three (3) weeks, or four (4) weeks, delay between administrations.
  • the administration with a delay follows a pattern where one of or both of or any combination thereof a Cdc7 inhibitor and/or the additional treatment are administered continuously for a given period of time from about ten (10) minutes to about three hundred and sixty five (365) days and then is not administered for a given period of time from about ten (10) minutes to about thirty (30) days.
  • the present disclosure provides for methods where either one of or any combination of a Cdc7 inhibitor and/or the additional treatment are administered intermittently while the other is given continuously.
  • the present disclosure provides for methods where the combination of the effective amount of a Cdc7 inhibitor is administered sequentially with the second effective amount of an additional treatment.
  • the present disclosure provides for methods where a Cdc7 inhibitor and the additional treatment are administered simultaneously.
  • the present disclosure provides for methods where the combination of the effective amount of th Cdc7 inhibitor is administered sequentially with the second effective amount of an additional treatment.
  • the combination is also said to be“coadministered” since the term includes any and all methods where the subject is exposed to both components in the combination.
  • such aspects are not limited to the combination being given just in one formulation or composition. It may be that certain concentrations of a Cdc7 inhibitor and the additional treatment are more advantageous to deliver at certain intervals and as such, the effective amount of Cdc7 inhibitor and the second effective amount of the additional treatment may change according to the formulation being administered.
  • the present disclosure provides for methods wherein a Cdc7 inhibitor and the additional treatment are administered simultaneously or sequentially. In some aspects, the present disclosure provides for methods where the effective amount of Cdc7 inhibitor is administered sequentially after the second effective amount of the additional treatment. In some aspects, the present disclosure provides for methods where the second effective amount of the additional treatment is administered sequentially after the effective amount of Cdc7 inhibitor. [0099] In some aspects, the present disclosure provides for methods where the combination is administered in one formulation. In some aspects, the present disclosure provides for methods where the combination is administered in two (2) compositions where the effective amount of Cdc7 inhibitor is administered in a separate formulation from the formulation of the second effective amount of the additional treatment.
  • the present disclosure provides for methods where the effective amount of Cdc7 inhibitor is administered sequentially after the second effective amount of the additional treatment. In some aspects, the present disclosure provides for methods where the second effective amount of the additional treatment is administered sequentially after the effective amount of Cdc7 inhibitor. In some aspects, the Cdc7 inhibitor and the additional treatment are administered; and subsequently both a Cdc7 inhibitor and the additional treatment are administered intermittently for at least twenty-four (24) hours. In some aspects, SRA141 and the additional treatment are administered on a non-overlapping every other day schedule. In some aspects, the additional treatment is administered on day 1, and a Cdc7 inhibitor is administered on days 2 and 3 of a weekly schedule.
  • the present disclosure provides for methods where the effective amount of Cdc7 inhibitor is administered no less than four (4) hours after the second effective amount of the additional treatment.
  • the present disclosure provides for methods where the effective amount of Cdc7 inhibitor is administered no less than ten (10) minutes, no less than fifteen (15) minutes, no less than twenty (20) minutes, no less than thirty (30) minutes, no less than forty (40) minutes, no less than sixty (60) minutes, no less than one (1) hour, no less than two (2) hours, no less than four (4) hours, no less than six (6) hours, no less than eight (8) hours, no less than ten (10) hours, no less than twelve (12) hours, no less than twenty four (24) hours, no less than two (2) days, no less than four (4) days, no less than six
  • the present disclosure provides for methods where the second effective amount of the additional treatment is administered no less than ten (10) minutes, no less than fifteen (15) minutes, no less than twenty (20) minutes, no less than thirty (30) minutes, no less than forty (40) minutes, no less than sixty (60) minutes, no less than one (1) hour, no less than two (2) hours, no less than four (4) hours, no less than six (6) hours, no less than eight (8) hours, no less than ten (10) hours, no less than twelve (12) hours, no less than twenty four (24) hours, no less than two (2) days, no less than four (4) days, no less than six (6) days, no less than eight (8) days, no less than ten (10) days, no less than twelve (12) days, no less than fourteen (14) days, no less than twenty one (21) days, or no less than thirty (30) days after the effective amount of a Cdc7 inhibitor.
  • Methods are disclosed for the treatment of cancer. Accordingly, the present disclosure provides for methods of treating a cancer in a subject in need thereof (i.e., a subject with cancer or a subject suffering from cancer), the method comprising administering an effective amount of a Cdc7 inhibitor to the subject with cancer, wherein the cancer is melanoma, uterine cancer, thyroid cancer, blood cancer, bladder cancer, breast cancer, cervical cancer, colorectal cancer (CRC), gastric cancer, endometrial cancer,
  • the cancer is a blood cancer, such as acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), chronic eosinophilic leukemia, and diffuse large B-cell lymphoma (DLBCL).
  • AML acute myeloid leukemia
  • CML chronic myelogenous leukemia
  • DLBCL diffuse large B-cell lymphoma
  • the cancer is AML.
  • the cancer is metastatic colorectal cancer (mCRC).
  • mCRC metastatic colorectal cancer
  • the mCRC can be categorized as not having a microsatellite instability high (MSI-H) status.
  • MSI-H microsatellite instability high
  • microsatellite instability refers to tumors that are characterized by having genomic instability in specific repetitive DNA sequences ( e.g ., short tandem repeats or simple sequence repeats).
  • Methods for detection of microsatellite instability include any methods known in the art including, but not limited to, those methods described in Wang M. et al ., Screening for Microsatellite Instability in Colorectal Cancer and Lynch Syndrome - A Mini Review; N A J Med Sci. 20l6;9(l):5— 11.
  • microsatellite instability is characterized by detection of mononucleotide repeat markers (e.g., BAT-25, BAT-26, NR-21, NR-24 and MONO-27, also known as the Promega MSI Multiplex System).
  • microsatellite instability is characterized by detection of quasimonomorphic markers and dinucleotide repeat markers (e.g, BAT25, BAT26, D2S123, D5S346, and D17S250, also known as the Bathesda Panel).
  • MSI can be classified as MSI-high (MSI-H), MSI-Low (MSI-L), and MS-Stable (MSS) using where instability in two or more markers is classified as MSI-H, instability in one only as MSI-L, and no observed instability in the five markers as MSS.
  • MSI can be also classified as MSI-H if greater than 30% of markers tested demonstrate instability (MSI-H), MSI-L if 10-30% of markers tested demonstrate instability, and MSS if less than 10% of markers tested demonstrate instability.
  • MSI status can be tested using 5, 6, 7, 8, 9, 10, or greater than 10 markers.
  • the mCRC can also have an unknown MSI status, i.e., the tumor is not known to have an MSI-H status.
  • microsatellite instability in certain embodiments also refers to tumors that are characterized by having one or more repetitive DNA sequences known in the art to be correlated with loss of mismatch repair (MMR) compared to at least one reference sample.
  • MMR loss of mismatch repair
  • the present disclosure provides for methods of inhibiting the growth of a tumor wherein the tumor is from a cancer that is melanoma, uterine cancer, thyroid cancer, blood cancer, bladder cancer, breast cancer, cervical cancer, colorectal cancer (CRC), gastric cancer, endometrial cancer, cholangiocarcinoma, hepatocellular cancer, leukemia, lymphoma, myeloma, non-small cell lung cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain cancer, sarcoma, small cell lung cancer, neuroblastoma, or head and neck cancer.
  • a cancer that is melanoma, uterine cancer, thyroid cancer, blood cancer, bladder cancer, breast cancer, cervical cancer, colorectal cancer (CRC), gastric cancer, endometrial cancer, cholangiocarcinoma, hepatocellular cancer, leukemia, lymphoma, myeloma, non-small cell lung cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain
  • the tumor can be from a blood cancer, such as acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), chronic eosinophilic leukemia, and diffuse large B- cell lymphoma (DLBCL).
  • AML acute myeloid leukemia
  • CML chronic myelogenous leukemia
  • DLBCL diffuse large B- cell lymphoma
  • the tumor can be from an AML cancer.
  • the tumor can be from a metastatic colorectal cancer (mCRC).
  • mCRC metastatic colorectal cancer
  • the tumor can be from a metastatic colorectal cancer (mCRC) not categorized as having an MSI-H status.
  • the tumor can have a Wnt signaling pathway mutation, such as an Adenomatous polyposis coli (APC) gene mutation, a FAT1 mutation, a FAT4 mutation, or combinations thereof.
  • the tumor can have a FBXW7 mutation.
  • the tumor associated with the cancer comprises a phenotype that is prospectively screened by using either an archival or a fresh tumor biopsy and examining the pattern of chromosome separation by employing histochemical staining of tumor sections with hematoxylin and eosin (or similar stains) and determining whether the tumor exhibits aberrant mitosis (e.g ., lagging chromosomes, anaphase bridges, multipolar spindles), such as upon examination under a microscope either manually or by automated methods.
  • a pharmacodynamic marker of aberrant mitosis is determined by histochemical staining using post-treatment tumor biopsies after Cdc7 inhibitor administration. Post treatment biopsies can be obtained several days after Cdc7 inhibitor administration, such as the same day biopsies are obtained for pMCM2 monitoring (see below).
  • the present disclosure provides for administering an effective amount of a Cdc7 inhibitor to a subject that is in need thereof.
  • the present disclosure provides for administering an effective amount of Cdc7 inhibitor in a combination therapy with a additional treatment to a subject that is in need thereof.
  • the tumor from a subject is screened with genetic testing and/sequencing prior to administration.
  • the tumor from a subject is screened with genetic testing and/sequencing after administration.
  • the tumor from a subject is screened both after and before administration.
  • healthy cells from the subject are screened with genetic testing and/sequencing prior to administration, after administration, or both.
  • the tumor from a subject is screened with other biological tests or assays to determine the level of expression of certain biomarkers, such as microsatellites or repetitive DNA. In some aspects, the tumor from a subject is screened with both genetic testing and/sequencing and other biomarker tests or assays.
  • the present disclosure provides for methods wherein the subject is a mammal. In some aspects, the present disclosure provides for methods wherein the subject is a primate.
  • the present disclosure provides for methods wherein the subject is a mouse.
  • the present disclosure provides for methods wherein the subject is a human.
  • the present disclosure provides for methods wherein the tumor is in a human suffering from cancer that is selected from the group consisting of: melanoma, uterine cancer, thyroid cancer, blood cancer, bladder cancer, breast cancer, cervical cancer, colorectal cancer (CRC), gastric cancer, endometrial cancer, cholangiocarcinoma, hepatocellular cancer, leukemia, lymphoma, myeloma, non-small cell lung cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain cancer, sarcoma, small cell lung cancer, neuroblastoma, or head and neck cancer.
  • cancer is selected from the group consisting of: melanoma, uterine cancer, thyroid cancer, blood cancer, bladder cancer, breast cancer, cervical cancer, colorectal cancer (CRC), gastric cancer, endometrial cancer, cholangiocarcinoma, hepatocellular cancer, leukemia, lymphoma, myeloma, non-small cell lung cancer, ovarian cancer, prostate cancer
  • the cancer is a blood cancer, such as acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), chronic eosinophilic leukemia, and diffuse large B-cell lymphoma (DLBCL).
  • AML acute myeloid leukemia
  • CML chronic myelogenous leukemia
  • DLBCL diffuse large B-cell lymphoma
  • the cancer is AML.
  • the cancer is metastatic colorectal cancer (mCRC). The mCRC can be categorized as not having a MSI-H status.
  • subjects have tumors that harbor genomic alterations expected to confer sensitivity to Cdc7 inhibition, such as not having a MSI-H status, having a
  • CIN chromosome instability
  • MSI microsatellite unstable
  • Wnt signaling pathway mutation e.g., an Adenomatous polyposis coli (APC) gene mutation, a FAT1 mutation, a FAT4 mutation, or combinations thereof
  • the methods described herein for the treatment of cancer can result in a plasma Cmax greater than 400 ng/mL, greater than 600 ng/mL, greater than 1000 ng/mL, or greater than 1400 ng/mL of the Cdc7 inhibitor.
  • the methods described herein for the treatment of cancer can result in a in a plasma Cmax of at least 500 ng/mL, at least 600 ng/mL, at least 700 ng/mL, at least 800 ng/mL, at least 900 ng/mL, at least 1000 ng/mL, at least 1100 ng/mL, at least 1100 ng/mL, at least 1200 ng/mL, at least 1300 ng/mL, at least 1400 ng/mL, or at least 1500 ng/mL of the Cdc7 inhibitor.
  • the methods described herein for the treatment of cancer can result in a in an AUCiast greater than 3000 ng.h/mL, greater than 5800 ng.h/mL, greater than 11900 ng.h/mL, or greater than 16400 ng.h/mL of the Cdc7 inhibitor.
  • the methods described herein for the treatment of cancer can result in a in an AUCiast of at least 5000 ng.h/mL, at least 6000 ng.h/mL, at least 7000 ng.h/mL, at least 8000 ng.h/mL, at least 9000 ng.h/mL, at least 10000 ng.h/mL, at least 11000 ng.h/mL, at least 12000 ng.h/mL, at least 13000 ng.h/mL, at least 14000 ng.h/mL, at least 15000 ng.h/mL, or at least 16000 ng.h/mL of the Cdc7 inhibitor.
  • the methods described herein for the treatment of cancer can result in a in an intra-tumoral concentration of greater than 500 ng/mL, greater than 600 ng/mL, greater than 900 ng/mL, or greater than 1300 ng/mL of the Cdc7 inhibitor.
  • the methods described herein for the treatment of cancer can result in a in an intra-tumoral concentration of at least 500 ng/mL, at least 600 ng/mL, at least 700 ng/mL, at least 800 ng/mL, at least 900 ng/mL, at least 1000 ng/mL, at least 1100 ng/mL, at least 1100 ng/mL, at least 1200 ng/mL, at least 1300 ng/mL, at least 1400 ng/mL, or at least 1500 ng/mL of the Cdc7 inhibitor.
  • the methods described herein result in in vivo kinase inhibition, e.g., inhibition of MCM2 phosphorylation.
  • MCM2 mini-chromosome maintenance proteins
  • pre-RC pre-replication complex
  • the in vivo inhibition of MCM2 phosphorylation can be in a tumor associated with the cancer.
  • the in vivo inhibition of MCM2 phosphorylation in a tumor can be at least 50% relative to an untreated tumor sample.
  • the in vivo inhibition of MCM2 phosphorylation in a tumor can be at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% relative to an untreated tumor sample.
  • the in vivo inhibition of MCM2 phosphorylation can be in skin of a subject. In vivo inhibition of MCM2 phosphorylation can be measured by assays known to those skilled in the art including, but not limited to, Western blot analysis,
  • IHC immunohistochemistry
  • LC/MS liquid chromatography-mass spectrometry
  • LC/MS/MS LC/MS/MS
  • the methods described herein for the treatment of cancer may not result in (i.e., may avoid) specific inhibition of an off target kinase in the subject, wherein the off target kinase is selected from the group consisting of: WEE1, CDK7, CDK8, CDK9, and LATS2.
  • the methods described herein for the treatment of cancer can result in less than 90% inhibition of an off-target kinase.
  • the present disclosure is directed to methods using an effective amount of a Cdc7 inhibitor, e.g., SRA141, to inhibit the progression of, reduce the size in aggregate of aggregation of, reduce the volume of, reduce the diameter of, and/or otherwise inhibit the growth of a tumor.
  • a Cdc7 inhibitor e.g., SRA141
  • the methods described herein for the treatment of cancer can result in growth inhibition of a tumor associated with a cancer.
  • the methods described herein for the treatment of cancer can result in cytotoxicity of a tumor associated with a cancer.
  • the methods described herein for the treatment of cancer can result in growth inhibition and cytotoxicity of a tumor associated with a cancer.
  • the methods described herein for the treatment of cancer can result in growth inhibition or cytotoxicity of a tumor associated with a cancer.
  • the methods described herein for the treatment of cancer can result in a regression of a tumor associated with the cancer, including a complete regression or a partial regression. Also provided herein are methods of treating the underlying disease, e.g., cancer, and extending the survival of the subject.
  • a method of inhibiting the growth of a tumor in a subject in need thereof comprising administering to the subject an effective amount of a Cdc7 inhibitor.
  • the disclosure provides for a method of administering to the subject an effective amount of Cdc7 inhibitor to inhibit growth of a tumor, wherein tumor growth is reduced by at least 47%.
  • the disclosure provides for a method of administering to the subject an effective amount of Cdc7 inhibitor to inhibit growth of a tumor, wherein tumor growth is reduced by at least 93%.
  • the disclosure provides for a method of administering to the subject an effective amount of Cdc7 inhibitor to inhibit growth of a tumor, wherein tumor growth is reduced by 1%, 2%,
  • the disclosure provides for a method of administering to the subject an effective amount of Cdc7 inhibitor to inhibit growth of a tumor, wherein tumor growth is reduced by 1%, 2%, 3%, 4%, 5%, 6%,
  • the disclosure provides for a method of administering to the subject an effective amount of Cdc7 inhibitor to inhibit the growth of a tumor, wherein tumor growth is reduced by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, 30%, 32%, 34%,
  • the present disclosure is also directed to methods using an effective amount of the Cdc7 inhibitor and a second effective amount of an additional treatment to inhibit the progression of, reduce the size in aggregation of, reduce the volume of, reduce the diameter of, and/or otherwise inhibit the growth of a tumor. Also provided herein are methods of treating the underlying disease, e.g., cancer, and extending the survival of the subject.
  • a method of inhibiting the growth of a tumor in a subject in need thereof comprising administering to the subject an effective amount of Cdc7 inhibitor and a second effective amount of an additional treatment.
  • the disclosure provides for a method of administering to the subject an effective amount of Cdc7 inhibitor and a second effective amount of an additional treatment to inhibit growth of a tumor, wherein tumor growth is reduced by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, 30%, 32%, 34%, 36%, 38%, 40%, 42%, 44%, 46%, 48%, 50%, 52%, 54%, 56%, 58%, 60%, 62%, 64%, 66%, 68%, 70%,
  • the disclosure provides for a method of administering to the subject an effective amount of Cdc7 inhibitor and a second effective amount of an additional treatment to inhibit growth of a tumor, wherein tumor growth is reduced by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, 30%, 32%, 34%, 36%, 38%, 40%, 42%, 44%, 46%, 48%, 50%, 52%, 54%,
  • the disclosure provides for a method of administering to the subject an effective amount of Cdc7 inhibitor and a second effective amount of an additional treatment to inhibit growth of a tumor, wherein tumor growth is reduced by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, 30%, 32%, 34%, 36%, 38%, 40%, 42%, 44%, 46%, 48%, 50%, 52%, 54%, 56%, 58%, 60%, 62%, 64%, 66%,
  • kits for treating cancer e.g., inhibiting the growth of a tumor in a subject and/or inhibiting growth of a tumor cell, wherein the method results in a clinically relevant endpoint.
  • Tumor growth occurs when one or more biological cells grow and divide much more rapidly resulting in an increase in the number of cells in comparison to the normal and healthy process of cells division. This phenomenon is an indication that the cells are in a disease state such as cancer or pre-cancer. Moreover, tumor growth oftentimes comes about in discrete stages prior to the agglomerated cells forming a tumor.
  • the overall metabolic activity inside a cell can be measured via a labeled biological product.
  • a labeled biological product for example, there are several commercially available dyes (e.g., MTT) that can penetrate the cell and interact with certain enzymes and other factors to produce a detectable product.
  • cellular biomarkers can be measured in a cell.
  • a BrdU assay can incorporate a thymidine derivative into cellular DNA and be detected with an antibody.
  • Proliferating cell nuclear antigen (PCNA) is another such biomarker for detection.
  • PCNA Proliferating cell nuclear antigen
  • Besides tagging techniques the skilled artisan can also use for example, microscopy or flow cytometry to allow for cell counts.
  • cellular replication is measured by a clinical endpoint that includes: a quality of life (QOL) score, duration of response (DOR, clinical benefit rate (CBR), patient reported outcomes (PRO), an objective response rate (ORR) score, a disease-free survival (DFS) or progression-free survival (PFS) , a time to progression (TTP), an Overall Survival, a time-to-treatment failure (TTF), RECIST criteria, Partial Response (PR), Stable Disease (SD), Progressive Disease (PD) and/or a Complete Response (CR).
  • QOL quality of life
  • DOR clinical benefit rate
  • PRO patient reported outcomes
  • ORR objective response rate
  • DFS disease-free survival
  • PFS progression-free survival
  • TTP time to progression
  • TTF time-to-treatment failure
  • RECIST criteria Partial Response
  • PR Partial Response
  • SD Stable Disease
  • PD Progressive Disease
  • CR Complete Response
  • CR is disappearance of all target lesions wherein any
  • pathological lymph nodes are reduced in short axis to ⁇ 10 mm.
  • PR is at least a 30% decrease in the sum of diameters of target lesions in reference to the baseline sum diameters.
  • PD is at least a 20% increase in the sum of diameters of target lesions, taking as reference the smallest sum (including the baseline sum if it is the smallest).
  • the sum may demonstrate an absolute increase of at least 5 mm in addition to the relative increase of 20%.
  • SD is neither sufficient shrinkage to qualify of PR nor sufficient increase to qualify for PD, taking as reference the smallest sum diameters.
  • the present disclosure provides methods wherein the growth of the tumor is reduced at least about 5, 10, 20, 30, 40, 50, 60, 80, 90, 95, 97, 99, or 99.9% after administration of the effective amount of Cdc7 inhibitor.
  • the present disclosure provides methods wherein the % reduction is calculated based on measurement s) of one or more clinical endpoints.
  • the present disclosure provides methods wherein the growth of the tumor is reduced as measured by an increase or a decrease in total cell count in a MTT assay, or by change in genetic profile as measured by a ctDNA assay, by no more than or at least 5, 10, 20, 30, 40, 50, 60, 80, 90, 95, 97, 99, or 99.9% after administration of the effective amount of a Cdc7 inhibitor.
  • the present disclosure provides methods wherein the growth of the tumor is reduced at least 5, 10, 20, 30, 40, 50, 60, 80, 90, 95, 97, 99, or 99.9% after administration of the effective amount of Cdc7 inhibitor.
  • the present disclosure provides methods wherein the growth of the tumor is reduced as measured by an increase or a decrease in total cell count in a MTT assay, or by change in genetic profile as measured by a ctDNA assay, by at least 5, 10, 20, 30, 40, 50, 60, 80, 90, 95, 97, 99, or 99.9% after administration of the effective amount of Cdc7 inhibitor.
  • the present disclosure provides methods wherein administration results in an ICso value below 10 mM and/or a GIso value below 1 mM. In some aspects, the present disclosure provides methods wherein administration results in an ICso value below 10 pM and/or a GIso value below 1 pM at twenty-four (24) hours after administration. In some aspects, the present disclosure provides methods wherein administration results in an ICso value below 10 pM and/or a GIso value below 1 pM at forty-eight (48) hours after administration.
  • the present disclosure provides methods wherein the
  • administration results in an AUC of at least 1, 10, 25, 50, 100, 200, 400, 600, 800, or 1000.
  • the present disclosure provides methods wherein the
  • administration results in an ICso value of no more than 0.001, 0.005, 0.01, 0.05, 0.1, 1, 3, 5, 10, 20, 40, 50, 60, 80, 90, 100, 200, 250, 300, 350, or 400 pM.
  • the present disclosure provides methods wherein the
  • administration results in an ECso value of at least 0.01, 0.1, 1, 3, 5, 10, 20, 40, 50, 60, 80, 90, 100, 200, 250, 300, 350, or 400 pM.
  • the present disclosure provides methods wherein the
  • TI therapeutic index
  • the present disclosure provides methods wherein the
  • administration results in an GIso value of at least 0.1 pM, 0.3 pM, 0.5 pM, 0.7 pM, 1 pM, 1.5 pM, 2 pM, 2.5 pM, 3 pM, 4 pM, 5 pM, or 10 pM.
  • the present disclosure provides methods wherein the
  • Tumor growth can be expressed in terms of total tumor volume.
  • formulas both generally speaking and specific to certain tumor models, that the skilled artisan can use to calculate tumor volume based upon the assumption that solid tumors are more or less spherical.
  • the skilled artisan can use experimental tools such as: ultrasound imaging, manual or digital calipers, ultrasonography, computed tomographic (CT), microCT, 18 F-FDG-microPET, or magnetic resonance imaging (MRI) to measure tumor volume.
  • CT computed tomographic
  • microCT microCT
  • MRI magnetic resonance imaging
  • the present disclosure provides methods wherein administration results in a reduction in tumor volume of at least 5, 10, 20, 30, 40, 50, 60, 80, 90, 95, 97, 99 or 99.9% after administration of the effective amount of SRA141. In some aspects, the present disclosure provides methods wherein administration results in a reduction in tumor size, as measured by medical imaging techniques, of at least 5, 10, 20, 30, 40, 50, 60, 80, 90, 95, 97, 99 or 99.9% after administration of the effective amount of SRA141.
  • the present disclosure provides methods wherein administration results in method where administration results in a reduction in tumor volume of at least 5% after one (1), two (2), three (3), four (4), six (6), eight (8), twelve (12), sixteen (16), twenty (20), twenty four (24), thirty six (36), or fifty two (52) weeks.
  • the overall tumor burden at baseline may be estimated and used as a comparator for subsequent measurements.
  • Measurable disease may be defined by the presence of at least one measurable lesion.
  • the present disclosure provides methods wherein when more than one measurable lesion is present at baseline all lesions up to a maximum of five lesions total (and a maximum of two lesions per organ) representative of all involved organs are identified as target lesions and are recorded and measured at baseline.
  • target lesions are selected on the basis of their size (lesions with the longest diameter) and are
  • target lesions are those that lend themselves to reproducible repeated measurements.
  • the largest lesion does not lend itself to reproducible measurement in which circumstance the next largest lesion which can be measured reproducibly is selected, as exemplified in Fig. 3 Eisenhauer, et al. (2009).
  • Pathological lymph nodes may be defined as measurable and in some cases identified as target lesions when the node has a short axis of 3l5 mm by CT scan. In some embodiments the short axis of the nodes contributes to the baseline sum. In some
  • the short axis of the node is the diameter used by radiologists to judge if a node is involved by solid tumor.
  • nodal size is reported as two dimensions in the plane in which the image is obtained (e.g ., the axial plane for a CT scan; the axial, sagital, or coronal plane for an MRI scan, depending on the plane of acquisition of the scan).
  • the smaller of the measures is the short axis.
  • pathological nodes with a short axis 3l0 mm but ⁇ 15 mm are considered non-target lesions.
  • nodes that have a short axis ⁇ 10 mm are considered non-pathological and are not recorded or followed.
  • a sum of the diameters (longest for non-nodal lesions, short axis for nodal lesions) for all target lesions is calculated and reported as the baseline sum diameters.
  • lymph nodes are to be included in the sum, then only the short axis is added into the sum.
  • the baseline sum diameters are used as reference to characterize objective tumor growth or regression.
  • all other lesions or sites of disease are identified as non-target lesions and are recorded at baseline. In some cases measurements are not required and the lesions are followed as‘present,’‘absent,’ or ‘unequivocal progression.’ In some cases, multiple non-target lesions involving the same organ may be recorded (e.g.,‘multiple enlarged pelvic lymph nodes’ or‘multiple liver metastases’).
  • the methods described herein include administration of an effective amount of a Cdc7 inhibitor, and optionally, one or more additional treatments, e.g., an antineoplastic agent.
  • An effective amount can readily be determined by routine experimentation, as can the most effective and convenient route of administration and the most appropriate formulation.
  • Various formulations and drug delivery systems are available in the art. See, e.g., Gennaro, A.R., ed. (1995) Remington’s Pharmaceutical Sciences, 18th ed., Mack Publishing Co.
  • An effective amount can be estimated initially using a variety of techniques well- known in the art. Initial doses used in animal studies may be based on effective concentrations established in cell culture assays. Dosage ranges appropriate for human subjects can be determined, for example, using data obtained from animal studies and cell culture assays.
  • an effective amount or a therapeutically effective amount or dose of an agent refers to that amount of the agent that results in amelioration of symptoms or a prolongation of survival in a subject.
  • Toxicity and therapeutic efficacy of such molecules can be determined by standard pharmaceutical procedures in cell cultures or experimental animals and the minimum effective concentration (MEC) or minimum effective dose (MED).
  • MEC minimum effective concentration
  • MED minimum effective dose
  • the dose ratio of toxic to therapeutic effects is therapeutic index, which can be expressed as the ratio of MTD or highest non-severely toxic dose (HNSTD) to MED. Agents that exhibit high therapeutic indices are preferred.
  • the effective amount or therapeutically effective amount is the amount of the compound or pharmaceutical composition that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician. Dosages particularly fall within a range of circulating concentrations that includes the EDso with little or no toxicity. Dosages may vary within this range depending upon the dosage form employed and/or the route of administration utilized. The exact formulation, route of administration, dosage, and dosage interval should be chosen according to methods known in the art, in view of the specifics of a subject’s condition.
  • Dosage amount and interval may be adjusted individually to provide plasma levels of the active moiety that are sufficient to achieve the desired effects; i.e., the minimal effective concentration (MEC).
  • MEC minimal effective concentration
  • the MEC will vary for each compound but can be estimated from, for example, in vitro data and animal experiments. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.
  • the amount of agent or composition administered may be dependent on a variety of factors, including the sex, age, and weight of the subject being treated, the severity of the affliction, the manner of administration, and the judgment of the prescribing physician.
  • a therapeutically effective amount can be the same or different than either one of, or both of, the effective amount of Cdc7 inhibitor and the second effective amount of the additional treatment. This is because the present disclosure provides that the methods, as described herein, are effective even where neither the effective amount of Cdc7 inhibitor nor the second effective amount of the additional treatment must be an amount that, alone, will ameliorate a symptom of a disease (e.g ., the amount of the Cdc7 inhibitor and/or the additional treatment may be considered a“sub-therapeutic” amount if administered as an individual therapy). However, the present disclosure does provide that a therapeutically effective amount of the combination must be provided, i.e., the combination does at least affect a treatment of a symptom of a disease.
  • a unit dose form is a term that is generally understood by the skilled artisan.
  • a unit dose forms is a pharmaceutical drug product that is marketed for a specific use.
  • the drug product includes the active ingredient(s) and any inactive components, most often in the form of pharmaceutically acceptable carriers or excipients. It is understood that multiple unit dose forms are distinct drug products. Accordingly, one unit dose form may be e.g., the combination of a Cdc7 inhibitor and an additional treatment of 250 mg at a certain ratio of each component, while another completely distinct unit dose form is e.g, the combination of Cdc7 inhibitor and an additional treatment of 750 mg at the same certain ratio of each component referred to above.
  • the effective amount of a Cdc7 inhibitor and the second effective amount of the additional treatment may both remain the same.
  • the unit dose form is distinct.
  • the effective amount is unique to the Cdc7 inhibitor, i.e., it is different than the second effective amount of the additional treatment.
  • the effective amount of Cdc7 inhibitor is an amount that is equivalent to a“therapeutically effective amount” or an amount that brings about a therapeutic and/or beneficial effect.
  • the effective amount of Cdc7 inhibitor is a“therapeutically effective amount”.
  • the second effective amount of the additional treatment is a“therapeutically effective amount”.
  • both the effective amount of Cdc7 inhibitor and second effective amount of the additional treatment are not a“therapeutically effective amount”.
  • the second effective amount is unique to the additional treatment, i.e., the second effective amount is a different amount for different additional treatments.
  • the Cdc7 inhibitor and the additional treatment combination is formulated in one (1) unit dose form.
  • the same unit dose form is
  • the Cdc7 inhibitor and the additional treatment combination is formulated in at least two (2) separately distinct unit dose forms.
  • the first effective amount is different in the first unit dose form than in the second unit dose form.
  • the effective amount of Cdc7 inhibitor is the same in the first unit dose form as it is in the second unit dose form.
  • the first unit dose form is the same as the second unit dose form. In some aspects, the first unit dose form is the same as the second and third unit dose forms. In some aspects, the first unit dose form is the same as the second, third, and fourth unit dose forms.
  • the present disclosure provides for methods of use of Cdc7 inhibitors, e.g., compounds that inhibit the activity of a Cdc7 kinase.
  • Cdc7 inhibitors e.g., compounds that inhibit the activity of a Cdc7 kinase. Examples are well known to one of skill in the art and include small molecules.
  • the Cdc7 inhibitor is a furanone derivative represented by the following formula (I):
  • A represents -COOR1 or a hydrogen atom
  • Rl represents a hydrogen atom, a hydrocarbon group optionally having a substituent, or a heterocyclic ring optionally having a substituent
  • R2 and R3 are the same or different and each represent a hydrogen atom, a hydrocarbon group optionally having a substituent, a phenyl group optionally having a substituent, a heterocyclic ring optionally having a substituent, a heterocyclic fused ring optionally having a substituent, or an amino group optionally having a substituent, or R2 and R3 optionally form a heterocyclic ring optionally having a substituent or a heterocyclic fused ring optionally having a substituent, together with the nitrogen atom bonded thereto; and R4 represents a hydrogen atom or a halogen atom, provided that when A represents -COOR1, R2 and R3 do not represent the amino group optionally having a substituent at the same time, and when A
  • examples of the hydrocarbon group optionally having a substituent include a) a linear or branched alkyl group having 1 to 6 carbon atoms (e.g., methyl, ethyl, isopropyl, tert-butyl, and hexyl), b) a linear or branched alkenyl group having 1 to 6 carbon atoms (e.g., vinyl, allyl, isopropenyl, and 2-butenyl), c) an alkynyl group having 2 to 6 carbon atoms (e.g., ethynyl, propargyl, and 2-butynyl), d) a cycloalkyl group having 3 to 8 carbon atoms (e.g., cyclopropyl, cyclopentyl, cyclohexyl, and cycloheptyl),e) a linear or branched alkyl group having 1 to 6 carbon atoms (e.g., methyl,
  • cycloalkenyl group having 3 to 8 carbon atoms e.g., cyclohexenyl and cycloheptenyl
  • an aralkyl group whose aryl moiety is aryl having 6 to 14 carbon atoms e.g., phenyl, naphthyl, and indenyl
  • alkylene moiety is a group in which one hydrogen atom has been removed from the alkyl group.
  • heterocyclic moiety of the heterocyclic ring optionally having a substituent examples include an alicyclic heterocyclic group and an aromatic heterocyclic group.
  • Examples of the alicyclic heterocyclic group include a 3- to 8-membered heterocyclic group containing at least one heteroatom selected from a nitrogen atom, a sulfur atom, and an oxygen atom. Specific examples thereof include pyrrolidinyl, piperidyl, piperazinyl, morpholinyl, and thiomorpholinyl.
  • Examples of the aromatic heterocyclic group include a 5- or 6-membered monocyclic aromatic heterocyclic group containing at least one heteroatom selected from a nitrogen atom, a sulfur atom, and an oxygen atom. Specific examples thereof include imidazolyl, pyrazolyl, thienyl, thiazolyl, and pyridyl.
  • heterocyclic fused ring moiety of the heterocyclic fused ring optionally having a substituent examples include a 3- to 8-membered ring-fused bicyclic heterocyclic group containing at least one heteroatom selected from a nitrogen atom, a sulfur atom, and an oxygen atom.
  • Specific examples thereof include benzothiophenyl, benzimidazolyl, indazolyl, benzoxazolyl, benzothiazolyl, indolyl, isoquinolyl, and phthalimide.
  • Examples of the amino group optionally having a substituent include an amino group having a linear, branched, or cyclic alkyl group having 1 to 6 carbon atoms, an aryl group, a heteroaryl group, or the like which is substituted or unsubstituted and include an amino group bonded to an alkyl group, an alkylamino group, an aryl group, a heteroaryl group, a heterocyclic group, a heterocyclic fused ring group, or the like which has one or two or more substituents or is unsubstituted.
  • The“one or two or more substituents” in these groups bonded to an amino group may be one or two or more identical or different arbitrary groups, unless otherwise specified.
  • Examples thereof include a halogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted alkenyl group, a substituted or unsubstituted alkynyl group, a substituted or unsubstituted alkoxy group, an amino group, a nitro group, a cyano group, a hydroxy group, a substituted or unsubstituted alkylamino group, a carbamoyl group, a carboxyl group, a formyl group, an acetyl group, and a benzoyl group.
  • The“substituent” in the hydrocarbon group optionally having a substituent, the heterocyclic ring optionally having a substituent, the phenyl group optionally having a substituent, or the heterocyclic fused ring optionally having a substituent may be one or two or more arbitrary types of substituents that are located at arbitrary chemically possible positions, unless otherwise specified. In the case of two or more substituents, these substituents may be the same as or different from each other.
  • Examples thereof include a halogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted alkoxy group, a substituted or unsubstituted amino group, a nitro group, a cyano group, a hydroxy group, a substituted or unsubstituted alkylamino group, a carbamoyl group, a carboxyl group, a formyl group, an acetyl group, and a benzoyl group.
  • Examples of the heterocyclic group in the heterocyclic ring optionally having a substituent or the heterocyclic fused ring optionally having a substituent which is formed by R2 and R3 together with the nitrogen atom bonded thereto include a 3- to 8-membered heterocyclic group containing at least one heteroatom selected from a nitrogen atom, a sulfur atom, and an oxygen atom or a 3 -to 8-membered ring-fused bicyclic alicyclic heterocyclic group containing at least one heteroatom selected from a nitrogen atom, a sulfur atom, and an oxygen atom and specifically include pyrrolidinyl, piperidyl, morpholinyl, thiomorpholinyl, azepinyl, diazepinyl, dihydroisoquinolyl, tetrahydroisoquinolyl, tetrahydroquinolyl, isoindolinyl, indolinyl, tetrahydrobenzazepinyl
  • benzoxazepinyl and benzothiazepinyl.
  • halogen atom include fluorine, chlorine, and bromine.
  • the compound of formula (I) can include isomers, for example, depending on the types of substituents. These isomers may be indicated by only one form of a chemical structure. Also encompassed are all structurally possible isomers (geometric isomers, optical isomers, tautomers, etc.), and mixtures thereof. Also encompassed are stereoisomers specifically represented by the formulas (Z)-(I) and (E)-(I), and mixtures thereof.
  • Examples of the pharmaceutically-acceptable salt of the compound of formula (I) include: salts of inorganic acids such as hydrochloric acid, sulfuric acid, carbonic acid, and phosphoric acid; and salts of organic acids such as formic acid, acetic acid, fumaric acid, maleic acid, methanesulfonic acid, and p-toluenesulfonic acid. Also encompasseed are: salts of alkali metals such as sodium and potassium; salts of alkaline earth metals such as magnesium and calcium; salts of organic amines such as lower alkylamine and lower alcohol amine; salts of basic amino acids such as lysine, arginine, and ornithine; and other salts such as ammonium salt.
  • salts of inorganic acids such as hydrochloric acid, sulfuric acid, carbonic acid, and phosphoric acid
  • organic acids such as formic acid, acetic acid, fumaric acid, maleic acid, methanesulfonic acid, and p
  • the compound of formula (I) and the pharmaceutically-acceptable salt thereof can be produced by a method described in, for example, US Patent No. 8,742,113.
  • the defined groups may vary under conditions of the implemented method or may be inappropriate for carrying out the method.
  • the compound (I) and the pharmaceutically acceptable salt thereof can be readily produced by a method usually used in organic synthetic chemistry, for example, approaches of the protection and deprotection of functional groups [T.W. Greene, Protective Groups in Organic Synthesis 3rd Edition, John Wiley & Sons, Inc., 1999] If necessary, the order of reaction steps including the introduction of substituents may be changed.
  • the compound of the formula (I) is preferably a compound wherein A is -COOR1 or a hydrogen atom and more preferably has the structure of the following compound (I-A), compound (I-B), compound (I-C), compound (I-D), or compound (I-E).
  • the compound (I- A) is a compound of Example 245 in the specification of ETS Patent No. 8,742,113; the compound (I-B) is a compound of Example 244 in the specification of ETS Patent No. 8,742, 113; the compound (I-C) is a compound of Example 351 in the specification of ETS Patent No. 8,742, 113; the compound (I-D) is a compound of Example 246 in ETS Patent No. 8,742, 113; and the compound (I-E) is a compound of Example 347 in US Patent No. 8,742, 113.
  • the Cdc7 inhibitor is compound (I-F): 2- (pyridin-4-yl)-l,5,6,7-tetrahydro-4H-pyrrolo[3,2-c]pyridinone (Merck KGaA, catalog No.: 217707-5MGCN, 217707)
  • the Cdc7 inhibitor and the additional treatment can each be formulated in pharmaceutical compositions.
  • These pharmaceutical compositions may comprise, in addition to the active compound(s), a pharmaceutically acceptable excipient, carrier, buffer, stabilizer or other materials well known to those skilled in the art. Such materials should be non-toxic and should not interfere with the efficacy of the active ingredient.
  • the precise nature of the carrier or other material can depend on the route of administration, e.g., oral, intravenous, cutaneous or subcutaneous, nasal, intramuscular, intraperitoneal routes.
  • compositions for oral administration can be in tablet, capsule, powder or liquid form.
  • a tablet can include a solid carrier such as gelatin.
  • compositions generally include a liquid carrier such as water, petroleum derivative, animal or vegetable oils, mineral oil or synthetic oil.
  • a liquid carrier such as water, petroleum derivative, animal or vegetable oils, mineral oil or synthetic oil.
  • Physiological saline solution, dextrose or other saccharide solution or glycols such as ethylene glycol, propylene glycol or polyethylene glycol can be included.
  • the active ingredient will be in the form of a parenterally acceptable aqueous solution which is pyrogen-free and has suitable pH, isotonicity and stability.
  • a parenterally acceptable aqueous solution which is pyrogen-free and has suitable pH, isotonicity and stability.
  • isotonic vehicles such as Sodium Chloride Injection, Ringer’s Injection, Lactated Ringer’s Injection.
  • Preservatives, stabilizers, buffers, antioxidants and/or other additives can be included, as required.
  • a composition can be administered alone or in combination with other treatments, either simultaneously or sequentially dependent upon the condition to be treated.
  • compositions will be administered as pharmaceutical compositions by any one of the following routes: oral, systemic (e.g., transdermal, intranasal or by suppository), or parenteral (e.g., intramuscular, intravenous or subcutaneous) administration.
  • routes e.g., oral, systemic (e.g., transdermal, intranasal or by suppository), or parenteral (e.g., intramuscular, intravenous or subcutaneous) administration.
  • parenteral e.g., intramuscular, intravenous or subcutaneous
  • the preferred manner of administration is oral using a convenient daily dosage regimen that can be adjusted according to the degree of affliction.
  • Compositions can take the form of tablets, pills, capsules, semisolids, powders, sustained release formulations, solutions, suspensions, elixirs, aerosols, or any other appropriate compositions.
  • Another preferred manner for administering compounds of the present technology is inhalation.
  • the choice of formulation depends on various factors such as the mode of drug administration and bioavailability of the drug substance.
  • the compound can be formulated as liquid solution, suspensions, aerosol propellants or dry powder and loaded into a suitable dispenser for administration.
  • suitable dispenser for administration There are several types of pharmaceutical inhalation devices-nebulizer inhalers, metered dose inhalers (MDI) and dry powder inhalers (DPI).
  • MDI metered dose inhalers
  • DPI dry powder inhalers
  • Nebulizer devices produce a stream of high velocity air that causes therapeutic agents (which are formulated in a liquid form) to spray as a mist that is carried into the subject’s respiratory tract.
  • MDI’s typically are formulation packaged with a compressed gas.
  • the device Upon actuation, the device discharges a measured amount of therapeutic agent by compressed gas, thus affording a reliable method of administering a set amount of agent.
  • DPI dispenses therapeutic agents in the form of a free flowing powder that can be dispersed in the subject’s inspiratory air-stream during breathing by the device.
  • therapeutic agent is formulated with an excipient such as lactose.
  • a measured amount of therapeutic agent is stored in a capsule form and is dispensed with each actuation.
  • compositions of the present technology can include one or more physiologically acceptable inactive ingredients that facilitate processing of active molecules into preparations for pharmaceutical use.
  • compositions are comprised of in general, a compound of the present technology in combination with at least one pharmaceutically acceptable excipient.
  • Acceptable excipients are non-toxic, aid administration, and do not adversely affect therapeutic benefit of the claimed compounds.
  • excipient may be any solid, liquid, semisolid or, in the case of an aerosol composition, gaseous excipient that is generally available to one of skill in the art.
  • Solid pharmaceutical excipients include starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, dried skim milk and the like.
  • Liquid and semisolid excipients may be selected from glycerol, propylene glycol, water, ethanol and various oils, including those of petroleum, animal, vegetable or synthetic origin, e.g., peanut oil, soybean oil, mineral oil, sesame oil, etc.
  • Preferred liquid carriers, particularly for injectable solutions include water, saline, aqueous dextrose, and glycols.
  • Compressed gases may be used to disperse a compound of the present technology in aerosol form.
  • Inert gases suitable for this purpose are nitrogen, carbon dioxide, etc.
  • Other suitable pharmaceutical excipients and their formulations are described in Remington's Pharmaceutical Sciences, edited by E. W. Martin (Mack Publishing Company, 18th ed.,
  • the pharmaceutical compositions include a
  • pharmaceutically acceptable salt refers to salts derived from a variety of organic and inorganic counter ions well known in the art that include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, and tetraalkylammonium, and when the molecule contains a basic functionality, salts of organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, and oxalate. Suitable salts include those described in Stahl and Wermuth (Eds.), Handbook of Pharmaceutical Salts Properties, Selection, and Lise; 2002.
  • compositions may, if desired, be presented in a pack or dispenser device containing one or more unit dosage forms containing the active ingredient.
  • a pack or device may, for example, comprise metal or plastic foil, such as a blister pack, or glass, and rubber stoppers such as in vials.
  • the pack or dispenser device may be
  • compositions comprising a compound of the present technology formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.
  • the amount of the compound in a formulation can vary within the full range employed by those skilled in the art. Typically, the formulation will contain, on a weight percent (wt %) basis, from about 0.01-99.99 wt % of a compound of the present technology based on the total formulation, with the balance being one or more suitable pharmaceutical excipients. Preferably, the compound is present at a level of about 1-80 wt %. Representative pharmaceutical formulations are described below.
  • a composition can be administered alone or in combination with other treatments, either simultaneously or sequentially dependent upon the condition to be treated.
  • the present disclosure also provides for a kit comprising the combination of a Cdc7 inhibitor and, optionally, an additional treatment and instructions for use.
  • the present disclosure further provides for a kit comprising one or more pharmaceutical compositions where the pharmaceutical composition(s) comprise a Cdc7 inhibitor and an additional treatment, and instructions for use, optionally the combination includes at least one pharmaceutically acceptable carrier or excipient.
  • kits can be packaged in separate containers and, associated with such containers, can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale.
  • the kit may optionally contain instructions or directions outlining the method of use or administration regimen for the antigen-binding construct.
  • kits comprising a combination of SRA141 and a additional treatment and at least one pharmaceutically acceptable carrier or excipient.
  • the container means may itself be an inhalant, syringe, pipette, eye dropper, or other such like apparatus, from which the solution may be administered to a subject or applied to and mixed with the other components of the kit.
  • the components of the kit may also be provided in dried or lyophilized form and the kit can additionally contain a suitable solvent for reconstitution of the lyophilized components.
  • the kits described herein also may comprise an instrument for assisting with the administration of the composition to a patient.
  • Such an instrument may be an inhalant, nasal spray device, syringe, pipette, forceps, measured spoon, eye dropper or similar medically approved delivery vehicle.
  • an article of manufacture containing materials useful for the treatment, prevention and/or diagnosis of the disorders described herein, e.g., inhibition of tumor growth comprises a container and a label or package insert on or associated with the container.
  • Suitable containers include, for example, bottles, vials, syringes, iv. solution bags, etc.
  • the containers may be formed from a variety of materials such as glass or plastic.
  • the contained s) holds a composition which is by itself or combined with another composition effective for treating, preventing and/or diagnosing the disorder and may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle).
  • a sterile access port for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle.
  • the article of manufacture in this embodiment described herein may further comprise a label or package insert indicating that the compositions can be used to treat a particular condition.
  • the article of manufacture may further comprise a second (or third) container comprising a pharmaceutically-acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution.
  • BWFI bacteriostatic water for injection
  • phosphate-buffered saline such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution.
  • BWFI bacteriostatic water for injection
  • phosphate-buffered saline such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution.
  • BWFI bacteriostatic water for injection
  • phosphate-buffered saline such as
  • polypeptide and nucleic acid sequences of genes e.g., genes for Cdc7.
  • polypeptide and nucleic acid sequences are at least 95, 96, 97, 98, or 99% identical to sequences described herein or referred to herein by a database accession number.
  • polypeptide and nucleic acid sequences are 100% identical to sequences described herein or referred to herein by a database accession number.
  • the term“percent identity,” in the context of two or more nucleic acid or polypeptide sequences, refer to two or more sequences or subsequences that have a specified percentage of nucleotides or amino acid residues that are the same, when compared and aligned for maximum correspondence, as measured using one of the sequence comparison algorithms described below (e.g., BLASTP and BLASTN or other algorithms available to persons of skill) or by visual inspection.
  • the percent“identity” can exist over a region of the sequence being compared, e.g., over a functional domain, or, alternatively, exist over the full length of the two sequences to be compared.
  • sequence comparison typically one sequence acts as a reference sequence to which test sequences are compared.
  • test and reference sequences are input into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated.
  • sequence comparison algorithm then calculates the percent sequence identity for the test sequence(s) relative to the reference sequence, based on the designated program parameters.
  • Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l.
  • Cdc7 inhibitor compound I-D, also named SRA141
  • SRA141 an acute myeloid leukemia cell line
  • vehicle control 0.2 M HC1 / 0.5% MC400
  • SRA141 30 or 60 mg/kg BID for 3 weeks
  • SRA141 120 mg/kg QD 3 weeks
  • n 8 per treatment group.
  • the body weights of mice were recorded once per week before grouping and mice conditions were recorded daily during the treatment. After 21 days of dosing, mice were sacrificed for tumor weight measurement. Tumor growth inhibition was calculated on Day 21, the final day of the study.
  • the average tumor volumes of the four groups at day 21 were 1612.66 mm 3 , 655.68mm 3 , 300.77 mm 3 and 530.83 mm 3 , at vehicle and 30 mg/kg BID, 60 mg/kg BID and 120 mg/kg QD mg/kg dose levels, respectively.
  • Tumor weight for individual mice was also calculated (Fig. IB).
  • SRA141 administration resulted in tumor growth inhibition of 59%, 81% and 67% at the 30 mg/kg BID, 60 mg/kg BID and 120 mg/kg QD mg/kg dose levels, respectively.
  • Tumor volume measurements were taken biweekly and body weight measurements were taken biweekly to day 15, then daily to end of study. Tumor growth inhibition was calculated on Day 42, the final day of the study. [00213] As shown in Fig. 2A, significant tumor growth inhibition (TGI) was observed in rats treated with 75 mg/kg SRA141 (86. l%TGI) and 150 mg/kg SRA141 (86.7%TGI).
  • the concentration-dependent effect of SRA141 on cell viability was determined in a panel of 235 cancer cell lines, utilizing the CellTiter-Glo TM luminescence assay (Promega) with a 72-hour drug incubation period.
  • the 50% inhibition concentration (IC50) was determined in the cancer cell lines using Cell Titer-Glo ® luminescent cell viability assay after incubation with SRA141 at different concentrations.
  • SRA141 demonstrated potent inhibitory activity (IC50 ⁇ 3 mM) in a range of solid tumor cell lines, including those of bladder (5673, RT112/84), sarcoma (143B, CADO-ES1, RD-ES, A673), colon and cecum (Colo-205, DLD-l, LS1034, SK-CO-l, SNU-C1), renal (U0.31), head-and-neck (CNE-2Z, RPMI-2650), melanoma (A2058, A875), and gastric (HGC-27) lineages.
  • bladder 5673, RT112/84
  • sarcoma 143B, CADO-ES1, RD-ES, A673
  • colon and cecum Colo-205, DLD-l, LS1034, SK-CO-l, SNU-C1
  • renal U0.31
  • CNE-2Z head-and-neck
  • RPMI-2650 melanoma
  • HGC-27 gastric lineages
  • hematologic-derived cell lines The results for 59 hematologic cancer cell lines tested are shown in Fig. 3A. These results demonstrate that SRA141 alone as a monotherapy can provide effective therapy for the treatment of hematologic cancers, such as acute myeloid leukemia (Fig. 3A, cell lines denoted“C”) and chronic eosinophilic leukemia (Fig. 3A, cell lines denoted“L”). Significant activity was also observed in i ALL lines (RS411, SUP-B15, Reh) and other leukemias and lymphomas (KHYG-l and JeKo-l).
  • IC50 values A summary of determined IC50 values is summarized in Fig. 3B, and broken down by cancer type. Overall, hematologic cancer lines showed higher sensitivity than solid tumor lines, and within the latter, colorectal cancer lines were among the most sensitive.
  • the antiproliferative activity of SRA141 was further characterized in seven cell lines (Colo-205, SW620, SNU-398, NCI-H716, MDA-MB-231, NCI-H1573 and SW1116) using four orthogonal assays designed to measure ATP levels (CTG); metabolic activity (CellTiter-Blue (CTB)); DNA content of the cells (CyQuant); and esterase activity (Calcein AM).
  • CCG ATP levels
  • CellTiter-Blue CellTiter-Blue
  • CaQuant DNA content of the cells
  • esterase activity Calcein AM
  • TGI tumor growth inhibition
  • Example 5 SRA141 Treatment of Patient Derived Xenograft Rat Models of CRC
  • Cdc7 inhibitor compound SRA141
  • a monotherapy was tested in a rat xenograft model of colorectal cancer (CRC) (Fig. 7).
  • Female Rowett nude rats were inoculated sub-cutaneously with 2 X 10 7 COLO-205 human colorectal carcinoma cells. All rats were gamma-irradiated (4Gy) 24 hours before tumor cell injection.
  • the COLO-205 cells were injected in 0.2 mL PBS in 1 : 1 Matrigel ® .
  • Fig. 7A administration of SRA141 resulted in significant tumor growth inhibition of 50% and 93% at the 150 mg/kg QD and 75 mg/kg BID dose levels, respectively.
  • Complete tumor regressions were observed in 4/7 rats.
  • complete tumor regressions (defined as no measurable tumor for 3 consecutive measurements) were observed in 4 of 7 animals and a partial regression (tumor volume ⁇ 50% of initial volume for 3 consecutive measurements) in 1 of 7 animals at the 75 mg/kg BID dose. All 4 complete regressions persisted to study completion.
  • tumor SRA141 concentration of approximately 1.2 mM resulted in an approximate 50% decrease in phosphorylated MCM2 (Serine 53) as compared to rats treated with vehicle, suggesting that selective inhibition of Cdc7 is incompatible with tumor cell survival.
  • Colo-205 tumor-bearing rats were treated with a single dose of SRA141 or vehicle and the tumors were collected 12 hours later. Tumor homogenates prepared from three animals per treatment group were analyzed by LC/MSMS or Western blot. The results demonstrate that SRA141 as a monotherapy can provide effective therapy for the treatment of colorectal cancer.
  • the combination screen was performed using the co-treatment dosing schedule for SRA141 and the partner compound. Both enhancee (SRA141) and enhancer (partner compound) were added at time zero (Oh). Cells were exposed to SRA141 and the enhancer for the entire 72-hour treatment time. All data points were collected via automated processes and were subject to quality control and analyzed using proprietary software. Assay plates were accepted if they pass the following quality control standards: relative raw values were consistent throughout the entire experiment, Z-factor scores were greater than 0.6 and untreated/vehicle controls behaved consistently on the plate.
  • GI Growth Inhibition
  • a GI 100% represents complete growth inhibition (cytostasis) and in this case cells treated with compound for 72 or 96 hours would have the same endpoint reading as TO control cells.
  • a GI of 200% represents complete death (cytotoxicity) of all cells in the culture well and in this case the T reading at 72 or 96 hours will be lower than the To control (values near or at zero).
  • Synergy Score Analysis To measure combination effects in excess of Loewe additivity, a scalar measure was used to characterize the strength of synergistic interaction termed the Synergy Score. The Synergy Score is calculated as:
  • the fractional inhibition for each component agent and combination point in the matrix is calculated relative to the median of all vehicle-treated control wells.
  • the Synergy Score equation integrates the experimentally-observed activity volume at each point in the matrix in excess of a model surface numerically derived from the activity of the component agents using the Loewe model for additivity. Additional terms in the Synergy Score equation (above) are used to normalize for various dilution factors used for individual agents and to allow for comparison of synergy scores across an entire experiment.
  • the inclusion of positive inhibition gating or an Idata multiplier removes noise near the zero effect level, and biases results for synergistic interactions at that occur at high activity levels.
  • GI Growth Inhibition
  • Potency shifting was evaluated using an isobologram, which demonstrates how much less drug is required in combination to achieve a desired effect level, when compared to the single agent doses needed to reach that effect.
  • the isobologram was drawn by identifying the locus of concentrations that correspond to crossing the indicated inhibition level. This is done by finding the crossing point for each single agent concentration in a dose matrix across the concentrations of the other single agent. Practically, each vertical concentration CY is held fixed while a bisection algorithm is used to identify the horizontal concentration Cx in combination with that vertical dose that gives the chosen effect level in the response surface
  • Loewe Volume Score is used to assess the overall magnitude of the combination interaction in excess of the Loewe additivity model. Loewe Volume is particularly useful when distinguishing synergistic increases in a phenotypic activity (positive Loewe Volume) versus synergistic antagonisms (negative Loewe Volume). When antagonisms are observed, as in the current dataset, the Loewe Volume is assessed to examine if there is any correlation between antagonism and a particular drug target-activity or cellular genotype. This model defines additivity as a non- synergistic combination interaction where the combination dose matrix surface is indistinguishable from either drug crossed with itself.
  • adenocarcinoma cells and acute myeloid leukemia cells either with sequential treatment or simultaneous treatment with the Cdc7 inhibitor, SRA141, in combination with an anti neoplastic agent.
  • SRA141 Cdc7 inhibitor
  • an anti neoplastic agent an anti neoplastic agent.
  • mTOR mammalian target of rapamycin
  • RTK receptor tyrosine kinases
  • MAPK mitogen activated protein kinase pathway
  • PI3K phosphatidylinositol-4,5-bisphosphate 3 kinase
  • Flasks of Colo-205 cells were pre-treated with SRA141 (1 mM and 200 nM) and DMSO control for 48 hrs; cells were washed and seeded into 384-well plates, and plates were left to incubate for 6 hrs, followed by treatment with everolimus, aphidicolin or midostaurin six hours after plating as single agents. After 72 hrs, ATP levels were measured using ATPlite.
  • Pre-treatment of COLO-205 cells with SRA141 demonstrated an enhancer potency shift towards increase in sensitivity for midostaurin, and a 3.19 ECso fold change was observed with 200 nM SRA141 pre-treatment compared to DMSO pre-treatment.
  • a 4.60 ECso fold change was observed with 1 mM SRA141 pre-treatment compared to DMSO pre-treatment.
  • a 3.3 EC50 fold change was observed for 200 nM SRA141 pre-treatment compared to DMSO pre- treatment.
  • Colo-205 cells were simultaneously treated with the MAPK pathway inhibitor, trametinib, a PI3K pathway inhibitor, copanlisib, and an anti-metabolite, (anti-folte), methotrexate (Figs. 9 and 10A).
  • Cells were seeded into 384-well plates on day 0; plates were left to incubate for 24 hrs; cells were dosed with trametinib and copanlisib in combination with SRA141, and after 72 hours, ATP levels were measured using ATPlite. Strong synergies were observed in the COLO-205 cells with simultaneous combination treatment with inhibitors of the MAPK pathway and PI3K pathway and the antifolate antimetabolite, methotrexate.
  • a Cdc7 inhibitor to sensitize cancer cells when used in combination with an inhibitor of the mitogen activated protein kinase pathway (MAPK), a regulator of the retinoid pathway, an apoptosis regulator, a PARP inhibitor, or an inhibitor of the mammalian target of rapamycin (mTOR) pathway
  • MAPK mitogen activated protein kinase pathway
  • mTOR mammalian target of rapamycin
  • Flasks of MOLM-13 cells were pre treated with SRA141 (1 mM and 200 nM) and DMSO control for 48 hrs; cells were washed and seeded into 384-well plates; plates left to incubate for 6 hrs, followed by treatment with trametinib or bexarotene six hours after plating as single agents. After 72 hrs, ATP levels were measured using ATPlite. For trametinib, a 2.63 EC50 fold change was observed for 200 nM SRA141 pre-treatment compared to DMSO pre-treatment. For bexarotene, a 2.63 EC50 fold change was observed for 200 nM SRA141 pre-treatment compared to DMSO pre treatment.
  • MOLM-13 cells demonstrated strong synergies for BMN673 (PARP inhibitor), ABT-199 (BCL-2 inhibitor), bexarotene and tretinoin (regulators of the Retinoid pathway).
  • combination therapies comprising use of a Cdc7 inhibitor in combination with an inhibitor of the mTOR pathway, an inhibitor of DNA polymerase, and inhibitor of receptor tyrosine kinases, and inhibitor of the MAPK pathway, a regulator of the retinoid pathway, a regulator of apoptosis, or a PARP inhibitor, are effective for killing cancer cells, either with sequential or concurrent administration with a Cdc7 inhibitor and demonstrates that administration of a Cdc7 inhibitor in combination with an inhibitor of the mTOR pathway, an inhibitor of DNA polymerase, and inhibitor of receptor tyrosine kinases, and inhibitor of the MAPK pathway, a regulator of the retinoid pathway, a regulator of apoptosis, or a PARP inhibitor, are regulator of the retinoid pathway, a regulator of apoptosis, or a PARP inhibitor, are effective cancer treatment regimens.
  • SRA141 for Cdc7 was assessed using the DiscoverX kinome screening assay, containing approximately 430 native and mutant kinases. Minimal off-target kinase activity was detected at a compound concentration of 500 nM. Selectivity of SRA141 for Cdc7 was compared to TAK-931. Results of the kinome screening assay (Fig. 12), demonstrate that SRA141 has less off-target activity compared to TAK-931 at a compound concentration of 500 nM.
  • Colo-205 cells were treated with SRA141 (at concentrations between 0.033 and 3.3 mM) for 8 to 24 hours with subsequent assessment of the phosphorylation status of the downstream targets for Cdc7 (MCM2, Ser40/4l and Ser53), and the putative off-target kinases CDK8 (STAT1, Ser727); CDK7 and 9 (RNA pol II, Ser2); LATS2 (YAP1, Serl27) as well as the levels of MCL-l, known to be indirectly controlled by CDK9 (Gregory, 2015).
  • MCM2 Cdc7
  • Ser40/4l and Ser53 putative off-target kinases
  • CDK8 STAT1, Ser727
  • CDK7 and 9 RNA pol II, Ser2
  • LATS2 YAP1, Serl27
  • MV411 cells were also treated with SRA141 in a dose-dependent manner (at concentrations between 0.5 and 3.3 pM) and were subsequently assessed for phosphorylation status of MCM2 at Ser40/4l and Ser53.
  • the mean SRA141 IC50 value for the normal bone marrow preparations was 0.25 mM, while the mean IC50 value for the AML samples was approximately 0.16 mM (with the resistant progenitor sample AML810 censored; Table 2).
  • the positive control, cytosine arabinoside (Ara C) had equipotent IC50 values (approximately 0.004 mM) for both the normal bone marrow and AML-blast progenitor samples.
  • the Therapeutic Index was also calculated for each sample [(Primary AML ICso) ⁇ (BMNC control ICso)]. As shown in Table 2, 5 out of 9 primary AML samples demonstrated response to SRA141 with a TI
  • Table 2 SRA141 Activity in Normal Bone Marrow and AML-Blast Progenitors Grown in 3D
  • FIG. 15 flow cytometry analysis of cells labelled with propidium iodide demonstrated that a sub-Gl population, indicative of apoptotic cells, accumulated as the cells progressed through M phase provided they were treated with SRA141 during the preceding S phase (Fig. 15 A), but not if the treatment started after S phase completion (beginning of M phase) (Fig. 15B). If SRA141 was added first at M phase, the sub-Gl accumulation was delayed and required that cells progress through a subsequent S phase in the presence of SRA141 before showing signs of apoptosis.
  • FIG. 15C Colo-205 cells treated with 0.1 mM SRA141 for 48 hours were assessed for cell cycle and DNA damage markers by western blot. Results demonstrate the presence of mitotic markers of G2/M phase following treatment with SRA141. These results are consistent with data obtained by use of high content imaging that demonstrates that Colo-205 and SW620 cell populations treated with SRA141 for 48 hours have an accumulation of cells in mitosis (Fig. 15D). The percent of cells in mitosis is greater in populations treated with SRA141 compared to other Cdc7 inhibitors.
  • Example 13 Colo-205 Colorectal Xenograft Study (Mouse)
  • mice bearing Colo-205 tumor xenografts were orally
  • SRA141 administration resulted in tumor growth inhibition of 15%, 58% and 37% at the 30 mg/kg BID, 60 mg/kg BID and 120 mg/kg QD dose levels, respectively.
  • mice treated with SRA141 at 60 mg/kg TID experienced a maximum body weight loss greater than 20%. Dosing to these animals was suspended until the body weight loss recovered to less than 20%. None of the mice treated with SRA141 120 mg/kg QD met the 20% weight loss threshold.
  • Example 15 MV-4-11 Human Leukemia Systemic Survival Study
  • Example 16 A20 Immunocompetent Lymphoma Xenograft Model (Mouse)
  • Example 17 MDA-MB-486 Breast Xenograft Study (Mouse)
  • Example 18 Mouse SW620 Colorectal Xenograft Model PK/PD Assessment
  • pMCM2 levels decreased following a single SRA141 administration, peaking between 2-4 hours for 30mg/kg and 60 mg/kg doses and between 2-8 hours for the 120 mg/kg dose (pMCM2 levels shown in Fig. 21A, % inhibition quantified and normalized to actin in Fig. 21B).
  • Maximal SRA141 plasma concentrations of ca 1, 1.5, and 2 pg/mL were also observed at 2 to 4 hours after administration at the 30, 60 and 120 mg/kg doses, respectively. Tumoral concentrations of drug were similar.
  • Correlation of PK and PD data from this study suggested that a circulating plasma concentration and intra-tumoral tissue concentration of approximately 1.1 pM is required to inhibit pMCM2 by 50%.
  • Example 19 Rat Colo-205 Colorectal Xenograft Model PK/PD Assessment
  • SRA141 presented as free base suspension
  • moderate absolute oral bioavailability (%F) in the fasted mouse, rat and dog
  • Further studies in the dog suggested absolute oral bioavailability of the bis- hydrochloride salt in suspension may be improved following post-prandial administration.
  • Systemic exposure (Cmax, AUC) following oral dosing generally increased with increasing dose, but in a less than dose proportional manner.
  • the vehicle was 0.5% CMC-Na/l% Lutrol in water.
  • SRA141 plasma concentrations were determined by LC-MS/MS following last dose. As shown in Fig. 23, systemic exposure increased with an increase in the oral dose from 50 to 150 mg/kg.
  • Biliary and Kupffer cell changes were the only findings noted following 28-day repeat dose administration of SRA141 to the rat, with incidental hematologic and clinical chemistry changes.
  • changes in other organs and tissues were identified including liver (hepatocellular degeneration), spleen (reduced red pulp cellularity) and stomach (inflammation and epithelial hyperplasia).
  • liver hepatocellular degeneration
  • spleen reduced red pulp cellularity
  • stomach inflammation and epithelial hyperplasia
  • SRA141 was negative for both mutagenicity and clastogenicity in in vitro bacterial reverse mutation and human lymphocyte chromosomal aberration assays.
  • the results from these studies indicate that the toxicity findings after administration of SRA141 are generally monitorable in a clinical setting, supportive therapies are available and/or are less relevant in an advanced oncology study population.
  • MTD and hi ghest-non-severely -toxic dose (HNSTD) following a 5 days on / 2 days off dosing schedule for 4 weeks were considered to be 100 mg/kg/day (600 mg/m2/day) and 10 mg/kg/day (200 mg/m2/day) in rats and dogs, respectively.
  • Example 22 Immunohistochemistry assessment of SRA141 treatment in rat MV-4-11 xenograft model of biphenotypic B myelomonocytic leukemia
  • Rats bearing subcutaneous biphenotypic B myelomonocytic leukemia MV-4-11 tumors were treated with SRA141 at 75 mg/kg or vehicle, BID, or with SRA141 at 100 mg/kg, QD, for 5 days. Tumors were collected from the animals 12 hours after
  • Tumor cells in xenograft tumors (but not stroma or surrounding rat tissue) were scored. Scoring is performed semi-quantitatively with the main components to scoring tumor cells for total MCM2, pMCM2-S53, pMCM2-S40, and gH2AX reactivity as percentages at differential intensities and H-Scores. Percentage and intensity measures are estimated by a scientist in the same relative region of tumor (one 40X-field from the bottom edge) within each xenograft sample.
  • the H-Score is calculated by summing the percentage of cells with intensity of expression (brown staining) multiplied by their corresponding differential intensity on a four- point semi-quantitative scale (0, 1+, 2+, 3+). Thus, scores range from 0 to 300.
  • H-Score [ (% at ⁇ l) x 0 ] + [ (% at 1+) x 1 ] + [ (% at 2+) x 2 ] + [ (% at 3+) x 3] [00303] Evaluation of Rat Skin
  • Epithelial cells of the epidermis in rat skin samples were scored jointly by two scientists. Scoring was performed quantitatively with the main component to scoring epidermal cells for total MCM2, pMCM2-S53, pMCM2-S40, and gH2AX reactivity as counts of positive cells. For each skin sample, four evenly distributed regions were marked for review. The regions were aligned to the same approximate position in each skin sample. For pMCM2-S53 and pMCM2-S40, the number of positive cells in a 10X field at the four representative regions was counted. For gH2Ax, the number of positive cells in a 20X field at the four representative regions was counted. Cells were counted if staining was readily observed at 10X or 20X, regardless of intensity. Cells were only counted within the flat epidermis and not in regions of invagination in order to consistently capture the same total area.
  • the number of positive skin cells counted across the regions analyzed for each skin sample were averaged for each biomarker.
  • the average number of positive skin cells for pMCM2-S53 and pMCM2-S40 were then represented as a percentage relative to the average number of positive skin cells for total MCM2.
  • the average number of positive skin cells for gH2AX was multiplied by 2 to account for its assessment at 20X versus 10X (normalized) and was then also represented as a percentage relative to the average for total MCM2.
  • Skin cell counts were performed by recording the number of positive cells (at any intensity) counted by two independent scientists in multiple regions of skin (two regions for total MCM2, four regions for pMCM2 and gH2AX) that are in the same approximate location for each sample. The number of positive cells recorded at each skin region for each biomarker were averaged for each sample. The average cell counts for pMCM2-S53, pMCM2-S40, and gH2AX (normalized for magnification) were compared as percentages of total MCM2. Cell counts were performed at 20X for gH2AX and 10X for all other markers.
  • Avg. Count [region 1 pos cells] + [region 2 pos cells] + [region 3 pos cells] + [region 4 pos cells]/4
  • Avg. Count (total MCM2) [region 1 pos cells] + [region 4 pos cells]/2
  • Results shown in Fig. 25A and Fig. 25B demonstrate a dose-dependent decrease in pMCM2-S40 in both tumor and skin in the rat MV-4-11 xenograft model.
  • Epithelial cells of the epidermis in human skin samples were scored jointly by two scientists. Scoring was performed quantitatively with the main component to scoring epidermal cells for total MCM2, pMCM2-S53, pMCM2-S40, and gH2AX reactivity as counts of positive cells. For each skin sample, two evenly distributed regions were marked for review. The regions were aligned to the same approximate position in each skin sample and assessed at 10X for MCM2, and pMCM2. For gH2AX, the number of positive cells in a 20X filed at the two regions was counted. Cells were counted if staining was readily observed at 10X or 20X, regardless of intensity. Cells were only counted within the flat epidermis and not in regions of invagination in order to consistently capture the same total area.
  • the number of positive skin cells counted across the regions analyzed for each skin sample were averaged for each biomarker.
  • the average number of positive skin cells for pMCM2-S53 and pMCM2-S40 were then represented as a percentage relative to the average number of positive skin cells for total MCM2.
  • the average number of positive skin cells for gH2AX was multiplied by 2 to account for its assessment at 20X versus 10X (normalized) and was then also represented as a percentage relative to the average for total MCM2.
  • Avg. Count [region 1 pos cells] + [region 2 pos cells] /2
  • Phase I the 20% inhibition concentration (IC20) and 50% inhibition concentration (IC50) were determined for each of the following nine agents in all six cell lines: ABT-199, the ATM kinase inhibitor KEG-60019, the Aurora B kinase inhibitor
  • Results of combination treatments in Colo-205 cells demonstrate that SRA141 and Barasertib act synergistically, particularly in the presence of 0.012 mM Barasertib.
  • the combination of SRA141 and Bexarotene appear to have an antagonistic effect.
  • Trametinib and Copanlisib appear to have a slight additive effect with SRA141 in Colo-205 cells.
  • Barasertib, Trametinib, and Copanlisib all have slight additive effects on SRA141.
  • Results of combination treatments in A375 (Fig. 27C) and KG-l (Fig. 27D) cells show that Barasertib, but not Trametinib or Copanlisib have slight additive effects with SRA141.
  • Barasertib (Fig. 27H), Trametinib, Copanlisib, Bexarotene, and Tretinoin all demonstrate slight additive effects with SRA141 in MOLM-13 cells.
  • Anti-apoptotic genes BCL-XL, BCL-2, and MCL-l were inhibited in HCT116 and Hela cells using RNAi knockdown.
  • Cells were plated in 96-well plates at 2000 cells/well and were transfected with an RNAi/Lipofectamine RNAimax solution at a final concentration of 10 nM.
  • Transfected cells were treated with SRA141 at a range of doses for 72 hours.
  • Cell viability of treated cells transfected with RNAi against CTRL (non-toxic control RNAi), BCL-2, BCL-XL, or MCL-l was measured using CTBlue assays as previously described (Example 3).
  • Data shown in Fig. 28A demonstrate that anti-apoptotic genes inhibited by RNAi synergize with SRA141.
  • the anti-apoptotic gene BCL-2 was inhibited in Molm-l3 cells by treatment with 0.1 pM ABT-199 as previously described (see Examples 6 and 24). Cells were treated with 0.1 pM ABT-199 and SRA141 for 72 hours at SRA141 concentrations of 0.04 pM to 3.30 pM. Synergy with BCL-2 inhibition and SRA141 was demonstrated at the following concentrations of SRA141 : 0.12 pM, 0.37 pM, 1.10 pM and 3.30 pM (Fig. 28B) as indicated by Combination Index values less than 1 (see Example 24)
  • Example 26 Clinical Trial Evaluation of Cdc7 Inhibitor
  • Suitable patients include patients diagnosed with cancer (e.g., breast cancer, colon cancer, lung cancer; blood cancers such as leukemia, lymphoma, myeloma, acute myeloid leukemia (AML) and chronic myelogenous leukemia (CML), melanoma, uterine cancer, thyroid cancer, chronic eosinophilic leukemia, diffuse large B-cell lymphoma (DLBCL), bladder cancer, cervical cancer, colorectal cancer (CRC), gastric cancer, endometrial cancer, hepatocellular cancer, non-small cell lung cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain cancer, sarcoma, small cell lung cancer, neuroblastoma and head and neck cancer).
  • cancer e.g., breast cancer, colon cancer, lung cancer
  • blood cancers such as leukemia, lymphoma, myeloma, acute myeloid leukemia (AML) and
  • Dosages of SRA-141 can be 100-5,000 mg; 100-1,000 mg; 1,000-2,000 mg; 2,000-3,000 mg; 3,000-4,000 mg; 4,000-5,000 mg; 1,500-2,500 mg; 500-1,000 mg; 1,000- 1,500 mg; 1,500-2,000 mg; 2,000 mg-2,500 mg; 2,500-3,000 mg; 3,000-3,500 mg; 3,500- 4,000 mg; 4,000-4,500 mg or 4,500-5,000 mg.
  • SRA-141 can be administered daily, twice daily, thrice daily, every other day, weekly, monthly, daily dosing for 1-7 days followed by 1- 28 days of non-dosing, 1-28 days of daily dosing followed by 1-28 days of non-dosing.
  • Suitable patients include patients diagnosed with cancer (e.g., breast cancer, colon cancer, lung cancer; blood cancers such as leukemia, lymphoma, myeloma, acute myeloid leukemia
  • cancer e.g., breast cancer, colon cancer, lung cancer
  • blood cancers such as leukemia, lymphoma, myeloma, acute myeloid leukemia
  • AML and chronic myelogenous leukemia
  • CML chronic myelogenous leukemia
  • melanoma uterine cancer
  • thyroid cancer chronic eosinophilic leukemia
  • DLBCL diffuse large B-cell lymphoma
  • bladder cancer cervical cancer
  • colorectal cancer CRC
  • gastric cancer endometrial cancer
  • hepatocellular cancer non-small cell lung cancer, ovarian cancer, prostate cancer, pancreatic cancer
  • brain cancer sarcoma
  • small cell lung cancer neuroblastoma and head and neck cancer
  • Patients are orally administered the Cdc7 inhibitor, SRA-141.
  • Patients are concurrently administered an inhibitor of the mTOR pathway, an inhibitor of DNA polymerase, and inhibitor of receptor tyrosine kinases, and inhibitor of the MAPK pathway, a regulator of the retinoid pathway, a regulator of apoptosis, a PARP inhibitor, or an antifolate antimetabolite in human patients.
  • Maximum effective dosages to be administered are determined using techniques that are known to those of ordinary skill of the art. Patients are monitored for disease progression.

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Abstract

La présente invention concerne des procédés de traitement par administration d'un inhibiteur de Cdc7 en monothérapie ou en polythérapie utile pour inhiber la croissance de tumeurs telles que celles de patients atteints de cancer.
PCT/US2019/019676 2018-02-26 2019-02-26 Procédés de traitement du cancer comprenant des inhibiteurs de cdc7 Ceased WO2019165473A1 (fr)

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JP2021516638A JP7579781B2 (ja) 2018-09-24 2019-08-28 Ｃｄｃ７阻害剤を含む癌の治療方法
AU2019350581A AU2019350581B2 (en) 2018-09-24 2019-08-28 Methods of treatment of cancer comprising Cdc7 inhibitors
US17/275,732 US20210393620A1 (en) 2018-09-24 2019-08-28 Methods of Treatment of Cancer Comprising CDC7 Inhibitors
CN201980069832.XA CN113348020A (zh) 2018-09-24 2019-08-28 包括cdc7抑制剂的治疗癌症的方法
PCT/US2019/048657 WO2020068347A1 (fr) 2018-09-24 2019-08-28 Méthodes de traitement du cancer comprenant des inhibiteurs de cdc7
CA3113621A CA3113621A1 (fr) 2018-09-24 2019-08-28 Methodes de traitement du cancer comprenant des inhibiteurs de cdc7
KR1020217010445A KR20210064252A (ko) 2018-09-24 2019-08-28 Cdc7 억제제를 포함하는 암의 치료 방법
CN202511057693.2A CN121081469A (zh) 2018-09-24 2019-08-28 包括cdc7抑制剂的治疗癌症的方法
EP19866944.2A EP3856352A4 (fr) 2018-09-24 2019-08-28 Méthodes de traitement du cancer comprenant des inhibiteurs de cdc7

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WO2020005063A1 (fr) * 2018-06-26 2020-01-02 Stichting Het Nederlands Kanker Instituut-Antoni van Leeuwenhoek Ziekenhuis Polythérapie et son utilisation pour le traitement du cancer
WO2020068347A1 (fr) * 2018-09-24 2020-04-02 Sierra Oncology, Inc. Méthodes de traitement du cancer comprenant des inhibiteurs de cdc7
CN114544811A (zh) * 2022-02-17 2022-05-27 南京正济医药研究有限公司 一种cdc7抑制剂有关物质的检测方法
WO2022167999A1 (fr) * 2021-02-08 2022-08-11 Takeda Pharmaceutical Company Limited Polythérapie pour le traitement du cancer
EP4275686A1 (fr) * 2022-05-11 2023-11-15 National University of Ireland Galway Thérapie combinée pour le cancer comprenant un inhibiteur de cdc7 et un inhibiteur de cdk8 ou un inhibiteur de ccnc
CN117563005A (zh) * 2023-10-26 2024-02-20 浙江大学 一种治疗结直肠癌的药物组合物及其应用

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US20150021813A1 (en) * 2011-12-28 2015-01-22 Bridgestone Corporation Tire mold, tire, and tire manufacturing method
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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020005063A1 (fr) * 2018-06-26 2020-01-02 Stichting Het Nederlands Kanker Instituut-Antoni van Leeuwenhoek Ziekenhuis Polythérapie et son utilisation pour le traitement du cancer
WO2020068347A1 (fr) * 2018-09-24 2020-04-02 Sierra Oncology, Inc. Méthodes de traitement du cancer comprenant des inhibiteurs de cdc7
AU2019350581B2 (en) * 2018-09-24 2025-07-10 Carna Biosciences, Inc. Methods of treatment of cancer comprising Cdc7 inhibitors
WO2022167999A1 (fr) * 2021-02-08 2022-08-11 Takeda Pharmaceutical Company Limited Polythérapie pour le traitement du cancer
CN114544811A (zh) * 2022-02-17 2022-05-27 南京正济医药研究有限公司 一种cdc7抑制剂有关物质的检测方法
CN114544811B (zh) * 2022-02-17 2023-10-20 南京正济医药研究有限公司 一种cdc7抑制剂有关物质的检测方法
EP4275686A1 (fr) * 2022-05-11 2023-11-15 National University of Ireland Galway Thérapie combinée pour le cancer comprenant un inhibiteur de cdc7 et un inhibiteur de cdk8 ou un inhibiteur de ccnc
WO2023217980A1 (fr) * 2022-05-11 2023-11-16 National University Of Ireland, Galway Polythérapie pour le cancer comprenant un inhibiteur de cdc7 et un inhibiteur de cdk8 ou un inhibiteur de ccnc
CN117563005A (zh) * 2023-10-26 2024-02-20 浙江大学 一种治疗结直肠癌的药物组合物及其应用

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