Phospholipid ether conjugates as cancer-targeted drug vehicles

Abstract

The present invention discloses therapeutic compounds capable of targeting a wide range of tumor cells. The disclosure is further directed to compositions comprising the therapeutic compounds, methods for producing the therapeutic compounds, and methods for treating cancer comprising administering the therapeutic compounds.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 62 / 899,611, filed September 12, 2019, U.S. Provisional Patent Application No. 62 / 899,615, filed September 12, 2019, U.S. Provisional Patent Application No. 62 / 899,618, filed September 12, 2019, U.S. Provisional Patent Application No. 62 / 946,870, filed December 11, 2019, U.S. Provisional Patent Application No. 62 / 956,844, filed January 3, 2020, and U.S. Provisional Patent Application No. 62 / 956,907, filed January 3, 2020, the contents of which are incorporated herein by reference in their entireties.

[0002] Field The present disclosure relates to therapeutic compounds that can target a wide range of tumor cells. The present disclosure is further directed to compositions comprising the therapeutic compounds, methods for making the therapeutic compounds, and methods for treating cancer that include administering the therapeutic compounds. [Background technology]

[0003] Introduction In 2018, 18 million people worldwide were diagnosed with cancer and 9.6 million died from the disease. In the United States, approximately 40% of the total population will be diagnosed with cancer during their lifetime. As of 2018, lung cancer (2.09 million cases), breast cancer (2.09 million cases), colorectal cancer (1.8 million cases), prostate cancer (1.28 million cases), skin cancer (non-melanoma) (1.04 million cases), and stomach cancer (1.03 million cases) were the most common types of cancer. Despite many available treatments, cancer remains the second leading cause of death worldwide.

[0004] Cancer is the result of cell division, but is not limited to this. Healthy cells have checkpoints that prevent unlimited cell division. Some examples of these checkpoints include nutrient availability, DNA damage, and contact inhibition (i.e., a cell coming into contact with another cell). Furthermore, most cells can only replicate a finite number of times and are therefore programmed to die after a certain number of cell divisions.

[0005] Cancer is the result of cells overcoming these inherent checkpoints and growing uncontrollably. This uncontrolled growth leads to the formation of tumors. There are two types of tumors: benign and malignant. Benign tumors cannot overcome the natural boundaries between tissue types. On the other hand, malignant tumors can invade nearby tissues or enter the bloodstream and metastasize to different sites. Only malignant tumors are considered cancer. It is this ability to invade and metastasize that makes cancer such a deadly disease. Furthermore, lipid metabolism can play a major role in cancer metastasis. Cancer cells often exhibit radically altered cellular metabolism. However, the role of lipid metabolism in the development of malignant cancer remains unclear.

[0006] Further complicating the fight against cancer is the fact that malignant tumors contain distinct cell types. One particularly troubling type is cancer stem cells ("CSCs"). CSCs are capable of self-renewal and differentiation into the different cancer cell types found in malignant tumors. Thus, CSCs are a major factor in a tumor's metastatic potential. CSCs often survive radiation and chemotherapy. It is hypothesized that cancer recurrence after radiation and chemotherapy is the result of a combination of the inability of radiation and chemotherapy to kill all CSCs and the ability of CSCs to form new tumors.

[0007] Chemotherapy is a term used to describe certain types of cancer treatment that involve the use of cytotoxic anticancer drugs. The cytotoxic drugs used during chemotherapy can be divided into several major categories, including alkylating agents, antimetabolites, antitumor antibiotics, topoisomerase inhibitors, and mitotic inhibitors. Cytotoxic anticancer drugs typically stop cell division, thus affecting both healthy and cancerous tissues. Alkylating agents stop cancer cell division by damaging the DNA of cancer cells. Some common alkylating agents used in cancer treatment include nitrogen mustards (e.g., cyclophosphamide (Cytoxan®; Cytoxan is a registered trademark of Baxter International), nitrosoureas, alkylsulfonates, triazeines, and ethyleneimines. Platinum drugs such as cisplatin and carboplatin act similarly to alkylating agents. Antimetabolites stop cancer cells from dividing by inhibiting DNA and RNA synthesis. Some common antimetabolites used in cancer treatment include 6-mercaptopurine, gemcitabine (Gemzar®; Gemzar is a registered trademark of EliLilly and Company), methotrexate, and pemetrexed (Alimta®; Alimta is a registered trademark of EliLilly and Company). Company). Topoisomerase inhibitors stop cancer cells from dividing by preventing the topoisomerase enzyme from separating DNA for replication. Some common topoisomerase inhibitors include topotecan, irinotecan, etoposide, and teniposide. Mitotic inhibitors stop cancer cells from dividing by inhibiting key cell division enzymes. Some common mitotic inhibitors include taxanes (e.g., paclitaxel (Taxol®; Taxol is a registered trademark of Bristol-Myers Squibb Company) and docetaxel (Taxotere®; Taxotere is a registered trademark of Aventis Pharma SA)), epothilones, and vinca alkaloids.

[0008] One drawback of all these anticancer drugs is that they also damage healthy tissues. Because drugs treat cancer by inhibiting the function of normal cells, they can also severely damage healthy tissues that rely on constant cell division, such as blood cells, mucosal surfaces, and skin. This damage can result in significant morbidity and limit the amount of chemotherapy that can be safely delivered. Side effects that occur during chemotherapy treatment include low blood counts, hair loss, muscle and joint pain, nausea, vomiting, diarrhea, mouth sores, fever, and chills. To overcome this problem, novel drugs with unique mechanisms of action that provide increased targeting and affect proteins and cellular functions that occur only in cancer cells continue to be developed. For example, antibody-drug conjugates (ADCs) are designed to bind to specific epitopes on the surface of tumor cells, providing an alternative method of targeting tumor cells to reduce associated toxicity. Although highly selective, they achieve only low cellular uptake (<1% of the injected drug) and have limited cell-killing activity, rendering few ADCs therapeutically useful. Some specific anticancer drugs include imatinib (Gleevec®; Gleevec is a registered trademark of Novartis AG), gefitinib (Iressa®; Iressa is a registered trademark of AstraZeneca UK Limited), sunitinib (Sutent®; Sutent is a registered trademark of CP Pharmaceuticals, International CV), and bortezomib (Velcade®; Velcade is a registered trademark of Millennium Pharmaceuticals, Inc.). However, these drugs are not approved for all cancer types and are universally associated with the development of treatment resistance. In addition, many of these compounds still lack absolute tumor selectivity, and off-target effects continue to limit their therapeutic utility.

[0009] Recently, phospholipid ether ("PLE") analogs have been demonstrated to be effective molecular platforms for anticancer drug delivery. See U.S. Patent No. 9,480,754 and Weichertetal et al. (Sci TranslMed, 2014, 6(240), 240ra75), each of which is incorporated herein by reference in its entirety. As seen therein, the majority of clinically used anticancer drugs have limited utility due to their toxicity to all proliferating cells and / or their inability to exert their effects in all tumor cells. Therefore, there remains a need in the art for alternative anticancer drug delivery vehicles that can deliver potent, effective, broad-spectrum anticancer drugs to cancer cells, including CSCs, while avoiding substantial drug uptake by healthy cells. Furthermore, anticancer drug delivery vehicles should be able to cross barriers such as the blood-brain barrier (BBB). Summary of the Invention

[0010] overview In one aspect, the present disclosure provides a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof, where: n is 2 to 20; Q 1 is a bond or the following chemical formula [ka] where m is between 0 and 100; L is the following chemical formula [ka] where R x is H or a halogen; Q 2 is a bond or self-immolative spacer; and Z is an anticancer drug.

[0011] In another embodiment, the present disclosure provides a method of treating a subject in need thereof, comprising administering an effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof.

[0012] The present disclosure provides other aspects and embodiments that will become apparent in light of the following detailed description and accompanying drawings. [Brief explanation of the drawings]

[0013] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1A] Figures 1A-1B show the uptake of phospholipid drug conjugates (PDCs) into tumor cell lines. Figure 1A shows phospholipid ether (PLE) + BODIPY, with green fluorescence. [Figure 1B] FIG. 1B shows the ratio of MFI to autofluorescence ratio of CLR1502 uptake.

[0014] [Figure 2A] Figures 2A-2B show the uptake of PDCs into tumor cell lines (A375 and A549 cell lines). Figure 2A shows the concentration of fully conjugated PLE in the cytoplasm. [Figure 2B] FIG. 2B shows the concentration of released payload in the cytoplasm.

[0015] [Figure 3] Figure 3 shows the uptake of CLR1502 and CLR1501 via lipid rafts in tumor cells and primary tumor samples, respectively. CLR1502 is a near-infrared molecule bound to PLEs (Figure 3A, Figure 3B). Blue is Hoechst nuclear staining. Red is cholera toxin subunit B, indicating lipid rafts.

[0016] [Figure 4] FIG. 4 shows the in vitro efficacy of PDC-SM2 against melanoma (A375) and lung cancer (A549) cells.

[0017] [Figure 5] Figure 5 shows the cytotoxicity of PDCs tolerated in vivo. At a payload dose of 0.5 mg / kg, the circles indicate when mice died or were sacrificed. The arrows indicate when the dose was administered.

[0018] [Figure 6] FIG. 6 shows the in vitro uptake of CLR2000045 in MCF-7 and NHDF cell lines.

[0019] [Figure 7] FIG. 7 shows the in vitro cytotoxicity of CLR2000045 in breast cancer cell lines.

[0020] [Figure 8] FIG. 8 shows the in vivo antitumor activity in a chick embryo chorioallantoic membrane model (MCF-7).

[0021] [Figure 9] FIG. 9 shows the in vivo antitumor effect in a xenograft model transplanted with TNBC (HCC70).

[0022] [Figure 10] FIG. 10 shows Kaplan-Meier survival curves in TNBC (HCC70) xenograft model mice.

[0023] [Figure 11] Figures 11A and 11B show the changes in body weight of (HCC70) xenograft model mice after treatment. Figure 11A shows the administration of 1 mg / kg three times a week. Figure 11B shows the administration of 1 mg / kg twice a week.

[0024] [Figure 12] FIG. 12 shows the in vitro uptake of CLR180099A and CLR180099B in A549 and NHDF cells.

[0025] [Figure 13] Figure 13 shows the in vitro uptake of CLR180095 in A549 (black line) and HCT116 (gray line) cells. Cells were incubated for 48 hours, with an initial incubation concentration of 100 nM. Uptake was assessed by LC / LC / MS.

[0026] [Figure 14] FIG. 14 shows the in vitro release of the payload in A549 cells.

[0027] [Figure 15] FIG. 15 shows the in vitro cytotoxicity of CLR180099A in lung cancer, breast cancer and melanoma cells.

[0028] [Figure 16] FIG. 16 shows the in vivo antitumor effect of CLR180099A in an implanted colon cancer xenograft model.

[0029] [Figure 17] FIG. 17 shows Kaplan-Meier survival curves for CLR180099A in a colon cancer xenograft model.

[0030] [Figure 18] Figure 18 shows the in vivo tolerability of CLR180099A.

[0031] [Figures 19A-19C]Figures 19A-19F show the selective uptake of CLR1502 in intestinal tumors. Figure 19A shows the whole colon removed at necropsy 96 hours after administration of 50 μg of CLR1502 per mouse. Figure 19B shows the terminal portion of the small intestine removed at necropsy 96 hours after administration of 50 μg of CLR1502 per mouse. Using IVIS Spectrum, we observed areas of increased signal intensity. These areas represent non-invasive (colon Figure 19C; terminal portion of small intestine Figure 19F) and invasive (colon Figure 19D; terminal small intestine Figure 19E) tumors. Figures 19C, 19D, 19E, and 19F are magnified as indicated by the black boxes. Arrows point to malignant glands within the intestinal musculature. Bar: 1 mm. [Figures 19D-19E] Using IVIS Spectrum, areas of increased signal intensity were observed. These areas represent non-invasive (colon Figure 19C; terminal small intestine Figure 19F) and invasive (colon Figure 19D; terminal small intestine Figure 19E) tumors. Figures 19C, 19D, 19E, and 19F are enlarged as indicated by the black boxes. Arrows point to malignant glands within the intestinal musculature. Bar: 1 mm. [Figure 19F] Using IVIS Spectrum, areas of increased signal intensity were observed. These areas represent non-invasive (colon Figure 19C; terminal small intestine Figure 19F) and invasive (colon Figure 19D; terminal small intestine Figure 19E) tumors. Figures 19C, 19D, 19E, and 19F are enlarged as indicated by the black boxes. Arrows point to malignant glands within the intestinal musculature. Bar: 1 mm.

[0032] [Figure 20A]Figures 20A-20F show CLR1501 uptake in the brain. Figures 20A, 20B, and 20C show U251-derived orthotopic brain tumors labeled with CLR1501 (Figure 20C) along with ToPro3 nuclear counterstain (Figure 20B), verified by magnetic resonance imaging (MRI; Figure 20A, T2-weighted image). Figures 20D, 20E, and 20F are histological analyses of the brain tumor border in 22T glioblastoma multiforme-derived orthotopic xenografts labeled with CLR1501 (green). Figure 20D is an epifluorescence visualization of the xenograft brain border with blue DAPI nuclear counterstain. Figure 20E is a confocal image of a CLR1501-labeled xenograft. Figure 20F is a confocal and brightfield image of the xenograft and adjacent normal brain. N indicates normal brain; RFU indicates relative fluorescence units; T indicates tumor. [Figure 20B] Figures 20A-20F show CLR1501 uptake in the brain. Figures 20A, 20B, and 20C show U251-derived orthotopic brain tumors labeled with CLR1501 (Figure 20C) along with ToPro3 nuclear counterstain (Figure 20B), verified by magnetic resonance imaging (MRI; Figure 20A, T2-weighted image). Figures 20D, 20E, and 20F are histological analyses of the brain tumor border in 22T glioblastoma multiforme-derived orthotopic xenografts labeled with CLR1501 (green). Figure 20D is an epifluorescence visualization of the xenograft brain border with blue DAPI nuclear counterstain. Figure 20E is a confocal image of a CLR1501-labeled xenograft. Figure 20F is a confocal and brightfield image of the xenograft and adjacent normal brain. N indicates normal brain; RFU indicates relative fluorescence units; T indicates tumor. [Figure 20C]Figures 20A-20F show CLR1501 uptake in the brain. Figures 20A, 20B, and 20C show U251-derived orthotopic brain tumors labeled with CLR1501 (Figure 20C) along with ToPro3 nuclear counterstain (Figure 20B), verified by magnetic resonance imaging (MRI; Figure 20A, T2-weighted image). Figures 20D, 20E, and 20F are histological analyses of the brain tumor border in 22T glioblastoma multiforme-derived orthotopic xenografts labeled with CLR1501 (green). Figure 20D is an epifluorescence visualization of the xenograft brain border with blue DAPI nuclear counterstain. Figure 20E is a confocal image of a CLR1501-labeled xenograft. Figure 20F is a confocal and brightfield image of the xenograft and adjacent normal brain. N indicates normal brain; RFU indicates relative fluorescence units; T indicates tumor. [Figure 20D] Figures 20A-20F show CLR1501 uptake in the brain. Figures 20A, 20B, and 20C show U251-derived orthotopic brain tumors labeled with CLR1501 (Figure 20C) along with ToPro3 nuclear counterstain (Figure 20B), verified by magnetic resonance imaging (MRI; Figure 20A, T2-weighted image). Figures 20D, 20E, and 20F are histological analyses of the brain tumor border in 22T glioblastoma multiforme-derived orthotopic xenografts labeled with CLR1501 (green). Figure 20D is an epifluorescence visualization of the xenograft brain border with blue DAPI nuclear counterstain. Figure 20E is a confocal image of a CLR1501-labeled xenograft. Figure 20F is a confocal and brightfield image of the xenograft and adjacent normal brain. N indicates normal brain; RFU indicates relative fluorescence units; T indicates tumor. [Figure 20E]Figures 20A-20F show CLR1501 uptake in the brain. Figures 20A, 20B, and 20C show U251-derived orthotopic brain tumors labeled with CLR1501 (Figure 20C) along with ToPro3 nuclear counterstain (Figure 20B), verified by magnetic resonance imaging (MRI; Figure 20A, T2-weighted image). Figures 20D, 20E, and 20F are histological analyses of the brain tumor border in 22T glioblastoma multiforme-derived orthotopic xenografts labeled with CLR1501 (green). Figure 20D is an epifluorescence visualization of the xenograft brain border with blue DAPI nuclear counterstain. Figure 20E is a confocal image of a CLR1501-labeled xenograft. Figure 20F is a confocal and brightfield image of the xenograft and adjacent normal brain. N indicates normal brain; RFU indicates relative fluorescence units; T indicates tumor. [Figure 20F] Figures 20A-20F show CLR1501 uptake in the brain. Figures 20A, 20B, and 20C show U251-derived orthotopic brain tumors labeled with CLR1501 (Figure 20C) along with ToPro3 nuclear counterstain (Figure 20B), verified by magnetic resonance imaging (MRI; Figure 20A, T2-weighted image). Figures 20D, 20E, and 20F are histological analyses of the brain tumor border in 22T glioblastoma multiforme-derived orthotopic xenografts labeled with CLR1501 (green). Figure 20D is an epifluorescence visualization of the xenograft brain border with blue DAPI nuclear counterstain. Figure 20E is a confocal image of a CLR1501-labeled xenograft. Figure 20F is a confocal and brightfield image of the xenograft and adjacent normal brain. N indicates normal brain; RFU indicates relative fluorescence units; T indicates tumor.

[0033] [Figure 21A] FIG. 21A shows a CLR1502-treated brain in vivo with visible light (left) and CLR1502 fluorescence of an in vivo 22CSC-derived orthotopic xenograft (right).

[0034] [Figure 21B]Figure 21B shows visible light (top left) and CLR1502 fluorescence (top right) of 22 CSC-derived xenografts ex vivo demonstrating excellent macroscopic tumor delineation from normal brain. The figure also shows histological verification of tumor (T) (hematoxylin and eosin; bottom left) and normal brain (hematoxylin and eosin; bottom right).

[0035] [Figure 22] Figure 22 shows that tumor thickness does not account for the increased signal intensity seen in intestinal cancer. Figure 22A shows the layers of the colon. Figure 22B shows the total radiant efficiency of each layer.

[0036] [Figure 23] Figure 23 shows in vivo optical scanning of CLR1502 uptake in a colon cancer model. Fluorescence intensity (indicated by color bar) and biodistribution were measured in vivo over time.

[0037] [Figure 24] Figure 24 shows in vivo optical scanning of CLR1502 uptake in a breast cancer model. Orthotopic breast cancer xenografted athymic nude mice (MDA-MB-231) were imaged daily for 7 days (168 hours) using Fluoptics Fluobeam® and IVIS® Spectrum systems (yellow and green arrows indicate Fluobeam and IVIS Spectrum, respectively).

[0038] [Figure 25] Figure 25 shows an athymic nude mouse intravenously injected with CLR1502 with a lung cancer xenograft (H226 lung) in each flank, imaged using IVIS Spectrum. At 96 hours, the difference in radiative efficiency between malignant and normal tissue creates sufficient contrast for the tumor margin, as indicated by the black arrows. DETAILED DESCRIPTION OF THE INVENTION

[0039] Detailed Description Described herein are therapeutic compounds that can target a wide range of tumor cells. The compounds disclosed herein can target special structures in tumor cell membranes, such as lipid rafts. Therefore, the compounds disclosed herein can be used to target tumor cells with high specificity. In particular, the compounds disclosed herein can be used in cancer treatment.

[0040] 1.Definition Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. In case of conflict, the present document, including definitions, will control. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and are not intended to be limiting.

[0041] As used herein, the terms "comprise(s)," "include(s)," "having," "has," "can," and "contain(s)," and variations thereof, are intended to be open-ended transitional phrases, terms, or words that do not exclude the possibility of additional acts or structures. The singular forms "a," "and," and "the" include plural references unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments that "comprising," "consisting of," and "consisting essentially of" the embodiments or elements presented herein, whether expressly stated or not.

[0042] For the recitation of numerical ranges herein, each intervening value is expressly contemplated with the same degree of precision. For example, in the range of 6 to 9, the values ​​7 and 8 are contemplated in addition to 6 and 9, and in the range of 6.0 to 7.0, the values ​​6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are expressly contemplated.

[0043] As used herein, the term "about" or "approximately" as applied to one or more values ​​of interest refers to a value similar to a stated reference value or within an acceptable error range of a particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, such as the limitations of the measurement system. In certain embodiments, the term "about" refers to a range of values ​​that fall within 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in any direction (greater or less than) of the stated reference value, unless otherwise specified or otherwise clear from the context (unless such number would exceed 100% of the possible values). Alternatively, "about" can mean within 3 standard deviations or more, as is customary in the art. Alternatively, such as with respect to biological systems or processes, the term "about" can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold of a value.

[0044] The definitions of specific functional groups and chemical terms are described in more detail below.For the purpose of this disclosure, chemical elements are identified according to the Periodic Table of Elements (CAS version, Handbook of Chemistry and Physics, 75th Edition, inside cover), and specific functional groups are generally defined as described therein.In addition, the general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March March's Advanced Organic Chemistry, 5th Edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; Carruthers, Some Modern Methods of Organic Synthesis, 3rd Edition, Cambridge University Press, Cambridge, 1987; the entire contents of each of these are incorporated herein by reference.

[0045] The term "cancer" as used herein refers to any disease resulting from uncontrolled cell division capable of metastasis. The term "cancer" as used herein refers to breast cancer, including male breast cancer; digestive / gastrointestinal cancer, including anal cancer, appendix cancer, extrahepatic bile duct cancer, gastrointestinal carcinoid, colon cancer, esophageal cancer, gallbladder cancer, gastric cancer, gastrointestinal stromal tumor ("gist"), pancreatic islet cell tumor, adult primary liver cancer, pediatric liver cancer, pancreatic cancer, rectal cancer, small intestine cancer, and stomach (gastric) cancer; pancreatic adenocarcinoma, adrenocortical carcinoma, pancreatic neuroendocrine tumor, Merkel cell carcinoma, non-small cell lung neuroendocrine tumor, small cell lung neuroendocrine tumor, parathyroid carcinoma, pheochromocytoma, pituitary tumor, and thyroid tumor. endocrine and neuroendocrine cancers, including carcinoma; eye cancer, including intraocular melanoma and retinoblastoma; genitourinary cancers, including bladder cancer, kidney (renal cell) cancer, penile cancer, prostate cancer, transitional cell renal pelvis and ureter cancer, testicular cancer, urethral cancer, and Wilms' tumor; germ cell cancers, including pediatric central nervous system cancer, pediatric extracranial germ cell tumors, extragonadal germ cell tumors, ovarian germ cell tumors, and testicular cancer; gynecological cancers, including cervical cancer, endometrial cancer, gestational trophoblastic tumor, epithelial ovarian cancer, ovarian germ cell tumors, uterine sarcoma, vaginal cancer, and vulvar cancer; hypopharyngeal cancer, laryngeal cancer, lip and oral cavity cancer, metastatic cancer with occult primary Head and neck cancers, including metastatic squamous cell carcinoma of the neck, oral cancer, nasopharyngeal cancer, oropharynx cancer, paranasal sinus and nasal cavity cancer, parathyroid cancer, pharyngeal cancer, salivary gland cancer, and throat cancer; leukemias, including adult acute lymphoblastic leukemia, childhood acute lymphoblastic leukemia, adult acute myeloid leukemia, childhood acute myeloid leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell leukemia; multiple myeloma, including malignant plasma cell lymphoma; AIDS-related lymphoma, cutaneous T-cell lymphoma, adult Hodgkin lymphoma, childhood Hodgkin lymphoma, Hodgkin lymphoma during pregnancy, mycosis fungoides, and adult non-Hodgkin's lymphoma Lymphomas, including childhood non-Hodgkin's lymphoma, non-Hodgkin's lymphoma during pregnancy, primary central nervous system lymphoma, Sézary syndrome, and Waldenstrom's macroglobulinemia; musculoskeletal cancers, including Ewing's sarcoma, osteosarcoma and malignant fibrous histiocytoma of bone, childhood rhabdomyosarcoma, and soft tissue sarcoma; nervous system cancers, including adult brain tumors, childhood brain tumors, astrocytoma, brain stem glioma, central nervous system atypical teratoid / rhabdoid tumor, central nervous system embryonal tumors, craniopharyngioma, ependymoma, neuroblastoma, and primary central nervous system (CNS) malignant lymphoma;This refers to various types of cancer, including, but not limited to, respiratory / thoracic cancers, including non-small cell lung cancer, small cell lung cancer, malignant mesothelioma, thymoma, and thymic carcinoma; and skin cancers, including Kaposi's sarcoma, melanoma, and squamous cell carcinoma.

[0046] As used herein, the term "cancer stem cell" refers to a cancer cell that is capable of self-renewal and differentiation into the different types of cancer cells found in malignant tumors.

[0047] The terms "chemotherapeutic agent," "anti-cancer agent," and "anti-tumor agent" are used interchangeably throughout this specification.

[0048] Generally, reference to a "circulating tumor cell" is intended to refer to a single cell, while reference to a "circulating tumor cell" or a "cluster of circulating tumor cells" is intended to refer to one or more cancer cells. However, those skilled in the art will understand that reference to a "circulating tumor cell" is intended to include a population of circulating tumor cells comprising one or more circulating tumor cells, while reference to a "circulating tumor cell" can include one or more circulating tumor cells. As used herein, the term "circulating tumor cell" or "circulating tumor cells" refers to any cancer cell or cluster of cancer cells found in a subject's blood or serum sample. CTCs can also comprise or consist of cancer stem cells or clusters of cancer stem cells found in a subject's blood or serum sample.

[0049] As used herein, the term "composition" is intended to encompass a product containing specified ingredients in specified amounts, as well as any product that results, directly or indirectly, from combining specified ingredients in specified amounts.

[0050] The terms "control," "reference level," and "reference" are used interchangeably herein. A reference level is used as a benchmark for evaluating measurement results and can be a predetermined value or range. As used herein, a "control group" refers to a group of control subjects. A predetermined level can be a cutoff value from the control group. A predetermined level can be the mean from the control group. A cutoff value (or a predetermined cutoff value) can be determined by Adaptive Index Model (AIM) methodology. A cutoff value (or a predetermined cutoff value) can be determined by receiver operating curve (ROC) analysis from biological samples from a patient group. As commonly known in the biological field, ROC analysis determines the ability of a test to distinguish one condition from another, for example, determining the performance of each marker in identifying ideal patients for IL-1Ra therapy. A description of ROC analysis is provided by PJ Heagertyetal et al. (Biometrics 2000, 56, 337-44), the disclosure of which is incorporated herein by reference in its entirety. Alternatively, the cutoff value can be determined by quartile analysis of biological samples from a patient group. For example, the cutoff value can be determined by selecting a value corresponding to any value in the 25th to 75th percentile range, preferably the 25th, 50th, or 75th percentile, and more preferably the 75th percentile. Such statistical analysis can be performed using any method known in the art and can be implemented through any number of commercially available software packages (e.g., from Analyse-it Software Ltd., Leeds, UK; StataCorp LP, College Station, TX; SAS Institute Inc., Cary, NC). Healthy or normal levels or ranges for target or protein activity can be defined according to standard practice. The control can be a tumor-free subject or cell, as described in detail herein.The control may be a subject, or a sample from a subject, whose condition is known. The subject, or sample from a subject, may be healthy, diseased, diseased before treatment, diseased during treatment, diseased after treatment, or a combination thereof.

[0051] The term "dose" as used herein means any form of active ingredient formulation or composition containing at least an amount sufficient to produce a therapeutic effect upon single administration. "Formulation" and "compound" are used interchangeably herein.

[0052] As used herein, the term "dosage" refers to the administration of any amount, number, and frequency of doses over a particular period of time.

[0053] As used herein, the term "effective amount" or "therapeutically effective amount" refers to any quantity of a drug or pharmaceutically acceptable composition or compound administered that will relieve to some extent one or more symptoms of the disease or condition being treated. The result can be a reduction and / or alleviation of the signs, symptoms, or causes of a disease, or other desired alteration of a biological system. For example, an "effective amount" for therapeutic use is the amount of a composition comprising a compound disclosed herein that is required to provide a clinically significant reduction in a disease symptom. An appropriate "effective" amount in any individual case can be determined using techniques, such as a dose escalation study.

[0054] As used herein, the term "halogen" means Cl, Br, I, F, At, or a synthetic halogen such as tennessine (Ts).

[0055] The term "heterocycloalkyl," as used herein, refers to a cyclic group of 3 to 24 atoms (C3-C24) selected from carbon, nitrogen, sulfur, phosphate, and oxygen, where at least one atom is carbon.

[0056] As described herein, the term "isomer" includes, but is not limited to, optical isomers and analogs, structural isomers and analogs, conformational isomers and analogs, etc. In one embodiment, the present disclosure encompasses the use of different optical isomers as detailed herein. It will be understood by those skilled in the art that the anti-cancer compounds useful in the present invention may contain at least one asymmetric center (steriogenic center). Thus, the compounds used in the methods of the present invention may exist and be isolated in optically active or racemic forms. Some compounds may also exhibit polymorphism.

[0057] The terms "malignant tumor cell," "tumor cell," and "cancer cell" are used interchangeably throughout this specification. The terms "malignant tumor stem cell," "tumor stem cell," and "cancer stem cell" are used interchangeably throughout this specification.

[0058] As used herein, a "sample" or "test sample" can refer to any sample in which the presence and / or level of a target is detected or measured. A sample can include a liquid, solution, emulsion, or suspension. A sample can include a medical sample. A sample can include any biological fluid or tissue, such as blood, whole blood, blood fractions such as plasma and serum, cartilage, ligaments, tendons, muscles, interstitial fluid, sweat, saliva, urine, tears, synovial fluid, synovial membrane, meniscus, bone marrow, cerebrospinal fluid, nasal secretions, sputum, amniotic fluid, bronchoalveolar lavage fluid, gastric lavage, vomit, feces, lung tissue, peripheral blood mononuclear cells, total white blood cells, lymph node cells, spleen cells, tonsil cells, cancer cells, tumor cells, bile, digestive fluid, skin, or a combination thereof. In some embodiments, a sample comprises an aliquot. In other embodiments, a sample comprises a biological fluid. A sample can be obtained by any means known in the art. The sample can be used directly as obtained from the patient, or it can be pretreated in several ways, such as those discussed herein or otherwise known in the art, by filtration, distillation, extraction, concentration, centrifugation, inactivation of interfering components, addition of reagents, etc., to modify the characteristics of the sample.

[0059] The terms "subject" and "patient," used interchangeably herein, refer to any vertebrate, including, but not limited to, a mammal, that desires or requires the compositions or methods described herein. A subject can be human or non-human. A subject can be a vertebrate. A subject can be a mammal. A mammal can be a primate or a non-primate. A mammal can be a non-primate, such as, for example, a cow, pig, camel, llama, hedgehog, anteater, platypus, elephant, alpaca, horse, goat, rabbit, sheep, hamster, guinea pig, cat, dog, rat, and mouse. A mammal can be a primate, such as a human. A mammal can be a non-human primate, such as, for example, a monkey, cynomolgous monkey, rhesus monkey, chimpanzee, gorilla, orangutan, or gibbon. A subject can be of any age or stage of development, such as, for example, an adult, adolescent, or infant. A subject can be male. A subject can be female. In some embodiments, the subject has a particular cancer. The subject may be undergoing other forms of treatment.

[0060] As used herein, the term "therapeutic compound" refers to any compound capable of providing cancer treatment.

[0061] The term "treat" or "treating" or "treatment" means suppressing, repressing, reversing, mitigating, ameliorating, or inhibiting the progression of a disease, or eliminating the disease entirely. Treatment can be performed in either acute or chronic conditions. The term also refers to reducing the severity of a disease or symptoms associated with such a disease.

[0062] Unless otherwise defined herein, scientific and technical terms used in connection with this disclosure shall have the meanings commonly understood by those skilled in the art. For example, any technical terms used in connection with cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization described herein, and the techniques thereof, are well known and commonly used in the art. The meaning and scope of terms should be clear; however, in the event of any potential ambiguity, the definitions provided herein shall prevail over any dictionary or external definitions. Furthermore, unless otherwise required by context, the singular shall include the plural, and the plural shall include the singular.

[0063] 2.Compound In one aspect, the present disclosure provides a compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, n is 2 to 20; Q 1 is a bond or the following chemical formula [ka] where m is between 0 and 100; L is the following chemical formula [ka] where R x is H or a halogen; Q 2 is a bond or self-immolative spacer; and Z is an anticancer drug.

[0064] The number "n" can be any integer from 2 to 20. In some embodiments, n is 2, 4, 6, 8, 10, 12, 14, 16, 18, or 20. In certain embodiments, n is 18.

[0065] The number "m" can be any integer between 0 and 100. In some embodiments, m is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, m is an integer between 10 and 20, an integer between 10 and 40, an integer between 10 and 60, or an integer between 10 and 80. In some embodiments, m is 0, and Q 1 is a bond or the following chemical formula [ka] is.

[0066] Q 2 may be any known self-immolative spacer, including, for example, para-aminobenzyloxycarbonyl (PABC).

[0067] In some embodiments, R x is H. In some embodiments, R x is Cl.

[0068] In some embodiments, n is 2 to 20, and Q 1 is a bond or the following chemical formula [ka] and LQ 2 Part [ka] and R x is H or a halogen, and Z is an anticancer agent.

[0069] Z can be any anti-cancer agent, including a variety of known chemotherapeutic agents.

[0070] In some embodiments, Z is a polo-like kinase 1 (PLK-1) inhibitor. Suitable PLK-1 inhibitors include, for example, diaminopyrimidine (DAP) derivatives such as BI2536, BI6727 (volasertib), DAP-81 and DAP-83, and compounds disclosed in Kumar et al. (Biomed Res Int. 2015, 2015: 705745) and Peters et al. (Nat Chem Biol. 2006, 2(11):618-26), the contents of which are incorporated herein in their entireties.

[0071] In some embodiments, Z is a tubulin polymerase inhibitor such as nocodazole.

[0072] In some embodiments, Z is a tubulin stabilizer such as a taccalonolide.

[0073] In some embodiments, Z is an anti-tumor drug such as monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), or monomethylauristatin D (MMAD).

[0074] In some embodiments, Z is a eukaryotic translation initiation factor 4 (EIF4) inhibitor, such as an EIF4A or EIF4E inhibitor. In some embodiments, Z is an EIF4E inhibitor. Suitable EIF4 inhibitors include, for example, ribavirin and the compounds disclosed by D'Abronzo et al. (Neoplasia, 2018, 20(6), 563-573) and U.S. Patent No. 10,577,378, the contents of which are incorporated herein by reference in their entirety.

[0075] In some embodiments, Z is a combretastatin A-4 analog, such as combretastatin A-4 phosphate or ombrabulin. Suitable combretastatin A-4 analogs also include, for example, compounds disclosed by Beilina et al. (Bioorganic & Medicinal Chemistry Letters 2006, 16(22), 5757-5762), the contents of which are incorporated herein in their entirety.

[0076] In some embodiments, Z is a flavoglin analog. Suitable flavoglin analogs include, for example, compounds disclosed in U.S. Patent Application Publication No. US 2018 / 0086729, the contents of which are incorporated herein in their entirety.

[0077] In certain embodiments, Z is one of other known anti-cancer agents, including, for example, (i) other antiproliferative / anti-tumor agents such as alkylating agents, antimetabolites, antitumor antibiotics, antimitotic agents, and topoisomerase inhibitors; (ii) cytostatic agents such as antiestrogens, antiandrogens, LHRH antagonists or agonists, progestogens, and aromatase inhibitors; (iii) anti-invasive agents (e.g., c-Src kinase family inhibitors); (iv) inhibitors of growth factor function such as tyrosine kinase inhibitors; (v) anti-angiogenic agents; (vi) vascular damaging agents; and (vii) endothelin receptor antagonists.

[0078] Examples of suitable anti-cancer agents include paclitaxel, irinotecan, topotecan, gemcitabine, cisplatin, geldanamycin, mertansine, abiraterone, afatinib, aminolevulinic acid, aprepitant, axitinib, azacitidine, belinstat, bendamustine, bexarotene, bleomycin, bortezomib, bosutinib, busulfan, cabazitaxel, cabozantinib, capecitabine, carboplatin, carfilzomib, carmustine, ceritinib, cetuximab, chlorambucil, clofarabine, crizotinib, cyclophosphamide, and the like. , cytarabine, dabrafenib, dacarbazine, dactinomycin, dasatinib, daunorubicin, decitabine, denosumab, dexrazoxane, docetaxel, dolastatins (e.g., monomethyl auristatin E), doxorubicin, enzalutamide, epirubicin, eribulin mesylate, erlotinib, etoposide, everolimus, floxuridine, fludarabine phosphate, fluorouracil, ganetespib, gefitinib, gemtuzumab ozogamicin, hexamethylmelamine, hydroxyurea, ibritumomab tiuxetan, ibrutinib Nib, idelalisib, ifosfamide, imatinib, ipilimumab, ixabepilone, lapatinib, leucovorin calcium, lomustine, maytansinoids, mechlorethamine, melphalan, mercaptopurine, mesna, methotrexate, mitomycin C, mitotane, mitoxantrone, nelarabine, nelfinavir, nilotinib, obinutuzumab, ofatumumab, omacetaxine mepesuccinate, oxaliplatin, panitumumab, pazopanib, pegasus propionate, pembrolizumab, pemetrexed, pentostatin, pertuzumab, prilostatin Plicanycin, pomalidomide, ponatinib hydrochloride, pralatrexate, procarbazine, radium-223 dichloride, ramucirumab, regorafenib, letaspimycin, ruxolitinib, semustine, siltuximab, sorafenib, streptozocin, sunitinib malate, tanespimycin, temozolomide, temsirolimus, teniposide, thalidomide, thioguanine, thiotepa, toremifene, trametinib, trastuzumab, vandetanib, vemurafenib, vinblastine, vincristine, vinorelbine,including, but not limited to, vismodegib, vorinostat, and zirbab-aflibercept.

[0079] In some embodiments, the compound of Formula (I) has the structure of Formula (Ia): 1 is the following chemical formula [ka] and LQ 2 is the following chemical formula [ka] and Z is a PLK-1 inhibitor, a tubulin polymerase inhibitor, a tubulin stabilizer, an anti-tumor agent, or a eukaryotic translation initiation factor 4 (EIF4) inhibitor. Specifically, formula (Ia) can be represented by the following formula: [ka] or a pharmaceutically acceptable salt thereof, wherein n is 2 to 20, and Z is a PLK-1 inhibitor, a tubulin polymerase inhibitor, a tubulin stabilizer, an anti-tumor agent, or a eukaryotic translation initiation factor 4 (EIF4) inhibitor.

[0080] In some embodiments, the compound has the structure of formula (Ia), where n is 18. In some embodiments, the compound has the structure of formula (Ia), where Z is a PLK-1 inhibitor or an anti-tumor agent. In some embodiments, the compound has the structure of formula (Ia-1), (Ia-2), or (Ia-3), or a pharmaceutically acceptable salt thereof, where Z is a PLK-1 inhibitor. For example, Z has the following formula: [ka] It could be.

[0081] In some embodiments, the compound has the structure of formula (Ia-1), (Ia-2), or (Ia-3), or a pharmaceutically acceptable salt thereof, where Z is an antitumor agent selected from the group consisting of monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), and monomethyl auristatin E (MMAD). In some embodiments, the compound has the structure of formula (Ia-3), or a pharmaceutically acceptable salt thereof, where Z is MMAE, MMAF, or MMAD (shown below). [ka] is.

[0082] In some embodiments, the compound of Formula (I) has the structure of Formula (Ib), or a pharmaceutically acceptable salt thereof, where n is 18 and Q 1 is the following chemical formula [ka] and LQ 2 is the following chemical formula [ka] and Z is a combretastatin A-4 analog. Specifically, formula (Ib) is [ka] or a pharmaceutically acceptable salt thereof, wherein Z is a combretastatin A-4 analog.

[0083] In some embodiments, the compound has the structure of formula (Ibl), (Ib-2), or (Ib-3), or a pharmaceutically acceptable salt thereof, wherein Z is [ka] and combretastatin A-4 analogs such as

[0084] In some embodiments, the compound of Formula (I) has the structure of Formula (Ic), or a pharmaceutically acceptable salt thereof, where n is 18 and Q 1 is a bond or the following chemical formula [ka] and LQ 2 is the following chemical formula [ka] and Z is a flavogrine analog. Specifically, the chemical formula (Ic) is [ka] or a pharmaceutically acceptable salt thereof, wherein Z is a flavoglin analog.

[0085] In some embodiments, the compound has the structure of formula (Ic-1), (Ic-2), or (Ic-3), or a pharmaceutically acceptable salt thereof, wherein Z is [ka] and other flavugrine analogs.

[0086] Suitable compounds disclosed herein include those having the following chemical formula: [ka] [ka] or a pharmaceutically acceptable salt thereof.

[0087] The disclosed compounds can exist as pharmaceutically acceptable salts. The term "pharmaceutically acceptable salt" refers to a water- or oil-soluble or dispersible salt or zwitterion of a compound that is effective for its intended use in treating disorders without undue toxicity, irritation, and allergic response, commensurate with a reasonable benefit / risk ratio. Representative salts include acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfite, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, formate, isethionate, fumarate, lactate, maleate, methanesulfonate, naphthylenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, oxalate, maleate, pivalate, propionate, succinate, tartrate, trichloroacetate, trifluoroacetate, glutamate, paratoluenesulfonate, undecanoate, hydrochloride, hydrobromide, sulfate, phosphate, and the like. The amino groups of the compounds may also be quaternized with alkyl chlorides, bromides, and iodides, such as methyl, ethyl, propyl, isopropyl, butyl, lauryl, myristyl, stearyl, and the like.

[0088] Base addition salts may be prepared during the final isolation and purification of the disclosed compounds by reaction of the carboxyl group with a suitable base, such as the hydroxide, carbonate, or bicarbonate of a metal cation, such as lithium, sodium, potassium, calcium, magnesium, or aluminum, or with a primary, secondary, or tertiary organic amine. Quaternary amine salts can be prepared, such as those derived from methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine, tributylamine, pyridine, N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, dicyclohexylamine, procaine, dibenzylamine, N,N-dibenzylphenethylamine, 1-ephenamine, and N,N'-dibenzylethylenediamine, ethylenediamine, ethanolamine, diethanolamine, piperidine, and piperazine.

[0089] Compounds may exist as stereoisomers where asymmetric or chiral centers are present. Stereoisomers are "R" or "S" depending on the configuration of substituents around the chiral carbon atom. As used herein, the terms "R" and "S" refer to the configurations described in the IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, in Pure Appl. Chem., 1976, 45:13-30. The present disclosure contemplates various stereoisomers and mixtures thereof, and these are specifically included within the scope of the present disclosure. Stereoisomers include enantiomers and diastereomers, and mixtures of enantiomers or diastereomers. Individual stereoisomers of compounds can be prepared synthetically from commercially available starting materials containing asymmetric or chiral centers, or by preparation of racemic mixtures followed by separation methods well known to those skilled in the art. These separation methods include (1) coupling the mixture of enantiomers to a chiral auxiliary, separating the resulting mixture of diastereomers by recrystallization or chromatography, and optionally liberating the optically pure product from the auxiliary, as described in Furniss, Hannaford, Smith, and Tatchell, "Vogel's Textbook of Practical Organic Chemistry," 5th edition (1989), Longman Scientific & Technical, Essex CM20 2JE, England; (2) direct separation of the mixture of optical enantiomers in a chiral chromatography column; or (3) fractional recrystallization. Compounds may have tautomeric forms, as well as geometric isomers, and these also constitute aspects of the present disclosure.

[0090] The present disclosure also includes isotopically labeled compounds that are identical to those set forth in formula (I) except that one or more atoms have been replaced with an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature. Examples of isotopes suitable for inclusion in the disclosed compounds include, but are not limited to, 2 H,3 H, 13 C. 14 C. 15 N, 18 0, 17 0, 31 P, 32 P, 35 S, 18 F, and 36 These include, but are not limited to, hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as Cl. 2 Substitution with heavier isotopes, such as H, can offer certain therapeutic advantages due to greater metabolic stability, for example, increased half-life in vivo or reduced dosage requirements, and may therefore be preferred in some circumstances. Compounds may also incorporate positron-emitting isotopes for medical imaging and positron emission tomography (PET) studies to measure receptor distribution. Suitable positron-emitting isotopes that can be incorporated into compounds of formula (I) include: 11 C. 13 N, 15 O, and 18 F. Isotopically labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples, substituting an appropriate isotopically labeled reagent for the non-isotopically labeled reagent.

[0091] The compounds can be prepared according to the synthesis schemes detailed herein. Compounds and intermediates can be isolated and purified by methods well known to those skilled in the art of organic synthesis. Examples of conventional methods for isolating and purifying compounds include, but are not limited to, chromatography on solid supports such as silica gel, alumina, or silica derivatized with alkylsilane groups, recrystallization at high or low temperatures, optionally with pretreatment with activated carbon, thin-layer chromatography, distillation at various pressures, sublimation under vacuum, and trituration, as described, for example, in "Vogel's Textbook of Practical Organic Chemistry" 5th edition (1989), Longman Scientific & Technical, Essex CM20 2JE, England.

[0092] The reaction conditions and reaction times for each individual step may vary depending on the specific reactants used and the substituents present in the reactants. Specific procedures are provided in the Examples section. The reactions can be worked up in conventional ways, for example, by removing the solvent from the residue, and further purified according to methodologies commonly known in the art, such as, but not limited to, crystallization, distillation, extraction, trituration, and chromatography. Unless otherwise specified, starting materials and reagents are commercially available or can be prepared by one skilled in the art from commercially available materials using methods described in the chemical literature. If the starting materials are not commercially available, they can be prepared by procedures selected from standard organic chemistry techniques, techniques analogous to the synthesis of known structurally similar compounds, or procedures analogous to those described in the above schemes or synthetic examples section.

[0093] Routine experimentation, including appropriate manipulation of reaction conditions, reagents, and sequence of synthetic routes, protection of any chemical functional groups incompatible with the reaction conditions, and deprotection at appropriate points in the reaction sequence of the method, is within the scope of this invention. Suitable protecting groups and methods for protecting and deprotecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples thereof can be found in P. G. M. Wuts and T. W. Greene, in Greene's book titled "Protective Groups in Organic Synthesis" (4 th ed.), John Wiley & Sons, NY (2006), which is incorporated herein by reference in its entirety. Synthesis of the compounds of the invention can be achieved by methods analogous to those described in the synthetic schemes and specific examples.

[0094] 3. Pharmaceutical Compositions In another aspect, the present disclosure provides a pharmaceutical composition comprising a compound disclosed herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0095] The pharmaceutical compositions may be manufactured by processes known in the art, for example, by conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes.

[0096] As described herein, pharmaceutically acceptable carriers include any and all solvents, diluents, or other liquid vehicles, dispersing or suspending aids, surfactants, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, etc., appropriate for the particular dosage form desired. The various carriers used in formulating pharmaceutically acceptable compositions and techniques for their preparation are known in the art (e.g., Remington's Pharmaceutical Sciences, Sixteenth Edition, E.W. Martin (Mack Publishing Co., Easton, Pa., 1980)).

[0097] Pharmaceutically acceptable carriers can be functional molecules such as vehicles, adjuvants, or diluents. Pharmaceutically acceptable carriers can be any type of non-toxic, inert solid, semi-solid, or liquid filler, diluent, encapsulating material, or formulation aid. Pharmaceutically acceptable carriers include, for example, diluents, lubricants, binders, disintegrating agents, colorants, flavorings, sweeteners, antioxidants, preservatives, glidants, solvents, suspending agents, wetting agents, surfactants, softeners, propellants, moisturizers, powders, pH adjusters, and combinations thereof.

[0098] Some examples of materials that can function as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as phosphates), glycine, sorbic acid, or potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts), colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes, polyethylene polyoxypropylene block polymers, wool fat, sugars (such as lactose, glucose, and sucrose), starches (such as corn starch and potato starch), cellulose and its derivatives (such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate), powdered tragacanth, malt, gelatin, talc, excipients (such as cocoa butter and suppository waxes), and the like. waxes), oils (peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, soybean oil, and the like), glycols (propylene glycol or polyethylene glycol, and the like), esters (ethyl oleate, ethyl laurate, and the like), agar, compatible non-toxic lubricants (such as sodium lauryl sulfate and magnesium stearate), coloring agents, releasing agents, coating agents, emulsifiers, sweeteners, flavoring agents, preservatives, antioxidants, and the like can also be present in the composition at the discretion of the formulator.

[0099] In some embodiments, the pharmaceutical composition consists essentially of a therapeutically effective amount of a compound as disclosed herein, or a pharmaceutically acceptable salt thereof.

[0100] Liquid dosage forms include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. Solid dosage forms include, but are not limited to, capsules, tablets, pills, powders, cements, putties, and granules. Dosage forms for topical or transdermal administration of the compound include, but are not limited to, ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches.

[0101] The liquid carrier or vehicle can be a solvent or liquid dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like), vegetable oils, non-toxic glyceryl esters, and suitable mixtures thereof.

[0102] The pharmaceutical composition may be in a dosage form suitable for injection or infusion, such as a sterile aqueous solution or dispersion or a sterile powder containing an active ingredient(s) suitable for the extemporaneous preparation of a sterile injectable or infusible solution or dispersion. The final dosage form should be sterile, fluid, and stable under the conditions of manufacture and storage. Sterile injectable solutions can be prepared by incorporating at least the compound disclosed herein or a pharmaceutically acceptable salt thereof in the required amount in an appropriate solvent, along with various other ingredients as needed, optionally followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, preparation methods may include vacuum drying and freeze-drying techniques, which can yield a powder of the active ingredient(s) plus any additional desired ingredients present in the sterile solution.

[0103] In some embodiments, the composition is a solution, such as a solution suitable for administration by infusion or injection.The solution can be prepared in water, optionally mixed with a non-toxic surfactant.Dispersions can also be prepared in glycerol, liquid polyethylene glycol, triacetin, and mixtures thereof, and oils.These preparations may contain preservatives to prevent the growth of microorganisms.Prevention of microbial activity can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc.

[0104] Injectable formulations can be prepared by forming microencapsule matrices of the compound(s) disclosed herein, or pharmaceutically acceptable salts thereof, in biodegradable polymers such as polylactide-polyglycolide. The rate of drug release can be controlled depending on the ratio of compound to polymer and the properties of the particular polymer. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Injectable formulations can also be prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

[0105] In some embodiments, a composition may comprise at least one compound described herein and at least one additional anti-cancer agent. Anti-cancer agents useful in the present disclosure include paclitaxel, irinotecan, topotecan, gemcitabine, cisplatin, geldanamycin, mertansine, abiraterone, afatinib, aminolevulinic acid, aprepitant, axitinib, azacitidine, belinstat, bendamustine, bexarotene, bleomycin, bortezomib, bosutinib, busulfan, cabazitaxel, cabozantinib, capecitabine, carboplatin, carfilzomib, carmustine, ceritinib, cetuximab, chlorambucil, and clofala. ib, crizotinib, cyclophosphamide, cytarabine, dabrafenib, dacarbazine, dactinomycin, dasatinib, daunorubicin, decitabine, denosumab, dexrazoxane, docetaxel, dolastatins (e.g., monomethyl auristatin E), doxorubicin, enzalutamide, epirubicin, eribulin mesylate, erlotinib, etoposide, everolimus, floxuridine, fludarabine phosphate, fluorouracil, ganetespib, gefitinib, gemtuzumab ozogamicin, hexamer Tilmelamine, hydroxyurea, ibritumomab tiuxetan, ibrutinib, idelalisib, ifosfamide, imatinib, ipilimumab, ixabepilone, lapatinib, leucovorin calcium, lomustine, maytansinoids, mechlorethamine, melphalan, mercaptopurine, mesna, methotrexate, mitomycin C, mitotane, mitoxantrone, nelarabine, nelfinavir, nilotinib, obinutuzumab, ofatumumab, omacetaxine mepesuxinate, oxaliplatin, panitumumab, Zopanib, pegaspargase, pembrolizumab, pemetrexed, pentostatin, pertuzumab, plicamycin (plicanycin), pomalidomide, ponatinib hydrochloride, pralatrexate, procarbazine, radium-223 dichloride, ramucirumab, regorafenib, letaspimycin, ruxolitinib, semustine, siltuximab, sorafenib, streptozocin, sunitinib malate, tanespimycin, temozolomide, temsirolimus, teniposide, thalidomide, thioguanine,Any compound currently known or capable of acting as an anti-cancer agent is also useful in the present disclosure, including, but not limited to, thiotepa, toremifene, trametinib, trastuzumab, vandetanib, vemurafenib, vinblastine, vincristine, vinorelbine, vismodegib, vorinostat, and dibenzo-aflibercept.

[0106] 4. Method The basis for the selective tumor targeting of the compounds detailed herein lies in differences between the cell membranes of cancer cells compared to those of most normal cells. Phospholipid ether (PLE) molecules exploit the metabolic shift that tumor cells undergo to generate the energy necessary for rapid cell division. Tumors promote the use of the beta-oxidation pathway to convert long-chain fatty acids (LCFAs) into energy. To increase LCFA uptake, tumor cells alter their cell membranes by forming specialized microdomains known as "lipid rafts." Lipid rafts form due to the metabolic shift and the need for phospholipids. Within tumor cells, these regions become abundant and stabilized, making them potential tumor-specific targets. In particular, cancer cell membranes are highly enriched in lipid rafts. In normal tissues, the presence of lipid rafts is limited and transient (~2 nanoseconds). In tumors, the presence of lipid rafts increases and stabilizes (up to 10 days). Cancer cells have 5-10 times more lipid rafts than normal cells. Furthermore, lipid rafts are highly abundant in nearly all tumor types, demonstrating their presence in 100% of individual cancer cells tested. Lipid rafts are highly organized and specialized regions of the phospholipid membrane bilayer that contain high concentrations of various signaling molecules, sphingolipids, glycosphingolipids, and cholesterol and serve to organize cell surface and intracellular signaling molecules (e.g., growth factor and cytokine receptors, phosphatidylinositol 3-kinase (PI3K) / Akt survival pathways). Data suggest that lipid rafts function as entry points for phospholipid ethers (PLEs). The remarkable selectivity of these compounds for cancerous versus non-cancer cells is due to the high affinity of PLEs for cholesterol and the abundance of cholesterol-rich lipid rafts in cancer cells. The crucial role played by lipid rafts is emphasized by the fact that disruption of lipid raft structure inhibits the uptake of PLEs into cancer cells. It has been shown that PLE uptake is reduced by 60% when lipid raft formation is blocked. These features, combined with lipid rafts providing rapid internalization of phospholipid-drug conjugates, make them ideal targets.

[0107] The compounds described herein, such as PLE analogs, may be LCFA mimetics. The molecules disclosed herein have undergone extensive structure-activity relationship (SAR) analysis related to targeting lipid rafts on tumor cells and have been shown to specifically bind to these regions. The molecules disclosed herein provide direct entry into the cytoplasm and translocate along the Golgi apparatus network within the cytoplasm to the endoplasmic reticulum and mitochondria. In some embodiments, the phospholipid drug conjugates (PDCs) disclosed herein comprise a uniquely designed phospholipid ether conjugated via a cleavable linker to a novel combretastatin A (CBA) analog. CBAs are potent cytotoxins that have demonstrated the ability to inhibit tubulin polymerization in tumor cells and disrupt the local vasculature surrounding and within tumors. In some embodiments, the compounds disclosed herein comprise a uniquely designed phospholipid ether conjugated via a cleavable linker to a flavagrine (FLV) analog. FLVs are potent cytotoxins that inhibit translation, cell cycle progression, and induce apoptosis.

[0108] The compounds detailed herein, or pharmaceutically acceptable salts thereof, or compositions comprising the compounds detailed herein, can be used to treat cancer. In one aspect, the present disclosure provides a method of treating cancer in a subject in need thereof, comprising administering an effective amount of a compound detailed herein, or a pharmaceutically acceptable salt thereof, or a composition comprising a compound detailed herein.

[0109] In another aspect, the present disclosure provides a compound disclosed herein, or a pharmaceutically acceptable salt thereof, for use in treating cancer in a subject in need thereof.

[0110] In another aspect, the present disclosure provides a compound disclosed herein, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of cancer in a subject in need thereof.

[0111] Cancers that may be treated with the compounds detailed herein, or pharmaceutically acceptable salts thereof, or compositions comprising the compounds detailed herein, include: breast cancer, including male breast cancer; digestive / gastrointestinal cancers, including anal cancer, appendix cancer, extrahepatic bile duct cancer, gastrointestinal carcinoid, colon cancer, esophageal cancer, gallbladder cancer, gastric cancer, gastrointestinal stromal tumors ("gist"), pancreatic islet cell tumors, adult primary liver cancer, pediatric liver cancer, pancreatic cancer, rectal cancer, small intestine cancer, and stomach (gastric) cancer; pancreatic adenocarcinoma, adrenocortical carcinoma, pancreatic neuroendocrine tumor, Merkel cell carcinoma, non-small cell lung neuroendocrine tumors. endocrine and neuroendocrine carcinomas, including small cell lung neuroendocrine tumors, parathyroid carcinoma, pheochromocytoma, pituitary tumors, and thyroid carcinoma; eye cancer, including intraocular melanoma and retinoblastoma; genitourinary cancer, including bladder cancer, kidney (renal cell) cancer, penile cancer, prostate cancer, transitional cell renal pelvis and ureter cancer, testicular cancer, urethral cancer, and Wilms' tumor; germ cell cancer, including pediatric central nervous system cancer, pediatric extracranial germ cell tumors, extragonadal germ cell tumors, ovarian germ cell tumors, and testicular cancer; women's cancer, including cervical cancer, endometrial cancer, gestational trophoblastic tumor, epithelial ovarian cancer, ovarian germ cell tumors, uterine sarcoma, vaginal cancer, and vulvar cancer Gynecological cancers; head and neck cancers, including hypopharyngeal cancer, laryngeal cancer, lip and oral cavity cancer, metastatic squamous cell neck cancer with occult primary, oral cancer, nasopharyngeal cancer, oropharynx cancer, paranasal sinus and nasal cavity cancer, parathyroid cancer, pharyngeal cancer, salivary gland cancer, and throat cancer; leukemias, including adult acute lymphoblastic leukemia, childhood acute lymphoblastic leukemia, adult acute myeloid leukemia, childhood acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, and hairy cell leukemia; AIDS-related lymphoma, cutaneous T-cell lymphoma, adult Hodgkin lymphoma, childhood Hodgkin lymphoma, Hodgkin lymphoma during pregnancy, mycosis fungoides, Lymphomas, including adult non-Hodgkin's lymphoma, childhood non-Hodgkin's lymphoma, non-Hodgkin's lymphoma during pregnancy, primary central nervous system lymphoma, Sézary syndrome, and Waldenstrom's macroglobulinemia; musculoskeletal cancers, including Ewing's sarcoma, osteosarcoma and malignant fibrous histiocytoma of bone, childhood rhabdomyosarcoma, and soft tissue sarcoma; nervous system cancers, including adult brain tumors, childhood brain tumors, astrocytoma, brain stem glioma, central nervous system atypical teratoid / rhabdoid tumor, central nervous system embryonal tumors, craniopharyngioma, ependymoma, neuroblastoma, and primary central nervous system (CNS) malignant lymphoma;Respiratory / thoracic cancers, including non-small cell lung cancer, small cell lung cancer, malignant mesothelioma, thymoma, and thymic carcinoma; and skin cancers, including Kaposi's sarcoma, melanoma, and squamous cell carcinoma. In certain embodiments, the cancer may be melanoma, lung cancer, colon cancer, breast cancer, or a combination thereof.

[0112] In another embodiment, the cancer may comprise one or more CTCs, which may be selected from the group consisting of breast cancer, lung cancer, thyroid cancer, cervical cancer, melanoma, squamous cell carcinoma, prostate cancer, pancreatic cancer, colon cancer, and cancer stem cells, and malignant plasma cells.

[0113] In another embodiment, the cancer may be metastatic. In certain embodiments, the metastatic cancer may be selected from the group consisting of breast cancer, lung cancer, melanoma, and colon cancer.

[0114] In another embodiment, the cancer may be cancer stem cells. In a particular embodiment, the cancer stem cells may be derived from the group consisting of breast cancer, lung cancer, melanoma, and colon cancer.

[0115] In some embodiments, the lung cancer may include small cell lung cancer, non-small cell lung cancer, or a combination thereof.

[0116] In some embodiments, the melanoma may include superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, acral lentiginous melanoma, amelanotic melanoma, nevoid melanoma, spitzoid melanoma, desmoplastic melanoma, or a combination thereof.

[0117] In some embodiments, the colon cancer may include adenocarcinoma.

[0118] In some embodiments, a compound of Formula (Ia), (Ia-1), (Ia-2), or (Ia-3) detailed herein, or a pharmaceutically acceptable salt thereof, or a composition comprising a compound detailed herein, can be used to treat melanoma, lung cancer, colon cancer, or a combination thereof.

[0119] In some embodiments, the breast cancer may include invasive ductal carcinoma, metastatic breast cancer, inflammatory breast cancer, triple-negative breast cancer, ductal carcinoma in situ, or a combination thereof. In further embodiments, the cancer is breast cancer, and the subject may have estrogen receptor-positive, both estrogen receptor-negative and progesterone receptor-negative, HER2-expressing (HER2+), no HER2-expressing (HER2-), or a combination thereof. In some embodiments, a compound of formula (Ib), (Ib-1), (Ib-2), or (Ib-3) detailed herein, or a pharmaceutically acceptable salt thereof, or a composition comprising a compound detailed herein, may be used to treat breast cancer.

[0120] In some embodiments, a compound of formula (Ic), (Ic-1), (Ic-2), or (Ic-3), or a pharmaceutically acceptable salt thereof, or a composition comprising a compound as described herein, can be used to treat melanoma, lung cancer, colon cancer, breast cancer, or a combination thereof.

[0121] In some embodiments, the subject is a human, such as an adult or infant. In some embodiments, the subject is an animal, such as a mammal.

[0122] The method can include administering a compound detailed herein, or a pharmaceutically acceptable salt thereof, or a composition comprising a compound detailed herein in an amount detailed herein. In some embodiments, the method includes administering about 0.0001 to about 1000 mg / kg of a compound detailed herein, or a pharmaceutically acceptable salt thereof.

[0123] Useful dosages of the compound(s) in the composition can be determined by comparing their in vitro activity and in vivo activity in animal models. Methods for estimating effective dosages for humans in rodents, pigs, and other animals are known in the art; see, for example, U.S. Patent No. 4,938,949.

[0124] The actual dosage levels of the compounds in the therapeutic compositions detailed herein can be varied to obtain the amount of compound(s) useful for achieving the desired therapeutic response for a particular patient, composition, and mode of administration. The selected dosage level and amount of the compound, or a pharmaceutically acceptable salt thereof, for therapeutic use can vary depending on the particular compound or salt selected, the route of administration, the disease or condition being treated, the age and condition of the subject being treated, the severity of the condition being treated, and the condition and prior medical history of the patient being treated. When administering a pharmaceutically acceptable salt, the dosage can be calculated as the free base. However, it is within the skill of one of ordinary skill in the art to begin administering the compound at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. In certain circumstances, the disclosed compounds may be administered in amounts exceeding the dosage ranges described herein to effectively and aggressively treat particularly aggressive diseases or conditions.

[0125] In some embodiments, the compounds disclosed herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof, can be administered orally or intravenously. Generally, however, suitable doses will often be in the range of about 0.0001 mg / kg to about 1000 mg / kg, such as about 0.001 mg / kg to about 10.0 mg / kg. For example, suitable dosages may range from about 0.01 mg / kg to about 1.0 mg / kg of the recipient's body weight per day, about 0.01 mg / kg to about 3.0 mg / kg of the recipient's body weight per day, about 0.1 mg / kg to about 5.0 mg / kg of the recipient's body weight per day, or about 0.2 mg / kg to 4.0 mg / kg of the recipient per day. The compounds can be administered in unit dosage forms; for example, containing 1 to 100 mg, 10 to 100 mg, or 5 to 50 mg of active ingredient per unit dosage form.

[0126] The desired dose may conveniently be presented in a single dose or as divided dose administered at appropriate intervals, for example, as two, three, four or more sub-doses per day, which sub-dose itself may be further divided, for example, into a number of discrete loosely spaced administrations.

[0127] The specific in vivo dosage and specific administration mode to be administered may vary depending on the age, weight, severity of the affliction, and mammalian species to be treated, the specific compound to be applied, and the specific application of these compounds.Determining the effective dosage level to achieve the desired results can be accomplished by known methods, such as human clinical trials, in vivo studies, or in vitro studies.For example, the effective dosage of the compounds disclosed herein or their pharmaceutically acceptable salts can be determined by comparing the in vitro activity and in vivo activity of animal models.Such comparisons may be made by comparison with established drugs.

[0128] Dosage amount and interval can be individually adjusted to provide plasma levels of the active moiety sufficient to maintain modulating effects, or the minimum effective concentration (MEC). The MEC will vary for each compound but can be estimated from in vivo and / or in vitro data. The dosage required to achieve the MEC will depend on individual characteristics and the route of administration. However, FIPLC assays or bioassays can be used to measure plasma concentrations. Dosage intervals can also be determined using the MEC value. Compositions should be administered using a regimen that maintains plasma levels above the MEC value for 10-90% of the time, preferably between 30-90%, and most preferably between 50-90%, preferably between 30-90% and more preferably between 50-90%. In cases of local administration or selective uptake, the effective local concentration of a drug may not be related to plasma concentration.

[0129] The compounds, salts, and compositions disclosed herein can be evaluated for efficacy and toxicity using known methods. For example, the toxicity of a particular compound, or a subset of compounds sharing a particular chemical moiety, can be established by measuring in vitro toxicity in mammalian, and preferably human, cell lines, such as cell lines. The results of such studies are often predictive of toxicity in mammalian, or more specifically, animal, such as human, cell lines. Alternatively, the toxicity of a particular compound in an animal model, such as a mouse, rat, rabbit, dog, or monkey, can be measured by known methods. The efficacy of a particular compound can be established using several recognized methods, such as in vitro methods, animal models, or human clinical trials. To select a model for measuring efficacy, one skilled in the art can be guided by the state of the art to select the appropriate model, dose, route of administration, and / or regime.

[0130] The compound(s) detailed herein, or pharmaceutically acceptable salts thereof, or compositions comprising the compound(s) detailed herein, can be administered to humans and other mammals by a variety of known routes, including, but not limited to, oral, rectal, parenteral, intracisternally, intravaginally, transdermally (e.g., using a patch), transmucosally, sublingually, pulmonary, intraperitoneal, topical (by powder, ointment, or drops), buccal, or as an oral or nasal spray. As used herein, the terms "parenteral" or "parenterally" refer to modes of administration, including intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous, and intraarticular injection and infusion.

[0131] The compositions described herein can be administered with additional compositions to prolong the stability, delivery, and / or activity of the compositions, or can be combined with additional therapeutic agents, or can be administered before or after the administration of additional therapeutic agents.Combined therapy includes the administration of a single pharmaceutical dosage form containing one or more compounds described herein and one or more additional pharmaceutical agents, as well as the administration of a compound and each additional pharmaceutical agent in its own separate pharmaceutical dosage form.For example, the compounds described herein can be administered to a subject together with additional anti-cancer agents described herein.

[0132] The compounds described herein, or pharmaceutically acceptable salts thereof, can also be administered in the form of liposomes. As known in the art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by mono- or multilamellar hydrated liquid crystals dispersed in an aqueous medium. Optionally, physiologically acceptable and metabolizable lipids capable of forming liposomes can be used. The present composition in liposome form can contain, in addition to the compounds described herein, anticancer agents, stabilizers, preservatives, additives, etc. Preferred lipids include natural and synthetic phospholipids and phosphatidylcholines (lecithins), used separately or together. Methods for forming liposomes are known in the art. See, for example, Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, New York, NY (1976), p. 33 and subsequent pages. Such compositions will affect the physical state, solubility, stability, in vivo release rate, and in vivo clearance rate.

[0133] In one method of the present disclosure, the pharmaceutically acceptable composition can be delivered in a controlled-release system. For example, the drug can be administered using intravenous infusion, an implantable osmotic pump, a transdermal patch, liposomes, or other modes of administration. In one embodiment, a pump can be used (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201(1987); Buchwald et al., Surgery 88:507 (1980); Saudek et al., N. Engl. J. Med. 321:574(1989)). In another embodiment, a polymeric material can be used. In yet another embodiment, the controlled-release system can be placed near the therapeutic target, for example, the liver, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138 (1984)). Other controlled-release systems are discussed in the review article by Langer (Science 249:1527-1533 (1990)). [Example]

[0134] 5. Working Example The foregoing may be better understood by reference to the following examples, which are presented for purposes of illustration and are not intended to limit the scope of the invention. The present disclosure has multiple aspects and embodiments, which are illustrated by the accompanying non-limiting examples.

[0135] Example 1. Materials and Methods In vitro uptake of CLR2000045 was assessed using MCF-7 breast cancer cells and normal human dermal fibroblast (NHDF) cells and measured via LC / MS / MS. Breast cancer cells were maintained in minimal essential medium supplemented with 10% FBS. All cells were maintained at 37°C and 5% CO2. Cells were incubated with 1 μM drug, and reported values ​​are the average of triplicate evaluations. In vitro cytotoxicity was measured by Cell Titer-Glo® assay using MCF-7 breast cancer cells and Hs578T triple-negative breast cancer cells.

[0136] The in vitro uptake and release of CLR180099 was evaluated using A549 tumor cells, HCT116 tumor cells, and normal human dermal fibroblast (NHDF) cells and measured via LC / MS / MS. Cells were incubated with 1 μM drug, and reported values ​​were the average of triplicate evaluations. In vitro cytotoxicity was measured by the Cell Titer-Glo® assay.

[0137] In an in vivo efficacy screening model using chicken embryos, CLR2000045 was administered at 72 μM to measure efficacy against MCF-7 tumors and compared with a vehicle control and a paclitaxel positive control at 50 μM. CLR2000045 was applied topically to the embryo casing. Fertilized White Leghorn eggs were cultured at 37.5°C and 50% relative humidity for 9 days. At that time (E9), a small hole was made in the eggshell to drop the chorioallantoic membrane (CAM) onto the air sac, and a 1 cm eggshell was placed on the CAM. 2 A window of 1000 cells was opened. At least 20 eggs were used for each group (more than 20 eggs per group may be present depending on the embryonic survival rate after 9 days of development). Because some embryonic deaths may occur after tumor implantation or may be related to defective tumor implantation, data may be collected with fewer than 20 eggs per group (a minimum of 15 eggs per group). Tumor cells were cultured in DMEM supplemented with 10% FBS and 1% penicillin / streptomycin. On day E9, cells were detached with trypsin, washed with complete medium, and then suspended in transplantation medium. 3 × 10 6An inoculum of cells was added appropriately to the CAM of each egg (E10) per group, and then the eggs were randomly divided into groups.

[0138] Embryo viability was monitored daily. The number of embryos that died on E18 was counted along with the final observation of visible gross abnormalities to assess treatment-induced embryonic toxicity. Final mortality and Kaplan-Meier curves were calculated for all groups. Any visible abnormalities observed were also noted. On E18, the upper part of the CAM (with tumor) was removed from all surviving tumor-bearing embryos, washed with PBS buffer, and then directly transferred to PFA (fixed for 48 hours). The tumors were then carefully separated from the normal CAM tissue and weighed.

[0139] In vivo efficacy was further evaluated in R2G2 mice bearing HCC70 triple-negative breast cancer (TNBC) xenografts. CLR2000045 was evaluated at three doses (1 mg / kg) administered once, twice, or three times weekly for two weeks. CLR2000045 was administered systemically via tail vein injection. Each group contained 10 mice. Tumor volume was recorded for efficacy assessment, and body weight was recorded for tolerability assessment. Survival rates were also recorded.

[0140] CLR180099 was administered intravenously (IV) to healthy C57BL / 6 mice to determine the maximum tolerated dose (MTD) compared to the FLV molecule alone. In this case, the vehicle used to administer CLR180099 was PBS, but any pharmaceutically appropriate vehicle can be used. Each group contained five mice. In vivo efficacy was evaluated in HCT116 xenografted athymic nude mice. Mice received approximately 1 x 10 FLV resuspended in 5 ml of 1.2% methylcellulose. 6 The flank model was developed by injecting 100 cells into the hind flank of mice. The study was conducted with a mean tumor volume of approximately 120 mm for the group. 3The study was initiated when tumor volume reached 1000 mg / kg / day. Tumor volume was measured using calipers, and measurements of tumor length, width, and depth were used to calculate tumor volume. Two doses of CLR180099 were evaluated (2 mg / kg twice or 2 mg / kg three times). Each group contained 10 mice. Tumor volume was recorded for efficacy assessment, and body weight for tolerability assessment. Total conjugated CLR180099 and free FLV were measured by mass spectrometry.

[0141] Example 2. Phospholipid lipid ether delivery vehicles demonstrate specificity for a broad range of tumor cells To demonstrate PDC uptake in various tumor cell lines, various tumor cell lines, including MCT-116, MeS SA / Dx5, MIa PaCa-2, Ovcar-3, and U-87MG, were incubated with 5 μM CLR1501 (PLE + BODIPY fluorescent payload) in complete medium for 24 h at 37 °C. Each cell line may have a slightly different medium to optimize growth, and any appropriate medium known in the art can be used for each cell line. All cells were maintained at 37 °C in the appropriate medium supplemented with 10% FBS and 5% CO2. CLR1501 was excited and then detected with an Alexa-Fluor 488 filter. CLR1501 was highly localized in all different tumor cell lines (Figure 1A and Figure 1B). Similar results have been reported in over 100 tumor cell lines, including MM.IS, MM.IR, RPM18226, U266, and NCIH929, Panc-1, A375, PC-3, Caki-2, HCT-116, A549, metastatic PC-3, MDA-MB-231, HT-29, SV-40, CNS-1, BxPC3, MCF-7, Lucap, LNCap, MES SA / Dx5, Capanl, HTB-77, Lan5, CHLA-20, NB1691, and SK-N-AS. CLR1501 was administered in vitro to various cancer cell lines and a normal human skin fibroblast cell line. After 24 hours, CLR1501 was shown to be preferentially taken up by these cancer cell lines 5-9 times more frequently than normal fibroblasts. The retained CLR1501 was associated with plasma and organelle membranes.

[0142] In vitro uptake and release of cytotoxic payloads were measured in A375 and A549 cell lines by incubating them with 2 μM of the cytotoxic small molecule PDC with a semistable linker (CLR2208, "PDC-SM1") in complete medium at 37°C for 48 hours. PDC-SM1 uptake was measured by LC / MS / MS. PDC-SM1 demonstrated uptake beginning within 30 minutes. 20–40% of the conjugate exposed to cells was measured in the tumor cell cytosol within 24 hours (Figure 2A). Subsequently, CLR2206 ("PDC-SM2," similar to PDC-SM1 but without the cleavable linker) was utilized to measure payload release within tumor cells. CLR2200 ("PDC-SM3") was also studied. Measurable release of the small molecule payload occurred between 1 and 2 hours after incubation (Figure 2B). Minor release of the payload occurred in the medium (<1 nM). These results demonstrated that phospholipid ether molecules have the ability to target a wide range of tumors, and that PDCs have the ability to achieve 20-40% uptake of exposed drugs into tumor cell lines.

[0143] To measure lipid raft-mediated uptake on tumor cells, multiple myeloma cells were incubated with CLR1502 (a near-infrared molecule conjugated to PLE) for 24 hours at 37 °C. The next day, cells were washed and co-stained with a nuclear stain (Hoechst 33342). Cholera toxin subunit B was used to further stain for the presence of lipid rafts. Cells were incubated with cholera toxin subunit B for 24 hours. Furthermore, to measure lipid raft-mediated uptake in primary tumor samples, patient-derived multiple myeloma cells were stained with Hoechst 33342 and incubated with CLR1501 (Figure 3). These results demonstrate that PDC uptake is associated with lipid rafts on the tumor cell membrane in both cell lines and primary tumor samples.

[0144] In vitro efficacy of the cytotoxic payload was measured. PDC-SM2 demonstrated submicromolar activity (measured based on the concentration of the complete conjugate incubated on the cells) against melanoma (A375) and lung cancer (A549) cells. PDC-SM2 showed lower activity against melanoma than lung cancer (IC50 0.131 vs. 0.016), but was more potent (0% vs. 12% viable cells remaining, Figure 4). PDC-SM2 also demonstrated similar activity and potency against colon cancer (HCT-116) cells, but no activity against normal fibroblasts. Thus, PDCs demonstrated payload release and potent nanomolar activity against tumor cells.

[0145] To determine whether the cytotoxicity of PDCs was tolerated in vivo, C57BL / 6 mice were administered the following doses: PDC-SM2 was administered at dose levels of 0.5 mg / kg, 1.0 mg / kg, or 2.0 mg on days 0, 3, and 7; payload alone was administered at 0.25 mg / kg, 0.4 mg / kg, or 0.5 mg / kg on day 0; and vehicle was administered on days 0, 3, and 7. PDCs and vehicle controls showed no toxicity or adverse events during repeated administration, as measured by changes in body weight (no weight loss). Payload doses of 0.25 and 0.4 mg / kg were tolerated, although some toxicity was observed in the skin and integument of mice. The 0.5 mg / kg payload dose was not tolerated; two mice died by day 4 after a single injection, and all mice were sacrificed on day 5 (Figure 5). These PDCs demonstrated good plasma stability in human plasma. Plasma stability was measured using a Cyprotex plasma stability assay. Samples were incubated at 1 μM for 0, 15, 30, 60, and 120 minutes. A positive control compound that undergoes degradation in plasma was used. The percentage of compound remaining at each incubation time point was measured. PDC-SM2 exhibited some instability in mouse plasma and may cause some toxicity (Table 1). These PDCs were well tolerated in vivo. Overall, PDCs offer a novel and original approach for targeting small molecules to tumor cells.

[0146] [Table 1]

[0147] Selective uptake of CLR1502 was also measured in vivo in intestinal tumors. At necropsy 96 hours after administration of 50 μg of CLR1502, the entire colon and the terminal portion of the small intestine were removed (Figures 19A and 19B). CLR1502 was administered via tail vein injection. Areas of increased signal intensity were observed using an IVIS Spectrum, which allows direct visualization of CLR1502 through the animal's skin. After euthanasia, tissues identified by microdissection using the IVIS system were excised and histology was performed to confirm tumor versus nontumor tissue and the location of near-infrared labeling. These areas represented noninvasive (colon Figure 19C; terminal small intestine Figure 19F) and invasive (colon Figure 19D; terminal small intestine Figure 19E) tumors.

[0148] In other studies, CLR1502 accumulated in metastatic lesions and regional lymph nodes. After intestinal removal, the mesenteric fat, pancreas, and spleen were isolated en bloc. In one case, two metastatic tumor deposits measuring ~4 mm in size were observed within the mesentery. These lesions were easily visualized with a Fluobeam near-infrared imaging device. H&E confirmed these lesions as metastatic malignant lesions. Regional lymphadenopathy was also shown to accumulate CLR1502 using Fluobeam. No malignant cells were observed within these hyperplastic lymph nodes.

[0149] Tumor thickness does not account for the increased signal intensity seen in intestinal cancer (Figures 22A and 22B). Autopsies were performed on mice 96 hours after injection with 50 μg of CLR1502 per mouse. To examine the effect of tissue thickness, normal-appearing colon sections were overlaid on top of each other. Radiation efficiency was measured to compare signal intensity between one, two, and three layers of normal colon and intestinal tumor. One layer of normal colon was found to be approximately 1 mm thick. Tissue thickness may account for the increased intensity seen in adenomas, but does not account for the differences seen in adenocarcinomas.

[0150] In vivo optical scanning of CLR1502 uptake in a colon cancer model demonstrated preferential retention in malignant compared to normal tissue. Athymic nude mice bearing colon cancer (HCT-116) xenografts were intravenously injected with 1 mg of CLR1502 and imaged using a Li-COR Pearl® Impulse system (Figure 23). Fluorescence intensity (shown by the color bar) and biodistribution were measured in vivo over time.

[0151] In vivo optical scanning of CLR1502 uptake in a mouse breast cancer model demonstrated preferential retention in malignant compared to normal tissue. Athymic nude mice bearing orthotopic breast cancer xenografts (MDA-MB-231) were intravenously injected with approximately 80 μg of CLR1502 and imaged daily for 7 days (168 hours) in vivo using the Fluoptics Fluobeam® and IVIS® Spectrum systems (Figure 24). Study results demonstrated selective uptake and prolonged retention within tumors (yellow and green arrows in Fluobeam and IVIS Spectrum, respectively), as well as a relative increase in clearance from normal tissue over time.

[0152] Athymic nude mice bearing lung cancer xenografts (H226 lung) in each flank were injected intravenously with approximately 50 μg of CLR1502 and imaged in epifluorescence mode using an IVIS Spectrum (Figure 25). Note that at 96 hours, differences in radiative efficiency between malignant and normal tissue create sufficient contrast at the edge of the tumor, as indicated by the black arrow.

[0153] Example 3. CLR2000045 with Combretastatin A-4 Analogue Improves Breast Cancer Treatment CLR2000045 showed significant uptake into tumor cells with minimal uptake into normal tissues. Drug release showed approximately 50% release at each time point. A steady state between drug uptake and release was achieved between 24 and 48 hours (Figure 6). CLR2000045 demonstrated excellent activity and potency against two breast cancer cell lines (MCF-7 and Hs578T), with IC50 values ​​of 76 nM and 51 nM, respectively (Figure 7). The molecule also demonstrated activity against several other solid tumors, including lung cancer, melanoma, and colorectal cancer. The half-maximal inhibitory concentration (IC50) was measured in the cell lines (Table 2). The plasma stability of CLR2000045 was also measured (Table 3).

[0154] [Table 2]

[0155] [Table 3]

[0156] Fertilized eggs from White Leghorn chickens (20 eggs / treatment group) were incubated at 37.5°C for 9 days. MCF-7 cells were cultured under standard conditions before implantation. On day 10, 3 × 10 6 An inoculum of MCF-7 eggs was added to the chorion. Eggs were then randomly assigned to treatment groups and treated four times (days 11, 13, 15, and 17) with vehicle, paclitaxel at 50 μM per dose, and CLR2000045 at 72 μM per dose. CLR2000045 demonstrated activity similar to paclitaxel in this screening model (Figure 8).

[0157] This study was conducted with a mean tumor volume of 200 mm 3Treatment was initiated when tumor volume reached 100% (day 4). CLR2000045 was administered IV at the following doses: 1 mg / kg on either days 5 and 12, or days 5, 8, 12, and 15, or days 5, 7, 9, 12, 14, and 16. CLR2000045 demonstrated a dose-response reduction in tumor volume from dose group 1 to dose group 3 (three times weekly for 2 weeks), with the highest dose tested showing nearly 100% eradication of tumors. The two highest dose groups showed statistically significant reductions in tumor volume compared to vehicle controls (p≦0.05 and p≦0.01, respectively) (Figure 9). Kaplan-Meier curves show that treatment with CLR2000045 at 1 mg / kg three times weekly for 2 weeks resulted in a significant increase in survival compared to vehicle and weekly administration (p≦0.001 and p≦0.05, respectively). 1 mg / kg twice weekly for 2 weeks resulted in a significant increase compared to vehicle (p≦0.05; FIG. 10). Body weight changes after treatment were measured in a (HCC70) mouse xenograft model (FIGS. 11A and 11B).

[0158] CLR2000045 demonstrated significant payload uptake and release (20-40% of the exposed drug) in tumor cell lines, while showing minimal uptake in normal cells. CLR200045 exhibits potent in vitro activity against multiple breast cancer cell lines. CLR2000045 demonstrated potent in vivo activity against a triple-negative breast cancer model (HCC70) and a metastatic adenocarcinoma breast cancer model (MCF-7). CLR200045 provided a significant survival benefit in a TNBC (HCC70) model and was well tolerated at the two highest doses, as measured by weight loss. Collectively, these data demonstrate the potent in vitro and in vivo activity of CLR200045 against various breast cancer cell lines and animal models, supporting the continued development of this PDC.

[0159] Example 4. CLR180099 improves the safety and efficacy of antitumor drugs against colon tumors CLR180099 demonstrated excellent activity and efficacy against breast cancer and lung cancer, with IC50 values ​​of 0.024 and 0.011, respectively (Figure 12). The compound also demonstrated activity against several other solid tumors, including melanoma and colon cancer. The plasma stability of CLR1800095, CLR180099A, and CLR180099B was measured in mice and humans (Table 4). CLR1800095 exhibited some instability in mouse plasma, which may cause some toxicity.

[0160] [Table 4]

[0161] The study showed that the mean tumor volume in the group was ~120 mm 3 Treatment was initiated when the tumor reached a normal normal (day 1). CLR180099 was administered IV at 2 mg / kg on either days 1 and 4 or days 1, 3, and 5. Docetaxel was administered at 10 mg / kg on days 1 and 4. CLR180099 demonstrated tumor volume reduction comparable to or greater than docetaxel and demonstrated a dose-dependent effect. In the docetaxel group, multiple deaths occurred from the start of day 18 to the end of day 26 (Figure 16). Kaplan-Meier curves showed that treatment with CLR180099 at 2 mg / kg on days 1 and 4 or days 1, 3, and 5 resulted in a significant increase in survival compared to docetaxel (Figure 17, log-rank test, p≦0.001). Weight loss was measured, and all mice treated with CLR180099 (both doses) demonstrated normal weight gain throughout the study (Figure 18). Five mice per group were administered at each dose level. Both PDCs were tolerated at a dose of 10 mg / kg with all mice surviving and showing no peripheral organ toxicity (Table 5). Payload alone was not tolerated at doses above 0.5 mg / kg (all mice died at 0.5 mg / kg).

[0162] [Table 5]

[0163] CLR180099 demonstrated significant payload uptake and release (20-40% of the exposed drug) in tumor cell lines, while showing minimal uptake in normal cells. CLR180099 demonstrated potent in vitro activity against various solid tumors, including lung cancer (A549), breast cancer (MCF7), and melanoma (A375), as well as other tumor types. Two or three in vivo doses of CLR180099 demonstrated activity equivalent to or superior to docetaxel in colorectal cancer. Furthermore, CLR180099 significantly improved survival compared to docetaxel at both doses. Tolerability assessment demonstrated that CLR180099 was well tolerated in both tumor-bearing and normal animals, while the FLV payload was toxic in both normal and tumor-bearing mice. CLR180099 demonstrated no toxic effects compared to the FLC analog payload alone, indicating that the payload may be effective for targeted delivery via phospholipid ethers (PLEs).

[0164] Example 5. Synthesis of Compounds The chemical synthesis steps were carried out as follows: The products were isolated using known techniques such as HPLC, and the resulting structures were verified by NMR and MS.

[0165] CLR2208 is synthesized by the following scheme 1 [ka] It was synthesized by

[0166] CLR2206 was synthesized according to Scheme 2. Compound 2 was prepared from compound 1 using hypophosphate chloride 1A (2.5 equivalents) in EtN (10 equivalents) and THF at −40° C. for 3 hours. Compound 2 was reacted with 2A (1 equivalent) in EtN (1 equivalent), CDI (1.5 equivalents), ZnCl (2.6 equivalents), and DMF at 15° C. for 12 hours to give compound 3. Compound 3 was deprotonated in piperidine (5 equivalents DMF, 15° C., 3 hours) to provide compound 4. Compound 4 was reacted with 4A (1 equivalent) in EtN (4 equivalents), COMU (1.15 equivalents), and CHCl at 15° C. for 2 hours to give CLR2206. [ka]

[0167] CLR2200 was synthesized according to Scheme 3. Compound 5 was reacted with pyridine (Py, 20 equivalents), HOBt (0.5 equivalents), and MMAE (0.8 equivalents) in DMF at room temperature for 12 hours to give compound 6. Compound 6 was deprotonated in piperidine (10 equivalents) and DMF:AcN (1:1) at room temperature for 12 hours to give compound 7. Compound 7 was reacted with 7A (1 equivalent), TEA (4.5 equivalents), COMU (1.2 equivalents), and CHCl3 at room temperature for 12 hours to give CLR2200. [ka]

[0168] CLR200045 was prepared according to Scheme 4, or alternatively according to Scheme 5. [ka] [ka]

[0169] CLR2013 was prepared according to Scheme 6. [ka]

[0170] CLR1800095 was prepared according to Scheme 7. [ka]

[0171] CLR180099B was prepared according to Scheme 8 (LCMS purity 97%). [ka]

[0172] CLR180099A was prepared according to Scheme 9. [ka]

[0173] For completeness, the various aspects of the invention are set out in the following numbered paragraphs:

[0174] Item 1. A compound of the following chemical formula (I): [ka] or a pharmaceutically acceptable salt thereof; During the ceremony, n is 2 to 20; Q 1 is a bond or the following chemical formula [ka] where m is between 0 and 100; L is the following chemical formula [ka] where R x is H or a halogen; Q 2is a bond or self-immolative spacer; and Z is an anticancer drug.

[0175] Item 2. The compound of item 1, or a pharmaceutically acceptable salt thereof, wherein: Q 1 is a bond or the following chemical formula [ka] and LQ 2 teeth [ka] is.

[0176] Item 3. The compound according to any one of Items 1 and 2, or a pharmaceutically acceptable salt thereof, wherein Z is a polo-like kinase 1 (PLK-1) inhibitor, a tubulin polymerase inhibitor, a tubulin stabilizer, an antitumor drug, a eukaryotic translation initiation factor 4 (EIF4) inhibitor, a combretastatin A-4 analog, or a flavagrine analog.

[0177] Item 4. A compound according to any one of items 1 to 3, having the structure of chemical formula (Ia), or a pharmaceutically acceptable salt thereof, wherein Q 1 is the following chemical formula [ka] and LQ 2 is the following chemical formula [ka] and; and Z is a PLK-1 inhibitor, a tubulin polymerase inhibitor, a tubulin stabilizer, an anti-tumor agent, or a eukaryotic translation initiation factor 4 (EIF4) inhibitor.

[0178] Item 5. The compound according to item 4, wherein Z is a PLK-1 inhibitor or an antitumor agent selected from the group consisting of monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), and monomethyl auristatin E (MMAD).

[0179] Item 6. A compound according to any one of items 1 to 3, having the structure of chemical formula (Ib), or a pharmaceutically acceptable salt thereof, wherein n is 18; Q 1 is the following chemical formula [ka] and LQ 2 is the following chemical formula [ka] and Z is a combretastatin A-4 analog.

[0180] Item 7. A compound according to any one of items 1 to 3, having the structure of chemical formula (Ic), or a pharmaceutically acceptable salt thereof, wherein n is 18; Q 1 is a bond or the following chemical formula [ka] and; LQ 2 is the following chemical formula [ka] and Z is a flavogrine analogue.

[0181] Item 8. The following chemical formula [ka] [ka] [ka] 2. The compound according to item 1, selected from the group consisting of:

[0182] Item 9. A pharmaceutical composition comprising the compound according to any one of items 1 to 8, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0183] Item 10. A method for treating cancer in a subject in need thereof, comprising administering an effective amount of the compound according to any one of items 1 to 8, or a pharmaceutically acceptable salt thereof.

[0184] Item 11. The method according to item 10, wherein the cancer is melanoma, lung cancer, colon cancer, breast cancer, or a combination thereof.

[0185] Item 12. The method according to any one of Items 10 to 11, wherein The lung cancer includes small cell lung cancer, non-small cell lung cancer, or a combination thereof; The melanoma includes superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, acral lentiginous melanoma, amelanotic melanoma, nevoid melanoma, spitzoid melanoma, desmoplastic melanoma, or a combination thereof; the colorectal cancer comprises adenocarcinoma; or The breast cancer includes invasive ductal carcinoma, metastatic breast cancer, inflammatory breast cancer, triple-negative breast cancer, ductal carcinoma in situ, or a combination thereof.

[0186] Item 13. The method according to any one of Items 10 to 12, wherein the cancer comprises cancer stem cells.

[0187] Item 14. The method according to any one of Items 10 to 13, wherein the cancer comprises metastatic cancer cells.

[0188] Item 15. The method according to any one of items 10 to 14, wherein the cancer comprises circulating tumor cells.

[0189] Item 16. The method according to any one of Items 10 to 15, wherein the cancer is melanoma, lung cancer, colon cancer, or a combination thereof, and the compound is a compound of formula (Ia) or a pharmaceutically acceptable salt thereof.

[0190] Item 17. The method according to any one of items 10 to 15, wherein the cancer is breast cancer, and wherein the subject (1) is estrogen receptor positive, (2) is both estrogen receptor negative and progesterone receptor negative, (3) expresses HER2 (HER2+), and (4) does not express HER2 (HER2-), or a combination thereof.

[0191] Item 18. The method according to any one of Items 10 to 15, wherein the cancer is breast cancer, and the compound is a compound of formula (Ib) or a pharmaceutically acceptable salt thereof.

[0192] Item 19. The method according to any one of Items 10 to 15 and 17, wherein the cancer is melanoma, lung cancer, colon cancer, breast cancer, or a combination thereof, and wherein the compound is compound (Ic) or a pharmaceutically acceptable salt thereof.

[0193] The foregoing description of specific embodiments sufficiently reveals the general nature of the present invention so that others, by applying knowledge within the art, can readily modify and / or adapt such specific embodiments for various uses without undue experimentation and without departing from the general concepts of the present disclosure. Such adaptations and modifications are therefore intended to be within the meaning and range of equivalents of the disclosed embodiments, based on the embodiments and guidance presented herein. It should be understood that the terminology or phraseology used herein is for the purpose of illustration and not of limitation, as interpreted by those skilled in the art in light of the teachings and guidance.

[0194] The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.

[0195] All publications, patents, patent applications, and / or other documents cited in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, and / or other document was individually indicated to be incorporated by reference.

Claims

1. A compound of the following formula (I): 【Chemical 1】 or a pharmaceutically acceptable salt thereof, wherein n is 2 to 20; Q 1 is a bond or the following chemical formula 【Chemistry 2】 where m is 0 to 100; L-Q 2 is the following chemical formula 【Chemistry 3】 where R x is H or halogen; Z is a PLK-1 inhibitor, a tubulin polymerase inhibitor, a tubulin stabilizer, an antitumor agent, a eukaryotic translation initiation factor 4 (EIF4) inhibitor, or a combretastatin A-4 analog. However, it is not a conjugate selected from the following: 【Chemistry 4】 and 【Chemistry 5】 A compound of formula (I), or a pharmaceutically acceptable salt thereof:

2. Q 1 is bonded or has the following chemical formula 【Chemistry 6】 2. The compound of claim 1, wherein:

3. Q 1 is the following chemical formula 【Chemistry 7】 and L-Q 2 is the following chemical formula 【Chemistry 8】 and and 3. The compound of claim 1 or 2, having the structure of formula (I-a), or a pharmaceutically acceptable salt thereof, wherein Z is a PLK-1 inhibitor, a tubulin polymerase inhibitor, a tubulin stabilizer, an anti-tumor agent, or a eukaryotic translation initiation factor 4 (EIF4) inhibitor.

4. 4. The compound of claim 3, wherein Z is a PLK-1 inhibitor or an antitumor agent selected from the group consisting of monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), and monomethyl auristatin D (MMAD).

5. n is 18; Q 1 is the following chemical formula 【Chemistry 9】 and L-Q 2 is the following chemical formula 【Chemistry 10】 and Z is a combretastatin A-4 analog; 3. The compound of claim 1 or 2, having the structure of chemical formula (Ib), or a pharmaceutically acceptable salt thereof.

6. The following chemical formula 【Chemistry 11-1】 【Chemistry 11-2】 【Chemistry 11-3】 【Chemistry 11-4】 2. The compound of claim 1, selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

7. 10. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier thereof.

8. 10. A pharmaceutical composition for the treatment of cancer in a subject in need thereof, comprising an effective amount of a compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof.

9. 9. The pharmaceutical composition of claim 8, wherein the cancer is melanoma, lung cancer, colon cancer, breast cancer, or a combination thereof.

10. the lung cancer comprises small cell lung cancer, non-small cell lung cancer, or a combination thereof; the melanoma comprises superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, acral lentiginous melanoma, amelanotic melanoma, nevoid melanoma, spitzoid melanoma, desmoplastic melanoma, or a combination thereof; the colon cancer comprises adenocarcinoma; or the breast cancer comprises invasive ductal carcinoma, metastatic breast cancer, inflammatory breast cancer, triple-negative breast cancer, ductal carcinoma in situ, or a combination thereof; The pharmaceutical composition according to claim 8 or 9.

11. The pharmaceutical composition of claim 8 , wherein the cancer comprises cancer stem cells.

12. The pharmaceutical composition of claim 8 , wherein the cancer comprises metastatic cancer cells.

13. 13. The pharmaceutical composition of any one of claims 8 to 12, wherein the cancer comprises circulating tumor cells.

14. 14. The pharmaceutical composition of any one of claims 8 to 13, wherein the cancer is melanoma, lung cancer, colon cancer, or a combination thereof, and the compound is a compound of formula (Ia) or a pharmaceutically acceptable salt thereof.

15. 14. The pharmaceutical composition of any one of claims 8 to 13, wherein the cancer is breast cancer, and wherein the subject (1) is estrogen receptor positive, (2) is both estrogen receptor negative and progesterone receptor negative, (3) expresses HER2 (HER2+), and (4) does not express HER2 (HER2-), or a combination thereof.

16. 16. The pharmaceutical composition of any one of claims 8 to 13 and 15, wherein the cancer is breast cancer and the compound is a compound of formula (Ib), or a pharmaceutically acceptable salt thereof:

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