WO2022120242A1 - Y box binding protein 1 inhibitors - Google Patents

Y box binding protein 1 inhibitors Download PDF

Info

Publication number
WO2022120242A1
WO2022120242A1 PCT/US2021/061906 US2021061906W WO2022120242A1 WO 2022120242 A1 WO2022120242 A1 WO 2022120242A1 US 2021061906 W US2021061906 W US 2021061906W WO 2022120242 A1 WO2022120242 A1 WO 2022120242A1
Authority
WO
WIPO (PCT)
Prior art keywords
pharmaceutically acceptable
acceptable salt
group
cancer
compound
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2021/061906
Other languages
English (en)
French (fr)
Inventor
Sanjay V. MALHOTRA
Dhanir TAILOR
Arpit DHEERAJ
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Oregon Health and Science University
Leland Stanford Junior University
Original Assignee
Oregon Health and Science University
Leland Stanford Junior University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Oregon Health and Science University, Leland Stanford Junior University filed Critical Oregon Health and Science University
Priority to US18/255,827 priority Critical patent/US20240016784A1/en
Priority to CA3201032A priority patent/CA3201032A1/en
Priority to AU2021391931A priority patent/AU2021391931A1/en
Priority to CN202180092273.1A priority patent/CN116761602A/zh
Priority to EP21901582.3A priority patent/EP4255426A4/en
Priority to JP2023533819A priority patent/JP2023552368A/ja
Publication of WO2022120242A1 publication Critical patent/WO2022120242A1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • A61K31/4355—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 oxygen as a ring hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
    • 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
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4741—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having oxygen as a ring hetero atom, e.g. tubocuraran derivatives, noscapine, bicuculline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • Y BOX BINDING PROTEIN 1 INHIBITORS FIELD OF THE INVENTION
  • the present invention concerns novel compounds, pharmaceutical compositions, and their use as inhibitors of Y box protein 1 (YB1 or YBX1) to treat conditions including gynecological, breast, bladder, kidney, head and neck, neuronal, and prostate cancers, lymphomas, and leukemias.
  • Y box binding protein 1 YB ⁇ 1
  • TR treatment resistance
  • This first ⁇ in ⁇ class YB ⁇ 1 inhibitor inhibits cell proliferation, resistance to apoptosis in ovarian cancer (OC) cells and arrests in the G1 phase. Inhibitor treatment leads to enrichment of proteins associated with apoptosis and RNA degradation pathways while downregulating spliceosome pathway.
  • SU056 independently restrains OC progression and exerts a synergistic effect with paclitaxel to further reduce disease progression with no observable liver toxicity.
  • in vitro mechanistic studies showed delayed disease progression via inhibition of drug efflux and multi ⁇ drug resistance 1 (MDR1), and significantly lower neurotoxicity as compared to Etoposide.
  • MDR1 multi ⁇ drug resistance 1
  • Y box binding protein 1 (YB ⁇ 1, YBX1) is a multifunctional cold shock protein that binds to DNA and RNA. It regulates DNA and RNA associated cellular events including mRNA transcription, splicing, packaging, stability, and translation (Lyabin et al., 2014). mRNA stabilization is an important event for sustained expression of any gene and YB ⁇ 1 robustly stabilizes the mRNA via blocking the 5' end from mRNA degradation (Evdokimova et al., 2001). It was first described by Didier et al.
  • YB ⁇ 1 is associated with the development of treatment resistance (TR) via its role in activating proliferation, promoting cancer cell stemness, responding to growth factors, cytokines, cellular stress responses, and promoting drug efflux via the membrane P ⁇ glycoprotein ATP ⁇ dependent efflux pump ABCB1 (MDR1) (Bargou et al., 1997, Saupe et al., 2015, Mo et al., 2016).
  • TR treatment resistance
  • MDR1 membrane P ⁇ glycoprotein ATP ⁇ dependent efflux pump ABCB1
  • Y ⁇ box ⁇ binding protein 1 (YB ⁇ 1), encoded by the YBX1 gene, has been noted as modulating or regulating cellular signaling pathways and may be seen as a molecular marker for cancer progression and as a target for cancer therapies. Lasham et al. describe in their review article YB ⁇ 1: oncoprotein, prognostic marker and therapeutic target?, Biochem. J.
  • YB ⁇ 1 regulates multiple proliferation pathways, overrides cell ⁇ cycle check points, promotes replicative immortality and genomic instability, may regulate angiogenesis, has a role in invasion and metastasis, and promotes inflammation. They further describe cell lines in which YB ⁇ 1 reduction induced apoptosis or inhibited cell proliferation, including melanoma, fibrosarcoma, liver cancer, lung cancer, bladder cancer, multiple myeloma, paediatric glioblastoma, breast cancer (ER ⁇ negative), breast cancer (ER ⁇ positive), prostate cancer, and colon cancer cell lines. Sobo ⁇ an et al.
  • Y ⁇ box ⁇ protein 1 mTOR
  • mTOR Y ⁇ box ⁇ protein 1
  • mTOR mTOR
  • Oncogenic Y ⁇ box binding protein ⁇ 1 as an effective therapeutic target in drug ⁇ resistant cancer Kuwano et al., Cancer Science, 2019, 110:1536 ⁇ 1543, describes the function of YBX2 in promoting transcriptional activation of the ABCB1 transporter gene, which has been associated as a transcriptional mechanism of how tumor multidrug resistance is acquired during chemotherapeutic treatments in human malignancies, including breast, lung, ovarian, prostate, colorectal, and gastric cancers.
  • YBX1 The relationship between increased expression of YBX1 and melanoma is discussed in the article The increased expression of Y box ⁇ binding protein 1 in melanoma stimulates proliferation and tumor invasion, antagonizes apoptosis and enhances chemoresistance, Schittek et al., Int. J. Cancer: 120, 2110 ⁇ 2118 (2007). YB1 overexpression has also been associated with radio ⁇ resistance in colorectal cancer cells, as discussed by Kim et al., Mol. Cancer Ther., 30 Oct 2019, 19(2), 479 ⁇ 89.
  • WO 2019/178091 A1 (Malholtra et al., The Board of Trustees of the Leland Stanford Junior University) teaches novel N ⁇ hydroxyethyl didehydroazapodophyllotoxins as GBP1 inhibitors and methods for their use in overcoming treatment resistance in cancers.
  • azopodophyllotoxin (AzP) small molecule SU056 potently inhibits YB ⁇ 1 and reduces OC progression while sensitizing to chemotherapy ⁇ mediated cytotoxicity.
  • X is selected from the group of: , F, C 1 ⁇ C 4 fluoroalkyl, SF 5 , Cl, Br, I, OH, C 1 ⁇ C 4 alkyl, C 1 ⁇ C 4 alkoxy, CN, NO 2 , and OH;
  • n is an integer selected independently in each instance from the group of 1, 2, 3, 4, 5, and 6;
  • R 6 is selected from the group of H, C 1 ⁇ C 6 alkyl, C 3 ⁇ C 6 cycloalkyl, ⁇ (CH 2 ) n ⁇ C 3 ⁇ C 6 cycloalkyl, 3 ⁇ 6 ⁇ membered heterocycle, ⁇ (CH 2 ) n ⁇ 3 ⁇ 6 ⁇ membered heterocycle, phenyl, and ⁇ (CH 2 ) n ⁇ phenyl; wherein the C 1 ⁇ C 6 alkyl group is substituted with 0, 1, 2, 3, or 4 substituents selected from F, Cl, Br, I, OH,
  • FIGURE 1A depicts conversion of SU093 to SU056.
  • FIGURE 1B charts IC 50 values of SU093 and SU056 on various ovarian cancer cells.
  • FIGURE 1C presents a photo of colony formation from respective wells.
  • FIGURE 1D graphs the number of colonies formed after SU093 and SU056 treatment.
  • FIGURE 1E provides a table representing % inhibition values of etoposide, SU093, and SU056 treatment in neuronal (SH ⁇ SY5Y, N27) and HEK293 cells.
  • FIGURE 1F presents a table of cell cycle distribution of propidium iodide (PI) ⁇ stained OVCAR8, SKOV3, and ID8 cells.
  • PI propidium iodide
  • FIGURE 1G graphs effects of SU093 and SU056 on apoptotic cell death.
  • FIGURE 2A presents representative images of tumor regression compared to control in mice after 42 days of drug treatment.
  • FIGURE 2B graphs tumor volume/mouse as a function of time.
  • FIGURE 2C graphs tumor weight/mouse at the end of the study.
  • FIGURE 2D provides a graph of liver toxicity parameters at the end of 42 days showing no significant difference between control, SU093, and SU056.
  • FIGURE 2E presents an image of H&E staining from a lung metastasis assay.
  • FIGURE 2F provides a graph of the number of lung metastatic nodules.
  • FIGURE 3A provides a heat map representation of the thermal stability of 804 soluble proteins in ovarian cancer cells treated with vehicle ⁇ DMSO (left) and SU056 (right).
  • FIGURE 3B graphs density distributions of protein Tm values calculated in SU056 treated cells and vehicle cells.
  • FIGURE 3C presents a graph of density distributions of Tm shifts between SU056 and vehicle treatment.
  • FIGURE 3D provides a scatter plot of Tm calculated in SU056 and vehicle treatment.
  • FIGURE 3E presents melting curves for six proteins with and without SU056 treatment.
  • FIGURE 3F provides a chart of change in melting temperature (T m ) of the top six proteins upon SU056 treatment.
  • FIGURE 4A depicts a Western blot analysis was performed for the top three targets identified by CETSA.
  • FIGURE 4B provides subset images for immunohistochemistry of tumor samples from ID8 tumor xenograft study.
  • FIGURE 4C shows a graph of % inhibition in respective OC cells treated with SU056 and YB ⁇ 1.
  • FIGURE 4D presents a graph of YB ⁇ 1 (IC50) of SU056 for OC cell.
  • FIGURE 4E presents graphs depicting YB ⁇ 1 inhibition time kinetics study for SU056 effect on OC cell lines.
  • FIGURE 5A presents the structure of biotinylated SU056.
  • FIGURE 5B depicts a pulldown assay using biotinylated SU056.
  • FIGURE 5C provides a representative sensogram for SU093.
  • FIGURE 5D provides a representative sensogram for S SU056.
  • FIGURE 5E presents a Western blot analysis conforming YB ⁇ 1 expression in transduced cells.
  • FIGURE 5F presents an image demonstrating the cellular effect of SU056 is dependent on YB ⁇ 1 expression.
  • FIGURE 5G provides a graph of IC 50 values of SU056 on different transduced OVCAR8 cells expressing SC, YBX1 shRNA1, YBX1 shRNA2.
  • FIGURE 6A depicts a cycloheximide chase assay to determine the effect of SU056 on YB ⁇ 1 protein stability and a graph of fold change over time.
  • FIGURE 6B depicts a SDS ⁇ PAGE and Western blot analysis performed for YB ⁇ 1, cell cycle, and apoptosis ⁇ associated markers.
  • FIGURE 6C and FIGURE 6D represent observed enrichment in the Apoptosis and RNA degradation pathway in proteins that increase in abundance upon treatment with SU056.
  • FIGURE 6E provides a graph of enrichment in the Spliceosome pathways observed in proteins that decrease in abundance upon treatment with SU056.
  • FIGURE 7A provides a chart representing the sensitizing effects of SU056 on the viability of OVCAR8 and SKOV3 cells in combination with paclitaxel treatment.
  • FIGURE 7B graphs an Alexa Fluor ⁇ 488 ⁇ tagged paclitaxel efflux assay showing SU056 cotreatment inhibits paclitaxel efflux.
  • FIGURE 7C presents an SDS ⁇ PAGE gel for immunoblotting of YB ⁇ 1 and MDR1.
  • OVCAR8 cells treated with either vehicle (C), paclitaxel, SU056 and paclitaxel + SU056.
  • FIGURE 7D provides microscopic images of spheroid at 10X magnification
  • FIGURE 7E provides graphs of spheroid formation quantified after 7 days of incubation
  • FIGURE 7F presents representative images of mice after 28 days of drug treatment showing tumor regression compared to control.
  • FIGURE 7G graphs tumor volume/mouse as a function of time.
  • FIGURE 7H tumor weight/mouse at the end of the study.
  • I Immunohistochemistry staining. Tumor sections were stained with Ki67, and slides were scored for KI67 staining. Data shown are mean ⁇ SD from 5 mice in each group. * P ⁇ 0.05, ** P ⁇ 0.01, *** P ⁇ 0.001 compared with respective control.
  • FIGURE 8A depicts a western blot analysis for cell lysates from control and SU056 treated cells exposed at 37 and 53°C temperature and analyzed for expression of YB ⁇ 1, TMSB10 and PSMB2.
  • FIGURE 8B depicts micrographs of cells imaged using confocal microscope for the mCherry ⁇ YB1 after 3 h treatment of SU056 (2.5 & 5 ⁇ M) at 10X magnification.
  • FIGURE 8C presents a graph representing 2500 cells treated with SU056
  • FIGURE 9 presents graphs representing the sensitizing effects of SU056 on the viability of OVCAR8 and SKOV3 cells in combination with paclitaxel treatment.
  • FIGURE 10 presents a graph representing pharmacokinetics of SU056.
  • FIGURE 11A provides graphs representing the growth inhibitory effect of SU056 evaluated using MTT assay.
  • FIGURE 11B provides an image representing colony formation from cells treated with SU056 and incubated further to 7 ⁇ 10 days.
  • FIGURE 11C depicts graphs representing the number of colonies formed after SU056 treatment.
  • FIGURE 11D provides graphs representing the effect of SU056 on cell cycle distribution in TNBC cells.
  • FIGURE 12A presents SDS ⁇ PAGE and western blot analysis for SU056 treatment inhibition of protein translation associated molecules among MDA ⁇ MB ⁇ 231b cells.
  • FIGURE 12B presents SDS ⁇ PAGE and western blot analysis for SU056 treatment inhibition of protein translation associated molecules among MDA ⁇ MB ⁇ 468 cells
  • FIGURE 12C presents SDS ⁇ PAGE and western blot analysis for SU056 treatment inhibition of protein translation associated molecules among SUM 159 cells.
  • FIGURE 13A presents a graph of tumor volume (MDA ⁇ MB ⁇ 231) as a function of time.
  • FIGURE 13B presents a graph of tumor weight (MDA ⁇ MB ⁇ 231) at the end of the study.
  • FIGURE 13C presents a graph of body weight (MDA ⁇ MB ⁇ 231) as a function of time.
  • FIGURE 13D presents a graph of tumor volume (MDA ⁇ MB ⁇ 468) as a function of time.
  • FIGURE 13E presents a graph of tumor weight (MDA ⁇ MB ⁇ 468) at the end of the study.
  • FIGURE 13F presents a graph of body weight (MDA ⁇ MB ⁇ 468) as a function of time.
  • FIGURE 13G provides representative images of tumor (MDA ⁇ MB ⁇ 231) at the end of study.
  • FIGURE 13H provides representative images of tumor (MDA ⁇ MB ⁇ 468) at the end of study.
  • FIGURE 13I presents a graph representing tumor volume (SUTI151 ⁇ PDX) as a function of time.
  • FIGURE 13J presents a graph representing tumor weight (SUTI151 ⁇ PDX) at the end of the study.
  • FIGURE 13K presents a graph representing Body weight (SUTI151 ⁇ PDX) as a function of time.
  • FIGURE 14A presents a graph of tumor volume (4T1) as a function of time in SU056 inhibition in a 4T1 tumor xenograft in BALB/c.
  • FIGURE 14B presents a graph of tumor weight (4T1) at the end of the study.
  • FIGURE 14C presents a graph of body weight (4T1) as a function of time.
  • FIGURE 14D provides representative images of tumor (4T1) at the end of study.
  • FIGURE 14E presents a graph of tumor volume (4T1) as a function of time.
  • FIGURE 14F presents a graph of tumor weight (4T1) at the end of the study.
  • FIGURE 14G presents a graph of body weight (4T1) as a function of time.
  • FIGURE 15A present a graph depicting SU056 treatment is well tolerated in mice and rat as reflected by change in body weight in mice.
  • FIGURE 15B present a graph depicting SU056 treatment is well tolerated in mice and rat as reflected by change in body weight in rat.
  • FIGURE 15C presents a table of data indicating SU056 treatment of different concentration did not cause death among mice.
  • FIGURE 15D presents a table of data indicating SU056 treatment of different concentration did not cause death among rats.
  • FIGURE 15E presents a graph showing SU056 had a mean half ⁇ life of 40 minutes.
  • FIGURE 16 presents normalized expression measurements of YBX1 in depicted cell lines.
  • FIGURES 17, 18, and 19 represent SDS ⁇ PAGE and western blots demonstrating SU056 treatment inhibited translation initiation factors in TNBC cells.
  • DETAILED DESCRIPTION OF THE INVENTION It is understood that, in the proviso above, that the rotational nature of phenyl rings that a compound in which R 2 is F or CF 3 and R 1 , R 3 , R 4 , and R 5 are each H indicates the same substitution pattern as when R 4 is F or CF 3 and R 1 , R 2 , R 3 , and R 5 are each H.
  • all variables when present are as defined for Formula (I) above, with the compound of Formula (Ia) having the proviso that, when R 2 is F or CF 3 , at least one of R 1 , R 3 , R 4 , and R 5 is not H.
  • Another embodiment comprises a compound of Formula (I) defined otherwise as above, wherein X, R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are as defined and subject to the provisos above, and n is an integer independently in each instance from the group of 1, 2, 3, 4, and 5.
  • Still another embodiment comprises a compound of Formula (I) defined otherwise as above, wherein X, R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are as defined and subject to the provisos above, and n is an integer independently in each instance from the group of 1, 2, 3, and 4.
  • a further embodiment comprises a compound of Formula (I) defined otherwise as above, wherein X, R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are as defined and subject to the provisos above, and n is an integer independently in each instance from the group of 1, 2, and 3.
  • a further embodiment comprises a compound of Formula (I) defined otherwise as above, wherein X, R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are as defined and subject to the provisos above, and n is an integer independently in each instance from the group of 2 and 3.
  • a further embodiment comprises a compound of Formula (I) defined otherwise as above, wherein X, R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are as defined and subject to the provisos above, and n is 2.
  • a further embodiment comprises a compound of Formula (I) defined otherwise as above, wherein X, R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are as defined and subject to the provisos above, and n is 3.
  • X, R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are as defined and subject to the provisos above, and n is 3.
  • R 6 is H.
  • R 1 , R 2 , R 3 , R 4 , and R 5 are each independently selected from the group of H, F, C 1 ⁇ C 4 fluoroalkyl, SF 5 , Cl, Br, I, OH, C 1 ⁇ C 4 alkyl, C 1 ⁇ C 4 alkoxy, CN, NO 2 , and OH; n is an integer selected independently in each instance from the group of 1, 2, 3, 4, 5, and 6; R 6 is selected from the group of H, C 1 ⁇ C 6 alkyl, C 3 ⁇ C 6 cycloalkyl, ⁇ (CH 2 ) n ⁇ C 3 ⁇ C 6 cycloalkyl, 3 ⁇ 6 ⁇ membered heterocycle, ⁇ (CH 2 ) n ⁇ 3 ⁇ 6 ⁇ membered heterocycle, phenyl, and ⁇ (CH 2 ) n ⁇ phenyl; wherein the C 1 ⁇ C 6 alkyl group is substituted with 0, 1,
  • a further embodiment comprises a compound of Formula (II), wherein R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are as defined and subject to the provisos above, and n is an integer independently in each instance from the group of 1, 2, and 3.
  • a further embodiment comprises a compound of Formula (II), wherein R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are as defined and subject to the provisos above, and n is an integer independently in each instance from the group of 2 and 3.
  • a further embodiment comprises a compound of Formula (II), wherein R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are as defined and subject to the provisos above, and n is 2.
  • a further embodiment comprises a compound of Formula (II), wherein R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are as defined and subject to the provisos above, and n is 3.
  • R 1 , R 2 , R 3 , R 4 , R 5 , and n are as defined for the particular embodiment, wherein R 6 is H.
  • a further embodiment provides a compound of Formula (III), wherein: n is an integer selected m the group of 1, 2, 3, 4, 5, and 6; R 1 , R 2 , R 3 , R 4 , and R 5 are each independently selected from the group of H, F, C 1 ⁇ C 4 fluoroalkyl, SF 5 , Cl, Br, I, OH, C 1 ⁇ C 4 alkyl, C 1 ⁇ C 4 alkoxy, CN, NO 2 , and OH; with the proviso that at least one of R 1 , R 2 , R 3 , R 4 , and R 5 is selected from the group of F, C 1 ⁇ C 4 fluoroalkyl, and SF 5 ; and with the proviso that, when R 2 is F or CF 3 , then at least one of R 1 , R 3 , R 4 , and R 5 is not H; or a pharmaceutically acceptable salt thereof.
  • R 1 , R 2 , R 3 , R 4 , and R 5 are each
  • n is an integer selected independently in each instance from the group of 1, 2, 3, 4, 5, and 6;
  • R 1 is selected from the group of F, C 1 ⁇ C 4 fluoroalkyl, and SF 5 ; and
  • R 2 , R 3 , R 4 , and R 5 are each independently selected from the group of H, F, C 1 ⁇ C 4 fluoroalkyl, SF 5 , Cl, Br, I, OH, C 1 ⁇ C 4 alkyl, C 1 ⁇ C 4 alkoxy, CN, NO 2 , and OH.
  • Yet another embodiment provides a compound of Formula (III), or a pharmaceutically acceptable salt thereof, wherein n is an integer selected independently in each instance from the group of 1, 2, 3, 4, 5, and 6; R 3 is selected from the group of F, C 1 ⁇ C 4 fluoroalkyl, and SF 5 ; and R 1 , R 2 , R 4 , and R 5 are each independently selected from the group of H, F, C 1 ⁇ C 4 fluoroalkyl, SF 5 , Cl, Br, I, OH, C 1 ⁇ C 4 alkyl, C 1 ⁇ C 4 alkoxy, CN, NO 2 , and OH.
  • a still further embodiment provides a compound of Formula (III), or a pharmaceutically acceptable salt thereof, wherein: n is an integer selected independently in each instance from the group of 1, 2, 3, 4, 5, and 6; R 1 , R 2 , R 3 , R 4 , and R 5 are each independently selected from the group of H, F, C 1 ⁇ C 4 fluoroalkyl, SF 5 , Cl, Br, I, OH, C 1 ⁇ C 4 alkyl, C 1 ⁇ C 4 alkoxy, CN, NO 2 , and OH; with the proviso that at least two of R 1 , R 2 , R 3 , R 4 , and R 5 are selected from the group of F, C 1 ⁇ C 4 fluoroalkyl, and SF 5 .
  • a still further embodiment provides a compound of Formula (III), or a pharmaceutically acceptable salt thereof, wherein: n is an integer selected independently in each instance from the group of 1, 2, 3, 4, 5, and 6; R 1 , R 2 , R 3 , R 4 , and R 5 are each independently selected from the group of H, F, C 1 ⁇ C 3 fluoroalkyl, SF 5 , Cl, Br, I, OH, C 1 ⁇ C 4 alkyl, C 1 ⁇ C 4 alkoxy, CN, NO 2 , and OH; with the proviso that at least two of R 1 , R 2 , R 3 , R 4 , and R 5 are selected from the group of F, C 1 ⁇ C 3 fluoroalkyl.
  • a still further embodiment provides a compound of Formula (III), or a pharmaceutically acceptable salt thereof, wherein: n is an integer selected independently in each instance from the group of 1, 2, 3, 4, 5, and 6; R 1 , R 2 , R 3 , R 4 , and R 5 are each independently selected from the group of H, F, CF 3 , SF 5 , Cl, Br, I, OH, C 1 ⁇ C 4 alkyl, C 1 ⁇ C 4 alkoxy, CN, NO 2 , and OH; with the proviso that at least two of R 1 , R 2 , R 3 , R 4 , and R 5 are selected from the group of F and CF 3 .
  • a still further embodiment provides a compound of Formula (III), or a pharmaceutically acceptable salt thereof, wherein: n is an integer selected independently in each instance from the group of 1, 2, 3, 4, 5, and 6; R 1 , R 2 , R 3 , R 4 , and R 5 are each independently selected from the group of H, F, CF 3 , SF 5 , Cl, Br, I, OH, C 1 ⁇ C 4 alkyl, C 1 ⁇ C 4 alkoxy, CN, NO 2 , and OH; with the proviso that at least two of R 1 , R 2 , R 3 , R 4 , and R 5 are F.
  • a further embodiment comprises a compound of Formula (III), wherein R 1 , R 2 , R 3 , R 4 , and R 5 are as defined and subject to the proviso above, and n is an integer independently in each instance from the group of 1, 2, and 3.
  • a further embodiment comprises a compound of Formula (III), wherein R 1 , R 2 , R 3 , R 4 , and R 5 are as defined and subject to the proviso above, and n is an integer independently in each instance from the group of 2 and 3.
  • a further embodiment comprises a compound of Formula (III), wherein R 1 , R 2 , R 3 , R 4 , and R 5 are as defined and subject to the proviso above, and n is 2.
  • a further embodiment comprises a compound of Formula (III), wherein R 1 , R 2 , R 3 , R 4 , and R 5 are as defined and subject to the proviso above, and n is 3.
  • compositions comprising a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, pharmaceutically acceptable co ⁇ crystal, ester, solvate, hydrate, isomer, tautomer, isotope, polymorph, or prodrug thereof, and a pharmaceutically acceptable carrier or excipient.
  • Methods of Treatment Each of the methods of treatment below will reference the use of a compound of Formula (I), or a pharmaceutically acceptable salt, pharmaceutically acceptable co ⁇ crystals, pharmaceutically acceptable esters, pharmaceutically acceptable solvates, hydrates, isomers (including optical isomers, racemates, or other mixtures thereof), tautomers, isotopes, polymorphs, and pharmaceutically acceptable prodrugs thereof.
  • each method provides additional embodiments wherein the compound or compounds used is of Formula (Ia), Formula (Ib), Formula (Ic), Formula (Id), Formula (II), Formula (III), SU056, etc., or a pharmaceutically acceptable salt, pharmaceutically acceptable co ⁇ crystal, ester, solvate, hydrate, isomer, tautomer, isotope, polymorph, or prodrug thereof.
  • a method of inhibiting YB1 protein activity in a subject comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, pharmaceutically acceptable co ⁇ crystal, ester, solvate, hydrate, isomer, tautomer, isotope, polymorph, or prodrug thereof.
  • a method of inhibiting YB1 protein activity in a subject experiencing a cancer comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, pharmaceutically acceptable co ⁇ crystal, ester, solvate, hydrate, isomer, tautomer, isotope, polymorph, or prodrug thereof.
  • the cancer experienced by the subject is selected from the group of a gynecological cancer (including ovarian, endometrial, fallopian tube, and cervical cancers), breast cancers, lung cancers, ovarian cancer, prostate cancer, colorectal cancer, and gastric cancer.
  • Methods of inhibiting YB1 protein activity may be used in treating other cancers that have been associated with YBX1 expression, including acute myeloid leukemia (Zhou et al., Journal of Experimental & Clinical Cancer Research (2021) 40:353), renal cell carcinoma (Ruan et al., Oncogene (2020) 39:6113 ⁇ 6128), bladder cancer (Xu et al., Oncotarget, 2017, Vol. 8, No. 39, pp. 65946 ⁇ 65956, osteosarcoma (Fujiwara ⁇ Okada et al., British Journal of Cancer (2013) 108, pp. 836 ⁇ 847), head and neck cancer (Kolk et al., British Journal of Cancer (2011) 105, pp.
  • a method of sensitizing cancer cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, pharmaceutically acceptable co ⁇ crystal, ester, solvate, hydrate, isomer, tautomer, isotope, polymorph, or prodrug thereof.
  • YBX1 YB1
  • a method of sensitizing cancer cells expressing YB1 (YBX1) protein in a subject to treatment with radiation comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, pharmaceutically acceptable co ⁇ crystal, ester, solvate, hydrate, isomer, tautomer, isotope, polymorph, or prodrug thereof.
  • YBX1 YBX1
  • the pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof may be administered in a regimen concurrently with an additional anticancer agent or radiation.
  • the pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof may be administered to a subject in need thereof in a dose or regimen prior to subsequent administration of a designated cancer agent or agents and/or radiation therapy.
  • the compound of Formula (I), or a pharmaceutically acceptable salt thereof may be administered for an initial period of time, such as from 1 to 7 days, followed in sequence by administration to the subject in need thereof of a designated cancer agent or agents and/or radiation therapy.
  • the compound of Formula (I), or a pharmaceutically acceptable salt thereof, and one or more designated cancer agent or agents and/or radiation therapy may be administered to the subject in need thereof in repeating sequential periods of time, such as from 1 to 14 days each, with or without a refractory period involving neither treatment in between each pair of administrations.
  • the compound of Formula (I) may be administered for an initial period of time, such as from 1 to 7 days, followed by a second period of co ⁇ administration to the subject in need thereof of both a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically effective amount of a designated cancer agent or agents and/or radiation therapy.
  • the cancer cells expressing YB1 (YBX1) protein in a subject sensitized to the treatments described herein are selected from the group of a gynecological cancer (including ovarian, endometrial, fallopian tube, and cervical cancers), leukemias, lymphomas, kidney cancer, bladder cancer, pancreatic cancer, head and neck cancer, breast cancers (including triple negative, ER ⁇ negative, ER ⁇ positive breast cancers, and progesterone ⁇ positive), lung cancers, ovarian cancer, prostate cancer, colorectal cancer, gastric cancer, and neuronal cancer (including gliomas).
  • a gynecological cancer including ovarian, endometrial, fallopian tube, and cervical cancers
  • leukemias including lymphomas, kidney cancer, bladder cancer, pancreatic cancer, head and neck cancer
  • breast cancers including triple negative, ER ⁇ negative, ER ⁇ positive breast cancers, and progesterone ⁇ positive
  • lung cancers ovarian cancer, prostate cancer, colore
  • Gynecological Cancers Provided is a method of treatment of gynecological cancers expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. Also provided is a method of treatment of gynecological cancers expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically effective amount of an mTOR inhibitor, or a pharmaceutically acceptable salt thereof.
  • the mTOR inhibitor is selected from the group of sirolimus, everolimus, deforolimus, and temsirolimus. Also provided is a method of enhancing the effect of an anticancer agent in a subject experiencing a gynecological cancer expressing YB1 (YBX1) protein, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, the treatment with a compound of Formula (I) sensitizes gynecological cancer cells expressing YB1 (YBX1) protein in the subject to the treatment of the anticancer agent.
  • the anticancer agent used to treat the gynecological cancer is an inhibitor or antagonist of phosphoinositide 3 ⁇ kinase (PI3K)/protein kinase B (Akt).
  • PI3K phosphoinositide 3 ⁇ kinase
  • Akt protein kinase B
  • the gynecological cancer expressing YB1 (YBX1) protein to be treated is ovarian cancer.
  • the gynecological cancer to be treated is endometrial cancer.
  • the gynecological cancer to be treated is cervical cancer.
  • a method of treatment of ovarian cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of cisplatin, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of ovarian cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of a taxane compound, or a pharmaceutically acceptable salt thereof.
  • the taxane compound used in the method of treatment of the gynecological cancers expressing YB1 (YBX1) protein discussed herein is selected from the group of paclitaxel, docetaxel, and cabazitaxel.
  • YB1 (YBX1) protein for each of the methods of sensitizing cancer cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent and/or radiation or inhibiting YB1 protein activity in a subject experiencing a cancer, there is a corresponding method with the initial step of detecting the presence or absence of expressed YB1 (YBX1) protein in sample cells of the cancer and, when YB1 (YBX1) protein is determined to be present in the sample cells, treating the subject experiencing the cancer in question as described for each method with a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt, pharmaceutically acceptable co ⁇ crystal, ester, solvate, hydrate, isomer, tautomer, isotope, polymorph, or prodrug thereof, and any other agent or agents indicated by the particular method.
  • a method of treatment of ovarian cancer expressing YB1 (YBX1) protein in a subject comprising the steps of: a) determining the presence or absence of expressed YB1 protein in an ovarian cancer tumor sample collected from the subject in need thereof; and b) when expressed YB1 protein is determined to be present in the ovarian cancer tumor sample, administering to the subject in need thereof: i) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and ii) a pharmaceutically effective amount of a taxane compound, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of fallopian tube cancer (fallopian tube carcinoma) expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of a taxane compound, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of fallopian tube cancer (fallopian tube carcinoma) expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; b) a pharmaceutically effective amount of a taxane compound, or a pharmaceutically acceptable salt thereof; and c) a pharmaceutically effective amount of carboplatin, or a pharmaceutically acceptable salt thereof.
  • the taxane compound in the methods of treating fallopian tube cancer is selected from the group of paclitaxel, albumin ⁇ bound paclitaxel, docetaxel, and cabazitaxel.
  • Prostate Cancer Also provided is a method of treatment of prostate cancer expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. Further provided is a method of inhibiting prostate cancer metastasis expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing prostate cancer expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • the administration of a compound of Formula (I) sensitizes the prostate cancer expressing YB1 (YBX1) protein in the subject to treatment with a taxane anticancer agent.
  • the taxane anticancer agent is selected from the group of paclitaxel, docetaxel, and cabazitaxel, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of prostate cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of a taxane compound selected from the group of paclitaxel, docetaxel, and cabazitaxel, or a pharmaceutically acceptable salt thereof.
  • the administration of a compound of Formula (I) sensitizes the prostate cancer expressing YB1 (YBX1) protein in the subject to treatment with an androgen receptor inhibitor anticancer agent.
  • the androgen receptor inhibitor is selected from the group of apalutamide, enzalutamide, darolutamide, and abiraterone acetate.
  • a method of treatment of prostate cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of an androgen receptor inhibitor compound selected from the group of apalutamide, enzalutamide, darolutamide, and abiraterone acetate , or a pharmaceutically acceptable salt thereof.
  • the apalutamide is administered to the subject in need thereof at a daily dosage of from about 100 mg to about 300 mg. In some embodiments, the apalutamide is administered at a dosage of about 240 mg per day.
  • the anticancer agent is a luteinizing hormone ⁇ releasing hormone (LHRH) agonist. In some embodiments, the LHRH agonist is selected from the group of leuprolide/leuprorelin, goserelin, triptorelin, buserelin, and histrelin.
  • a method of treatment of prostate cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of a luteinizing hormone ⁇ releasing hormone (LHRH) agonist compound selected from the group of leuprolide/leuprorelin, goserelin, triptorelin, buserelin, and histrelin, or a pharmaceutically acceptable salt thereof.
  • the anticancer agent is a luteinizing hormone ⁇ releasing hormone (LHRH) antagonist.
  • the LHRH agonist is degarelix.
  • a method of treatment of prostate cancer in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of degarelix, or a pharmaceutically acceptable salt thereof.
  • the anticancer agent is anti ⁇ androgen agent.
  • the anti ⁇ androgen agent is selected from the group of flutamide, bicalutamide, and nilutamide.
  • a method of treatment of prostate cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of an anti ⁇ androgen compound selected from the group of flutamide, bicalutamide, and nilutamide, or a pharmaceutically acceptable salt thereof.
  • the prostate cancer in question is an androgen ⁇ independent prostate cancer.
  • the prostate cancer in question is castration ⁇ sensitive prostate cancer.
  • the prostate cancer in question is metastatic castration ⁇ sensitive prostate cancer.
  • the prostate cancer expressing YB1 (YBX1) protein to be treated is non ⁇ metastatic castration ⁇ resistant prostate cancer.
  • the prostate cancer is hormone ⁇ refractory prostate cancer (HRPC).
  • HRPC hormone ⁇ refractory prostate cancer
  • Melanoma Also provided is a method of treatment of melanoma expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of inhibiting melanoma metastasis expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing melanoma cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of melanoma expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: c) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and d) a pharmaceutically effective amount of a PD ⁇ 1 inhibitor agent selected from the group of pembrolizumab and nivolumab, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of melanoma expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of atezolizumab, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of melanoma expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; b) a pharmaceutically effective amount of atezolizumab, or a pharmaceutically acceptable salt thereof; and c) a pharmaceutically effective amount of a third agent selected from the group of cobimetinib and vemurafenib, or a pharmaceutically acceptable salt thereof.
  • a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof a pharmaceutically effective amount of a CTLA ⁇ 4 inhibitor (such as ipilimumab).
  • a method of treatment of melanoma expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of interleukin ⁇ 2 (IL ⁇ 2).
  • Cisplatin resistance YB ⁇ 1 expression or overexpression has also been associated with resistance to cisplatin treatments in some cancers, including breast, bladder, and ovarian cancers.
  • the breast cancer to be treated is refractory to endocrine therapeutics, such as selective estrogen receptor modulators (SERMs), including tamoxifen and toremifene.
  • SERMs selective estrogen receptor modulators
  • the breast cancer is refractory to selective estrogen receptor degrader (SERDs), such as fulvestrant and elacestrant.
  • SESDs selective estrogen receptor degrader
  • the breast cancer to be treated is refractory to aromatase inhbitors, such as letrozole, anastrozole, exemestane, and testolactone.
  • YBX1 YBX1
  • a method of sensitizing a cancer expressing YB1 (YBX1) protein in a subject to treatment with a taxane compound comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of breast cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of inhibiting breast cancer metastasis expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing breast cancer cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • the method sensitizes breast cancer cells expressing YB1 (YBX1) protein in the subject in need thereof to treatment with one or more agents selected from the group of anthracyclines (such as doxorubicin, pegylated liposomal doxorubicin, and epirubicin), taxane compounds (such as paclitaxel, albumin ⁇ bound paclitaxel, docetaxel, and cabazitaxel), 5 ⁇ fluorouracil, capecitabine, cyclophosphamide, vinarelbine, gemcitabine, ixabepilone, eribulin, and platinum agents (such as carboplatin and cisplatin).
  • anthracyclines such as doxorubicin, pegylated liposomal doxorubicin, and epirubicin
  • taxane compounds such as paclitaxel, albumin ⁇ bound paclitaxel, docetaxel, and cabazitaxel
  • the method sensitizes breast cancer cells expressing YB1 (YBX1) to treatment with radiation therapy.
  • An embodiment provides a method of treatment of breast cancer expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anticancer agents selected from the group of doxorubicin, pegylated liposomal doxorubicin, epirubicin, paclitaxel, docetaxel, 5 ⁇ fluorouracil, capecitabine, cyclophosphamide, and carboplatin, or a pharmaceutically acceptable salt thereof.
  • Another embodiment provides a method of treatment of breast cancer expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anticancer agents selected from the group of paclitaxel, albumin ⁇ bound paclitaxel, docetaxel, doxorubicin, pegylated liposomal doxorubicin, epirubicin, cisplatin, carboplatin, vinorelbine, capecitabine, gemcitabine, ixabepilone, and eribulin, or a pharmaceutically acceptable salt thereof.
  • a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof selected from the group of paclitaxel, albumin ⁇ bound paclitaxel, docetaxel, doxorubicin, pegylated lip
  • Colorectal Cancer Also provided is a method of treatment of colorectal cancer expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. Further provided is a method of inhibiting colorectal cancer metastasis expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing colorectal cancer cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of colorectal cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of an anti ⁇ cancer agent selected from the group of 5 ⁇ fluorouracil, capecitabine, irinotecan, oxaliplatin, and trifluridine and tipiracil, or a pharmaceutically acceptable salt thereof.
  • Bladder Cancer Also provided is a method of treatment of bladder cancer expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. Further provided is a method of inhibiting bladder cancer metastasis expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing bladder cancer cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of bladder cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: d) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; e) a pharmaceutically effective amount of an anticancer agent selected from the group of cisplatin, cisplatin plus 5 ⁇ fluorouracil, and mitomycin with 5 ⁇ fluorouracil, or a pharmaceutically acceptable salt thereof; f) a therapeutically effective dose of radiation.
  • a method of treatment of bladder cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of an anticancer agent selected from the group of i) gemcitabine and cisplatin; ii) Dose ⁇ dense methotrexate, vinblastine, doxorubicin (Adriamycin), and cisplatin (DDMVAC); iii) Cisplatin, methotrexate, and vinblastine (CMV); and iv) Gemcitabine and paclitaxel
  • a method of treatment of bladder cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; b
  • Liver Cancer Also provided is a method of treatment of liver cancer expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. Further provided is a method of inhibiting liver cancer metastasis expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing liver cancer cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of liver cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of an anti ⁇ cancer agent selected from the group of gemcitabine, oxaliplatin, cisplatin, doxorubicin, 5 ⁇ fluorouracil, capecitabine, and mitoxantrone, or a pharmaceutically acceptable salt thereof.
  • an anti ⁇ cancer agent selected from the group of gemcitabine, oxaliplatin, cisplatin, doxorubicin, 5 ⁇ fluorouracil, capecitabine, and mitoxantrone, or a pharmaceutically acceptable salt thereof.
  • Lung Cancer Also provided is a method of treatment of small cell lung cancer expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. Further provided is a method of inhibiting small cell lung cancer metastasis expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing small cell lung cancer cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of small cell lung cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of an anti ⁇ cancer agent selected from the group of cisplatin and etoposide, carboplatin and etoposide, cisplatin and irinotecan, and carboplatin and irinotecan, or a pharmaceutically acceptable salt thereof.
  • an anti ⁇ cancer agent selected from the group of cisplatin and etoposide, carboplatin and etoposide, cisplatin and irinotecan, and carboplatin and irinotecan, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of non ⁇ small cell lung cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of inhibiting non ⁇ small cell lung cancer metastasis expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing non ⁇ small cell lung cancer cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of non ⁇ small cell lung cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anti ⁇ cancer agents selected from the group of cisplatin, carboplatin, paclitaxel, albumin ⁇ bound paclitaxel, docetaxel, gemcitabine, vinarelbine, etoposide, and premetrexed, or a pharmaceutically acceptable salt thereof.
  • Multiple Myeloma Also provided is a method of treatment of multiple myeloma expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. Further provided is a method of inhibiting multiple myeloma metastasis expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing multiple myeloma cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of multiple myeloma expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of an anti ⁇ cancer agent selected from the group of Melphalan, vincristine, cyclophosphamide, etoposide, doxorubicin, liposomal doxorubicin, and bendamustine, or a pharmaceutically acceptable salt thereof.
  • an anti ⁇ cancer agent selected from the group of Melphalan, vincristine, cyclophosphamide, etoposide, doxorubicin, liposomal doxorubicin, and bendamustine, or a pharmaceutically acceptable salt thereof.
  • Soft Tissue Sarcomas Also provided herein are methods of treatment of soft tissue sarcomas expressing YB1 (YBX1) protein, including angiosarcoma, dermatofibrosarcoma protuberans, epitheloid sarcoma, gastrointestinal stromal tumor (GIST), Kaposi’s sarcoma, Leiomyosarcoma, liposarcoma, malignant peripheral nerve sheath tumors, myxofibrosarcoma, rhabdomyosarcoma, solitary fibrous tumors, synovial sarcoma, and undifferentiated pleomorphic sarcoma.
  • YBX1 YB1
  • a method of treatment of soft tissue sarcomas expressing YB1 (YBX1) protein, such as fibrosarcoma expressing YB1 (YBX1) protein, in a subject comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of inhibiting soft tissue sarcomas expressing YB1 (YBX1) protein such as fibrosarcoma expressing YB1 (YBX1) protein, in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing soft tissue sarcoma cells expressing YB1 (YBX1) protein, such as fibrosarcoma cells expressing YB1 (YBX1) protein, cells in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of soft tissue sarcomas expressing YB1 (YBX1) protein, such as fibrosarcoma expressing YB1 (YBX1) protein, in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anti ⁇ cancer agents selected from the group of ifosfamide, doxorubicin, dacarbazine (DTIC), epirubicin, temozolomide, docetaxel, gemcitabine, vinorelbine, trabectedin, and eribulin, or a pharmaceutically acceptable salt thereof.
  • a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof selected from the group of ifosfamide, doxorubicin, dacarbazine (DTIC), epirubicin, temozolomide, docetaxe
  • the drug mesna is also given to protect the bladder from the toxic effects of ifosfamide.
  • the anti ⁇ cancer agent is a combination of mesna, Adriamycin [doxorubicin], ifosfamide, and dacarbazine, sometimes referred to by the acronym MAID.
  • the anti ⁇ cancer agent is a combination of Adriamycin [doxorubicin], ifosfamide, and mesna, sometimes referred to by the acronym AIM.
  • the anti ⁇ cancer agent or agents are administered to the subject in need thereof using isolated limb perfusion.
  • Osteosarcomas Also provided is a method of treatment of osteosarcoma expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. Further provided is a method of inhibiting osteosarcoma expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing osteosarcoma cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of osteosarcoma expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anti ⁇ cancer agents selected from the group of methotrexate, doxorubicin, cisplatin, carboplatin, Ifosfamide, cyclophosphamide, etoposide, and gemcitabine, or a pharmaceutically acceptable salt thereof.
  • the anti ⁇ cancer agent is a combination of High ⁇ dose methotrexate, doxorubicin, and cisplatin (the MAP regimen).
  • a combination of doxorubicin and cisplatin are administered.
  • a combination of ifosfamide and etoposide are used.
  • a combination is administered of ifosfamide and epirubicin with either cisplatin or carboplatin.
  • Ewing’s Sarcoma Also provided is a method of treatment of Ewing’s sarcoma expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. Further provided is a method of inhibiting Ewing’s sarcoma expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing Ewing’s sarcoma cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of Ewing’s sarcoma expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anti ⁇ cancer agents selected from the group of cyclophosphamide, doxorubicin, etoposide, Ifosfamide, and vincristine, or a pharmaceutically acceptable salt thereof.
  • the anti ⁇ cancer agent is a combination of vincristine, doxorubicin, and cyclophosphamide, alternating with ifosfamide and etoposide, the regimen referred to as VDC/IE.
  • Gastric cancers Also provided is a method of treatment of gastric (stomach) cancer expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of inhibiting gastric cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing gastric cancer cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of gastric cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anti ⁇ cancer agents selected from the group of 5 ⁇ fluorouracil, capecitabine, carboplatin, cisplatin, docetaxel, epirubicin, irinotecan, oxaliplatin, paclitaxel, and trifluridine + tipracil (LONSURF®) or a pharmaceutically acceptable salt thereof.
  • a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof selected from the group of 5 ⁇ fluorouracil, capecitabine, carboplatin, cisplatin, docetaxel, epirubicin, irinotecan, oxaliplatin, paclitaxel, and triflur
  • the anti ⁇ cancer agent is a combination of epirubicin, cisplatin, and 5 ⁇ fluorocil, sometimes referred to by the acronym ECF.
  • ECF epirubicin
  • the combination of docetaxel or paclitaxel with either 5 ⁇ FU or capecitabine sometimes combined with radiation.
  • cisplatin is administered with either 5 ⁇ FU or capecitabine, sometimes combined with radiation.
  • paclitaxel and carboplatin are administered, sometimes combined with radiation.
  • the combination of docetaxel, cisplatin, and 5 ⁇ fluoruracil (DCF) are administered.
  • irinotecan is administered along with cisplatin, 5 ⁇ flourouracil, or capecitabine.
  • oxaliplatin is administered with 5 ⁇ fluorouracil or capecitabine.
  • trifluridine + tipracil is given.
  • Glioblastoma Also provided is a method of treatment of glioblastoma multiforme (GBM or glioblastoma) expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of inhibiting glioblastoma expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing glioblastoma cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of glioblastoma expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anti ⁇ cancer agents selected from the group of temozolomide, bevacizumab, lomustine, carmustine, fluzoparil, pembrolizumab, nivolumab, ipilimumab, anlotinib, glasdegib, and bavituximab, or a pharmaceutically acceptable salt thereof.
  • the glioblastoma in the methods above is a pediatric glioblastoma expressing YB1 (YBX1) protein.
  • the glioblastoma is a primary glioblastoma expressing YB1 (YBX1) protein. In others, it is a secondary glioblastoma expressing YB1 (YBX1) protein.
  • Head and Neck Cancer refers to any of the cancers of the oral cavity, throat (pharynx, including the nasopharynx, oropharynx, and hypopharynx), larynx, paranasal sinuses, nasal cavity, and salivary glands.
  • the head and neck cancers include Hypopharyngeal cancer, laryngeal cancer, lip and oral cavity cancer, metastatic squamous neck cancer, nasopharyngeal cancer, oropharyngeal cancer, paranasal sinus and nasal cavity cancer, and salivary gland cancer. It is understood that, for each of the methods of treatment of head and neck cancer described herein, disclosed also is the corresponding method for each of the head and neck cancers listed in this paragraph. Also provided is a method of treatment of head and neck cancer expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • YBX1 YBX1
  • a method of inhibiting head and neck cancer metastasis expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing head and neck cancer expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • the anticancer agent used to treat the subject is radiation therapy.
  • the radiation utilized is external ⁇ beam radiation therapy.
  • the treatment comprises administering a pharmaceutically effective amount of one or more EGFR inhibitors to the subject in need thereof.
  • Other embodiments concern respectively administering a pharmaceutically effective amount of larotrectinib (Vitrakvi) and/or larotrectinib to the subject in need thereof.
  • Other methods of treating head and neck cancer comprise the use of immunotherapy, such as the administration a pharmaceutically effective amount of pembrolizumab and/or nivolumab to the subject in need thereof.
  • a method of treatment of head and neck cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of an anticancer agent selected from the group of paclitaxel, docetaxel, cisplatin, carboplatin, 5 ⁇ fluorouracil, methotrexate, and capecitabine, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of head and neck cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of a taxane compound selected from the group of paclitaxel and docetaxel, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of head and neck cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of a cisplatin, or a pharmaceutically acceptable salt thereof.
  • the cisplatin is administered to the subject in need thereof at a dose of from about 20 mg/m 2 to about 100 mg/m 2 delivered every 3 weeks ⁇ 3.
  • a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof a pharmaceutically effective amount of a carboplatin, or a pharmaceutically acceptable salt thereof
  • a pharmaceutically effective amount of a drug selected from the group of 5 ⁇ fluorouracil (5FU) and cetuximab selected from the group of 5 ⁇ fluorouracil (5FU) and cetuximab.
  • a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof a pharmaceutically effective amount of cisplatin, or a pharmaceutically acceptable salt thereof
  • a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof a pharmaceutically effective amount of hydroxyurea, or a pharmaceutically acceptable salt thereof
  • a method of treatment of nasopharyngeal cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of hydroxyurea, or a pharmaceutically acceptable salt thereof; and c) a pharmaceutically effective amount of a drug selected from the group of carboplatin, doxorubicin, epirubicin, paclitaxel, docetaxel, gemcitabine, bleomycin, and methotrexate.
  • a drug selected from the group of carboplatin, doxorubicin, epirubicin, paclitaxel, docetaxel, gemcitabine, bleomycin, and methotrexate selected from the group of carboplatin, doxorubicin, epirubicin, paclitaxel, docetaxel, gemcitabine, bleomycin, and
  • Pancreatic Cancer Provided is a method of treatment of pancreatic cancer expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. Further provided is a method of inhibiting pancreatic cancer metastasis expressing YB1 (YBX1) protein in a subject, the method comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing pancreatic cancer expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of pancreatic cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anticancer agents selected from the group of gemcitabine, 5 ⁇ fluoruracil, oxaliplatin, paclitaxel, albumin ⁇ bound paclitaxel, docetaxel, capecitabine, cisplatin, and irinotecan, or a pharmaceutically acceptable salt thereof.
  • the compounds of Formula (I) may also be used to treat and/or sensitize to treatment neuronal cancers (brain and spinal cord cancers), including medulloblastoma, glioblastoma multiforme (GBM), astrocytomas (anapastic astrocytomas and pilocytic astrocytomas), ependymomas, and oligodendrogliomas. It is understood for each of the following methods for treating a neuronal cancer or sensitizing a neuronal cancer to treatment described herein, included separate methods of each type for each of the neuronal cancers listed in this paragraph.
  • a method of treatment of neuronal cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of inhibiting neuronal cancer metastasis expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of sensitizing neuronal cancer expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • a method of treatment of neuronal cancer expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anticancer agents selected from the group of carboplatin, carmustine (BCNU), cisplatin, irinotecan, cyclophosphamide, etoposide, lomustine, methotrexate, procarbazine, temozolomide, and vincristine, or a pharmaceutically acceptable salt thereof.
  • BCNU carmustine
  • cisplatin irinotecan
  • cyclophosphamide etoposide
  • lomustine lomustine
  • methotrexate procarbazine
  • procarbazine temozolomide
  • vincristine or a pharmaceutically acceptable salt thereof.
  • the pharmaceutically effective amount of carmustine administered to the subject in need thereof is in the form of a carmustine wafer or implant, such as that in the GLIADEL® Wafer (carmustine implant) product available from Arbor Pharmaceuticals, LLC.
  • Leukemias Methods of the present invention also include those for the treatment of leukemias, wherein the leukemia cells in question express YB ⁇ 1 protein, including acute myeloid leukemia (AML), Chronic myelogenous leukemia (CML), acute lymphoblastic (or lymphocytic) leukemia (ALL), and chronic lymphocytic leukemia (CLL).
  • a method of sensitizing leukemia cells expressing YB1 (YBX1) protein in a subject to treatment with an anticancer agent comprising administering to the subject in need thereof a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.
  • the anticancer agent used to treat the subject is radiation therapy.
  • a method of treatment of acute myeloid leukemia expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of an anthracycline drug selected from the group of daunorubicin and idarubicin, or a pharmaceutically acceptable salt thereof; and c) a pharmaceutically effective amount of cytarabine, or a pharmaceutically acceptable salt thereof
  • a method of treatment of acute myeloid leukemia expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anticancer agents selected from the group of cladribine (2 ⁇ CdA), fludar
  • a method of treatment of chronic myeloid leukemia expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anticancer agents selected from the group of hydroxyurea, cytarabine (Ara ⁇ C), busulfan, cyclophosphamide (CYTOXAN ® ), and vincristine (ONCOVIN ® ), or a pharmaceutically acceptable salt thereof
  • a method of treatment of chronic myeloid leukemia expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more tyrosine kinase inhibitor
  • a method of treatment of chronic myeloid leukemia expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of interferon ⁇ alpha, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of acute lymphoblastic leukemia expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anticancer agents selected from the group of vincristine, dexamethasone, imatinib, prednisone, doxorubicin and daunorubicin, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of acute lymphoblastic leukemia expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anticancer agents selected from the group of methotrexate, 6 ⁇ mercaptopurine, vincristine, prednisone, and imatinib, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of acute lymphoblastic leukemia expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anticancer agents selected from the group of vincristine, dexamethasone, prednisone, doxorubicin, and daunorubicin, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of acute lymphoblastic leukemia expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anticancer agents selected from the group of methotrexate, 6 ⁇ mercaptopurine (6 ⁇ MP), vincristine, prednisone, and imatinib, or a pharmaceutically acceptable salt thereof.
  • a method of treatment of chronic lymphocytic leukemia expressing YB1 (YBX1) protein in a subject comprising administering to the subject in need thereof: a) a pharmaceutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically effective amount of one or more anticancer agents selected from the group of ibrutinib, acalabrutinib, idelalisib, and duvelisib, or a pharmaceutically acceptable salt thereof.
  • YB1 and YBX1 refer to Y box binding protein 1, also known as Y ⁇ box transcription factor or nuclease ⁇ sensitive element ⁇ binding protein 1, a protein that in humans is encoded by the YBX1 gene.
  • the wavy line ( ) in chemical structures indicates a bond through which the structure shown is bound to another chemical moiety or group.
  • a “heterocycle” or “heterocyclic group” herein refers to a chemical ring containing carbon atoms and at least one ring heteroatom selected from O, S, and N.
  • heterocycle and heterocyclic include groups with saturated rings, partially unsaturated rings, and aromatic rings (i.e., heteroaromatic rings).
  • Examples of 5 ⁇ membered and 6 ⁇ membered heterocycles include by way of example and not limitation pyridyl, dihydroypyridyl, tetrahydropyridyl (piperidyl), thiazolyl, tetrahydrothiophenyl, sulfur oxidized tetrahydrothiophenyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl, 4 ⁇ piperidinyl, pyrrolidinyl, 2 ⁇ pyrrolidonyl, pyrrolinyl, tetrahydrofuranyl, triazinyl, 6H ⁇ 1,2,5 ⁇ thiadiazinyl, 2H,6H ⁇ 1,5,2 ⁇ dithiazinyl, thienyl, thianthrenyl, pyranyl, 2H ⁇ pyrrolyl, isothiazolyl, isoxazolyl,
  • alkyl refers to a straight or branched hydrocarbon.
  • an alkyl group can include those having 1 to 6 carbon atoms (i.e, C 1 ⁇ C 6 alkyl), 1 to 4 carbon atoms (i.e., C 1 ⁇ C 4 alkyl), or 1 to 3 carbon atoms (i.e., C 1 ⁇ C 3 alkyl).
  • alkyl groups include, but are not limited to, methyl, ethyl, n ⁇ propyl, isopropyl ( ⁇ CH(CH 3 ) 2 ), 1 ⁇ butyl (n ⁇ Bu, n ⁇ butyl, ⁇ CH 2 CH 2 CH 2 CH 3 ), 2 ⁇ methyl ⁇ 1 ⁇ propyl (i ⁇ Bu, i ⁇ butyl, ⁇ CH 2 CH(CH 3 ) 2 ), 2 ⁇ butyl (s ⁇ Bu, s ⁇ butyl, ⁇ CH(CH 3 )CH 2 CH 3 ), 2 ⁇ methyl ⁇ 2 ⁇ propyl (t ⁇ Bu, t ⁇ butyl, ⁇ C(CH 3 ) 3 ), 1 ⁇ pentyl (n ⁇ pentyl, ⁇ CH 2 CH 2 CH 2 CH 2 CH 3 ), 2 ⁇ pentyl ( ⁇ CH(CH 3 )CH 2 CH 2 CH 3 ), 3 ⁇ pentyl ( ⁇ CH(CH 2 CH 3 ) 2 ), 2 ⁇ methyl ⁇ 2 ⁇ butyl ( ⁇ C(CH 3
  • alkoxy refers to a group having the formula ⁇ O ⁇ alkyl, in which an alkyl group, as defined above, is attached to the parent molecule via an oxygen atom.
  • the alkyl portion of an alkoxy group can have 1 to 6 carbon atoms (i.e., C 1 ⁇ C 6 alkoxy), 1 to 4 carbon atoms (i.e., C 1 ⁇ C 4 alkoxy), or 1 to 3 carbon atoms (i.e., C 1 ⁇ C 3 alkoxy).
  • alkoxy groups include, but are not limited to, methoxy ( ⁇ O ⁇ CH 3 or ⁇ OMe), ethoxy ( ⁇ OCH 2 CH 3 or ⁇ OEt), n ⁇ propoxy ( ⁇ CH 2 ⁇ CH 2 ⁇ CH 3 ), isopropoxy ( ⁇ CH(CH 3 ) 2 ), n ⁇ butyl ( ⁇ CH 2 ⁇ CH 2 ⁇ CH 2 ⁇ CH 3 ), isobutoxy ( ⁇ CH 2 ⁇ CH(CH 3 ) 2 ), sec ⁇ butoxy (–CH(CH 3 )CH 2 ⁇ CH 3 ), t ⁇ butoxy ( ⁇ O ⁇ C(CH 3 ) 3 or ⁇ OtBu), and the like.
  • cycloalkyl refers to a saturated ring having 3 to 6 carbon atoms as a monocycle, including cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl groups.
  • subject refers to an animal, such as a mammal, that has been or will be the object of treatment, observation or experiment. The methods described herein may be useful in both human therapy and veterinary applications.
  • the subject is a mammal; in some embodiments the subject is human; and in some embodiments the subject is chosen from cats and dogs.
  • Subject in need thereof or “human in need thereof” refers to a subject, such as a human, who may have or is suspected to have diseases or conditions that would benefit from certain treatment; for example treatment with a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt or co ⁇ crystal thereof, as described herein.
  • a “subject in need thereof” concerning a method of treatment herein is a patient from whom a tumor sample, such as from a tumor biopsy, is taken and the presence of expressed YBX1 protein is identified in the sampled material, such as through immunohistochemical or Western Blotting techniques known in the art.
  • the terms “effective amount,” “therapeutically effective amount,” or “pharmaceutically effective amount” refer to an amount that is sufficient to effect treatment, as defined below, when administered to a subject (e.g., a mammal, such as a human) in need of such treatment.
  • an “effective amount,” “therapeutically effective amount,” or a “pharmaceutically effective amount” of a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt or co ⁇ crystal thereof is an amount sufficient to modulate YXB1 expression or activity, and thereby treat a subject (e.g., a human) suffering an indication, or to ameliorate or alleviate the existing symptoms of the indication.
  • a therapeutically or pharmaceutically effective amount may be an amount sufficient to decrease a symptom of a disease or condition responsive to inhibition of YXB1 activity.
  • an “effective amount” is an amount of a subject compound that, when administered to an individual in one or more doses, in monotherapy or in combination therapy, is effective to inhibit YB ⁇ 1 by about 20% (20% inhibition), at least about 30% (30% inhibition), at least about 40% (40% inhibition), at least about 50% (50% inhibition), at least about 60% (60% inhibition), at least about 70% (70% inhibition), at least about 80% (80% inhibition), or at least about 90% (90% inhibition), compared to the YB ⁇ 1 activity in the individual in the absence of treatment with the compound, or alternatively, compared to the YB ⁇ 1 activity in the individual before or after treatment with the compound.
  • an “effective amount” is an amount of a subject compound that, when administered to an individual in one or more doses, in monotherapy or in combination therapy, is effective to decrease tumor burden in the subject by about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%, compared to tumor burden in the individual in the absence of treatment with the compound, or alternatively, compared to the tumor burden in the subject before or after treatment with the compound.
  • tumor burden refers to the total mass of tumor tissue carried by a subject with cancer.
  • an “effective amount” is an amount of a subject compound that, when administered to an individual in one or more doses, in monotherapy or in combination therapy, is effective to reduce the dose of radiotherapy required to observe tumor shrinkage in the subject by about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%, compared to the dose of radiotherapy required to observe tumor shrinkage in the individual in the absence of treatment with the compound.
  • an “effective amount” of a compound is an amount that, when administered in one or more doses to an individual having cancer, is effective to achieve a 1.5 ⁇ log, a 2 ⁇ log, a 2.5 ⁇ log, a 3 ⁇ log, a 3.5 ⁇ log, a 4 ⁇ log, a 4.5 ⁇ log, or a 5 ⁇ log reduction in tumor size.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof may be administered at a daily dose of from about 0.2 mg/kg to about 10 mg/kg. In other embodiments, it may be administered at a daily dose of from about 0.2 mg/kg to about 5 mg/kg.
  • an effective amount of a compound is an amount that ranges from about 50 ng/kg body weight to about 50 pg/kg body weight (e.g., from about 50 ng/kg body weight to about 40 pg/kg body weight, from about 30 ng/kg body weight to about 20 pg/kg body weight, from about 50 ng/kg body weight to about 10 pg/kg body weight, from about 50 ng/kg body weight to about 1 pg/kg body weight, from about 50 ng/kg body weight to about 800 ng/kg body weight, from about 50 ng/kg body weight to about 700 ng/kg body weight, from about 50 ng/kg body weight to about 600 ng/kg body weight, from about 50 ng/kg body weight to about 500 ng/kg body weight, from about 50 ng/kg body weight to about 400 ng/kg body weight, from about 60 ng/kg body weight to about 400 ng/kg body weight, from about 70 ng/kg body weight to about 300 ng/kg body weight
  • an effective amount of a compound is an amount that ranges from about 10 pg to about 100 mg, e.g., from about 10 pg to about 50 pg, from about 50 pg to about 150 pg, from about 150 pg to about 250 pg, from about 250 pg to about 500 pg, from about 500 pg to about 750 pg, from about 750 pg to about 1 ng, from about 1 ng to about 10 ng, from about 10 ng to about 50 ng, from about 50 ng to about 150 ng, from about 150 ng to about 250 ng, from about 250 ng to about 500 ng, from about 500 ng to about 750 ng, from about 750 ng to about 1 pg, from about 1 pg to about 10 pg, from about 10 pg to about 50 pg, from about 50 mg to about 150 gg, from about 150 gg to about 250 gg, from about 250 gg to about 500 gg, from about 500
  • the amount can be a single dose amount or can be a total daily amount.
  • the total daily amount can range from 10 pg to 100 mg, or can range from 100 mg to about 500 mg, or can range from 500 mg to about 1000 mg.
  • a single dose of a compound is administered.
  • multiple doses are administered. Where multiple doses are administered over a period of time, the compound can be administered twice daily (qid), daily (qd), every other day (qod), every third day, three times per week (tiw), or twice per week (biw) over a period of time.
  • a compound is administered qid, qd, qod, tiw, or biw over a period of from one day to about 2 years or more.
  • a compound is administered at any of the aforementioned frequencies for one week, two weeks, one month, two months, six months, one year, or two years, or more, depending on various factors.
  • Administration of an effective amount of a subject compound to an individual with cancer can result in one or more of: 1) a reduction in tumor burden; 2) a reduction in the dose of radiotherapy required to effect tumor shrinkage (e.g.
  • a treatment method e.g., a biological sample obtained from an individual who has been treated with a subject method can be assayed.
  • inhibiting indicates a decrease, such as a significant decrease, in the baseline activity of a biological activity or process.
  • “Inhibition of YB ⁇ 1 activity” refers to a decrease in YB ⁇ 1 activity as a direct or indirect response to the presence of a compound of Formula I, or a pharmaceutically acceptable salt or co ⁇ crystal thereof, relative to the activity of YB ⁇ 1 in the absence of such compound or a pharmaceutically acceptable salt or co ⁇ crystal thereof.
  • the decrease in activity may be due to the direct interaction of the compound with YB ⁇ 1, or due to the interaction of the compound(s) described herein with one or more other factors that in turn affect YB ⁇ 1 activity.
  • the presence of the compound(s) may decrease YB ⁇ 1 activity by directly binding to the YB ⁇ 1, by causing (directly or indirectly) another factor to decrease YB ⁇ 1 activity, or by (directly or indirectly) decreasing the amount of YB ⁇ 1 present in the cell or organism.
  • the inhibition of YB ⁇ 1 activity may be compared in the same subject prior to treatment, or other subjects not receiving the treatment.
  • the term “inhibitor” is understood to refer to a compound or agent that, upon administration to a human in need thereof at a pharmaceutically or therapeutically effective dose, provides the inhibition activity desired.
  • composition refers to a composition containing a pharmaceutically effective amount of one or more of the isotopic compounds described herein, or a pharmaceutically acceptable salt thereof, formulated with a pharmaceutically acceptable carrier, which can also include other additives, and manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal.
  • compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gelcap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or in any other formulation described herein.
  • unit dosage form e.g., a tablet, capsule, caplet, gelcap, or syrup
  • topical administration e.g., as a cream, gel, lotion, or ointment
  • intravenous administration e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use
  • pharmaceutically acceptable excipient is a pharmaceutically acceptable vehicle that includes, without limitation, any and all carriers, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutically active substances is well known in the art.
  • compositions can also be incorporated into the compositions.
  • pharmaceutically acceptable carrier refers to any ingredient in a pharmaceutical composition other than the disclosed pharmaceutically active or therapeutic compounds, or a pharmaceutically acceptable salt thereof (e.g., a carrier capable of suspending or dissolving the active isotopic compound) and having the properties of being nontoxic and non ⁇ inflammatory in a patient.
  • Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspensing or dispersing agents, sweeteners, or waters of hydration.
  • excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, B
  • salts include, for example, salts with inorganic acids and salts with an organic acid.
  • salts may include hydrochloride, phosphate, diphosphate, hydrobromide, sulfate, sulfinate, nitrate, malate, maleate, fumarate, tartrate, succinate, citrate, acetate, lactate, methanesulfonate (mesylate), benzenesuflonate (besylate), p ⁇ toluenesulfonate (tosylate), 2 ⁇ hydroxyethylsulfonate, benzoate, salicylate, stearate, and alkanoate (such as acetate, HOOC ⁇ (CH 2 ) n ⁇ COOH where n is 0 ⁇ 4).
  • the free base can be obtained by basifying a solution of the acid salt.
  • an addition salt particularly a pharmaceutically acceptable addition salt, may be produced by dissolving the free base in a suitable organic solvent and treating the solution with an acid, in accordance with conventional procedures for preparing acid addition salts from base compounds.
  • Those skilled in the art will recognize various synthetic methodologies that may be used to prepare nontoxic pharmaceutically acceptable addition salts.
  • a compound of Formula I are the pharmaceutically acceptable salts, pharmaceutically acceptable co ⁇ crystals, pharmaceutically acceptable esters, pharmaceutically acceptable solvates, hydrates, isomers (including optical isomers, racemates, or other mixtures thereof), tautomers, isotopes, polymorphs, and pharmaceutically acceptable prodrugs of such compounds.
  • crystal forms and related terms herein refer to the various crystalline modifications of a given substance, including, but not limited to, polymorphs, solvates, hydrates, co ⁇ crystals, and other molecular complexes, as well as salts, solvates of salts, hydrates of salts, other molecular complexes of salts, and polymorphs thereof. Crystal forms of a substance can be obtained by a number of methods, as known in the art.
  • Such methods include, but are not limited to, melt recrystallization, melt cooling, solvent recrystallization, recrystallization in confined spaces such as, e.g., in nanopores or capillaries, recrystallization on surfaces or templates, such as, e.g., on polymers, recrystallization in the presence of additives, such as, e.g., co ⁇ crystal counter ⁇ molecules, desolvation, dehydration, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, sublimation, grinding and solvent ⁇ drop grinding.
  • additives such as, e.g., co ⁇ crystal counter ⁇ molecules, desolvation, dehydration, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, sublimation, grinding and solvent ⁇ drop grinding.
  • refractory used herein in regard to a cancer refers to a cancer that does not respond to one or more treatments. In some embodiments, the cancer does not respond to one or more chemotherapeutic agents.
  • the cancer does not respond to radiation therapy.
  • the refractory cancer does not respond to one or more chemotherapeutic agents and radiation therapy.
  • a refractory cancer may be resistant at the beginning of such treatments or may become resistant over the course of one or more treatments.
  • refractory cancers may also be referred to as “chemotherapy resistant cancers” or “chemo ⁇ resistant cancers.”
  • “Androgen ⁇ independent prostate cancer” or “hormone ⁇ refractory prostate cancer (AIPC)” is prostate cancer that progresses after primary androgen ⁇ ablation therapy, either from orchiectomy or a gonadotropin ⁇ releasing hormone (LHRH) agonist, followed by addition and subsequent withdrawal of an antiandrogen.
  • LHRH gonadotropin ⁇ releasing hormone
  • hormone ⁇ refractory prostate cancer is defined as 2 ⁇ 3 consecutive rises in prostate ⁇ specific antigen (PSA) levels obtained at intervals of greater than 2 weeks and/or documented disease progression based on findings from CT scan and/or bone scan, bone pain, or obstructive voiding symptoms.
  • PSA prostate ⁇ specific antigen
  • the PSA level does not rise at diagnosis or throughout the entire course of the disease.
  • the prostate cancer is an advanced prostate cancer.
  • the prostate cancer or metastatic prostate cancer is resistant to treatments with hormone ⁇ blocking therapies, such as abiraterone (ZYTIGA®), enzalutamide (XTANDI®), bicalutamide (CASODEX®), flutamide (DROGENIL®), or cyproterone acetate (CYPROSTAT®).
  • hormone ⁇ blocking therapies such as abiraterone (ZYTIGA®), enzalutamide (XTANDI®), bicalutamide (CASODEX®), flutamide (DROGENIL®), or cyproterone acetate (CYPROSTAT®).
  • hormone ⁇ blocking therapies such as abiraterone (ZYTIGA®), enzalutamide (XTANDI®), bicalutamide (CASODEX®), flutamide (DROGENIL®), or cyproterone acetate (CYPROSTAT®).
  • the ranges of "from 2 to 10" and “2 ⁇ 10” are inclusive of the endpoints, 2 and 10, and all the intermediate values between in context of the units considered.
  • reference to “Claims 2 ⁇ 10” or “C 2 ⁇ C 10 alkyl” includes units 2, 3, 4, 5, 6, 7, 8, 9, and 10, as claims and atoms are numbered in sequential numbers without fractions or decimal points, unless described in the context of an average number.
  • the context of “pH of from 5 ⁇ 9” or “a temperature of from 5 ⁇ C to 9 ⁇ C”, on the other hand, includes whole numbers 5, 6, 7, 8, and 9, as well as all fractional or decimal units in between, such as 6.5 and 8.24.
  • AzP Treatment Inhibits Ovarian Cancer Cell Proliferation AzP analogs were screened against an ovarian cancer cell line panel (OVCAR ⁇ 3, OVCAR ⁇ 4, OVCAR ⁇ 5, OVCAR ⁇ 8, and SKOV ⁇ 3) with an additional screening on HEK293, SH ⁇ SY5Y, and N27 cell lines.
  • ovarian cancer cell line panel OVCAR ⁇ 3, OVCAR ⁇ 4, OVCAR ⁇ 5, OVCAR ⁇ 8, and SKOV ⁇ 3
  • HEK293, SH ⁇ SY5Y, and N27 cell lines We tested the relationship between AzPs dose and the viability of human ovarian cancer cell lines and the ID8 murine ovarian cancer cell line.
  • SU056 showed decreased IC 50 values after 48 h of treatment compared to SU093 (In OVCAR4, OVCAR5 and ID8 cells, this decrease was up to 2 fold), a trend which was also reflected in the clonogenic assay shown in Figures 1C and 1D, where both inhibitors decreased OC colony formation in a dose ⁇ dependent and significant manner in OVCAR ⁇ 8 and ID8 cells.
  • SU093 moderately affect SKOV ⁇ 3 at 0.5 & 1 ⁇ M concentrations.
  • Etoposide is natural podophyllotoxin and an approved chemotherapy drug used to treat many cancers.
  • a major limitation of etoposide is the potential for neuropathy and neurotoxicity in long ⁇ term treatment.
  • SU056 biophysically binds to YB ⁇ 1 and its cellular activity is YB ⁇ 1 dependent Previously, our group reported that SU093 interacts with GBP1 and inhibits the GBP1:PIM1 interaction (Andreoli et al., 2014).
  • SU056 is the second generation small molecule derivative of SU093 and, to confirm whether it has the same molecular target as SU093, we performed a pulldown assay using biotinylated SU056 (Figure 5A). Pulldown from cells treated with Biotinylated ⁇ SU056 and from protein lysate indicates that SU056 physically interacts with YB ⁇ 1 but not with GBP1 ( Figure 5B).
  • Cycloheximide chase (CHX) assay Cycloheximide is a protein translation inhibitor and is used as a molecular biology tool to determine the half ⁇ life of proteins.
  • OVCAR8 cells were treated with CHX and DMSO or SU056 (2.5 ⁇ M), and protein lysates were collected at different time points (0, 30, 60, 120, 180 min) followed by YB ⁇ 1 immunoblotting. Results suggest that SU056 treatment lead the proteasomal degradation and reduces the half ⁇ life of YB ⁇ 1 to from ⁇ 130 minutes to ⁇ 40 minutes ( Figure 6A).
  • YB ⁇ 1 is associated with the transcription and translation of many oncogenic proteins including CD44, ABCB1/MDR1, c ⁇ Myc, and Bcl ⁇ 2.
  • Immunoblotting of whole ⁇ cell lysates from DMSO ⁇ or SU056 ⁇ (2.5 ⁇ M, 12 h) treated SKOV3 and OVCAR8 cells indicate that SU056 treatment inhibits YB ⁇ 1 protein (Figure 6B). This was followed by a decrease in the expression of CDK2, CDC25A, MDR1, CD44, c ⁇ Myc, and Bcl ⁇ 2 and an increase in the expression of pro ⁇ apoptotic protein Bax ( Figure 6B).
  • YB ⁇ 1 protects and stabilizes the mRNA via 5' end capping (Evdokimova et al., 2001) and enhances exon splicing (ex. Alternative splicing of CD44) (Stickeler et al., 2001).
  • YB ⁇ 1 inhibition via SU056 treatment inhibits the Spliceosome pathway and induces RNA degradation and apoptosis.
  • YB ⁇ 1 Inhibition via SU056 Treatment Sensitizes OC to Paclitaxel YB ⁇ 1 is involved in the emergence of cisplatin and taxane drug resistance (Kang et al., 2013, Mo et al., 2016). Therefore, we tested whether SU056 synergizes with chemotherapies such as paclitaxel to modulate YB ⁇ 1. To measure this, we performed a dose dependent combination study of SU056 and paclitaxel. Using the MTT assay, we found that SU056 treatment significantly potentiates the cytotoxic effects of paclitaxel at 0.1, 0.5, and 1 nM doses ( Figure S2, 7A).
  • 3D spheroids are an alternative tumor model that closely mimic the TR phenotype via increased drug efflux.
  • OVCAR8 and SKOV3 cells in ultralow attachment plates with growth factor ⁇ defined media in the presence or absence of paclitaxel and SU056.
  • Results show that a combination of SU056 and paclitaxel significantly inhibits the formation by OC cell spheroids ( Figures 7D and 7E).
  • Combination ⁇ treated cells form 78% (OVACR8) and 83% (SKOV3) fewer spheroids than the vehicle ⁇ treated cells ( Figure 7E).
  • Ovarian cancer is the fifth most common cancer in women with an overall 5 ⁇ year survival rate of just 47.6% in the United States of America (USA).
  • Surgical resection and chemotherapy are the primary treatments for OC but are limited by surgical difficulties due to the abdominal spread of metastasizing OC and a high 2 ⁇ year relapse rate of 80 ⁇ 90% after taxane or platinum chemotherapy treatment (Jayson et al., 2014, Agarwal and Kaye, 2003).
  • paclitaxel offers some effect
  • relapsed disease is frequently TR, in which tumor cells bypass or overcome the molecular mechanisms of cytotoxicity despite ongoing treatment (Singh and Settleman, 2010, Blagosklonny and Fojo, 1999, Horwitz et al., 1986).
  • YB ⁇ 1 is a mRNA binding protein involved in nucleoprotein filament formation in cytoplasm (Kretov et al., 2019). Together with YB ⁇ 2 and YB ⁇ 3, YB ⁇ 1 binds with the cold ⁇ shock domain of single ⁇ stranded RNA/DNA (Graumann and Marahiel, 1998). It is involved in shuttling nucleic acids in both cytoplasm and nucleus (Matsumoto and Wolffe, 1998). In the cytoplasm, it regulates RNA stability, translation activity, and alternative splicing (Chansky et al., 2001).
  • SU056 ⁇ mediated YB ⁇ 1 inhibition significantly upregulates the RNA degradation pathway, while inhibiting the spliceosome pathway; a result which concords with the literatures about YB ⁇ 1 function.
  • SU056:YB ⁇ 1 interaction inhibits the YB ⁇ 1 activity in different OC cell line via proteasomal degradation and inhibits various downstream factors involved in tumor progression and TR.
  • ABCB1 also known as multidrug resistance 1 (MDR1), P ⁇ glycoprotein, ABCC1 (MRP1), and ABCG2 (BCRP/MXR) (Fletcher et al., 2010).
  • MDR1 multidrug resistance 1
  • MRP1 P ⁇ glycoprotein
  • BCRP/MXR BCRP/MXR
  • Paclitaxel is a substrate for ABCB1, and paclitaxel treatment upregulates its expression in various cancer types (Gottesman and Pastan, 1993).
  • ABC transporters are a hallmark of both cancer progression, and TR and has been viewed as a potential therapeutic target.
  • ABCB1 chemical inhibitors have been developed, but first, second, and third ⁇ generation compounds failed in clinical trials thus far.
  • Second ⁇ generation compounds had toxicity issues, whereas second ⁇ generation compounds such as Valspodar, showed no treatment benefit in combination with paclitaxel or carboplatin in ovarian or peritoneal cancer patients (Lingham et al., 2008).
  • the third ⁇ generation inhibitor Zosuquidar also failed to show any benefits (Ruff et al., 2009), leaving the inhibition of MDR1 and TR phenotype an unmet clinical goal.
  • paclitaxel, SU056, and their combination on the expression of MDR1 and YB ⁇ 1 we found that paclitaxel treatment upregulates the expression of both YB ⁇ 1 and MDR1, whereas SU056 alone or in combination with paclitaxel significantly decreases MDR1 and YB ⁇ 1.
  • paclitaxel efflux data shows that SU056 treatment inhibits MDR1 and leads to higher intracellular concentrations and activity of paclitaxel.
  • Tumor spheroids a common 3D cell culture model, have been used to examine stemness, efflux, and MDR1 expression changes after treatment (Wartenberg et al., 2005, Wartenberg et al., 1998, Chen et al., 2017).
  • treating OC cells with a combination of paclitaxel and SU056 significantly inhibited their spheroid formation capability, which indicates decreased tumorigenic potential.
  • YB ⁇ 1 is associated with transcriptional regulation of ABC transporters and epithelial ⁇ to ⁇ mesenchymal transition ⁇ associated proteins implicated in disease progression and TR (Lim et al., 2018, Wu et al., 2014, Evdokimova et al., 2009).
  • SU056 co ⁇ treatment with paclitaxel was sufficient to halt tumor progression in the OVCAR8 xenograft model.
  • the reaction mixture was stirred at rt for 18 h then diluted with water ( ⁇ 10 mL). The mixture was then extracted with ethyl acetate (2 X 25 mL). Combined organic phase was washed with water followed by brine solution. The organic phase was dried over Na2SO4, filtered, and evaporated to dryness.
  • the crude was purified with CombiFlash chromatography on silica gel using 0 ⁇ 10% of methanol in dichloromethane as an eluent. Combined pure fractions was evaporated to dryness to afford the desired product as an off ⁇ white solid (15.0 mg, 19%).
  • MRM Multiple reaction monitoring
  • a calibration curve was generated using known concentrations of SU056 and IS, and this curve was used to calculate unknown concentrations of SU056 in the plasma at different time points.
  • SU056 was injected at the dose of 20 mg/kg to each mouse at time zero intraperitonially. Blood was collected retro ⁇ orbitally at 5, 15, 30, 60, 120, 240 and 360 minutes after injection. Blood plasma was separated via centrifugation at 7,000 RPM for 10 minutes. A 5 ⁇ L plasma was taken from each sample and mixed with a 10 ⁇ L of IS solution and 990 ⁇ L of MS ⁇ grade acetonitrile then vortexed for 30 seconds followed by a 5 minutes incubation at RT.
  • KEGG Pathway Increase p ⁇ value Proteins in pathway detected (bold are enriched)
  • OVCAR3, OVCAR4, OVCAR5, OVCAR8, and SKOV ⁇ 3 cell lines were obtained from the NCI cell line repository (DTP).
  • SH ⁇ SY5Y and N27 cell lines were obtained from Dr. Manish Chamoli (Buck Institute, CA, USA).
  • ID8 and Luciferase ⁇ tagged ID8 cells were obtained from Dr. Erinn Rankin (Stanford University, CA, USA). Scrambled control (SC), YBX1 knockdown (1 & 2) and mCherry ⁇ YBX1 OVCAR8 cell lines were created using lentiviral based transduction and selected using puromycin resistance followed by cell shorting. All OVCAR cells were maintained in RPMI ⁇ 1640 (Corning, USA; #10 ⁇ 040 ⁇ CV) supplemented with 10% FBS (Corning, USA; #35 ⁇ 015 ⁇ CV) and 1% Antibiotic ⁇ Antimycotic solution (Gibco, USA; #15240062).
  • SKOV3 and ID8 cells were maintained in DMEM media (Corning, USA; #10 ⁇ 013 ⁇ CV) supplemented with 10% FBS and 1% Antibiotic ⁇ Antimycotic solution. All cells were maintained at 37 ⁇ C and 5% CO 2 .
  • SH ⁇ SY5Y and N27 cells were maintained in DMEM/F12 media (Hyclone, #SH30525.01) supplemented with 10% FBS and 1% Antibiotic ⁇ Antimycotic solution. All cells were maintained at 37 ⁇ C and 5% CO 2 .
  • OVCAR8 cells were tagged with Luciferase using lentiviral vector ⁇ based plasmid pLenti PGK Blast V5 ⁇ LUC (w528 ⁇ 1) (gifted by Eric Campeau & Paul Kaufman, Addgene # 19166) and selected using blasticidin. Luciferase ⁇ tagged cells were maintained as described above.
  • Cell viability assay Cell viability was assessed using the standard MTT assay protocol. In brief, 5,000 cells were plated in each well of 96 well plates (Corning ⁇ Costar, #3598) and allowed to attach for 24 h. Cells were treated with a respective concentration of compounds for respective time points. A stock solution of each compound was prepared in DMSO. DMSO concentration was kept constant and maintained below 0.1%.
  • the media was replaced after 24 h with media containing a respective concentration of the test compound and incubated for 5 ⁇ 8 days until visible colonies appeared in vehicle ⁇ treated wells. Cells were then washed and fixed with 2% paraformaldehyde, followed by washing and staining with 0.5% Crystal violet for 1h. Cells were de ⁇ stained using DI water and allowed to dry. Colonies were counted under the microscope at 100X magnification. Cell cycle analysis: 30,000 OC cells were plated in each well of 12 well plates and incubated for 24 h. Cells were treated with a respective concentration of test compounds for 6 h. Both live and dead cells were collected via trypsinization and cell pellets were fixed with 70% ethanol.
  • PI propidium iodide
  • RNase A 80 ⁇ g/mL RNase A and 50 ⁇ g/mL PI in saponin ⁇ EDTA solution
  • FlowJo software calculated the % cells in each cell cycle phase.
  • Apoptotic cell death assay Cells were plated and treated as described in cell cycle analysis assay above. Treated cells were incubated for 24 h and both live and dead cells were collected.
  • Cells were stained with Annexin V and PI using FITC Annexin V apoptosis detection kit (BD Pharmingen, San Jose, CA) by following the manufacturer’s protocol. Guava easyCyte Flow Cytometer was used to analyze the stained cells.
  • Cell migration assay 1 x 10 5 cells were plated in 60 mm cell culture dishes (Corning ⁇ Falcon, #353002) and incubated for 24 h. Cells were treated with a respective concentration of each compound for 12 h. Cells were trypsinized and collected in a conical tube for each plate. Pellets of cells were resuspended, and live cells were counted using a hemocytometer and trypan blue staining.
  • OVCAR8 cells were treated with vehicle (DMSO) or SU056 (2.5 ⁇ M) at 70 ⁇ 80% confluency for 1.5 h. Cells were harvested and washed with 1x PBS twice. Cells were pelleted and resuspended in PBS. 10 different PCR tubes with 1 x 10 6 cells/tube (in 100 ⁇ L PBS) were prepared for both the groups. Tubes were exposed to respective temperatures (37, 41, 44, 47, 50, 53, 56, 59, 63, 67 ⁇ C) for 3 min using a thermal cycler (Biorad, CA, USA) followed by 2 min incubation at room temperature. Each tube was snap ⁇ frozen in liquid nitrogen.
  • DMSO vehicle
  • SU056 2.5 ⁇ M
  • TMT Tandem Mass Tag
  • CETSA protein quantification normalization, curve fitting, estimation of slope, and melting point and statistical analysis: Proteins were quantified from individual peptide spectra by a sum ⁇ based bootstrap algorithm using each corresponding TMT reporter ion intensity after correcting for isotope impurities using MaxQuant (Cox and Mann, 2008) in both vehicle and SU056 treated samples. In each sample, the lowest temperature was used as reference to calculate the log2 ratio of signal of the soluble fraction in each temperature, and each signal was compared with the highest temperature to estimate the percentage of signal lost in the soluble fraction.
  • Immunoblotting Once 70% confluency of OC cells plated in 100 mm cell culture was obtained, dishes were treated with a respective concentration of SU056 for a respective time. At the end of treatment, cells were collected and lysed using M ⁇ PER TM lysis solution (Thermo Scientific, #78503), supplemented with Halt protease and phosphatase inhibitor cocktail (Thermo Scientific, #78440). An equal amount (40 ⁇ 60 ⁇ g) of proteins was resolved using 8%/10%/12% SDS ⁇ PAGE gel electrophoresis. Proteins were then transferred onto a PVDF membrane (BioRad, #162 ⁇ 01277).
  • YB ⁇ 1 Cell signaling technology (CST), #8475; 1:2000), TMSB10 (R&D Systems, #AF6429; 1:2000), SUMO2/3 (CST, #4971; 1:1000), PSMB2 (Bethyl Laboratories, #A303817AT; 1:1000), MDR1 (CST, #13978; 1:2000), CD44 (CST, #37259; 1:1000), c ⁇ Myc (Novusbio, #NB600 ⁇ 302SS; 1:2000), CDK2 (CST, #2546; 1:2000), CDC25A (CST, # 3652; 1:1000), Cyclin E (CST, #4132; 1:2000), Bax (CST, #5023; 1:1000), Bcl ⁇ 2 (CST, #2876; 1:1000), GBP1 (Abnova, #H00002633 ⁇ PW1, 1:2000), ⁇ actin (Novusbio, #NB600 ⁇ 501SS; 1:10000), anti ⁇ mouse IgG HR
  • Total YB ⁇ 1 sandwich ELISA Total YB ⁇ 1 protein level was analyzed using PathScan® Total YB1 Sandwich ELISA Kit (Cell Signaling, #12543) by following the manufacturer’s protocol.
  • Multidrug Resistance Assay The effect of SU056 on Multidrug Resistance of different OC cells was assayed by using the Multidrug Resistance Assay Kit (Fluorometric MDR Assay) (Sigma ⁇ Aldrich, #MAK161), following the manufacturer's protocol.
  • CD44 ELISA The effect of SU056 on CD44 expression of different OC cells was assayed by using Human CD44 ELISA Kit (Colorimetric) (Novusbio, #NBP1 ⁇ 86819), following the manufacturer’s protocol.
  • Pulldown assay using biotinylated SU056 protein pulldown assay was performed using biotinylated SU056.
  • OVCAR8 cells were treated with 2.5 ⁇ M biotinylated SU056 for 1.5 h. Treated cells were collected and lysed using M ⁇ PER TM lysis solution (Thermo Scientific, #78503), supplemented with Halt protease and phosphatase inhibitor cocktail (Thermo Scientific, #78440). 300 ⁇ g protein was incubated with magnetic conjugate streptavidin bead (CST, #5947) at 4°C on rocker overnight. 2.
  • OVCAR8 cells were collected and lysed using M ⁇ PER TM lysis solution (Thermo Scientific, #78503), supplemented with Halt protease and phosphatase inhibitor cocktail (Thermo Scientific, #78440). 1000 ⁇ g protein was incubated with 10 ⁇ M biotinylated SU056 at 4°C on rocker overnight followed by overnight incubation with magnetic conjugate streptavidin bead (CST, #5947) at 4°C. After both the pulldown, biotin ⁇ streptavidin conjugates were pulldown and washed using magnetic rack.
  • the His ⁇ tagged (N ⁇ Terminal) YB ⁇ 1 protein (Novusbio, # NBP2 ⁇ 30101) was captured via the His ⁇ tag on an NTA chip (GE Healthcare) and immobilized through amine coupling amine coupling chemistry using N ⁇ hydroxysuccinimide (NHS) and N′ ⁇ (3 ⁇ dimethylaminopropyl) carbodiimide hydrochloride (EDC) (GE Healthcare). All small molecule (SU093 and SU056) analysis experiments were performed in PBS (10 mM Phosphate buffer, 2.7 mM KCl, 0.137 NaCl) running buffer pH adjusted to yield pH 7.4 when supplemented with 5% DMSO.
  • PBS 10 mM Phosphate buffer, 2.7 mM KCl, 0.137 NaCl
  • Paclitaxel efflux assay OC cells were treated with 5 nM Oregon GreenTM 488 ⁇ conjugated paclitaxel (Molecular Probes, # P22310) for 1 h. Cells were washed and incubated in phenol red ⁇ free DMEM media supplemented with 10% FBS. After each respective time point (30, 60, 120, 180 minutes), the media was collected and centrifuged to remove floating cells. The fluorescence intensity of efflux paclitaxel in media was read at Ex 496 and Em 524 using a multimode plate reader.
  • Spheroid culture OC cells at a density of 100 per well were plated in an ultra ⁇ low attachment 24 well plate (Corning, #3473) in MEGM media (Lonza, #CC ⁇ 3150) supplemented with hEGF, insulin, hydrocortisone BPE, and 2 ⁇ mercaptoethanol. Cells were treated with each respective compound and incubated for 6 ⁇ 8 days. The number of spheroids was counted under the microscope at 40X magnification. 5 different fields from each well were imaged at 100X magnification. In vivo xenograft model and drug efficacy study: All animal experiments were reviewed and approved by the Animal Care and Use Committee of Stanford University, CA, USA.
  • Luciferase tagged ID8 (2 x 10 6 ) or OVCAR8 (5 x 10 6 ) cells were implanted into the right flank of 6 ⁇ 7 ⁇ week ⁇ old female C57BL/6 mice and NOD/SCID mice, respectively. Respective treatment began after tumors grew to 100 ⁇ 200 mm 3 diameters.
  • the ID8 syngeneic mice model was treated with a vehicle (30% PEG ⁇ 300 in saline), 20 mg/kg SU093, and 20 mg/kg SU056 intraperitoneally (IP) daily for 42 days.
  • ALT Alanine Aminotransferase
  • AST Aspartate Aminotransferase
  • AKP Alkaline Phosphatase
  • OVCAR8 xenograft mice model was treated with vehicle (30% PEG ⁇ 300 in saline), 5 mg/kg paclitaxel (once a week), 10 mg/kg SU056 (daily), a combination of paclitaxel (5 mg/kg, once a week) and SU056 (10 mg/kg, daily) for 4 weeks.
  • Biotinylated horse anti ⁇ mouse IgG (Vector Labs, CA) and horse anti ⁇ rabbit IgG (Vector Labs, CA) were used as secondary antibodies.
  • the following primary antibodies were used: YB ⁇ 1 (Cell signaling, #8475; 1:100) and anti ⁇ MDR1 (Cell signaling, #13978; 1:500) and Ki67 (Biolegend, #350502; 1:500).
  • Statistical analysis Each set of data was analyzed for its statistical significance using GraphPad Prism 6 software. Each result is represented in Mean ⁇ SD. P ⁇ values are denoted using * as follows: * P ⁇ 0.05, ** P ⁇ 0.01, *** P ⁇ 0.001.
  • Presence of expressed YBX1 protein in cancer cells referenced herein may be identified by methods known in the art.
  • Non ⁇ limiting examples include the YBX1 ELISA Kit available from antibodies ⁇ online Inc, Jones Blvd 321, Limerick, PA 19464 (Catalog No. ABIN6975583).
  • Y box binding protein 1 (YB1 or YBX1) is the multifunction protein which binds to the DNA and RNA and is associated with tumor progression and the emergence of TR.
  • YB ⁇ 1 plays the important role in transcription, translation and RNA stabilization of various oncogenic proteins. Role of YB ⁇ 1 is very well established in various cancers but there is no small molecule inhibitor available/reported so far.
  • AzP novel azopodophyllotoxin
  • This first ⁇ in class YB ⁇ 1 inhibitor potently inhibits the ovarian cancer (OC) cell proliferation and resistance to apoptosis and arrests the cells in G1 phase.
  • This treatment leads to an enrichment of proteins associated with apoptosis and RNA degradation pathway and downregulates the spliceosome pathway.
  • SU056 independently restrains ovarian cancer progression and exerts a synergistic effect with paclitaxel to further reduce disease progression with no liver toxicity.
  • in vitro mechanistic studies showed delayed disease progression via inhibition of drug efflux and multi ⁇ drug resistance 1 (MDR1) and significantly low neural toxicity as compared to Etoposide.
  • MDR1 multi ⁇ drug resistance 1
  • MaxQuant enables high peptide identification rates, individualized p.p.b. ⁇ range mass accuracies and proteome ⁇ wide protein quantification. Nat Biotechnol, 26, 1367 ⁇ 72. DE DONATO et al., 2012. Class III beta ⁇ tubulin and the cytoskeletal gateway for drug resistance in ovarian cancer. J Cell Physiol, 227, 1034 ⁇ 41. DIDIER et al., 1988. Characterization of the cDNA encoding a protein binding to the major histocompatibility complex class II Y box. Proc Natl Acad Sci U S A, 85, 7322 ⁇ 6. DIETL, J. 2014. Revisiting the pathogenesis of ovarian cancer: the central role of the fallopian tube.
  • YB ⁇ 1 an abundant core mRNA ⁇ binding protein, has the capacity to form an RNA nucleoprotein filament: a structural analysis. Nucleic Acids Res, 47, 3127 ⁇ 3141. KUWANO et al., 2004. The role of nuclear Y ⁇ box binding protein 1 as a global marker in drug resistance. Mol Cancer Ther, 3, 1485 ⁇ 92. KUZNETSOVA et al.,2012. Synthesis and Biological Evaluation of Novel 3' ⁇ Difluorovinyl Taxoids. J Fluor Chem, 143, 177 ⁇ 188. LAIRD ⁇ OFFRINGA et al., 1990. Poly(A) tail shortening is the translation ⁇ dependent step in c ⁇ myc mRNA degradation.
  • YB ⁇ 1 protein functions and regulation. Wiley Interdiscip Rev RNA, 5, 95 ⁇ 110. MAITY et al., 2011. Class III beta ⁇ tubulin (TUBB3): more than a biomarker in solid tumors? Curr Mol Med, 11, 726 ⁇ 31. MATSUMOTO et al., P. 1998. Gene regulation by Y ⁇ box proteins: coupling control of transcription and translation. Trends Cell Biol, 8, 318 ⁇ 23. MO et al., 2016. Human Helicase RECQL4 Drives Cisplatin Resistance in Gastric Cancer by Activating an AKT ⁇ YB1 ⁇ MDR1 Signaling Pathway. Cancer Res, 76, 3057 ⁇ 66. RUFF et al., 2009.
  • EMT cancer stem cells and drug resistance: an emerging axis of evil in the war on cancer. Oncogene, 29, 4741 ⁇ 51. SOBOCAN et al., 2020. The Communication Between the PI3K/AKT/mTOR Pathway and Y ⁇ box Binding Protein ⁇ 1 in Gynecological Cancer. Cancers (Basel), 12. STICKELER et al., 2001. The RNA binding protein YB ⁇ 1 binds A/C ⁇ rich exon enhancers and stimulates splicing of the CD44 alternative exon v4. EMBO J, 20, 3821 ⁇ 30. SUN et al., 2015.
  • YB ⁇ 1 is a Transcription/Translation Factor that Orchestrates the Oncogenome by Hardwiring Signal Transduction to Gene Expression. Transl Oncogenomics, 2, 49 ⁇ 65. WU et al., 2014. Cell fate factor DACH1 represses YB ⁇ 1 ⁇ mediated oncogenic transcription and translation. Cancer Res, 74, 829 ⁇ 39. ZENG et al., 2018. Targeting MYC dependency in ovarian cancer through inhibition of CDK7 and CDK12/13. Elife, 7. ZHAO et al., 2016. YBX1 regulates tumor growth via CDC25a pathway in human lung adenocarcinoma. Oncotarget, 7, 82139 ⁇ 82157.
  • Figure 1 A) Lead optimization of SU093 to obtain SU056.
  • E % inhibition values of etoposide, SU093, and SU056 treatment at 10 ⁇ M concentration for 48 h in neuronal (SH ⁇ SY5Y, N27) and HEK293 cells.
  • F Cell cycle distribution of propidium iodide (PI) ⁇ stained OVCAR8, SKOV3, and ID8 cells. Effects of SU093 and SU056 on cell cycle distribution showing G1 phase arrest after 12 h treatment.
  • G Effects of SU093 and SU056 on apoptotic cell death analyzed by Annexin ⁇ FITC staining. Both the compounds induce apoptotic cell death in ovarian cancer cells after 24 h treatment.
  • H Cell migration assay.
  • FIG. 1 No. of lung metastatic nodules. Data shown are mean ⁇ SD from 5 mice in each group. * P ⁇ 0.05, ** P ⁇ 0.01, *** P ⁇ 0.001, compared with respective control.
  • Figure 3 Target identification using Cellular Thermal Shift Assay (CETSA). Differential profiling of SU056 on the thermal proteome profile of OVCAR8 cells. OVCAR8 cells were treated with DMSO or SU056 (2.5 ⁇ M) for 1.5 h. Cells were collected and 10 6 cells in each PCR tube incubated at different temperatures (37, 41, 44, 47, 50, 53, 56, 59, 63, 67 ⁇ C).
  • E Melting curves for identified top six proteins (YB ⁇ 1, TMSB10, SUMO ⁇ 2, PSMB2, TMSB4X, and CALM3) with and without SU056 treatment.
  • F Change in melting temperature (T m ) of the top six proteins upon SU056 treatment.
  • Figure 4 SU056 inhibits YB ⁇ 1.
  • A) OVCAR8 cells were treated with SU056 (1, 2.5 and 5 ⁇ M) for 12 h and total cell lysates were prepared as described in the ‘Methods’ section. SDS ⁇ PAGE and Western blot analysis was performed for the top three targets identified by CETSA (YB ⁇ 1, TMSB10, SUMO ⁇ 2 and PSMB2).
  • YB ⁇ 1 50% inhibitory concentration (IC50) of SU056 for OC cell lines was determined after 12 h of treatment using PathScan® Total YB1 Sandwich ELISA Kit.
  • E Western blot analysis was performed to conform the YB ⁇ 1 expression in transduced cells.
  • F 500 cells (SC, shRNA1, shRNA2) were plated/well of 12 well plate and allow to attach for 24h. Transduced cells were treated with SU056 and incubated further for 7 days. Each well was stained with crystal violet and colonies were counted under 10X microscope. Representative colony formation from respective wells.
  • G IC 50 values of SU056 on different transduced OVCAR8 cells expressing SC, YBX1 shRNA1, YBX1 shRNA2.
  • Figure 6 SU056 modulates the YB ⁇ 1 associated proteins and pathways.
  • Proteins were resolved on SDS ⁇ PAGE gel and blotted for respective antibodies as described in the method section. Membranes were stripped and re ⁇ probed for loading control actin. Spheroid formation assay. 500 cells were cultured in ultra ⁇ low attachment plates and treated with each drug and their combinations. The cells incubated for 7 days to form spheroids. D) Microscopic images of spheroid at 10X magnification. Scale bar, 250 ⁇ m E) Spheroid formation was quantified after 7 days of incubation. Data are shown as mean ⁇ SD of triplicate samples. * P ⁇ 0.05, ** P ⁇ 0.01, *** P ⁇ 0.001, significantly different compared with respective controls by one ⁇ way ANOVA followed by Dunnett's test.
  • Figure 11 A). The growth inhibitory effect of SU056 was evaluated using MTT assay.
  • TNBC cell lines (MDA ⁇ MB ⁇ 231, MDA ⁇ MB ⁇ 468, SUM159, 4T1, E0771 and EMT6) were plated in 96 well plate. Next day, treated with vehicle (DMSO) alone or 0.005 ⁇ 50 ⁇ M of SU056 in fresh medium. After 48 hr of treatments, cell viability was measured using MTT assay.
  • SU056 treatment inhibited the protein translation associated molecules among all TNBC cells
  • Figure 13. SU056 inhibit tumor xenograft of TNBC models. Mice were subcutaneously injected with MDA ⁇ MB ⁇ 231 (2 x 10 6 cells), MDA ⁇ MB ⁇ 468 (5 x 10 6 cells) and patient derived xenograft SUTI151 ⁇ PDX (2 x 10 6 cells) and drug treatment started when tumor reached 100 mm 3 . Mice were given either vehicle (40% PEG in saline) or SU056 (50 mg/kg) through oral route using oral gavage.
  • Figure 14. SU056 inhibit 4T1 tumor xenograft in BALB/c. Mice were subcutaneously injected with 4T1 cells and drug treatment started after three day of implantation. Mice were given either vehicle (40% PEG in saline) or SU056 (50 mg/kg) through oral route using oral gavage.
  • Figure 16 depicts reference ⁇ normalized expression levels of YBX1 for the cancer cell lines: a) Lung – IMR90, A549, MRC5, H1299, NHBE, NCIH460, and BEAS2; b) Lymphocyte – JURKAT, MT4, BJAB, HEL CELLS, HL60, and RAJI; c) Breast/Mammary – MCF7, MCF10, MDAMB231, SKBR3, MDAMB468, and MDAMB453; d) Fibroblast – BJ, KB, HT1080, NHDF, and TIG; e) Prostate – LNCAP, PC3, DU145, and C42; f) Other – NALM6, DAOY, JEG3, and BEWO; g) Kidney – 293T, HEK293, 293F, and FLPIN TREX 293; h) Blood – THP1, PLB985, CEM, and HELAT4;
  • Figure 17 Assay of SU056 in NCI ⁇ 60 cell line panel. SU056 was assayed against the NCI ⁇ 60 cell line panel at five dose (100 to 0.01 ⁇ M) for 48 hours, providing the IC50 values in the table below. Growth inhibition was measured and displayed as a heatmap from no inhibition (black), to growth inhibition (white), as seen in Figure 17.
  • NCI60 ⁇ IC50 values Cell Lines IC50 ( ⁇ M) CCRF ⁇ CEM 237 SF ⁇ 268 3.2 SF ⁇ 295 12 TNBC cells were treated for 12 and 24 hr and total cell lysate were prepared. SDS ⁇ PAGE and western blot analyses were performed for translation associated molecules. Beta ⁇ actin were probed to ensure equal protein loading. SU056 treatment inhibited the translation initiation factors in TNBC cells. STAR Methods Key Resources Table Reagent or Resource Source Identifier Antibodies Cell Lines OVCAR3 NCI cell line repository (DTP) Human YB ⁇ 1 protein (His Tag) (Novus Biological Catalog No.
  • NBP2 ⁇ 30101 is a recombinant protein with a N ⁇ terminal His ⁇ tag and corresponding to the amino acids 1 ⁇ 324 of Human YB1.
  • Source E. Coli.
  • Gene YBX1.

Landscapes

  • Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
PCT/US2021/061906 2020-12-04 2021-12-03 Y box binding protein 1 inhibitors Ceased WO2022120242A1 (en)

Priority Applications (6)

Application Number Priority Date Filing Date Title
US18/255,827 US20240016784A1 (en) 2020-12-04 2021-12-03 Y box binding protein 1 inhibitors
CA3201032A CA3201032A1 (en) 2020-12-04 2021-12-03 Y box binding protein 1 inhibitors
AU2021391931A AU2021391931A1 (en) 2020-12-04 2021-12-03 Y box binding protein 1 inhibitors
CN202180092273.1A CN116761602A (zh) 2020-12-04 2021-12-03 Y盒结合蛋白质1抑制剂
EP21901582.3A EP4255426A4 (en) 2020-12-04 2021-12-03 Y-BOX BINDING PROTEIN 1 INHIBITORS
JP2023533819A JP2023552368A (ja) 2020-12-04 2021-12-03 Ｙボックス結合タンパク質１阻害剤

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US202063121674P 2020-12-04 2020-12-04
US63/121,674 2020-12-04
US202163256609P 2021-10-17 2021-10-17
US63/256,609 2021-10-17

Publications (1)

Publication Number Publication Date
WO2022120242A1 true WO2022120242A1 (en) 2022-06-09

Family

ID=81853609

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2021/061906 Ceased WO2022120242A1 (en) 2020-12-04 2021-12-03 Y box binding protein 1 inhibitors

Country Status (6)

Country Link
US (1) US20240016784A1 (https=)
EP (1) EP4255426A4 (https=)
JP (1) JP2023552368A (https=)
AU (1) AU2021391931A1 (https=)
CA (1) CA3201032A1 (https=)
WO (1) WO2022120242A1 (https=)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024015793A3 (en) * 2022-07-12 2024-03-14 Oregon Health & Science University Enantiomer of azopodophyllotoxin derivative su056

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170342086A1 (en) * 2015-02-26 2017-11-30 Ajay Kumar Compounds and methods for the treatment of drug resistance in cancer cells against paclitaxel
US11098052B2 (en) * 2016-03-02 2021-08-24 Arizona Board Of Regents On Behalf Of Arizona State University 4-azapodophylotoxins compounds
US11771692B2 (en) * 2018-03-13 2023-10-03 The Board Of Trustees Of The Leland Stanford Junior University N-hydroxyethyl didehydroazapodophyllotoxins as GBP1 inhibitors and methods of overcoming treatment resistance in cancer
CN116761602A (zh) * 2020-12-04 2023-09-15 俄勒冈健康与科学大学 Y盒结合蛋白质1抑制剂

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DATABASE PubChem substance 8 December 2016 (2016-12-08), ANONYMOUS : "SCHEMBL18056670", XP055941189, retrieved from NCBI Database accession no. 319685906 *
See also references of EP4255426A4 *
SETOGUCHI KIYOKO, CUI LIN, HACHISUKA NOBUTAKA, OBCHOEI SUMALEE, SHINKAI KENTARO, HYODO FUMINORI, KATO KIYOKO, WADA FUMITO, YAMAMOT: "Antisense Oligonucleotides Targeting Y-Box Binding Protein-1 Inhibit Tumor Angiogenesis by Downregulating Bcl-xL-VEGFR2/-Tie Axes", MOLECULAR THERAPY-NUCLEIC ACIDS, CELL PRESS, US, vol. 9, 1 December 2017 (2017-12-01), US , pages 170 - 181, XP055941192, ISSN: 2162-2531, DOI: 10.1016/j.omtn.2017.09.004 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024015793A3 (en) * 2022-07-12 2024-03-14 Oregon Health & Science University Enantiomer of azopodophyllotoxin derivative su056

Also Published As

Publication number Publication date
JP2023552368A (ja) 2023-12-15
EP4255426A4 (en) 2024-10-23
US20240016784A1 (en) 2024-01-18
EP4255426A1 (en) 2023-10-11
AU2021391931A1 (en) 2023-06-29
CA3201032A1 (en) 2022-06-09

Similar Documents

Publication Publication Date Title
WO2022135346A1 (en) Pharmaceutical combinations comprising a kras g12c inhibitor and uses of a kras g12c inhibitor for the treatment of cancers
DK2694072T3 (en) COMBINATION OF ACT-INHIBITOR RELATIONSHIP AND ABIRATERON FOR USE IN THERAPEUTIC TREATMENTS
US20160002222A1 (en) Prodrug forms of kinase inhibitors and their use in therapy
He et al. Targeting cullin neddylation for cancer and fibrotic diseases
CA3133688A1 (en) Compositions and methods for treating cancer
ES2780382T3 (es) Derivados de pirazolopirimidina
US9533969B2 (en) Materials and method for inhibiting replication protein A and uses thereof
Bharadwaj et al. STAT3 inhibitors in cancer: a comprehensive update
US20190262319A1 (en) Novel methods for treating cancer
WO2019171268A1 (en) New active principles for the treatment of tumours
Gu et al. Discovery of a highly potent and selective MYOF inhibitor with improved water solubility for the treatment of gastric cancer
CA2739250C (en) Selective proton coupled folate transporter and folate receptor, and garftase inhibitor compounds and methods of using the same
WO2022120242A1 (en) Y box binding protein 1 inhibitors
Chen et al. Design, synthesis and biological evaluation of novel 9-N-substituted-13-alkylberberine derivatives from Chinese medicine as anti-hepatocellular carcinoma agents
CN116761602A (zh) Y盒结合蛋白质1抑制剂
EP4132657A1 (en) Compositions and methods for treatment of anticancer-drug resistant cancers
EP4495121A1 (en) Camptothecin derivatives that bind to ddx5 protein and prodrugs thereof
KR20210060642A (ko) 성장 관련 질환 및/또는 그것의 임상적 상태를 억제 및/또는 치료하기 위한 조성물 및 방법
ES3034869T3 (en) Tg02 for use in treating gliomas in pediatric subjects
KR20130130802A (ko) 보르트만닌 유사체의 결정질 형태의 조성물 및 이의 사용 방법
US20260000665A1 (en) Enantiomer of azopodophyllotoxin derivative su056
Han et al. Pharmacological activation of the p53 pathway by a new compound CYZ2017 exerts anti-tumor effects
WO2025132831A1 (en) N-heteroaryl derivatives and uses thereof for treating cancer
KR20260056169A (ko) Rna 헬리카제 억제제
WO2024100402A1 (en) Proteolysis targeting chimera compounds

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21901582

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 3201032

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 2023533819

Country of ref document: JP

ENP Entry into the national phase

Ref document number: 2021391931

Country of ref document: AU

Date of ref document: 20211203

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2021901582

Country of ref document: EP

Effective date: 20230704

WWE Wipo information: entry into national phase

Ref document number: 202180092273.1

Country of ref document: CN