Phospholipid ether conjugates as cancer-targeting drug vehicles

Abstract

To provide therapeutic compounds capable of targeting a broad range of tumor cells.SOLUTION: The present disclosure is directed to compositions comprising therapeutic compounds, methods of manufacturing therapeutic compounds, and methods of treating cancer comprising administering therapeutic compounds.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims priority to U.S. Provisional Patent Application No. 62 / 899,611, filed on September 12, 2019; U.S. Provisional Patent Application No. 62 / 899,615, filed on September 12, 2019; U.S. Provisional Patent Application No. 62 / 899,618, filed on September 12, 2019; U.S. Provisional Patent Application No. 62 / 946,870, filed on December 11, 2019; U.S. Provisional Patent Application No. 62 / 956,844, filed on January 3, 2020; and U.S. Provisional Patent Application No. 62 / 956,907, filed on January 3, 2020, the contents of which are hereby incorporated by reference in their entirety.

[0002] Field The present disclosure relates to therapeutic compounds that can target a wide range of tumor cells. The present disclosure further relates to compositions containing the therapeutic compounds, methods of manufacturing the therapeutic compounds, and methods of treating cancer including administering the therapeutic compounds.

Background Art

[0003] Introduction In 2018, 18 million people were diagnosed with cancer and 9.6 million people died from cancer worldwide. 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) are the most common types of cancer. Despite many available treatments, cancer remains the second leading cause of death worldwide.

[0004] Cancer is a result of cell division, but 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., cells coming into contact with other cells). Additionally, most cells are programmed to die after a certain number of cell divisions because they can only replicate a finite number of times.

[0005] Cancer is a cell that has overcome these inherent checkpoints and proliferates uncontrollably. This uncontrolled proliferation leads to the formation of tumors. There are two types of tumors: benign and malignant. Benign tumors cannot cross 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. Additionally, lipid metabolism can play a major role in cancer metastasis. Cancer cells often exhibit fundamentally altered cell metabolism. However, the role of lipid metabolism in the development of malignant cancer remains unclear.

[0006] What further complicates the fight against cancer is that malignant tumors have different types of cells. One particularly troublesome type is cancer stem cells ("CSC's"). CSC's can self-renew and differentiate into different types of cancer cells found in malignant tumors. Therefore, CSC's are a major factor in the metastatic ability of tumors. CSC's often survive radiation and chemotherapy. It is hypothesized that the recurrence of cancer after radiation and chemotherapy is the result of a combination of the inability of radiation and chemotherapy to kill all CSC's and the ability of CSC's to form new tumors.

[0007] Chemotherapy is a term used to describe a specific type of cancer treatment that involves the use of cytotoxic anti-cancer agents. The cytotoxic drugs used during chemotherapy can be classified into several major categories, including alkylating agents, antimetabolites, antitumor antibiotics, topoisomerase inhibitors, and mitotic inhibitors. Cytotoxic anti-cancer agents typically halt cell division and thus affect both healthy tissues and cancerous tissues. Alkylating agents stop the division of cancer cells by damaging their DNA. Some common alkylating agents used in cancer treatment include nitrogen mustards (e.g., cyclophosphamide (Cytoxan®; Cytoxan is a registered trademark of Baxter International), nitrosoureas, alkyl sulfonates, triazenes, and ethyleneimines. Platinum drugs such as cisplatin and carboplatin act in a similar manner to alkylating agents. Antimetabolites stop the division of cancer cells by inhibiting DNA and RNA synthesis. Some common antimetabolites used in cancer treatment include 6-mercaptopurine, gemcitabine (Gemzar®; Gemzar is a registered trademark of Eli Lilly and Company), methotrexate, and pemetrexed (Alimta®; Alimta is a registered trademark of Eli Lilly and Company). Topoisomerase inhibitors stop the division of cancer cells by inhibiting the topoisomerase enzyme from separating DNA for replication. Some common topoisomerase inhibitors include topotecan, irinotecan, etoposide, and teniposide. Mitotic inhibitors stop the division of cancer cells by inhibiting the major 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 of the drawbacks of all these anticancer agents is that they also damage healthy tissues. Since drugs treat cancer by inhibiting the function of normal cells, they can also cause significant damage to healthy tissues that rely on a certain degree of cell division, such as blood cells, mucosal surfaces, and the skin. This damage can lead to serious medical conditions and may limit the amount of chemotherapy that can be safely delivered. Examples of side effects that occur during chemotherapy treatment include low blood cell counts, hair loss, muscle and joint pain, nausea, vomiting, diarrhea, mouth sores, fever, and chills. To overcome this problem, new drugs with unique mechanisms of action that provide increased targeting and affect proteins and cell functions that occur only in cancer cells are continuously being developed. For example, antibody-drug conjugates (ADCs) are designed to bind to specific epitopes on the surface of tumor cells and provide an alternative method of targeting tumor cells to reduce associated toxicity. Although highly selective, there are few therapeutically useful ADCs because they achieve only minimal intracellular uptake (<1% of the injected drug) and have limited cell death activity. Some specific anticancer agents 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 C.P. Pharmaceuticals, International CV), and bortezomib (Velcade®; Velcade is a registered trademark of Millennium Pharmaceuticals, Inc.). However, these agents are not approved for all cancer types and are commonly associated with the development of treatment resistance. In addition, many of these compounds still lack absolute tumor selectivity, and their therapeutic use continues to be limited due to off-target effects.

[0009] In recent years, phospholipid ether ("PLE") analogs have been demonstrated to be effective molecular platforms for anti-cancer drug delivery. See U.S. Patent No. 9,480,754 and Weichert et al. (Sci TranslMed, 2014, 6(240), 240ra75), each of which is hereby incorporated by reference in its entirety. As seen therein, most of the anti-cancer drugs used clinically have limited utility because they cannot exert their toxicity against all proliferating cells and / or their effects in all tumor cells. Thus, there remains a need in the art for alternative anti-cancer drug delivery vehicles that can deliver potent, effective, broad-spectrum anti-cancer drugs to cancer cells, including CSCs, while avoiding substantial uptake of the drug by healthy cells. Additionally, the anti-cancer drug delivery vehicle should be able to cross barriers such as the blood-brain barrier (BBB). SUMMARY OF THE INVENTION

[0010] Summary In one aspect, the present disclosure provides a compound of the following chemical formula (I)

Chemical formula

Chemical formula

Chemical formula

[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 the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Brief Description of the Drawings

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DETAILED DESCRIPTION OF THE INVENTION

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

[0040] 1. Definitions Unless otherwise indicated, all technical and chemical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document including the definitions will control. Preferred methods and materials are described below, but 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 not intended to be limiting.

[0041] As used herein, the terms "comprise(s)", "include(s)", "having", "has", "can", "contain(s)" and variations thereof are intended to be open-ended transitional phrases, terms, or words that do not preclude 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 contemplates other embodiments that "comprise", "consist of", and "consist essentially of" the embodiments or elements presented herein, whether or not expressly stated.

[0042] Regarding the description of numerical ranges in this specification, each intermediate value therebetween is clearly contemplated with the same degree of precision. For example, in the range of 6 to 9, in addition to 6 and 9, the numerical values of 7 and 8 are contemplated, and in the range of 6.0 to 7.0, the numerical values of 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9 and 7.0 are clearly contemplated.

[0043] The term “about” or “approximately” as used in this specification when applied to one or more values of interest refers to a value similar to the recited reference value, or a value within an acceptable error range of a particular value 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 of the recited reference value in either direction (above or below), provided such numbers do not exceed 100% of the possible value. Alternatively, “about” can mean within or more than 3 standard deviations, according to the convention of one of ordinary skill in the art. Alternatively, with respect to biological systems or processes, etc., the term “about” can mean within one order of magnitude, preferably within fivefold, more preferably within twofold of a value.

[0044] The definitions of specific functional groups and chemical terms are described in more detail below. For the purposes of this disclosure, chemical elements are identified by the Periodic Table of the Elements (CAS version, Handbook of Chemistry and Physics, 75th Ed., inside cover), and specific functional groups are generally defined as described therein. Further, general principles of organic chemistry, as well as specific functional moieties and reactivities, 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 hereby incorporated by reference into this specification.

[0045] As used herein, the term "cancer" refers to any disease resulting from uncontrolled cell division that can metastasize. The term "cancer" as used herein includes breast cancer, including male breast cancer; gastrointestinal / gastric cancers, including anal cancer, appendiceal cancer, extrahepatic bile duct cancer, gastrointestinal carcinoid, colon cancer, esophageal cancer, gallbladder cancer, gastric cancer, gastrointestinal stromal tumor ("GIST"), islet cell tumor, adult primary liver cancer, pediatric liver cancer, pancreatic cancer, rectal cancer, small intestine cancer, and stomach (stomach) (gastric) cancer; endocrine and neuroendocrine cancers, including pancreatic adenocarcinoma, adrenocortical carcinoma, pancreatic neuroendocrine tumor, Merkel cell carcinoma, non-small cell lung neuroendocrine tumor, small cell lung neuroendocrine tumor, parathyroid cancer, pheochromocytoma, pituitary tumor, and thyroid cancer; eye cancers, including intraocular melanoma and retinoblastoma; genitourinary cancers, including bladder cancer, kidney (renal cell) cancer, penile cancer, prostate cancer, transitional cell carcinoma of the renal pelvis and ureter, testicular cancer, urethral cancer, and Wilms tumor; germ cell cancers, including pediatric central nervous system cancer, pediatric extracranial germ cell tumor, extragonadal germ cell tumor, ovarian germ cell tumor, and testicular cancer; gynecologic cancers, including cervical cancer, endometrial cancer, gestational trophoblastic tumor, epithelial ovarian cancer, ovarian germ cell tumor, uterine sarcoma, vaginal cancer, and vulvar cancer; head and neck cancers, including hypopharyngeal cancer, laryngeal cancer, lip and oral cavity cancer, metastatic squamous neck cancer with occult primary, oral cavity cancer, nasopharyngeal cancer, oropharyngeal cancer, paranasal and nasal cavity cancer, parathyroid cancer, pharyngeal cancer, salivary gland cancer, and throat cancer; leukemias, including adult acute lymphoblastic leukemia, pediatric acute lymphoblastic leukemia, adult acute myeloid leukemia, pediatric acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, and hairy cell leukemia; multiple myeloma, including malignant plasma cells; lymphomas, including AIDS-related lymphoma, cutaneous T-cell lymphoma, adult Hodgkin lymphoma, pediatric Hodgkin lymphoma, Hodgkin lymphoma during pregnancy, mycosis fungoides, adult non-Hodgkin lymphoma, pediatric non-Hodgkin lymphoma, non-Hodgkin lymphoma during pregnancy, primary central nervous system lymphoma, Sézary syndrome, and Waldenström macroglobulinemia; musculoskeletal cancers, including Ewing sarcoma, osteosarcoma, and malignant fibrous histiocytoma of bone, pediatric rhabdomyosarcoma, and soft tissue sarcoma; nervous system cancers, including adult brain tumor, pediatric brain tumor, astrocytoma, brainstem glioma, central nervous system atypical teratoid / rhabdoid tumor, central nervous system fetal tumor, craniopharyngioma, ependymoma, neuroblastoma, and primary central nervous system (CNS) primary malignant lymphoma;Refers to various types of cancer, including but not limited to respiratory / thoracic cancers such as 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 capable of self-renewal and differentiation into different types of cancer cells found in malignant tumors.

[0047] The terms "chemotherapeutic agent", "anticancer agent" and "antineoplastic agent" are used interchangeably throughout this specification.

[0048] Generally, reference to "circulating tumor cell" is intended to refer to a single cell, while reference to "circulating tumor cells" or "cluster of circulating tumor cells" is intended to refer to one or more cancer cells. However, one of ordinary skill in the art will understand that reference to "circulating tumor cells" is intended to include a population of circulating tumor cells containing one or more circulating tumor cells, while reference to "circulating tumor cell" may 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 blood or serum sample of a subject. CTCs may also include or be composed of cancer stem cells or clusters of cancer stem cells found in a blood or serum sample of a subject.

[0049] As used herein, the term "composition" is intended to encompass a product containing a specific amount of a specific ingredient, as well as any product resulting directly or indirectly from a combination of a specific amount of specific ingredients.

[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 to be controlled. A predetermined level can be a cut-off value from a control group. A predetermined level can be an average from a control group. A cut-off value (or a predetermined cut-off value) can be determined by an Adaptive Index Model (AIM) methodology. A cut-off value (or a predetermined cut-off value) can be determined by receiver operating characteristic (ROC) analysis of biological samples from a patient group. As is generally known in the biological field, ROC analysis is the determination of the ability of a test to distinguish one state from another, for example, determining the performance of each marker in identifying ideal patients receiving IL-1Ra therapy. An explanation of ROC analysis is provided by P.J. Heagerty et al. (Biometrics 2000, 56, 337-44), the disclosure of which is incorporated herein by reference in its entirety. Alternatively, a cut-off value can be determined by quartile analysis of biological samples from a patient group. For example, a cut-off value can be determined by selecting a value corresponding to any value in the 25-75 percentile range, preferably the 25th percentile, 50th percentile, or 75th percentile, and more preferably the value corresponding to 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.). A healthy or normal level or range for a target or protein activity can be defined according to standard practice. A control can be a subject or cell without a tumor, as detailed herein.The control can be a subject whose medical condition is known, or a sample from them. The subject, or a sample from them, can be healthy, diseased, diseased before treatment, diseased during treatment, or diseased after treatment, or a combination thereof.

[0051] As used herein, the term "dose" means any form of active ingredient formulation or composition in an amount sufficient to produce a therapeutic effect with at least a 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 specific period of time.

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

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

[0055] As used herein, the term "heterocycloalkyl" refers to a cyclic group of 3 to 24 atoms (C3-C24) selected from carbon, nitrogen, sulfur, phosphate, and oxygen, wherein 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 (steriogenic) center. Thus, the compounds used in the methods of the present invention may exist as enantiomers or racemates and may be isolated. Some compounds may also exhibit polymorphism.

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

[0058] As used herein, the term "sample" or "test sample" can mean any sample in which the presence and / or level of a target is detected or measured. Samples can include liquids, solutions, emulsions, or suspensions. Samples can include medical samples. Samples can include any biological fluid or tissue such as blood, whole blood, blood fractions such as plasma and serum, cartilage, ligaments, tendons, muscle, interstitial fluid, sweat, saliva, urine, tears, synovial fluid, synovium, meniscus, bone marrow, cerebrospinal fluid, nasal mucus, 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 fluids, skin, or combinations thereof. In some embodiments, the sample includes an aliquot. In other embodiments, the sample includes a biological fluid. Samples can be obtained by any means known in the art. Samples can be used directly as obtained from a patient or can be pretreated by filtration, distillation, extraction, concentration, centrifugation, inactivation of interfering components, addition of reagents, etc., by several methods as discussed herein or otherwise known in the art, to alter the properties of the sample.

[0059] As used interchangeably herein, the terms "subject" and "patient" refer to any vertebrate, including but not limited to mammals, that desire or require the compositions or methods described herein. The subject can be human or non-human. The subject can be a vertebrate. The subject can be a mammal. The mammal can be a primate or non-primate. The mammal can be, for example, a non-primate such as a cow, pig, camel, llama, guinea pig, agouti, duckbill platypus, elephant, alpaca, horse, goat, rabbit, sheep, hamster, guinea pig, cat, dog, rat, and mouse. The mammal can be a primate such as a human. The mammal can be, for example, a non-human primate such as a monkey, cynomolgous monkey, rhesus monkey, chimpanzee, gorilla, orangutan, and baboon. The subject can be of any age or developmental stage, such as, for example, an adult, adolescent, or infant. The subject can be male. The subject can be female. In some embodiments, the subject has a particular cancer. The subject may also 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 terms "treat", "treating", or "treatment" mean suppressing, repressing, reversing, alleviating, improving, or inhibiting the progression of a disease, or eliminating the disease completely. Treatment can be effected in either an acute or chronic state. The term also refers to reducing the severity of a disease or symptoms associated with such disease.

[0062] Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings as commonly understood by one of ordinary skill in the art. For example, any specialized 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 meanings and scopes of the terms should be clear; provided, however, that when any potential ambiguity arises, the definitions provided herein shall prevail over any dictionary or external definition. Further, unless the context otherwise requires, the singular forms shall include the plural forms and the plural forms shall include the singular forms.

[0063] 2. Compound In one aspect, the present disclosure provides a compound of the following chemical formula (I)

Chemical formula

Chemical formula

Chemical formula

[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 from 0 to 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 from 10 to 20, an integer from 10 to 40, an integer from 10 to 60, or an integer from 10 to 80. In some embodiments, m is 0, and Q 1 is a bond or the following chemical formula

Chemical formula

[0066] Q 2 may be any known self-immolative spacer, 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 from 2 to 20, and Q 1 is a bond or the following chemical formula

Chemical formula

Chemical formula

[0069] Z may be any anticancer agent, including various known chemotherapeutic drugs.

[0070] In some embodiments, Z is a polo-like kinase 1 (PLK-1) inhibitor. Suitable PLK-1 inhibitors include, for example, diamino pyrimidine (DAP) derivatives such as BI2536, BI6727 (volasertib), DAP-81 and DAP-83, and the compounds disclosed by 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 by reference in their entirety.

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

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

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

[0074] In some embodiments, Z is an eukaryotic translation initiation factor 4 (EIF4) inhibitor such as an EIF4A and 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, the compounds disclosed by Beilina et al. (Bioorganic & Medicinal Chemistry Letters 2006, 16(22), 5757-5762), the content of which is incorporated herein by reference in its entirety.

[0076] In some embodiments, Z is a flavagline analog. Suitable flavagline analogs include, for example, the compounds disclosed in U.S. Patent Application Publication US 2018 / 0086729, the content of which is incorporated herein by reference in its entirety.

[0077] In certain embodiments, Z is, for example, (i) another anti-proliferative / anti-tumor drug such as an alkylating agent, antimetabolite, antitumor antibiotic, antimitotic agent; and a topoisomerase inhibitor; (ii) a cell cycle inhibitor such as an anti-estrogen agent, anti-androgen agent, LHRH antagonist or LHRH agonist, progestogen, and an aromatase inhibitor; (iii) an anti-invasive agent (e.g., a c-Src kinase family inhibitor); (iv) a growth factor function inhibitor such as a tyrosine kinase inhibitor; (v) an anti-angiogenic agent; (vi) a vascular damaging agent; and (vii) an endothelin receptor antagonist, one of other known anti-cancer agents.

[0078] Examples of suitable anti-cancer agents include paclitaxel, irinotecan, topotecan, gemcitabine, cisplatin, geldanamycin, melastatin, abiraterone, afatinib, aminolevulinic acid, aprepitant, axitinib, azacitidine, belinostat, bendamustine, bexarotene, bleomycin, bortezomib, bosutinib, busulfan, cabazitaxel, cabozantinib, capecitabine, carboplatin, carfilzomib, carmustine, ceritinib, cetuximab, chlorambucil, clofarabine, crizotinib, cyclophosphamide, cytarabine, dabrafenib, dacarbazine, dactinomycin, dasatinib, daunorubicin, decitabine, denosumab, dexrazoxane, docetaxel, dolastatin (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, idelalisib, ifosfamide, imatinib, ipilimumab, ixabepilone, lapatinib, leucovorin calcium, lomustine, maytansinoid, mechlorethamine, melphalan, mercaptopurine, mesna, methotrexate, mitomycin C, mitotane, mitoxantrone, nelarabine, nelfinavir, nilotinib, obinutuzumab, ofatumumab, omacetaxine mepesuccinate, oxaliplatin, panitumumab, pazopanib, pegaspargase, pembrolizumab, pemetrexed, pentostatin, pertuzumab, plicamycin, pomalidomide, ponatinib hydrochloride, pralatrexate, procarbazine, radium 223 dichloride, ramucirumab, regorafenib, retaspimycin, luxolitinib, semustine, siltuximab, sorafenib, streptozocin, sunitinib malate, tanespimycin, temozolomide, temsirolimus, teniposide, thalidomide, thioguanine, thiotepa, tamoxifen, trametinib, trastuzumab, vandetanib, vemurafenib, vinblastine, vincristine, vinorelbine,including, but not limited to, bismodigib, volinostat, and ziv-aflibercept.

[0079] In some embodiments, the compound of formula (I) has the structure of formula (I-a), or a pharmaceutically acceptable salt thereof, where Q 1 is the following chemical formula

Chemical formula

Chemical formula

Chemical formula

[0080] In some embodiments, the compound has the structure of formula (I-a), where n is 18. In some embodiments, the compound has the structure of formula (I-a), where Z is a PLK-1 inhibitor or an antitumor agent. In some embodiments, the compound has the structure of formula (I-a-1), (I-a-2), or (I-a-3), or a pharmaceutically acceptable salt thereof, where Z is a PLK-1 inhibitor. For example, Z can be the following chemical formula

Chemical formula

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

Chemical formula

[0082] In some embodiments, the compound of chemical formula (I) has the structure of chemical formula (I-b), or a pharmaceutically acceptable salt thereof, wherein n is 18 and Q 1 is the following chemical formula

Chemical formula

Chemical formula

Chemical formula

[0083] In some embodiments, the compound has the structure of chemical formula (I-b-1), (I-b-2), or (I-b-3), or a pharmaceutically acceptable salt thereof, wherein Z is a combretastatin A-4 analog such as the following chemical formula

Chemical formula

[0084] In some embodiments, the compound of formula (I) has the structure of formula (I-c), or a pharmaceutically acceptable salt thereof, wherein n is 18 and Q 1 is a bond or the following chemical formula

Chem.

Chem.

Chem.

[0085] In some embodiments, the compound has the structure of formula (I-c-1), (I-c-2), or (I-c-3), or a pharmaceutically acceptable salt thereof, wherein Z is the following chemical formula

Chem.

[0086] Suitable compounds disclosed herein include the following chemical formula:

Chem.

Chem.

[0087] The disclosed compounds may exist as pharmaceutically acceptable salts. The term "pharmaceutically acceptable salts" refers to salts or zwitterions of compounds that are water-soluble or oil-soluble or dispersible, suitable for the treatment of disorders without excessive toxicity, irritation, and allergic reactions, and effective for the intended use 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, naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, tartrate, maleate, pivalate, propionate, succinate, tartrate, trichloroacetate, trifluoroacetate, glutamate, paratoluenesulfonate, undecanoate, hydrochloride, hydrobromide, sulfate, and phosphate, etc. 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, etc.

[0088] During the final isolation and purification of the disclosed compounds, basic addition salts may be prepared 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 a primary, secondary, or tertiary organic amine. Quaternary amine salts can be prepared from quaternary amines 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, etc.

[0089] Compounds may exist as stereoisomers having asymmetric or chiral centers. Stereoisomers are either “R” or “S” depending on the arrangement of substituents around a chiral carbon atom. The terms “R” and “S” as used herein are the arrangements described in the lUPAC 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 the compounds can be prepared synthetically from commercially available starting materials containing asymmetric or chiral centers, or by the preparation of a racemic mixture followed by separation methods well known to those skilled in the art. These separation methods include (1) binding a mixture of enantiomers to a chiral 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, separating the resulting mixture of diastereomers by recrystallization or chromatography, and optionally liberating the optically pure product from the auxiliary, or (2) direct separation of a mixture of optical enantiomers on a chiral chromatography column, or (3) fractional recrystallization methods. Compounds may also have tautomers, 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 described by formula (I), except that one or more atoms are replaced with atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the disclosed compounds include, respectively 2 H,3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F, and 36 Cl, etc., include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, but are not limited thereto. Substitution with heavier isotopes such as deuterium, i.e., 2 H, etc., can provide certain therapeutic advantages due to higher metabolic stability, such as an extended half-life in vivo or a reduced required dose, and thus may be preferred in some situations. The compound may incorporate a positron-emitting isotope for medical imaging and positron emission tomography (PET) studies for measuring receptor distribution. Suitable positron-emitting isotopes that can be incorporated into the compound of formula (I) are 11 C, 13 N, 15 O, and 18 F. The isotope-labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by similar procedures as described in the appended examples using appropriate isotope-labeled reagents instead of non-isotope-labeled reagents.

[0091] This compound can be prepared by the synthetic schemes detailed herein. The 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, for example, chromatography on solid supports such as silica gel, alumina, or silica derivatized with alkylsilane groups, recrystallization at high or low temperature with optional pretreatment with activated carbon, thin layer chromatography, distillation at various pressures, sublimation under vacuum, and trituration, as described in "Vogel's Textbook of Practical Organic Chemistry", 5th edition (1989), Longman Scientific & Technical, Essex CM20 2JE, England, but are not limited thereto.

[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 used. Specific procedures are provided in the Examples section. The reaction can be worked up by conventional methods, for example, by removing the solvent from the residue, and can be further purified according to methodologies generally known in the art such as crystallization, distillation, extraction, trituration, and chromatography, but are not limited thereto. Unless otherwise stated, the starting materials and reagents are either commercially available or can be prepared by those 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 chemical techniques, techniques similar to the synthesis of known structurally similar compounds, or techniques similar to the procedures described in the schemes or Examples of synthesis above.

[0093] Routine experiments, including appropriate manipulation of reaction conditions, reagents, and the order of synthetic routes, protection of any chemical functional groups that are not compatible with the reaction conditions, and deprotection at the appropriate points in the reaction sequence of the method, are included within the scope of the present invention. Appropriate protecting groups and methods for protecting and deprotecting different substituents using such appropriate protecting groups are well known to those skilled in the art; examples thereof can be found in PGM Wuts and TW Greene, in Greene’s book titled Protective Groups in Organic Synthesis (4 th ed.), John Wiley & Sons, NY (2006), which is hereby incorporated by reference in its entirety. The synthesis of the compounds of the present invention can be achieved by methods similar 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 composition can be manufactured by processes known in the art, such as conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilization processes.

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

[0097] A pharmaceutically acceptable carrier can be a functional molecule such as a vehicle, adjuvant, or diluent. A pharmaceutically acceptable carrier can be any type of non-toxic and inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation aid. Pharmaceutically acceptable carriers include, for example, diluents, lubricants, binders, disintegrants, colorants, flavors, sweeteners, antioxidants, preservatives, lubricants, solvents, suspending agents, wetting agents, surfactants, softeners, propellants, humectants, 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, etc.), colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, polyacrylate, waxes, polyethylene polyoxypropylene block polymers, lanolin, sugars (such as lactose, glucose, and sucrose), starches (such as com starch and potato starch), cellulose and its derivatives (such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate), powdered tragacanth, malt, gelatin, talc, excipients (such as cocoa butter and suppository waxes), oils (such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, soybean oil, etc.), glycols (such as propylene glycol or polyethylene glycol), esters (such as ethyl oleate, ethyl laurate, etc.), agar, non-toxic compatible lubricants (such as sodium lauryl sulfate and magnesium stearate), colorants, release agents, coating agents, emulsifiers, sweeteners, flavors, preservatives, antioxidants, and 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 the 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 formulations include, but are not limited to, capsules, tablets, pills, powders, cements, pastes, and granules. Dosage forms for topical or transdermal administration of the present 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, for example, water, ethanol, polyols (such as glycerol, propylene glycol, liquid polyethylene glycol, etc.), vegetable oils, non-toxic glyceryl esters, and suitable mixtures thereof.

[0102] The pharmaceutical composition can be in a dosage form suitable for injection or infusion, such as a sterile aqueous solution or dispersion or sterile powder containing the 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. The sterile injectable solution can be prepared by incorporating the compound or its pharmaceutically acceptable salt as disclosed herein, optionally together with various other ingredients, in a suitable solvent in the required amounts and then, optionally, filter sterilizing. In the case of sterile powders for the preparation of sterile injectable solutions, the methods of preparation may include vacuum drying and freeze-drying techniques to obtain a powder of the active ingredient(s) + 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. The dispersion can also be prepared in glycerol, liquid polyethylene glycol, triacetin, and mixtures thereof, and in oils. These formulations may contain preservatives to prevent the growth of microorganisms. Prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.

[0104] Injectable formulations can be prepared by forming a microcapsule matrix of the disclosed compound(s) or a pharmaceutically acceptable salt thereof in a biodegradable polymer such as polylactic acid - polyglycolide. Depending on the ratio of the compound to the polymer and the nature of the particular polymer, the drug release rate can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Injectable formulations can also be prepared by encapsulating the drug within liposomes or microemulsions that are compatible with body tissues.

[0105] In some embodiments, the composition may include 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, melastatin, abiraterone, afatinib, aminolevulinic acid, aprepitant, axitinib, azacitidine, belinostat, bendamustine, bexarotene, bleomycin, bortezomib, bosutinib, busulfan, cabazitaxel, cabozantinib, capecitabine, carboplatin, carfilzomib, carmustine, ceritinib, cetuximab, chlorambucil, clofarabine, crizotinib, cyclophosphamide, cytarabine, dabrafenib, dacarbazine, dactinomycin, dasatinib, daunorubicin, decitabine, denosumab, dexrazoxane, docetaxel, dolastatin (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, idelalisib, ifosfamide, imatinib, ipilimumab, ixabepilone, lapatinib, leucovorin calcium, lomustine, maytansinoid, mechlorethamine, melphalan, mercaptopurine, mesna, methotrexate, mitomycin C, mitotane, mitoxantrone, nelarabine, nelfinavir, nilotinib, obinutuzumab, ofatumumab, omacetaxine mepesuccinate, oxaliplatin, panitumumab, pazopanib, pegaspargase, pembrolizumab, pemetrexed, pentostatin, pertuzumab, plicamycin, pomalidomide, ponatinib hydrochloride, pralatrexate, procarbazine, radium 223 dichloride, ramucirumab, regorafenib, retaspimycin, luxorutinib, semustine, siltuximab, sorafenib, streptozocin, sunitinib malate, tanespimycin, temozolomide, temsirolimus, teniposide, thalidomide, thioguanine,Including, but not limited to, thiotepa, toremifene, trametinib, trastuzumab, vandetanib, vemurafenib, vinblastine, vincristine, vinorelbine, besimodegib, vorinostat, and ziv-aflibercept. Any compound currently known or capable of acting as an anti-cancer agent is also useful in the present disclosure.,

[0106] 4. Method The basis for the selective tumor targeting of the compounds detailed in this specification lies in the differences between the cell membranes of cancer cells compared to those of most normal cells. Phospholipid ether (PLE) molecules utilize the metabolic shift that tumor cells undergo to generate the energy required for rapid cell division. Tumors promote the utilization of the beta-oxidation pathway to convert long-chain fatty acids (LCFAs) into energy. To increase the uptake of LCFAs, tumor cells form special microdomains known as "lipid rafts" that alter the cell membrane. Lipid rafts are formed due to the metabolic shift and the need for phospholipids. Within tumor cells, these regions become excessive and stabilized, capable of serving as potential tumor-specific targets. In particular, cancer cell membranes are highly enriched in lipid rafts. In normal tissues, the presence of lipid rafts is restricted 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 have been demonstrated to be highly abundant in almost all tumor types, being present in 100% of the individual cancer cells tested. Lipid rafts are highly organized and specialized regions of the phospholipid membrane bilayer, containing high concentrations of various signaling molecules, sphingolipids, glycosphingolipids, and cholesterol, and serving to organize cell surface and intracellular signaling molecules (e.g., growth factor and cytokine receptors, phosphatidylinositol 3-kinase (PI3K) / Akt survival pathway). Data suggests that lipid rafts function as an entry point for phospholipid ethers (PLEs). The significant selectivity of these compounds for cancer cells over 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 extremely important role played by lipid rafts is emphasized by the fact that disruption of the lipid raft structure inhibits the uptake of PLEs into cancer cells. It has been shown that when the formation of lipid rafts is blocked, the uptake of PLEs decreases by 60%. These functions, combined with lipid rafts that provide rapid internalization of phospholipid drug conjugates, make them ideal targets.

[0107] Compounds described herein, such as PLE analogs, can 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 move along the Golgi apparatus network in the cytoplasm to the endoplasmic reticulum and mitochondria. In some embodiments, the phospholipid drug conjugates (PDCs) disclosed herein include uniquely designed phospholipid ethers conjugated to novel combretastatin A (CBA) analogs via cleavable linkers. CBAs are potent cytotoxins and have been demonstrated to inhibit tubulin polymerization within tumor cells and disrupt the local vasculature around / within tumors. In some embodiments, the compounds disclosed herein include uniquely designed phospholipid ethers conjugated to flavagline (FLV) analogs via cleavable linkers. FLVs are potent cytotoxins that inhibit translation, progression of the cell cycle, and induce apoptosis.

[0108] The compounds detailed herein, or pharmaceutically acceptable salts thereof, or compositions containing 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 containing the 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 use in manufacturing a medicament for treating cancer in a subject in need thereof.

[0111] The cancers that can be treated with the compounds detailed herein, or pharmaceutically acceptable salts thereof, or compositions comprising the compounds detailed herein are: breast cancer including male breast cancer; anal cancer, appendiceal cancer, extrahepatic bile duct cancer, gastrointestinal carcinoid, colon cancer, esophageal cancer, gallbladder cancer, gastric cancer, gastrointestinal stromal tumor ("GIST"), islet cell tumor, adult primary liver cancer, pediatric liver cancer, pancreatic cancer, rectal cancer, small intestine cancer, and digestive / gastrointestinal cancers including stomach (gastric) cancer; endocrine and neuroendocrine cancers including pancreatic adenocarcinoma, adrenocortical carcinoma, pancreatic neuroendocrine tumor, Merkel cell carcinoma, non-small cell lung neuroendocrine tumor, small cell lung neuroendocrine tumor, parathyroid cancer, pheochromocytoma, pituitary tumor and thyroid cancer; eye cancers including intraocular melanoma and retinoblastoma; genitourinary cancers including bladder cancer, kidney (renal cell) cancer, penile cancer, prostate cancer, transitional cell carcinoma of the renal pelvis and ureter, testicular cancer, urethral cancer, and Wilms tumor; germ cell cancers including pediatric central nervous system cancer, pediatric extracranial germ cell tumor, extragonadal germ cell tumor, ovarian germ cell tumor, and testicular cancer; gynecological cancers including cervical cancer, endometrial cancer, gestational trophoblastic neoplasm, epithelial ovarian cancer, ovarian germ cell tumor, uterine sarcoma, vaginal cancer, and vulvar cancer; head and neck cancers including hypopharyngeal cancer, laryngeal cancer, lip and oral cavity cancer, metastatic squamous neck cancer with occult primary, oral cavity cancer, nasopharyngeal cancer, oropharyngeal cancer, paranasal and nasal cavity cancer, parathyroid cancer, pharyngeal cancer, salivary gland cancer and laryngeal cancer; leukemias including adult acute lymphoblastic leukemia, pediatric acute lymphoblastic leukemia, adult acute myeloid leukemia, pediatric acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, and hairy cell leukemia; lymphomas including AIDS-related lymphoma, cutaneous T cell lymphoma, adult Hodgkin lymphoma, pediatric Hodgkin lymphoma, Hodgkin lymphoma during pregnancy, mycosis fungoides, adult non-Hodgkin lymphoma, pediatric non-Hodgkin lymphoma, non-Hodgkin lymphoma during pregnancy, primary central nervous system lymphoma, Sézary syndrome and Waldenström macroglobulinemia; musculoskeletal cancers including Ewing sarcoma, osteosarcoma and malignant fibrous histiocytoma of bone, pediatric rhabdomyosarcoma and soft tissue sarcoma; nervous system cancers including adult brain tumor, pediatric brain tumor, astrocytoma, brainstem glioma, central nervous system atypical teratoid / rhabdoid tumor, central nervous system fetal tumor, craniopharyngioma, ependymoma, neuroblastoma, primary central nervous system (CNS) primary malignant lymphoma;Respiratory / chest 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, but not limited thereto. In certain embodiments, the cancer can be melanoma, lung cancer, colorectal cancer, breast cancer, or a combination thereof.;

[0112] In another embodiment, the cancer can include one or more CTCs. The one or more CTCs can be selected from the group consisting of breast cancer, lung cancer, thyroid cancer, cervical cancer, melanoma, squamous cell carcinoma, prostate cancer, pancreatic cancer, colorectal cancer, and cancer stem cells, and malignant plasma cells.;

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

[0114] In another embodiment, the cancer can be cancer stem cells. In certain embodiments, the cancer stem cells can be derived from the group consisting of breast cancer, lung cancer, melanoma, and colorectal cancer.;

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

[0116] In some embodiments, the melanoma can 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 colorectal cancer can include adenocarcinoma.;

[0118] In some embodiments, the compounds of formula (I-a), (I-a-1), (I-a-2), or (I-a-3) detailed herein, or pharmaceutically acceptable salts thereof, or compositions comprising the compounds detailed herein can be used to treat melanoma, lung cancer, colorectal cancer, or a combination thereof.

[0119] In some embodiments, breast cancer can include invasive ductal carcinoma, metastatic breast cancer, inflammatory breast cancer, triple-negative breast cancer, non-invasive ductal carcinoma, or a combination thereof. In further embodiments, the cancer is breast cancer and the subject can be estrogen receptor positive, both estrogen receptor negative and progesterone receptor negative, HER2-expressing (HER2+), HER2 non-expressing (HER2-), or a combination thereof. In some embodiments, the compounds of formula (I-b), (I-b-1), (I-b-2), or (I-b-3) detailed herein, or pharmaceutically acceptable salts thereof, or compositions comprising the compounds detailed herein can be used to treat breast cancer.

[0120] In some embodiments, the compounds of formula (I-c), (I-c-1), (I-c-2), or (I-c-3) detailed herein, or pharmaceutically acceptable salts thereof, or compositions comprising the compounds detailed herein can be used to treat melanoma, lung cancer, colorectal cancer, breast cancer, or a combination thereof.

[0121] In some embodiments, the subject is a human such as an adult and an 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 the compound detailed herein in an amount detailed herein. In some embodiments, the method includes administering a compound, or a pharmaceutically acceptable salt thereof, at about 0.0001 to about 1000 mg / kg as detailed herein.

[0123] The useful dosage of the compound(s) in the composition can be determined by comparing their in vitro and in vivo activities in an animal model. Methods for estimating effective dosages in 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 so as to obtain an amount of the compound(s) useful in achieving the desired therapeutic response for a particular patient, composition, and mode of administration. The selected dosage level and the amount of the compound, or a pharmaceutically acceptable salt thereof, to be used in therapy will vary depending upon the particular compound or salt selected, the route of administration, the disease or condition to be 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 the art to initiate administration of the compound at a level lower than that required to obtain the desired therapeutic effect and to 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, particularly for the effective and aggressive treatment of particularly aggressive diseases or conditions.

[0125] In some embodiments, the compounds disclosed herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions can be administered by oral administration or intravenous administration. However, generally, appropriate dosages 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, appropriate volumes can be in the range of about 0.001 mg / kg to about 5.0 mg / kg body weight per day of the recipient, such as about 0.01 mg / kg to about 1.0 mg / kg body weight per day of the recipient, about 0.01 mg / kg to about 3.0 mg / kg body weight per day of the recipient, about 0.1 mg / kg to about 5.0 mg / kg body weight per day of the recipient, about 0.2 mg / kg to 4.0 mg / kg per day of the recipient. The compound can be administered in unit dosage form; for example, it contains 1 to 100 mg, 10 to 100 mg, or 5 to 50 mg of the active ingredient per unit dosage form.

[0126] The desired dosage can conveniently be presented as a single dose, or as divided doses administered at appropriate intervals, for example, as 2, 3, 4, or more sub-doses per day. The sub-doses themselves can, for example, be further divided for administration at many individual, spaced intervals.

[0127] The specific in vivo dosage administered and the specific mode of administration can vary depending on the age, weight, severity of pain, and mammalian species being treated, the specific compound being applied, and the specific use to which these compounds are applied. Determination of an effective dosage level to achieve the desired result 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 pharmaceutically acceptable salts thereof, can be determined by comparing the in vitro activity and in vivo activity of animal models. Such comparison may be made by comparing with established drugs.

[0128] The dosage and intervals can be adjusted individually to provide a plasma level of the active moiety sufficient to maintain the 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 the individual characteristics and the route of administration. However, an FIPLC assay or a bioassay can be used for the measurement of plasma concentrations. The dosing interval can also be determined using the MEC value. The composition should be administered using regimens in which the plasma level is maintained above the MEC value for between 10 - 90% of the time, preferably 30 - 90%, and most preferably 50 - 90% of the time, preferably between 30 - 90% and more preferably between 50 - 90%. In the case of topical administration or selective uptake, the effective local concentration of the drug may not be related to the 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 that compound sharing a particular chemical moiety, can be established by measuring the in vitro toxicity against cell lines such as mammalian, and preferably human, cell lines. The results of such studies often predict toxicity in animals such as mammals, or more specifically, humans. Alternatively, the toxicity of a particular compound in animal models such as mice, rats, rabbits, dogs or monkeys 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 of ordinary skill in the art can be guided by the state of the art to select an appropriate model, dosage, route of administration and / or regime.

[0130] The compounds(s) detailed herein, or a pharmaceutically acceptable salt thereof, or a composition 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, by inhalation, intraperitoneally, topically (by powder, ointment or drops), buccally or by oral or nasal spray. As used herein, the term "parenteral" or "parenterally" refers to a mode of administration that includes intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intra-articular injection and infusion.

[0131] The compositions described herein can be administered with additional compositions, or in combination with, or before or after the administration of, additional therapeutic agents to extend the stability, delivery, and / or activity of the composition. A combination therapy can include the administration of a single pharmaceutical dosage form comprising one or more of the compounds described herein and one or more additional pharmaceuticals, as well as the administration of the compounds and each additional pharmaceutical in separate pharmaceutical dosage forms by themselves. For example, the compounds detailed herein can be administered to a subject in combination with additional anti-cancer agents detailed herein.

[0132] The compounds detailed in this specification, or pharmaceutically acceptable salts thereof, can also be administered in the form of liposomes. As is known in the art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by single or multiple layer hydrated liquid crystals dispersed in an aqueous medium. Optionally, physiologically acceptable and metabolizable lipids that can form liposomes can be used. The composition in liposome form can contain, in addition to the compounds described herein, anti-cancer agents, stabilizers, preservatives, additives, etc. Preferred lipids include natural and synthetic phospholipids and phosphatidylcholine (lecithin) 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, N.Y. (1976), p. 33 and the following 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, a pharmaceutically acceptable composition can be delivered in a controlled release system. For example, the agent can be administered using intravenous injection, 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 in the vicinity of a therapeutic target, such as the liver, and thus requires only a fraction of the systemic dose (see, for example, 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. Example The foregoing can 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 a plurality of aspects and embodiments, which are illustrated by the accompanying non-limiting examples.

[0135] Example 1. Materials and Methods The in vitro uptake of CLR2000045 was evaluated using MCF-7 breast cancer cells and normal human dermal fibroblast (NHDF) cells and measured via LC / MS / MS. The breast cancer cells were maintained in minimum essential medium supplemented with 10% FBS. All cells were at 37°C and 5% CO 2It was maintained. Cells were incubated with 1 μM of the drug, and the reported values were the average of three evaluations. In vitro cytotoxicity was measured by the 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 were 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 of the drug, and the reported values were the average of three evaluations. In vitro cytotoxicity was measured by the Cell Titer-Glo® assay.

[0137] In an in vivo efficacy screening model using chicken embryos, 72 μM of CLR2000045 was administered to measure its efficacy against MCF-7 tumors and compared with a vehicle control and a paclitaxel positive control at 50 μM. CLR2000045 was applied locally to the embryo's casing. Fertilized white leghorn eggs were incubated at 37.5 °C and 50% relative humidity for 9 days. At that moment (E9), a small hole was made in the eggshell to drop the chorioallantoic membrane (CAM) into the air sac, and a 1 cm 2 window was opened in the eggshell on the CAM. At least 20 eggs (there may be more than 20 eggs per group depending on the survival rate of the embryos after 9 days of development) were used for each group. Since some embryo deaths may occur after tumor transplantation or be related to defects in tumor transplantation, data can be collected with less 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 6 cell inoculum was appropriately added onto the CAM of each egg (E10) for each group, and then the eggs were randomly divided into groups.

[0138] The viability of the embryos was checked daily. The number of embryos that died at E18 was counted in conjunction with the observation of final visible gross abnormalities to assess the embryo toxicity induced by the treatment. The final mortality rate and Kaplan–Meier curves were calculated for the entire group. Any visible abnormalities observed were also noted. On day E18, the upper part of the CAM (with the tumor) was removed from all surviving embryos with tumors, washed with PBS buffer, and then transferred directly to PFA (fixed for 48 h). Subsequently, the tumors were carefully dissected from the normal CAM tissue and weighed.

[0139] In vivo efficacy was further evaluated in HCC70 triple-negative breast cancer (TNBC) xenograft R2G2 mice. CLR2000045 was evaluated at three doses (1 mg / kg) administered once, twice, or three times weekly for 2 weeks. CLR2000045 was administered systemically via tail vein injection. Each group contained 10 mice. Tumor volume was recorded for efficacy evaluation and body weight was recorded for tolerance evaluation. Survival rate was also recorded.

[0140] CLR180099 was intravenously administered (IV) to healthy C57BL / 6 mice to measure the maximum tolerated dose (MTD) compared to FLV molecule alone. In this case, the vehicle used to administer CLR180099 was PBS, although any pharmaceutically appropriate vehicle could be used. Each group contained 5 mice. In vivo efficacy was evaluated in HCT116 xenograft athymic nude mice. The mice were developed in a flank model by injecting approximately 1×10 6 cells resuspended in 5 ml of 1.2% methylcellulose into the posterior flanks of the mice. The study was initiated when the mean tumor volume of the group was approximately 120 mm 3It was initiated when [the specified condition] was reached. Tumor volume was measured using calipers, and the measured values of tumor length, width, and depth were used to calculate the tumor volume. Two doses of CLR180099 (administered 2 mg / kg twice or 2 mg / kg three times) were evaluated. Each group included 10 mice. Tumor volume was recorded for efficacy evaluation, and body weight was recorded for tolerance evaluation. All conjugated CLR180099 and free FLV were measured by mass spectrometry.

[0141] Example 2. Phospholipid lipid ether delivery vehicles are specific for a wide range of tumor cells To demonstrate the uptake of PDCs in various tumor cell lines, various tumor cell lines such as MCT-116, MeS SA / Dx5, MIa PaCa-2, Ovcar-3, and U-87MG were incubated with 5 μM of CLR1501 (PLE + BODIPY fluorescent payload) in complete medium at 37 °C for 24 hours. Each cell line can have a slightly different medium to optimize growth, and any suitable medium known in the art can be used for each cell line. All cells were cultured in 10% FBS and 5% CO 2It was maintained at 37 °C in an appropriate medium supplemented with [substance not specified]. CLR1501 was excited and then detected with an Alexa-Fluor488 filter. CLR1501 was highly localized in all different tumor cell lines (Figures 1A and 1B). Similar results have been reported in over 100 tumor cell lines such as 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 different cancer cell lines and normal human skin fibroblast cell lines. After 24 hours, in these cancer cell lines in vitro, CLR1501 was shown to be preferentially taken up 5 - 9 times more than normal fibroblasts. The retained CLR1501 was associated with plasma and organelle membranes.

[0142] The in vitro uptake and release of the cytotoxic payload was measured by incubating 2 μM of the cytotoxic small molecule PDC with the semi-stable linker (CLR2208, “PDC-SM1”) in complete medium at 37 °C for 48 h in A375 and A549 cell lines. The uptake of PDC-SM1 was measured by LC / MS / MS. PDC-SM1 was demonstrated to initiate uptake within 30 min. 20–40% of the conjugate exposed to cells was measured in the cytoplasm of tumor cells within 24 h (Figure 2A). Subsequently, CLR2206 (“PDC-SM2”, similar to PDC-SM1 except for the cleavable linker) was utilized to measure the release of the payload within tumor cells. CLR2200 (“PDC-SM3”) was also studied. Measurable release of the small molecule payload occurred between 1 and 2 h after incubation (Figure 2B). Slight release of the payload occurred in the medium (<1 nM). These results demonstrated that phospholipid ether molecules have the ability to target a broad range of tumors and that PDCs have the ability to achieve 20–40% uptake of the drug exposed to tumor cell lines.

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

[0144] The in vitro efficacy with the cytotoxic payload was measured. PDC-SM2 demonstrated submicromolar activity (concentration measured based on the concentration of the complete conjugate incubated on 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 showed similar activity and efficacy against colorectal cancer (HCT-116) cells as lung cancer, which was not active against normal fibroblasts. Thus, PDCs demonstrated payload release and potent nanomolar activity against tumor cells.

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

[0146]

Table 1

[0147] The 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 by tail vein injection. Regions with increased signal intensity were observed using an IVIS Spectrum, which allows direct visualization of CLR1502 through the animal's skin. Next, after euthanizing the animals, tissues identified with the IVIS system were excised by microdissection and histology was performed to confirm tumor tissue versus non-tumor tissue and the location where near-infrared labeling occurred. These regions showed non-invasive (colon Figure 19C; terminal small intestine Figure 19F) and invasive (colon Figure 19D; terminal small intestine Figure 19E) tumors.

[0148] In other studies, CLR1502 accumulates in metastases and the draining lymph nodes. After removing the intestine, the mesenteric fat, pancreas, and spleen were isolated together. In one example, two metastatic tumor deposits ~4 mm in size were identified within the mesentery. These lesions were easily visualized with a Fluobeam near-infrared imaging device. These lesions were confirmed to be metastatic neoplastic lesions by H&E. Local lymph node swelling was also shown to accumulate CLR1502 using Fluobeam. No malignant cells were observed within these hyperplastic lymph nodes.

[0149] Tumor thickness does not contribute to the increase in signal intensity observed in intestinal cancer (Figures 22A and 22B). Autopsies were performed on mice 96 hours after injecting 50 μg of CLR1502 per mouse. To examine the effect of tissue thickness, sections of the apparently normal colon were overlaid on each other. The radiation efficiency was measured and the signal intensity between 1, 2, and 3 layers of normal colon and intestinal tumors was compared. One layer of the normal colon was found to be approximately 1 mm thick. Tissue thickness may contribute to the increase in intensity seen in adenomas, but not to the differences seen in adenocarcinomas.

[0150] In vivo optical scanning of CLR1502 uptake in a colorectal cancer model demonstrated preferential retention in malignant tissues compared to normal tissues. Thymus-deficient nude mice bearing colorectal cancer (HCT-116) xenografts were intravenously injected with 1 mg of CLR1502 and imaged using the 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 breast cancer model mouse demonstrated preferential retention in malignant tissues compared to normal tissues. Approximately 80 μg of CLR1502 was intravenously injected into thymus-deficient nude mice bearing orthotopic breast cancer xenografts (MDA-MB-231), and imaged in vivo daily for 7 days (168 hours) using the Fluoptics Fluobeam® and IVIS® Spectrum systems (Figure 24). The study results showed selective uptake and long-term retention in the tumor (yellow and green arrows for Fluobeam and IVIS Spectrum, respectively), and a relative increase in clearance from normal tissues over time.

[0152] Approximately 50 μg of CLR1502 was intravenously injected into thymus-deficient nude mice bearing lung cancer xenografts (H226 lung) in each flank, and imaged in epifluorescence mode using the IVIS Spectrum (Figure 25). It should be noted that, as indicated by the black arrow, at 96 hours, due to the difference in the radiation efficiency of malignant and normal tissues, sufficient contrast in light was created at the edge of the tumor.

[0153] Example 3. CLR2000045 with Combretastatin A-4 analog improves breast cancer treatment CLR2000045 showed minimal uptake into normal tissues and significant uptake into tumor cells. The drug release showed approximately 50% release at each time point. A steady state was achieved between 24 and 48 hours between drug uptake and release (Figure 6). CLR2000045 showed excellent activity and efficacy 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 that cell line (Table 2). The plasma stability of CLR2000045 was also measured (Table 3).

[0154]

Table 2

[0155]

Table 3

[0156] Fertilized eggs of white leghorn chickens (20 eggs / group) were incubated at 37.5 °C for 9 days. MCF-7 cells were cultured under standard conditions before transplantation. On day 10, 3×10 6 individual MCF-7 inocula were added to the chorioallantoic membrane. The eggs were then randomly divided into treatment groups and treated 4 times (on days 11, 13, 15, and 17) under the following conditions; vehicle, 50 μM paclitaxel per dose, and 72 μM CLR2000045 per dose. CLR2000045 showed similar activity to paclitaxel in this screening model (Figure 8).

[0157] This study was conducted when the mean tumor volume of the group was ~200 mm 3It was initiated when reaching the (4th day). CLR2000045 was administered IV at the following dosages: 1 mg / kg on the 5th and 12th days, or on the 5th, 8th, 12th, and 15th days, or on the 5th, 7th, 9th, 12th, 14th, and 16th days. CLR2000045 demonstrated a dose-response decrease in tumor volume from dose group 1 to dose group 3 (3 times a week for 2 weeks), and the highest dose tested showed almost 100% eradication of the tumor. The two highest dose groups showed a statistically significant decrease in tumor volume compared to the vehicle control (p ≤ 0.05 and p ≤ 0.01, respectively) (Figure 9). The Kaplan-Meier curves showed that treatment with CLR2000045 at 1 mg / kg 3 times a week for 2 weeks resulted in a significant increase in survival rate compared to the vehicle and once-weekly administration (p ≤ 0.001 and p ≤ 0.05, respectively). Administration twice a week at 1 mg / kg for 2 weeks resulted in a significant increase compared to the vehicle (p ≤ 0.05; Figure 10). Changes in body weight after treatment were measured in the (HCC70) mouse xenograft model (Figure 11A and Figure 11B).

[0158] CLR2000045 demonstrated minimal uptake into normal cells while showing significant payload uptake and release (20 - 40% of the exposed drug) in tumor cell lines. CLR200045 showed potent in vitro activity against multiple breast cancer cell lines. CLR2000045 demonstrated potent in vivo activity against the triple-negative breast cancer model (HCC70) and the metastatic adenocarcinoma breast cancer model (MCF-7). CLR200045 provided a significant life-prolonging effect in the TNBC (HCC70) model and was well tolerated even 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, warranting the continued development of this PDC.

[0159] Example 4. CLR180099 improves the safety and efficacy of anti-tumor drugs against colorectal tumors CLR180099 showed excellent activity and efficacy against breast and lung cancers, 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 colorectal cancer. The plasma stabilities of CLR1800095, CLR180099A, and CLR180099B were measured in mice and humans (Table 4). CLR1800095 showed some instability in mouse plasma and could cause some toxicity.

[0160] [Table 4]

[0161] The study was initiated when the mean tumor volume of the group reached ~120 mm 3 (day 1). CLR180099 was administered IV at 2 mg / kg either on days 1 and 4 or on days 1, 3, and 5. Docetaxel was administered at 10 mg / kg on days 1 and 4. CLR180099 demonstrated a reduction in tumor volume equivalent to or greater than that of docetaxel and a dose-dependent effect. In the docetaxel group, multiple deaths occurred from the start of day 18 until the end of day 26 (Figure 16). The Kaplan-Meier curves showed that treating with CLR180099 at 2 mg / kg on days 1 and 4 or on days 1, 3, and 5 resulted in a significant increase in survival compared to docetaxel (Figure 17, log-rank test, p ≤ 0.001). When body weight loss was measured, all mice treated with CLR180099 (both volumes) 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). The payload alone was not tolerated at doses exceeding 0.5 mg / kg (all mice died at 0.5 mg / kg).

[0162] [Table 5]

[0163] CLR180099 showed minimal uptake in normal cells, while demonstrating significant payload uptake and release (20 - 40% of the exposed drug) in tumor cell lines. CLR180099 showed 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 administrations of CLR180099 showed activity equivalent to or greater than docetaxel in colorectal cancer. Furthermore, CLR180099 significantly improved the survival effect at both doses compared to docetaxel. Tolerability evaluation demonstrated that CLR180099 showed sufficient tolerability in both tumor-bearing and normal animals, and the FLV payload was shown to be toxic in both normal and tumor-bearing mice. CLR180099 showed no toxic effects compared to the FLC analog payload alone, indicating that the payload may be effective for targeted delivery by phospholipid ether (PLE).

[0164] Example 5. Synthesis of Compounds The chemical synthesis process was carried out as follows. The product was isolated using known techniques such as HPLC, and the obtained structure was verified by NMR and MS.

[0165] CLR2208 was synthesized according to Scheme 1 below [Chemical formula] as follows.

[0166] CLR2206 was synthesized according to Scheme 2. Hypophosphoryl chloride 1A (2.5 equivalents) was used at -40 °C for 3 hours in Et 3 N (10 equivalents) and THF to prepare Compound 2 from Compound 1. Compound 2 was reacted with Et 3 N (1 equivalent), CDI (1.5 equivalents), ZnCl 2(2.6 equivalents), and reacted with 2A (1 equivalent) in DMF at 15 °C for 12 hours to obtain Compound 3. Compound 3 was deprotonated in piperidine (5 equivalents of DMF, 15 °C, 3 hours) to give Compound 4. Compound 4 was 3 reacted with Et 3 N (4 equivalents), COMU (1.15 equivalents), and in CHCl

Chemical formula

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

Chemical formula

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

Chemical formula

Chemical formula

[0169] CLR2013 was prepared according to Scheme 6.

Chemical formula

[0170] CLR1800095 was prepared according to Scheme 7.

Chem.

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

Chem.

[0172] CLR180099A was prepared according to Scheme 9.

Chem.

[0173] To complete, various aspects of the present invention are set forth in the following numbered items.

[0174] Item 1. A compound of the following chemical formula (I),

Chem.

Chem.

Chem.

[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

Chem.

Chem.

[0176] Item 3. The compound according to any one of Items 1 to 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 agent, a eukaryotic translation initiation factor 4 (EIF4) inhibitor, a combretastatin A-4 analog, or a flavagline analog.

[0177] Item 4. The compound according to any one of Items 1 to 3 having the structure of chemical formula (I-a), or a pharmaceutically acceptable salt thereof, wherein Q 1 is the following chemical formula

Chem.

Chem.

[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 D (MMAD).

[0179] Item 6. The compound according to any one of Items 1 to 3 having the structure of chemical formula (I-b), or a pharmaceutically acceptable salt thereof, wherein n is 18; Q 1 is the following chemical formula

Chemical formula

Chemical formula

[0180] Item 7. The compound according to any one of Items 1 to 3 having the structure of chemical formula (I-c), or a pharmaceutically acceptable salt thereof, wherein n is 18; Q 1 is a bond or the following chemical formula

Chemical formula

Chemical formula

[0181] Item 8. The following chemical formula

Chemical formula

Chemical formula

Chemical formula

[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, colorectal cancer, breast cancer, or a combination thereof.

[0185] Item 12. The method according to any one of items 10 to 11, wherein the above-mentioned lung cancer includes small cell lung cancer, non-small cell lung cancer, or a combination thereof; the above-mentioned melanoma includes superficial spreading melanoma, nodular melanoma, lentigo melanoma, acral lentiginous melanoma, amelanotic melanoma, nevoid melanoma, spitzoid melanoma, desmoplastic melanoma, or a combination thereof, the above-mentioned colorectal cancer includes adenocarcinoma; or the above-mentioned breast cancer includes invasive ductal carcinoma, metastatic breast cancer, inflammatory breast cancer, triple-negative breast cancer, non-invasive ductal carcinoma, 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, colorectal cancer, or a combination thereof, and wherein the compound is a compound of formula (I-a), 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+), (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 wherein the compound is a compound of formula (I-b), 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, colorectal cancer, breast cancer, or a combination thereof, and wherein the compound is a compound of (I-c), or a pharmaceutically acceptable salt thereof.

[0193] The foregoing description of the specific embodiments has been so thorough as to disclose the general nature of the invention, so that others may, by applying the knowledge within the skill of the art, readily modify and / or adapt such specific embodiments without undue experimentation and without departing from the general concept of the present disclosure. Accordingly, such adaptations and modifications are intended to be within the meaning and scope of the disclosed embodiments and equivalents thereof as based upon the embodiments and guidance presented herein. It is to be understood that the terminology or phrasing herein is for the purpose of description and not of limitation, as it is to be interpreted by those of ordinary skill 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 hereby 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 were individually indicated to be incorporated by reference.

Claims

1. A compound of the following formula (I): 【Chemistry 1】 or a pharma- ceutically 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 is the following chemical formula 【Chemistry 3】 where R x is H or a halogen; Q 2 is a bond or a self-immolative spacer; and Z is an anticancer drug); A compound of formula (I), or a pharma- ceutically acceptable salt thereof.

2. Q 1 is bonded or has the following chemical formula 【Chemistry 4】 and L-Q 2 is the following chemical formula 【Chemistry 5】 2. The compound of claim 1, wherein:

3. 3. The compound of claim 1, wherein Z is a polo-like kinase 1 (PLK-1) inhibitor, a tubulin polymerase inhibitor, a tubulin stabilizer, an antitumor agent, a eukaryotic translation initiation factor 4 (EIF4) inhibitor, a combretastatin A-4 analog, or a flavugin analog, or a pharma- ceutically acceptable salt thereof.

4. Q 1 is the following chemical formula 【Chemistry 6】 and L-Q 2 is the following chemical formula 【Chemistry 7】 and and 4. The compound of any one of claims 1 to 3, having the structure of formula (I-a), or a pharma- ceutically 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.

5. 5. The compound of claim 4, wherein Z is a PLK-1 inhibitor or antineoplastic agent selected from the group consisting of monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), and monomethylauristatin E (MMAD).

6. n is 18; Q 1 is the following chemical formula 【Chemistry 8】 and L-Q 2 is the following chemical formula 【Chemistry 9】 and Z is a combretastatin A-4 analog; 4. The compound of any one of claims 1 to 3, having the structure of formula (Ib), or a pharma- ceutically acceptable salt thereof.

7. n is 18; Q 1 is bonded or has the following chemical formula 【Chemistry 10】 and L-Q 2 is the following chemical formula 【Chemistry 11】 and Z is a flavugin analogue; 4. The compound of any one of claims 1 to 3, having the structure of formula (Ic), or a pharma- ceutically acceptable salt thereof.

8. The following chemical formula 【Chemistry 12-1】 【Chemistry 12-2】 【Chemistry 12-3】 2. The compound of claim 1, selected from the group consisting of:

9. 9. A pharmaceutical composition comprising a compound according to any one of claims 1 to 8, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier thereof.

10. 10. A method of treating cancer in a subject in need thereof comprising administering an effective amount of a compound according to any one of claims 1 to 8, or a pharma- ceutically acceptable salt thereof.

11. 11. The method of claim 10, wherein the cancer is melanoma, lung cancer, colon cancer, breast cancer, or a combination thereof.

12. 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; 12. The method according to any one of claims 10 to 11.

13. The method of any one of claims 10 to 12, wherein the cancer comprises cancer stem cells.

14. 14. The method of any one of claims 10 to 13, wherein the cancer comprises metastatic cancer cells.

15. 15. The method of any one of claims 10 to 14, wherein the cancer comprises circulating tumor cells.

16. 16. The method of any one of claims 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 pharma- ceutically acceptable salt thereof.

17. 16. The method of any one of claims 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.

18. 18. The method of any one of claims 10 to 15 and 17, wherein the cancer is breast cancer and the compound is a compound of formula (Ib), or a pharma- ceutically acceptable salt thereof.

19. 16. The method of any one of claims 10 to 15, wherein the cancer is melanoma, lung cancer, colon cancer, breast cancer, or a combination thereof, and the compound is a compound of formula (I-c), or a pharma- ceutically acceptable salt thereof.

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