WO2026036128A1 - Compounds and methods for iron-dependent drug release - Google Patents

Abstract

The present invention relates to compounds that are activated in the presence of Fe^II that can be used to treat ailments such as cancer. The invention further relates to pharmaceutical composition comprising the compounds as well as intermediates for the preparation of these compounds. In aspects, the pro-drug compounds provide treatments with lower toxicity than conventional compounds. The prodrugs can increase bioavailability and improve ADME (adsorption, distribution, metabolism and excretion) of medicaments/agents such as exatecan, pimasertib, rucaparib or ASN007.

Description

_{PATENT TT3-003WO} COMPOUNDS AND METHODS FOR IRON-DEPENDENT DRUG RELEASE RELATED APPLICATIONS [0001] This application claims priority to U.S. provisional patent application No. 63/681,761 filed August 9, 2024, U.S. provisional patent application No.63/711,526 filed October 24, 2024 and U.S. provisional patent application No.63/815,305 filed May 30, 2025. The contents of the aforementioned applications are incorporated herein by reference. FIELD OF THE INVENTION [0002] The invention relates generally to small molecule therapeutics, and more specifically, to prodrug compounds that are activated in the presence of iron. BACKGROUND OF THE INVENTION [0003] Cancer can be defined as a group of diseases that involve abnormal cell growth with the potential to invade or spread to other parts of the body. Despite advances in technology, cancer continues to be a significant cause of death and incalculable suffering. Cancer has been linked to several factors including smoking, obesity, poor diet, lack of physical activity and excessive consumption of alcohol. Other factors include certain infections, exposure to ionizing radiation and environmental pollutants. Certain cancers have been linked to infections such as Helicobacter pylori, hepatitis B, hepatitis C, human papillomavirus infection, Epstein–Barr virus and human immunodeficiency virus (HIV). Collectively, cancers are the second most common cause of death in the United States. [0004] Patients with cancer often have limited treatment options. Conventional cancer treatments are directed at removing cancerous tissue and preventing it from spreading. Treatment can include a combination of surgery, radiation therapy, chemotherapy, targeted therapy, immunotherapy, hormonal therapy and palliative care. Treatments are usually pursued based on the type, location and grade of the cancer as well as the patient's health and preferences. Despite advances in research, these treatments 1 PATENT TT3-003WO have remained relatively unchanged in recent decades. Although recognized as the current standard of care, these treatments can be ineffective, particularly when cancer has metastasized. Moreover, they often have harmful side effects. [0005] Because cancer cells divide faster than most normal cells, they can be sensitive to chemotherapy drugs. However, chemotherapy drugs will also attack other cells in the body, especially fast-dividing cells such as blood cells and the cells lining the mouth, stomach, and intestines. Radiation and chemotherapy typically lead to multiple side-effects, especially epithelial cell damage. Accordingly, there is a need for improved medications and methods of treating cancer that are more targeted and have less deleterious side effects. [0006] Cancer treatments would be improved with methods to selectively target cancer cells. Many approaches have been proposed and developed with the goal of achieving selective targeting of cancer treatment agents. For example, cytotoxic agents have been linked to monoclonal antibodies and antigen-specific fragments thereof which are capable of binding specifically to certain tumor antigens. The Applicants have recognized that cancer cells can be targeted based on their iron content. [0007] In normal cells and tissues, iron remains sequestered in forms that are non- toxic to the cell, bound to the iron carrying protein transferrin for example, or bound as heme within hemoglobin. However, diseased tissues and cells can contain higher than normal concentrations of iron. Many neoplastic cells for example over-express the transferrin receptor to increase their uptake of iron. Increased iron uptake has been proposed to explain the increased toxicity that iron-dependent endoperoxides (e.g., artemisinin) exhibit towards cancer cell lines (see, e.g., Efferth, T. Drug Resistance Updates, 2005, 8:85-97). In one study, the expression level of the transferrin receptor was shown to correlate with the cytotoxicity of an artemisinin derivative towards HeLa cells (see for example Disbrow, G. L., et al Cancer Research, 2005, 65, 10854-10861). Artemisinin and its derivatives are believed to exert their cytotoxic effect via reaction with Fe^II and the resulting generation of reactive oxygen and carbon centered radical species. The cytotoxicity of artemisinin derivatives towards leukemia, astrocytoma, and 2 PATENT TT3-003WO breast cancer cell lines can be potentiated by the addition of exogenous Fe^II salts or transferrin (Efferth, T. et al Free Radical Biology & Medicine, 2004, 37, 998-1009; Singh, N. P. et al Life Sciences, 2001, 70, 49-56). US patent 5,578,637 describes the use of an endoperoxide moiety (i.e., an artemisinin) to kill cancer cells under conditions that enhance intracellular iron concentrations. [0008] There is a need for improved formulations and methods of treating cancer. The Applicant describes herein compounds and methods to target cancer cells based in part on their ferrous iron content. Aspects of the present invention provide further related advantages as described in the following summary. SUMMARY OF THE INVENTION [0009] Aspects of the present disclosure teach certain benefits in construction and use which give rise to the exemplary advantages described below. [0010] The present invention includes compositions and methods to be used in treatment of ailments such as cancers. Further, a lower drug dose can be administered to achieve a desired effect while minimizing toxicity. [0011] Embodiments include compounds that are activated by elevated levels of iron (e.g., in cancer cells). Accordingly, embodiments include small molecule therapeutics that include an active agent (e.g., exatecan, pimasertib, rucaparib or ASN007) and a prodrug component. A therapeutic amount of the composition can be administered to treat an ailment such as cancer. [0012] Embodiments include a compound having the general formula: D – L – X In aspects, D is an N-linked drug moiety or an O-linked drug moiety; L is a bond, a C1 – C12 alkylene, or a 3 – 12 membered heteroalkylene; X is -C(=O)O-R¹; and 3 PATENT TT3-003WO R¹ is an optionally substituted 4 – 12 membered heterocyclyl. [0013] In embodiments the compound includes: (D) a drug moiety, (L) a bond or linker and (X) an iron-sensitive cage group. In aspects, the compounds act as prodrug compounds and are activated in the presence of increased Fe^II concentration (relative to normal levels). [0014] In aspects, the drug moiety (D) contains at least one of a nucleophilic oxygen (O) or nitrogen (N) by which a covalent bond is made to the linker. [0015] In aspects, D is a diagnostic agent. In aspects, D is an antibody or antibody fragment (e.g., Traztuzumab, Sacituzumab, Enfortumab, Mirvetuximab, Datopotamab, Samrotamab, Zolbetuximab and Sigvotagtug). In aspects, D is an ERK inhibitor (e.g., KO-947 or Temuterkib®). [0016] In aspects, D is drug moiety that is a medicament for treating an ailment such as cancer. In aspects, the drug moiety portion has no (or reduced) activity when complexed. [0017] Embodiments also include a compound having the general formula: or a pharmaceutically acceptable salt thereof, wherein: Y¹ is NH, O, S, or N linked to a C1-C10 straight or branched alkyl, Y² is NH, O, S, or N linked to a C1-C10 straight or branched alkyl, Z is O, or S, NH or N linked to a C1-C10 straight or branched alkyl, R¹, R², R³, R⁴, R⁵, R⁶ are independently H, C1-C9 straight or branched alkyl, W is O, S, NH or N linked to a C1-C10 straight or branched alkyl, and 4 PATENT TT3-003WO n is 0, 1, or 2. [0018] Embodiments also include a compound having the general formula: or a pharmaceutically acceptable salt thereof, wherein: wherein D is an N-linked Drug Moiety or an O-linked Drug Moiety; L is a bond, a C1 – C12 alkylene, or a 3 – 12 membered heteroalkylene; Y¹ – Y⁵ comprise of a carboxycylic or heterocyclic ring, Y¹, Y², Y⁴, Y⁵ are CR¹⁰, R¹⁰ is H, a halide or C1-C10 straight or branched alkyl, and Y³ is CR¹⁰, O, S(O), SO₂, CONH, SO₂ -alkyl or a substituted alkyl. [0019] Embodiments also include a compound having the general formula: or a pharmaceutically acceptable salt thereof, wherein: wherein D is an N-linked Drug Moiety or an O-linked Drug Moiety; L is a bond, a C1 – C12 alkylene, or a 3 – 12 membered heteroalkylene; Y¹, Y², Y⁴, Y⁵ are CR¹⁰, Y³ is O, CR¹⁰, S(O), SO₂, CONH, SO₂ -alkyl or a substituted alkyl, NS(O2)N(CH3)2, 5 PATENT TT3-003WO R¹⁰ is H, a halide or C1-C10 straight or branched alkyl, CF3, OCF3, (CH3)OH, (CH3)NHCH2(OH)OC(CH3)3, (CH3)CO(CH3), (F)CO(OH), NHS(O)₂N(CH₃)₂, CONHS(O)₂N(CH₃)₂ and optionally, wherein Y¹ and Y⁵ form a 5-membered ring. [0020] Embodiments also include a compound having the general formula: or a pharmaceutically acceptable salt thereof, wherein: wherein D is an N-linked Drug Moiety or an O-linked Drug Moiety; L is a bond, a C1 – C12 alkylene, or a 3 – 12 membered heteroalkylene; Y¹, Y², Y⁴, Y⁵ are CR¹⁰, Y³ is O, CR¹⁰, S(O), SO₂, CONH, SO₂ -alkyl or a substituted alkyl, NS(O2)N(CH3)2, R¹⁰ is H, a halide or C1-C10 straight or branched alkyl, CF₃, OCF₃, (CH₃)OH, (CH₃)NHCH₂(OH)OC(CH₃)₃, (CH₃)CO(CH₃), (F)CO(OH), NHS(O)2N(CH3)2, CONHS(O)2N(CH3)2, and wherein n is 0, 1, 2 or 3. [0021] Embodiments include a compound of Formula A, Formula B, Formula C and Formula D: 6 PATENT TT3-003WO F F H N OH [0022] In aspects, the compounds described herein provide treatments with lower toxicity than conventional compounds. In aspects, the compounds reduce hepatotoxicity. [0023] The compounds can increase bioavailability and improve ADME (adsorption, distribution, metabolism and excretion) of medicaments/agents such as exatecan, pimasertib, rucaparib or ASN007. [0024] In aspects, the compounds are inactive and/or have lower toxicity and/or side- effects than the drug moiety (i.e., when unconjugated). This allows higher dosing with fewer side-effects than conventional methods (i.e., non-prodrug administration). 7 PATENT TT3-003WO [0025] In embodiments the compounds protect the mucosae from effects of medicaments/agents. In embodiments, the compounds enable targeted delivery to tumor cells, where the medicaments/agents induce apoptosis of the tumor cells. In embodiments, the compounds can cross the blood-brain barrier. [0026] In embodiments, the compounds described herein are administered to treat one or more of leukemia, lung cancer, breast cancer, colon cancer, colorectal cancer, prostate cancer, renal cell carcinoma. [0027] Embodiments also include methods of treating an ailment. The method can include combining a medicament/agent (e.g., exatecan, pimasertib, rucaparib or ASN007) with another molecule to create a prodrug compound that has greater bioavailability and/or improved ADME (adsorption, distribution, metabolism and excretion). The prodrug compound can be administered to treat an ailment such as cancer. [0028] In embodiments above, one or more additional cancer therapies (e.g., chemotherapy, radiation therapy, immunotherapy, surgery or hormone therapy) are co- administered with a compound of the invention. In one embodiment, the chemotherapeutic reagent is an alkylating agent. In another embodiment the chemotherapeutic reagent is a topoisomerase inhibitor or a cytotoxic antibiotic such as doxorubicin, mitoxantrone, bleomycin, actinomycin, and mitomycin. [0029] In another embodiment, the compounds described herein are administered with one or more additional anti-neoplastic agents. Such anti-neoplastic agents can include, for example, anti-CTLA-4 monoclonal antibodies, anti-PD-1 and PD-L1 monoclonal antibodies (e.g., immune checkpoint inhibitors), antibiotics/antineoplastics, proteasome inhibitors and VEGF/VEGFR inhibitors. [0030] Embodiments also include methods of treating an ailment associated with a cell or organism having an increased Fe^II level compared to a control. The methods can 8 PATENT TT3-003WO include administering an effective amount of the compound described herein to the subject. [0031] The invention relates to a kit or package comprising the compound of the present invention or the pharmaceutical composition of the present invention. [0032] Other features and advantages of aspects of the present invention will become apparent from the following more detailed description, taken in conjunction with the accompanying drawings, which illustrate, by way of example, the principles of aspects of the invention. BRIEF DESCRIPTION OF THE FIGURES [0033] FIG 1A shows the chemical structures of the payloads (i.e., drug moieties) according to embodiments. Each includes a nucleophilic oxygen or nitrogen by which a covalent bond is made to a linker. [0034] FIG.1B shows linker variations according to embodiments. [0035] FIG.2A shows the results of an assay for TOP1 inhibition (DLX-1). [0036] FIG.2B shows the results of an assay for TOP1 inhibition (Conj-33). [0037] FIG.2C shows trioxolane compounds used as controls in the TOP1 inhibition assays. [0038] FIG.3 shows N-linked exatecan compounds according to embodiments. [0039] FIG.4 shows O-linked exatecan compounds according to embodiments. [0040] FIG.5A shows ASN007 compounds according to embodiments. [0041] FIG.5B shows rucaparib compounds according to embodiments. 9 PATENT TT3-003WO [0042] FIG.5C shows pimasertib compounds according to embodiments. [0043] FIG.5D shows additional exatecan compounds according to embodiments. [0044] FIG.6A depicts an antibody with a linker (maleimide conjugation) according to embodiments. [0045] FIG.6B depicts an antibody with a linker (bis-tosyl conjugation) according to embodiments. Definitions [0046] Reference in this specification to "one embodiment/aspect" or "an embodiment/aspect" means that a particular feature, structure, or characteristic described in connection with the embodiment/aspect is included in at least one embodiment/aspect of the disclosure. The use of the phrase "in one embodiment/aspect" or "in another embodiment/aspect" in various places in the specification are not necessarily all referring to the same embodiment/aspect, nor are separate or alternative embodiments/aspects mutually exclusive of other embodiments/aspects. Moreover, various features are described which may be exhibited by some embodiments/aspects and not by others. Similarly, various requirements are described which may be requirements for some embodiments/aspects but not other embodiments/aspects. Embodiment and aspect can in certain instances be used interchangeably. [0047] The terms used in this specification generally have their ordinary meanings in the art, within the context of the disclosure, and in the specific context where each term is used. Certain terms that are used to describe the disclosure are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner regarding the description of the disclosure. It will be appreciated that the same thing can be said in more than one way. 10 PATENT TT3-003WO [0048] Consequently, alternative language and synonyms may be used for any one or more of the terms discussed herein. Nor is any special significance to be placed upon whether or not a term is elaborated or discussed herein. Synonyms for certain terms are provided. A recital of one or more synonyms does not exclude the use of other synonyms. The use of examples anywhere in this specification including examples of any terms discussed herein is illustrative only, and is not intended to further limit the scope and meaning of the disclosure or of any exemplified term. Likewise, the disclosure is not limited to various embodiments given in this specification. [0049] Without intent to further limit the scope of the disclosure, examples of instruments, apparatus, methods and their related results according to the embodiments of the present disclosure are given below. Note that titles or subtitles may be used in the examples for convenience of a reader, which in no way should limit the scope of the disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. In the case of conflict, the present document, including definitions, will control. [0050] As applicable, the terms "about" or "generally", as used herein in the specification and appended claims, and unless otherwise indicated, means a margin of +/- 20%. Also, as applicable, the term "substantially" as used herein in the specification and appended claims, unless otherwise indicated, means a margin of +/- 10%. It is to be appreciated that not all uses of the above terms are quantifiable such that the referenced ranges can be applied. [0051] The term “subject” or "patient" refers to any single animal, more preferably a mammal (including such non-human animals as, for example, dogs, cats, horses, rabbits, zoo animals, cows, pigs, sheep, and non-human primates) for which treatment is desired. Most preferably, the patient herein is a human. In an embodiment, a “subject” of diagnosis or treatment is a prokaryotic or a eukaryotic cell, a tissue culture, a tissue or an animal, e.g. a mammal, including a human. 11 PATENT TT3-003WO [0052] The term “diagnostic agent” refers to a substance used to detect an impairment to normal (i.e., healthy) function. Chemical compounds can be specific in their reactions so that conclusions can be drawn, both qualitative and quantitative, regarding the cause of and disease itself. [0053] The term “an effective amount” generally refers to the amount of the defined component sufficient to achieve the desired therapeutic result. In an embodiment, that result can be effective cancer treatment. [0054] The terms “treating,” “treatment” and the like are used herein, without limitation, to mean obtaining a desired pharmacologic and/or physiologic effect. The effect may be prophylactic in terms of completely or partially preventing a disorder or sign or symptom thereof, and/or may be therapeutic in terms of amelioration of the symptoms of the disease or infection, or a partial or complete cure for a disorder and/or adverse effect attributable to the disorder. [0055] The term “neoplasia” refers to a disease that is caused by or results in inappropriately high levels of cell division, inappropriately low levels of apoptosis, or both. For example, cancer is an example of a neoplasia. Examples of cancers include, leukemia (e.g., acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myelocytic leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphoma (e.g., Hodgkin's disease, non-Hodgkin's disease), Waldenstrom's macroglobulinemia, heavy chain disease, and solid tumors such as sarcomas and carcinomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary 12 PATENT TT3-003WO carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, nile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular cancer, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodenroglioma, schwannoma, meningioma, melanoma, neuroblastoma, and retinoblastoma). Lymphoproliferative disorders are also considered to be proliferative diseases. [0056] The term “cancer" can further refer to human cancers and carcinomas, sarcomas, adenocarcinomas, etc., including solid tumors, kidney, breast, lung, kidney, bladder, urinary tract, urethra, penis, vulva, vagina, cervical, colon, ovarian, prostate, pancreas, stomach, brain, head and neck, skin, uterine, testicular, esophagus, and liver cancer. In any of the embodiments above, one or more cancer therapies, e.g., chemotherapy, radiation therapy, immunotherapy, surgery, or hormone therapy can be co-administered further with the methods described herein. Additional cancers include, for example, Hodgkin's Disease, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer, rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia, small-cell lung tumors, primary brain tumors, stomach cancer, colon cancer, malignant pancreatic insulanoma, malignant carcinoid, premalignant skin lesions, testicular cancer, thyroid cancer, neuroblastoma, esophageal cancer, genitourinary tract cancer, malignant hypercalcemia, cervical cancer, endometrial cancer, and adrenal cortical cancer. [0057] The term “glioma” refers to a type of tumor that starts in the glial cells of the brain or the spine. Gliomas comprise about 30 percent of all brain tumors and central nervous system tumors, and 80 percent of all malignant brain tumors. [0058] The term “glioblastoma” refers to an aggressive type of cancer that can occur in the brain or spinal cord. Glioblastoma forms from cells called astrocytes that support nerve cells. Glioblastoma can occur at any age but tends to occur more often in older 13 PATENT TT3-003WO adults. Initially, signs and symptoms of glioblastoma are nonspecific. They may include headaches, personality changes, nausea, and symptoms similar to those of a stroke. Symptoms often worsen rapidly and may progress to unconsciousness. The cause of most cases of glioblastoma is not known. Uncommon risk factors include genetic disorders, such as neurofibromatosis and Li–Fraumeni syndrome, and previous radiation therapy. Glioblastomas represent 15% of all brain tumors. They can either start from normal brain cells or develop from an existing low-grade astrocytoma. The diagnosis typically is made by a combination of a CT scan, MRI scan, and tissue biopsy. Treatment usually involves surgery, after which chemotherapy and radiation therapy are used. The medication temozolomide is frequently used as part of chemotherapy. High- dose steroids may be used to help reduce swelling and decrease symptoms. Surgical removal of the tumor is linked to increased survival but only for a duration of months. [0059] The term “astrocytoma” refers to a type of brain tumor. They originate in a particular kind of glial cells, star-shaped brain cells in the cerebrum called astrocytes. This type of tumor does not usually spread outside the brain and spinal cord and it does not usually affect other organs. Astrocytomas are the most common glioma and can occur in most parts of the brain and occasionally in the spinal cord. Within the astrocytomas, two broad classes are generally recognized, those with (a) narrow zones of infiltration (mostly noninvasive tumors; e.g., pilocytic astrocytoma, subependymal giant cell astrocytoma, pleomorphic xanthoastrocytoma), that often are clearly outlined on diagnostic images and (b) diffuse zones of infiltration (e.g., high-grade astrocytoma, anaplastic astrocytoma, glioblastoma), that share various features, including the ability to arise at any location in the central nervous system, but with a preference for the cerebral hemispheres; they occur usually in adults, and have an intrinsic tendency to progress to more advanced grades. People can develop astrocytomas at any age. The low-grade type is more often found in children or young adults, while the high-grade type is more prevalent in adults. Astrocytomas in the base of the brain are more common in young people and account for roughly 75% of neuroepithelial tumors. 14 PATENT TT3-003WO [0060] The term “anti-neoplastic compound” refers to medicaments that can be used to treat cancer. They can be categorized as Antifolates (e.g., Methotrexate, Pemetrexed, Pralatrexate, Trimetrexate), Purine Analogues (e.g., Azathioprine, Cladribine, Fludarabine, Mercaptopurine, Thioguanine) and Pyrimidine Analogues (e.g., Azacitidine, Capecitabine, Cytarabine, Decitabine, Floxuridine, Fluorouracil, Gemcitabine, Trifluridine/Tipracil). [0061] The term “camptothecin” or “CPT” refers to a topoisomerase inhibitor. It has been used clinically in China for the treatment of gastrointestinal tumors. CPT showed anticancer activity in preliminary clinical trials, especially against breast, ovarian, colon, lung, and stomach cancers. However, it has low solubility and adverse effects have been reported when used therapeutically, so synthetic and medicinal chemists have developed numerous syntheses of camptothecin and various derivatives to increase the benefits of the chemical, with good results. Four CPT analogues have been approved and are used in cancer chemotherapy: topotecan, irinotecan, belotecan, and trastuzumab deruxtecan. [0062] The term “exatecan” refers to a drug which is a structural analog of camptothecin with antineoplastic activity. A derivative is used in Trastuzumab deruxtecan. [0063] The term “ERK inhibitor” refers to a small molecule that blocks the activity of ERK (Extracellular Signal-Regulated Kinase), a crucial signaling pathway involved in cell growth, proliferation, and survival. They can be used to control ERK's biological activity, particularly in phosphorylation and dephosphorylation processes. In aspects, the payload can be an ERK inhibitor. Examples of ERK inhibitors include PD325901, LY3214996, and VX-11e (VTX-11e), ravoxertinib (GDC-0994). In aspects, the drug moiety or payload of a molecule described herein is an ERK inhibitor. [0064] The term “blood-brain barrier” or “BBB” refers to a highly selective semipermeable border of endothelial cells that prevents solutes in the circulating blood from non-selectively crossing into the extracellular fluid of the central nervous system 15 PATENT TT3-003WO where neurons reside. The blood–brain barrier is formed by endothelial cells of the capillary wall, astrocyte end-feet ensheathing the capillary, and pericytes embedded in the capillary basement membrane. This system allows the passage of some small molecules by passive diffusion, as well as the selective and active transport of various nutrients, ions, organic anions, and macromolecules such as glucose and amino acids that are crucial to neural function. [0065] The term “prodrug” refers to a derivative of a known direct acting drug, which derivative has enhanced delivery characteristics and therapeutic value as compared to the drug and is transformed into the active drug by an enzymatic or chemical process. After administration, a prodrug is metabolized (i.e., converted within the body) into a pharmacologically active drug. Instead of administering a drug directly, a corresponding prodrug can be used to improve how the drug is absorbed, distributed, metabolized, and excreted (ADME). Approximately 10% of all marketed drugs worldwide can be considered prodrugs. For example, aspirin was originally synthesized in the 19th century in attempt to produce a prodrug of salicylic acid that would be tolerable when taken orally. [0066] Since 2008, at least 30 prodrugs have been approved by the FDA. Prodrugs can be classified into two major types, based on how the body converts the prodrug into the final active drug form. Type I prodrugs are bioactivated inside the cells (i.e., intracellularly). Examples include anti-viral nucleoside analogs that must be phosphorylated and the lipid-lowering statins. Type II prodrugs are bioactivated outside cells (i.e., extracellularly) such as in digestive fluids or in the body's circulatory system. Examples of Type II prodrugs include salicin and certain antibody-, gene- or virus- directed enzyme prodrugs used in chemotherapy or immunotherapy. Both types can be further categorized into subtypes, based on factors such as (Type I) whether the intracellular bioactivation location is also the site of therapeutic action, or (Type 2) whether or not bioactivation occurs in the gastrointestinal fluids or in the circulation system. 16 PATENT TT3-003WO [0067] The term “linker” refers to a functional group that covalently bonds two or more moieties in a compound or material. For example, the linker can serve to covalently bond a prodrug moiety to an active agent. [0068] The term “bioavailability” refers to the fraction of an administered dose of unchanged drug that reaches the systemic circulation. For example, when a medication is administered intravenously, its bioavailability is 100%. However, when a medication is administered via other routes (such as orally), its bioavailability generally decreases due to incomplete absorption and first-pass metabolism. Bioavailability is one of the essential tools in pharmacokinetics, as bioavailability must be considered when calculating dosages for non-intravenous routes of administration. Bioavailability is calculated by comparing plasma levels of a drug given via a particular route of administration (for example, orally) with plasma drug levels achieved by IV injection. Absolute bioavailability = F = AUC oral / dose oral AUC i .v . / dose i .v. [0069] The term “formulation” as used herein refers to the molecules disclosed herein and excipients combined together that can be administered to produce the desired activity. The formulation can optionally comprise other agents. [0070] The term "pharmaceutically acceptable solvate" in accordance with this invention should be understood as meaning any form of the active compound in accordance with the invention in which said compound is bonded by a non-covalent bond to another molecule (normally a polar solvent), including especially hydrates and alcoholates. [0071] The term “trioxolane” refers to a five-membered ring molecule that contains two carbon atoms and three oxygen atoms, arranged in the order C-O-C-O-O. It is also known as an oxocycle or saturated heterocycle. Trioxolane can be an intermediate in ozonolysis and cycloaddition reactions, and can produce reaction products such as ozone, alkenes, molozonide, carbonyl oxide and carbonyl. 17 PATENT TT3-003WO [0072] The term "alkyl" as used herein is defined as a saturated monovalent hydrocarbon moiety having straight or branched moieties or combinations thereof and containing 1 to 6 carbon atoms, preferably 1 to 3 carbon atoms. Alkyl moieties can optionally be substituted with cycloalkyl groups, -OH groups, NH2 groups, and/or halogen. They can also be substituted by an ester group, a sulfide group or secondary or tertiary amine group. The term alkyl includes branched, linear and cycloalkyls. [0073] The term "cycloalkyl" as used herein refers to a monovalent or divalent group of 3 to 8 carbon atoms, preferably 5 to 6 carbon atoms of saturated cyclic hydrocarbon. Cycloalkyl groups may optionally be substituted by alkyl groups, -OH groups, -NH₂ groups, and/or halogen. [0074] As used herein the linear, cyclo or branched (C1-C6)-alkyl can be any branched, cyclo or linear C₁-, C₂-, C₃-, C₄, C₅ or C₆ alkyl. For example, C₁-alkyl can be methyl. C₂- alkyl can be ethyl.C3-alkyl can be selected from the group consisting of propyl and cyclopropyl, preferably propyl. C4-alkyl can be selected from the group consisting of n- butyl, isopropyl, butan-2-yl, 2-methylpropyl, tert-butyl, cyclobutyl and methylcyclopropyl, preferably n-butyl, isopropyl, butan-2-yl, 2-methylpropyl, tert-butyl, most preferably n- butyl. C5-alkyl can be selected from the group consisting of n-pentyl, 2-methylbutyl, 2,2- dimethylpropyl, 3-methylbutyl, pentan-2-yl, pentan-3-yl, 3-methylbutan-2-yl, 2- methylbutyl, cyclopentyl, methylcyclobutyl, 1,1-dimethylcyclopropyl and 1,2- dimethylcyclopropyl, preferably n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, 3- methylbutyl, pentan-2-yl, pentan-3-yl, 3-methylbutan-2-yl, 2-methylbutyl, most preferably n-pentyl. C₆-alkyl can be selected from the group consisting of n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, cyclohexyl, methylcyclopentyl, 1,2- dimethylcyclobutyl, 1,3-dimethylcyclobutyl and 1,2,3-trimethylcyclopropyl, preferably n- hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, most preferably n-hexyl. [0075] As used herein the linear or branched (C₁-C₆)-alkyl can be any branched or linear C₁-, C₂-, C₃-, C₄, C₅ or C₆ alkyl. For example, C₁-alkyl can be methyl. C₂-alkyl can be ethyl. C3-alkyl can be propyl. C4-alkyl can be selected from the group consisting of n- 18 PATENT TT3-003WO butyl, isopropyl, butan-2-yl, 2-methylpropyl, tert-butyl, preferably n-butyl. C5-alkyl can be selected from the group consisting of n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, 3- methylbutyl, pentan-2-yl, pentan-3-yl, 3-methylbutan-2-yl, 2-methylbutyl, preferably n- pentyl. C6-alkyl can be selected from the group consisting of n-hexyl, 2-methylpentyl, 3- methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, preferably n-hexyl. [0076] Branched, cyclo or linear (C₁-C₆)-alkyl-OH as used herein means any branched, cyclo or linear (C1-C6)-alkyl-OH. The OH group may be attached to C1, C2, C3, C4, C5 or C6. It is envisioned that the OH group is attached to C6. (C1)-alkyl-OH can be methyl-OH. (C₂)-alkyl-OH may be ethyl-OH. (C₃)-alkyl-OH can be selected from the group consisting of propyl-OH and cyclopropyl-OH. (C4)-alkyl-OH can be selected from the group consisting of n-butyl-OH, isopropyl-OH, butan-2-yl-OH, 2-methylpropyl-OH, tert-butyl-OH, cyclobutyl-OH and methylcyclopropyl-OH. (C₅)-alkyl-OH can be selected from the group consisting of n-pentyl-OH, 2-methylbutyl-OH, 2,2-dimethylpropyl-OH, 3- methylbutyl-OH, pentan-2-yl-OH, pentan-3-yl-OH, 3-methylbutan-2-yl-OH, 2- methylbutyl-OH, cyclopentyl-OH, methylcyclobutyl-OH, 1,1-dimethylcyclopropyl-OH and 1,2-dimethylcyclopropyl-OH. (C₆)-alkyl-OH can be selected from the group consisting of n-hexyl-OH, 2-methylpentyl-OH, 3-methylpentyl-OH, 2,2-dimethylbutyl-OH, 2,3- dimethylbutyl-OH, cyclohexyl-OH, methylcyclopentyl-OH, 1,2-dimethylcyclobutyl-OH, 1,3-dimethylcyclobutyl-OH and 1,2,3-trimethylcyclopropyl-OH. [0077] Branched, cyclo or linear (C1-C6)-alkyl-NH2 as used herein means any branched, cyclo or linear (C₁-C₆)-alkyl-NH_2. The NH₂ group may be attached to C₁, C₂, C₃, C₄, C₅ or C₆. It is envisioned that the NH₂ group is attached to C₆. (C₁)-alkyl-NH₂ can be methyl-NH2. (C2)-alkyl- NH2 can be ethyl-NH2. (C3)-alkyl- NH2 can be selected from the group consisting of propyl-NH₂ and cyclopropyl-NH_2. (C₄)-alkyl- NH₂ can be selected from the group consisting of n-butyl-NH₂, isopropyl-NH₂, butan-2-yl-NH₂, 2- methylpropyl-NH2, tert-butyl- NH2, cyclobutyl-NH2 and methylcyclopropyl-NH2. (C5)- alkyl-NH2 can be selected from the group consisting of n-pentyl-NH2, 2-methylbutyl-NH2, 2,2-dimethylpropyl-NH₂, 3-methylbutyl-NH₂, pentan-2-yl-NH₂, pentan-3-yl-NH₂, 3- methylbutan-2-yl-NH2, 2-methylbutyl-NH2, cyclopentyl-NH2, methylcyclobutyl-NH2, 1,1- 19 PATENT TT3-003WO dimethylcyclopropyl-NH2 and 1,2-dimethylcyclopropyl-NH2. (C6)-alkyl-NH2 can be selected from the group consisting of n-hexyl-NH2, 2-methylpentyl-NH2, 3-methylpentyl- NH₂, 2,2-dimethylbutyl-NH₂, 2,3-dimethylbutyl-NH₂, cyclohexyl-NH₂, methylcyclopentyl- NH2, 1,2-dimethylcyclobutyl-NH2, 1,3-dimethylcyclobutyl-NH2 and 1,2,3- trimethylcyclopropyl-NH2. [0078] The term “heterocyclyl” as used herein refers to any saturated or partially unsaturated cyclic alkyl group with one or more ring heteroatoms independently selected from nitrogen, oxygen and sulfur. The term heterocyclyl includes heteroalkenyl groups (i.e. the heterocyclyl group having at least one double bond), bicyclic heterocyclic groups such as bridged-heterocyclyl groups or fused heterocyclyl groups. Thus, a heterocyclyl may be a single ring or multiple rings wherein the multiple rings may be fused or bridged and may comprise one or more (e.g.1 to 3) oxygen (O), nitrogen (N) and/or sulfur (S) groups. The term heterocyclyl as used herein excludes heteroaryls. Exemplary heterocyclyls can thus be pyrrolidine, 3-pyrroline, 2-pyrroline, pyrazolidine, imidazolidine, 2-pyrazoline, 2-imidazoline, tetrahydrofuran, 1,3-dioxolane, tetrahydrothiophene, 1,2-oxathiolane, 1,3-oxathiolane, piperidine, piperazine, tetrahydropyran, 2H-pyran, 4H-pyran, 1,4-dioxane, 1,4-dioxine, 1,3- or 1,4-thiane, 1,3,5- trithiane, morpholine, 4H-1,2-oxazine, 2H-1,2-oxazine, 6H-1,2-oxazine, 4H-1,3-oxazine, 2H-1,3-oxazine, 6H-1,3-oxazine, 4H-1,4-oxazine, 2H-1,4-oxazine, thiomorpholine, 4H- 1,4-thiazine, 2H-1,4-thiazine, 6H-1,2-thiazine, 2H-1,4-thiazine, decahydroisoquinoline, decahydroquinoline and the like. [0079] The term “aryl” refers to an aromatic carbocyclic group having a single ring (i.e. monocyclic) or multiple rings (e.g. bicyclic or tricyclic) including fused systems. As used herein an aryl group can have 6 to 10 ring atoms. These atoms can be carbon atoms in the case of aryls and carbon atoms and heteroatoms in the case of heteroaryls. [0080] The term “heteroaryl” refers to an aromatic monocyclic or multicyclic ring system of 6 to about 14 ring atoms in which one or more of the atoms in the ring system is/are element(s) other than carbon, for example, nitrogen, oxygen or sulfur. Representative heteroaryl groups include pyridinyl, pyridazinyl and quinolinyl. 20 PATENT TT3-003WO [0081] The heteroaryl as used herein refers to any heteroaryl. Heteroaryl as used herein refers to an aromatic group having a single ring, multiple bridged rings or multiple fused rings with one or more ting heteroatoms independently selected from nitrogen, oxygen and sulfur. A heteroaryl can include 5 to 12 atoms, of which 1 to 4 are ring heteroatoms, wherein the heteroatom is independently nitrogen, oxygen and/or sulfur and 1 to 11 or 0 to 8 are carbon atoms. As used herein heteroaryl can also include 5 to 10 atoms, of which 1 to 4 are ring heteroatoms, wherein the heteroatom is independently nitrogen, oxygen and/or sulfur and 3 to 8 or 1 to 6 are carbon atoms. It is also envisioned that the heteroaryl can include 5 to 10 atoms, of which 1 to 3 are ring heteroatoms, wherein the heteroatom is independently nitrogen, oxygen and/or sulfur and 2 to 7 or 2 to 5 are carbon atoms. Any aromatic ring having a single or multiple fused rings, containing at least one heteroatom is considered a heteroaryl, regardless of the attachment to the remainder of the molecule. The term heteroaryl is also embraced by the term aryl as defined herein. Heteroaryls are known to the skilled person. [0082] Exemplary heteroaryls include 2H-pyrrole, 1H-pyrrole, pyrazole, imidazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, furan, thiophene, oxazole, isoxazole, isothiazole, thiazole, -, 2, 5-oxadiazole, 1,3,4-thiadiazole, 1,2,5-thiadiazole, pyridine, pyridiazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, 1,4,5,6- tetrahydrocyclopental[b]pyrrole, 1,4-dihydropyrrolo[3,2-b]pyrrole, 1,4-dihydropyrrolo[3,2- b]pyrrole, 6H-furo[2,3-b]pyrrole, 4H-furo[3,2-b]pyrrole, 6H-thieno [3,2-b]pyrrole, 6H- thieno[2,3-b]pyrrole, indene, 2,3-dihydro-1H-indene, indoline, 3H-indole, 1H-indole, 2H- isoindole, indolizine, 1H-indazole, benzimidazole, 7-azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindole, pyrazolo[1,5-a]pyrimidine, purine, benzofuran, isobenzofuran, benzo[c]thiophene, benzo[b]thiophene, 1,2-benzisoxazole, anthranil, 1,2- benzisothiazole, benzoxazole, benzthiazole, quinoline, quinoxaline, phthalazine, 2H- chromene and the like. Examples of five membered monocyclic heteroaryl groups include but are not limited to pyrrole, furan, thiophene, imidazole, furazan, oxazole, oxadiazole, oxatriazole, isoxazole, thiazole, isothiazole, pyrazole, triazole and tetrazole groups. 21 PATENT TT3-003WO [0083] It is further contemplated that the heteroaryl can be selected from indolyl, pyrrolyl, furanyl, thiophenyl, pyridyl, pyrimidyl, pyridazyl, pyrazinyl, quinolinyl, isoquinolinyl, acridinyl, 1,2-methylenedioxyphenyl (= benzo-1,4-dioxanyl) or 1,2- ethylenedioxyphenyl (= 1,3-Benzodioxolyl), preferably 1-H-indolyl, 1,2- methylenedioxyphenyl and 1,2-ethylenedioxyphenyl. [0084] The term "antibody" refers to a polypeptide comprising a framework region from an immunoglobulin gene or fragments thereof that specifically binds and recognizes an antigen. The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon, and mu constant region genes, as well as the myriad immunoglobulin variable region genes. Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively. Typically, the antigen-binding region of an antibody will be most critical in specificity and affinity of binding. [0085] The term “DAR” or “drug-to-antibody ratio” refers to the average number of drug molecules attached to a single antibody molecule in an antibody-drug conjugate (ADC). This ratio is a crucial parameter in ADC development as it impacts the efficacy, safety, and pharmacokinetics of the ADC. [0086] The term “Trastuzumab” or “Herceptin®” refers to a monoclonal antibody used to treat HER2-positive breast cancer and gastric cancer. It works by binding to the HER2 receptor, a protein that can promote the growth of certain cancer cells, and interfering with the cell's ability to grow and divide. Trastuzumab is often used in combination with other chemotherapy drugs. [0087] All numerical designations, e.g., pH, temperature, time, concentration, and molecular weight, including ranges, are to be understood as approximations in accordance with common practice in the art. When used herein, the term “about” may connote variation (+) or (-) 1%, 5% or 10% of the stated amount, as appropriate given 22 PATENT TT3-003WO the context. It is to be understood, although not always explicitly stated, that the reagents described herein are merely exemplary and that equivalents of such are known in the art. [0088] Many known and useful compounds and the like can be found in Remington’s Pharmaceutical Sciences (13^th Ed), Mack Publishing Company, Easton, PA—a standard reference for various types of administration. As used herein, the term “formulation(s)” means a combination of at least one active ingredient with one or more other ingredient, also commonly referred to as excipients, which may be independently active or inactive. The term “formulation” may or may not refer to a pharmaceutically acceptable composition for administration to humans or animals and may include compositions that are useful intermediates for storage or research purposes. DETAILED DESCRIPTION OF THE INVENTION [0089] The following detailed description of the present disclosure is merely exemplary in nature and is not intended to limit the present disclosure or the application and uses of the present disclosure. Furthermore, there is no intention to be bound by any theory presented in the preceding background or the following detailed description. [0090] In normal cells and tissues, iron remains sequestered in forms that are non- toxic to the cell, bound to the iron carrying protein transferrin for example, or bound as heme within hemoglobin. Diseased tissues and cells can contain higher than normal concentrations of iron. Many neoplastic cells for example over-express the transferrin receptor to increase their uptake of iron. An excess of iron, and ferrous iron in particular, is therefore a distinguishing characteristic of many neoplastic cells. The antimalarial drug artemisinin and its related synthetic derivatives are thought to confer their antiparasitic effect via reaction with ferrous iron and the resulting generation of reactive oxygen and carbon centered radical species. An excess of iron, and ferrous iron in particular, is therefore a distinguishing characteristic of many neoplastic cells and pathogenic parasites. 23 PATENT TT3-003WO [0091] Accordingly, embodiments of the invention include prodrug compounds that are activated in the presence of increased Fe^II concentration (relative to normal levels). In embodiments, the prodrugs described herein can be targeted to a specific tissue such as tumors. The compounds can be used to treat an ailment such as cancer. [0092] Compounds provided herein may be agents (e.g. compounds, proteins, drugs, detectable agents, therapeutic agents) in a prodrug form. Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under select physiological conditions (e.g., increased Fe^II concentration relative to normal physiological levels, increased reductant levels relative to normal physiological levels) to provide the final agents (e.g. compounds, proteins, drugs, detectable agents, therapeutic agents). Additionally, prodrugs can be converted to agents (e.g. compounds, proteins, drugs, detectable agents, therapeutic agents) by chemical or biochemical methods in an ex vivo environment. Prodrugs described herein include compounds that readily undergo chemical changes under select physiological conditions (e.g. increased Fe^II concentration relative to normal physiological levels, increased reductant levels relative to normal physiological levels) to provide agents (e.g. compounds, proteins, drugs, detectable agents, therapeutic agents) to a biological system (e.g. in a subject, in an infected cell, in a cancer cell, in the extracellular space near an infected cell, in the extracellular space near a cancer cell from the moieties (e.g. moiety of a protein, drug, detectable agent) attached to the prodrug moiety and included in the prodrug (e.g. compound of formula I, including embodiments, compound described herein, examples). Compounds for Iron-Dependent Drug Release [0093] Embodiments include compounds having the general formula: D – L – X [0094] In embodiments the compound includes (D) a drug moiety, (L) a bond or linker and (X) an iron-sensitive cage group. In aspects, the compounds act as prodrug 24 PATENT TT3-003WO compounds and are activated in the presence of increased Fe^II concentration (relative to normal levels). [0095] In aspects, D is an N-linked drug moiety or an O-linked drug moiety; L is a bond, a C1 – C12 alkylene, or a 3 – 12 membered heteroalkylene; X is -C(=O)O-R¹; and R¹ is an optionally substituted 4 – 12 membered heterocyclyl. [0096] In embodiments, drug moiety (D) is an anti-neoplastic agent. Anti-neoplastic agents can include, for example, exatecan, pimersertib, rucaparib and ASN007 (FIG. 1A). It is contemplated that the drug moiety can be other small molecule anti-neoplastic agents that are known in the art. As used herein, the terms “payload” and “drug moiety” can be used interchangeably when describing the compounds (e.g., Payload – Linker – Cage Group). [0097] Linkers include, for example L1, L2, L3, L4, L5, L6, L7, L8, L9, L10, L11, L12, L13, L14, L15, L16, L17, L18, L19, L20, L21, L22, L23, L24, L25, L26, L27, L28 and L29: PATENT TT3-003WO 26 PATENT TT3-003WO [0098] FIG.1B shows variations of linkers, including carbonates and esters. As shown, linkers are covalently bonded to exatecan via a nucleophilic oxygen (O). [0099] In embodiments, the cage group is a ring that includes nitrogen (e.g., amine or hydroxyl amine). Cage groups include, for example C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, C30, C31, C32, C33, C34, C35 and C36: PATENT TT3-003WO PATENT TT3-003WO O O O O N S Iron Dependent Drug Release [00100] In embodiments, the compounds described herein act as pro-drugs. Accordingly, aspects include compounds and methods for administering a pro-drug to treat an ailment. In aspects, the pro-drug is administered (e.g., orally) to a subject to treat cancer. Upon exposure to an elevated Fe^II level at or near a tumor, a series of reactions begins that results in release of the active payload. For payloads containing a nucleophilic amine, the mechanism of drug release follows the below series of steps: X _{O Fe+2} O a_d [00101] For payloads containing a nucleophilic hydroxy group, a conjugate would include a self-cleaving linker (L2-LXX). In aspects, the mechanism of drug release follows the below series of steps: X O R² R² a_d PATENT TT3-003WO [00102] In aspects, the prodrug compound is inactive and/or has lower toxicity and/or side- effects than the unconjugated payload. This allows higher dosing with fewer side- effects than conventional methods (i.e., non-prodrug administration). [00103] Certain compounds of the present disclosure can exist in un solvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present disclosure. Certain compounds of the present disclosure may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present disclosure and are intended to be within the scope of the present disclosure. [00104] The salt or solvate may be any suitable salt or solvate. For example, the salt may be a pharmaceutically acceptable salt. The term "pharmaceutically acceptable salt" refers to any pharmaceutically acceptable salt, which upon administration to the patient is capable of providing (directly or indirectly) a compound as described herein. Such salts preferably are acid addition salts with physiologically acceptable organic or inorganic acids, or alkali addition salts with acceptable organic or inorganic bases. Examples of the acid addition salts include mineral acid addition salts such as, for example, hydrochloride, hydrobromide, hydroiodide, sulphate, nitrate, phosphate, and organic acid addition salts such as, for example, lactate, acetate, trifluoroacetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate, methanesulphonate and p-toluenesulphonate. Examples of the alkali addition salts include inorganic salts such as, for example, sodium, potassium, calcium and ammonium salts, and organic alkali salts such as, for example, ethylenediamine, ethanolamine, N,N-dialkylenethanolamine, triethanolamine and basic aminoacids salts. However, it will be appreciated that non-pharmaceutically acceptable salts also fall within the scope of the invention since those may be useful in the preparation of pharmaceutically acceptable salts. Procedures for salt formation are conventional in the art. 30 PATENT TT3-003WO Antibody Prodrugs for Cancer [00105] Oncology has advanced with the introduction of biological therapies that target cancerous cells while reducing the effect on normal tissue. However, the toxicity of potent new biological therapies for cancer has limited their utility. By improving tumor specificity, antibody prodrugs can widen or even create a therapeutic window for anticancer agents that are difficult or impossible to use otherwise because of poor tolerability. As described herein, biologics (i.e., antibodies) can be linked to an iron- sensitive cage group. The compound can thereafter be activated in the presence of increased Fe^II concentration. This can widen the therapeutic window for therapies that are otherwise difficult to use because of poor tolerability and/or create a therapeutic window for undruggable targets. [00106] In embodiments, the drug moiety (D) is an antibody or antibody fragment (e.g., Traztuzumab, Sacituzumab, Enfortumab, Mirvetuximab, Datopotamab, Samrotamab, Zolbetuximab, Sigvotagtug. Targets and respective antibodies are shown below in Table 1. Table 1 Target Antibody Administration [00107] The pharmaceutical compositions of the invention can be used as a medicament. For example, the pharmaceutical compositions can be used in treating 31 PATENT TT3-003WO pancreatic cancer, lung cancer, colorectal cancer, thyroid cancer, testicle cancer, melanoma, bladder cancer, liver cancer, kidney cancer, myelodysplastic syndrome, or leukemia. [00108] It is further contemplated that the pharmaceutical composition further comprises an excipient, carrier or solvent. Such compositions can be included in a capsule, sachet, paper or other container. In making the compositions, conventional techniques for the preparation of pharmaceutical compositions may be used. For example, the compounds for use according to present invention may be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier that may be in the form of an ampoule, capsule, sachet, paper, or other container. When the carrier serves as a diluent, it may be solid, semi-solid, or liquid material that acts as a vehicle, excipient, or medium for the active compound. The compounds for use according to present invention can be adsorbed on a granular solid container for example in a sachet. Some examples of suitable carriers are water, salt solutions, alcohols, polyethylene glycols, polyhydroxyethoxylated castor oil, peanut oil, olive oil, lactose, terra alba, sucrose, cyclodextrin, amylose, magnesium stearate, talc, gelatin, agar, pectin, acacia, stearic acid or lower alkyl ethers of cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid monoglycerides and diglycerides, pentaerythritol fatty acid esters, polyoxyethylene, hydroxymethylcellulose, and polyvinylpyrrolidone. Similarly, the carrier or diluent may include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax. Said compositions may also include wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents or flavoring agents. The compositions for use according to the invention may be formulated so as to provide quick, sustained, or delayed release of the compounds for use according to present invention after administration to the patient by employing procedures well known in the art. [00109] The pharmaceutical compositions can be sterilized and mixed, if desired, with auxiliary agents, emulsifiers, salt for influencing osmotic pressure, buffers and/or 32 PATENT TT3-003WO coloring substances and the like, which do not deleteriously react with the compounds for use according to present invention. [00110] In addition to the compounds for use according to present invention, the compositions for use according to the invention may include, depending on the composition and mode of delivery desired, pharmaceutically acceptable, non-toxic carriers or diluents, which include vehicles commonly used to form pharmaceutical compositions for animal or human administration. The diluents are selected so as not to unduly affect the biological activity of the combination. [00111] The compound of the present invention and/or the pharmaceutical composition of the present invention may be administered to a subject. Preferably, the subject is a subject in need of such administration. The subject may be any suitable subject. The subject may be a human or an animal. Preferably the subject is a human. [00112] One preferred embodiment disclosed herein refers to the route of administration, that may be any route which effectively transports the compounds and compositions disclosed in present disclosure, to the appropriate or desired site of action, such as oral, nasal, topical, pulmonary, transdermal, intrathecal or parenteral, e. g., rectal, subcutaneous, intravenous, intraurethral, intramuscular, intranasal, ophthalmic solution or an ointment. [00113] For oral administration, either solid or fluid unit dosage forms can be prepared. For preparing solid compositions such as tablets, the compounds for use according to present invention, are mixed into formulations with conventional ingredients such as talc, magnesium stearate, di-calcium phosphate, magnesium aluminum silicate, calcium sulphate, starch, lactose, acacia, methylcellulose, and functionally similar materials as pharmaceutical diluents or carriers. [00114] Capsules are prepared by mixing the compounds for use according to present invention with an inert pharmaceutical diluent and filling the mixture into a hard gelatin capsule of appropriate size. Soft gelatin capsules are prepared by machine encapsulation of slurry of the compound with an acceptable vegetable oil, light liquid 33 PATENT TT3-003WO petrolatum or other inert oil. Fluid unit dosage forms for oral administration such as syrups, elixirs and suspensions can be prepared. The water-soluble forms can be dissolved in an aqueous vehicle together with sugar, aromatic flavoring agents and preservatives to form syrup. An elixir is prepared by using a hydroalcoholic (e. g., ethanol) vehicle with suitable sweeteners such as sugar and saccharin, together with an aromatic flavoring agent. Suspensions can be prepared with an aqueous vehicle with the aid of a suspending agent such as acacia, tragacanthin, methylcellulose and the like. [00115] Examples of such diluents that are especially useful for injectable formulations are water, the various saline, organic or inorganic salt solutions, Ringer's solution, dextrose solution, and Hank's solution. In addition, the pharmaceutical composition or formulation may include additives such as other carriers; adjuvants; or non-toxic, non- therapeutic, non-immunogenic stabilizers and the like. [00116] Furthermore, excipients can be included in the compositions disclosed. Examples include, but are not limited to, cosolvents, surfactants, oils, humectants, emollients, preservatives, stabilizers and antioxidants. Any pharmacologically acceptable buffer may be used, such as, tris or phosphate buffers. Effective amounts of diluents, additives, and excipients are those amounts that are effective to obtain a pharmaceutically acceptable formulation in terms of solubility, biological activity, etc. [00117] The pharmaceutical compositions comprising the compounds for use according to present invention may be incorporated into a microsphere. The compounds for use according to present invention can be loaded into albumin microspheres, from which it is possible to recover such microspheres in a dry powder for nasal administration. Other materials suitable for the preparation of microspheres include agar, alginate, chitosan, starch, hydroxyethyl starch, albumin, agarose, dextran, hyaluronic acid, gelatin, collagen, and casein. The microspheres can be produced by various processes known to the person skilled in the art such as a spray drying process or an emulsification process. 34 PATENT TT3-003WO [00118] Another preferred embodiment of the invention is the dosage scheme of the compounds for use according to present invention. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for subjects, e. g., mammalian subjects, e. g. humans, dogs, cats, and rodents, each unit containing a predetermined quantity of active material calculated to produce the desired pharmaceutical effect in association with the required pharmaceutical diluent, carrier or vehicle. The specifications for the unit dosage forms of this invention are dictated by and dependent on (a) the unique characteristics of the compounds and extracts disclosed above herein and the particular effect to be achieved and (b) the limitations inherent in the art wherein said compounds or extracts are used in humans and animals. Examples of unit dosage forms are tablets, capsules, pills, powder packets, wafers, suppositories, granules, cachets, teaspoonfuls, tablespoonfuls, dropperfuls, ampoules, vials, aerosols with metered discharges, segregated multiples of any of the foregoing, and other forms as herein described. The compositions disclosed herein can be included in kits, which can contain one or more-unit dosage forms of the composition and instructions for use. [00119] Slow or extended-release delivery systems, including any of a number of biopolymers (biological-based systems), systems employing liposomes, colloids, resins, and other polymeric delivery systems or compartmentalized reservoirs, can be utilized with the compositions described herein to provide a continuous or long-term source of the therapeutic compound. [00120] The present invention also relates to a kit or package comprising the compound of the present invention or the pharmaceutical composition of the present invention. The kit or package may further comprise instructions for the medical use of the compound of the present invention or of the pharmaceutical composition of the present invention. [00121] Alternatively, or additionally, the kit or package may comprise a unit dosage form of the compound of the present invention, or of the pharmaceutical composition of the present invention. 35 PATENT TT3-003WO [00122] The present invention also relates to a use of a compound of the present invention or a salt or solvate thereof, or the pharmaceutical composition of the present invention, for the preparation of a medicament. [00123] The present invention also concerns a use of a compound of the present invention or a salt or solvate thereof or the pharmaceutical composition of the present invention for the preparation of a medicament for treating pancreatic cancer, lung cancer, colorectal cancer, thyroid cancer, testicle cancer, melanoma, bladder cancer, liver cancer, kidney cancer, myelodysplastic syndrome, or leukemia. [00124] The present invention also relates to a method of treating a disease, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention or a salt or solvate thereof, or a pharmaceutical composition of the present invention. [00125] The present invention further concerns a method of treating pancreatic cancer, lung cancer, colorectal cancer, thyroid cancer, testicle cancer, melanoma, bladder cancer, liver cancer, kidney cancer, myelodysplastic syndrome, or leukemia, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention or a salt or solvate thereof, or a pharmaceutical composition of the present invention. [00126] Embodiments also include a kit or package comprising a compound or the pharmaceutical composition as described above. In aspects, the kit or package further comprises instructions for the medical use of the compound or of the pharmaceutical composition and/or a unit dosage form of the compound or of the pharmaceutical composition. [00127] Embodiments also include the use of a compound or a salt or solvate thereof or the pharmaceutical composition described above in the preparation of a medicament. 36 PATENT TT3-003WO [00128] Embodiments also include the use of a compound or a salt or solvate thereof or the pharmaceutical composition described above in the preparation of a medicament for treating pancreatic cancer, lung cancer, colorectal cancer, thyroid cancer, testicle cancer, melanoma, bladder cancer, liver cancer, kidney cancer, myelodysplastic syndrome, or leukemia. Methods of Treatment [00129] Another aspect of the present application relates to a method for treating a cell proliferative disorder such as cancer. The method can include administering to a subject in need thereof an effective amount of a compound described herein. In another aspect, a method for treating a cell proliferative disorder includes administering to a subject in need thereof an effective amount of one or more compounds described herein with an additional medicament (e.g., a chemotherapeutic). [00130] Embodiments also include a method of treating a disease/ailment. The method can include administering to a subject in need thereof a therapeutically effective amount of a compound salt or solvate thereof, or a pharmaceutical composition as described above. [00131] Embodiments also include a method of treating pancreatic cancer, lung cancer, colorectal cancer, thyroid cancer, testicle cancer, melanoma, bladder cancer, liver cancer, kidney cancer, myelodysplastic syndrome, or leukemia, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound a salt or solvate thereof, or a pharmaceutical composition as described above. [00132] In embodiments, a pharmaceutical composition (i.e., a composition containing a compound described herein) is administered to a patient with to treat cancer. A pharmaceutical composition disclosed herein can include a therapeutic compound in an amount sufficient to allow customary administration to an individual. In certain embodiments, a pharmaceutical composition disclosed herein includes, e.g., at least 5 mg, at least 10 mg, at least 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at 37 PATENT TT3-003WO least 35 mg, at least 40 mg, at least 45 mg, at least 50 mg, at least 55 mg, at least 60 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least 80 mg, at least 85 mg, at least 90 mg, at least 95 mg, or at least 100 mg of a therapeutic compound. In certain embodiments, a pharmaceutical composition disclosed herein may comprise, e.g., at least 5 mg, at least 10 mg, at least 20 mg, at least 25 mg, at least 50 mg, at least 75 mg, at least 100 mg, at least 200 mg, at least 300 mg, at least 400 mg, at least 500 mg, at least 600 mg, at least 700 mg, at least 800 mg, at least 900 mg, at least 1,000 mg, at least 1,100 mg, at least 1,200 mg, at least 1,300 mg, at least 1,400 mg, or at least 1,500 mg of a therapeutic compound. In yet other aspects of this embodiment, a pharmaceutical composition disclosed herein may comprise in the range of, e.g., about 5 mg to about 100 mg, about 10 mg to about 100 mg, about 50 mg to about 150 mg, about 100 mg to about 250 mg, about 150 mg to about 350 mg, about 250 mg to about 500 mg, about 350 mg to about 600 mg, about 500 mg to about 750 mg, about 600 mg to about 900 mg, about 750 mg to about 1,000 mg, about 850 mg to about 1,200 mg, or about 1,000 mg to about 1,500 mg. In still certain embodiments, a pharmaceutical composition disclosed herein may comprise in the range of, e.g., about 10 mg to about 250 mg, about 10 mg to about 500 mg, about 10 mg to about 750 mg, about 10 mg to about 1,000 mg, about 10 mg to about 1,500 mg, about 50 mg to about 250 mg, about 50 mg to about 500 mg, about 50 mg to about 750 mg, about 50 mg to about 1,000 mg, about 50 mg to about 1,500 mg, about 100 mg to about 250 mg, about 100 mg to about 500 mg, about 100 mg to about 750 mg, about 100 mg to about 1,000 mg, about 100 mg to about 1,500 mg, about 200 mg to about 500 mg, about 200 mg to about 750 mg, about 200 mg to about 1,000 mg, about 200 mg to about 1,500 mg, about 5 mg to about 1,500 mg, about 5 mg to about 1,000 mg, or about 5 mg to about 250 mg. [00133] In one embodiment, the dose of the composition may be administered daily, semi-weekly, weekly, bi-weekly, or monthly. The period of treatment may be for a week, two weeks, a month, two months, four months, six months, eight months, a year, or longer. The initial dose may be larger than a sustaining dose. In one embodiment, the dose ranges from a weekly dose of at least 0.01 mg/kg, at least 0.25 mg/kg, at least 0.3 mg/kg, at least 0.5 mg/kg, at least 0.75 mg/kg, at least 1 mg/kg, at least 2 mg/kg, at 38 PATENT TT3-003WO least 3 mg/kg, at least 4 mg/kg, at least 5 mg/kg, at least 6 mg/kg, at least 7 mg/kg, at least 8 mg/kg, at least 9 mg/kg, at least 10 mg/kg, at least 15 mg/kg, at least 20 mg/kg, at least 25 mg/kg, or at least 30 mg/kg In one embodiment, a weekly dose may be at most 1.5 mg/kg, at most 2 mg/kg, at most 2.5 mg/kg, at most 3 mg/kg, at most 4 mg/kg, at most 5 mg/kg, at most 6 mg/kg, at most 7 mg/kg, at most 8 mg/kg, at most 9 mg/kg, at most 10 mg/kg, at most 15 mg/kg, at most 20 mg/kg, at most 25 mg/kg, or at most 30 mg/kg. In a particular aspect, the weekly dose may range from 5 mg/kg to 20 mg/kg. In an alternative aspect, the weekly dose may range from 10 mg/kg to 15 mg/kg. [00134] In an embodiment, the period of administration of a cancer therapeutic is for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or more. In a further embodiment, a period of during which administration is stopped is for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or more. [00135] In aspects of this embodiment, a therapeutically effective amount of a cancer therapeutic disclosed herein reduces or maintains a cancer cell population and/or tumor cell size in an individual by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or at least 100%. In other aspects of this embodiment, a therapeutically effective amount of a cancer therapeutic disclosed herein reduces or maintains a cancer cell population and/or tumor cell size in an individual by, e.g., at most 10%, at most 15%, at most 20%, at most 25%, at most 30%, at most 35%, at most 40%, at most 45%, at most 50%, at most 55%, at most 60%, at most 65%, at most 70%, at most 75%, at most 80%, at most 85%, at most 90%, at most 95% or at most 100%. 39 PATENT TT3-003WO In yet other aspects of this embodiment, a therapeutically effective amount of a cancer therapeutic disclosed herein reduces or maintains a cancer cell population and/or tumor cell size in an individual by, e.g., about 10% to about 100%, about 10% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 20% to about 100%, about 20% to about 90%, about 20% to about 80%, about 20% to about 20%, about 20% to about 60%, about 20% to about 50%, about 20% to about 40%, about 30% to about 100%, about 30% to about 90%, about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, or about 30% to about 50%. [00136] In one aspect, a pharmaceutical composition disclosed herein reduces a symptom of a disorder associated with a cancer (e.g., tumor size and/or number of tumor cells). In aspects of this embodiment, a pharmaceutical composition disclosed herein reduces a symptom of a disorder associated with a cancer by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95%. In other aspects of this embodiment, a pharmaceutical composition disclosed herein reduces a symptom of a disorder associated with a cancer by, e.g., about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70%. [00137] In another aspect, a pharmaceutical composition disclosed herein reduces the frequency of a symptom of a disorder associated with a cancer incurred over a given time period. In aspects of this embodiment, a pharmaceutical composition disclosed 40 PATENT TT3-003WO herein reduces the frequency of a symptom of a disorder associated with a cancer (e.g., such as tumor size) incurred over a given time period by, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95%. In other aspects of this embodiment, a pharmaceutical composition disclosed herein reduces the frequency of a symptom of a disorder associated with a cancer incurred over a given time period by, e.g., about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70%. [00138] In any of the embodiments above, one or more cancer therapies (e.g., chemotherapy, radiation therapy, immunotherapy, surgery or hormone therapy) are co- administered further with a compound of the invention. [00139] In one embodiment, the chemotherapeutic reagent is an alkylating agent: nitrogen mustards, nitrosoureas, tetrazines, aziridines, cisplatins and derivatives, and non-classical alkylating agents. Nitrogen mustards include mechlorethamine, cyclophosphamide, melphalan, chlorambucil, ifosfamide and busulfan. Nitrosoureas include N-Nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU) and semustine (MeCCNU), fotemustine and streptozotocin. Tetrazines include dacarbazine, mitozolomide and temozolomide. Aziridines include thiotepa, mytomycin and diaziquone (AZQ). Cisplatin and derivatives include cisplatin, carboplatin and oxaliplatin. In one embodiment the chemotherapeutic reagent is an anti-metabolites: the anti-folates (e.g., methotrexate), fluoropyrimidines (e.g., fluorouracil and capecitabine), deoxynucleoside 41 PATENT TT3-003WO analogues and thiopurines. In another embodiment the chemoptheraputic reagent is an anti-microtubule agent such as vinca alkaloids (e.g., vincristine and vinblastine) and taxanes (e.g., paclitaxel and docetaxel). In another embodiment the chemotherapeutic reagent is a topoisomerase inhibitor or a cytotoxic antibiotic such as doxorubicin, mitoxantrone, bleomycin, actinomycin, and mitomycin. [00140] In another embodiment, the compounds described herein are administered with one or more additional anti-neoplastic agents. Such anti-neoplastic agents can include, for example, alkylating agents, anti-CTLA-4 monoclonal antibodies, anti-PD-1 and PD- L1 monoclonal antibodies (immune checkpoint inhibitors), antibiotics/antineoplastics, antimetabolites, antineoplastic combinations, antineoplastic detoxifying agents, antineoplastic interferons, BCR-ABL tyrosine kinase inhibitors, BTK inhibitors, CD19 monoclonal antibodies, CD20 monoclonal antibodies, CD30 monoclonal antibodies, CD33 monoclonal antibodies, CD38 monoclonal antibodies, CD52 monoclonal antibodies, CDK 4/6 inhibitors, EGFR inhibitors, hedgehog pathway inhibitors, HER2 inhibitors, histone deacetylase inhibitors, hormones/antineoplastics, malignancy photosensitizers, miscellaneous antineoplastics, mitotic inhibitors, mTOR inhibitors, multikinase inhibitors, PARP inhibitors, PI3K inhibitors, proteasome inhibitors and VEGF/VEGFR inhibitors. [00141] The invention also provides methods of treating an ailment such as cancer that includes administering one or more compounds described herein or a salt thereof, or a pharmaceutical composition of the same. In some embodiments, the subject is a subject in need of such treatment. In some embodiments, the compound is administered as a prodrug. Examples of cancer include melanoma, endometrium, lung, hematopoietic/lymphoid, ovarian, cervical, soft-tissue sarcoma, urinary tract, pancreas, thyroid, kidney, glioblastoma, breast cancer. [00142] In any of the embodiments, one or more cancer therapies (e.g., chemotherapy, radiation therapy, immunotherapy, surgery or hormone therapy) can be co-administered further with a medicament of the invention. In one embodiment, the chemotherapeutic reagent is an alkylating agent: nitrogen mustards, nitrosoureas, tetrazines, aziridines, 42 PATENT TT3-003WO cisplatins and derivatives, and non-classical alkylating agents. Nitrogen mustards include mechlorethamine, cyclophosphamide, melphalan, chlorambucil, ifosfamide and busulfan. Nitrosoureas include N-Nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU) and semustine (MeCCNU), fotemustine and streptozotocin. Tetrazines include dacarbazine, mitozolomide and temozolomide. Aziridines include thiotepa, mytomycin and diaziquone (AZQ). Cisplatin and derivatives include cisplatin, carboplatin and oxaliplatin. In one embodiment the chemotherapeutic reagent is an anti- metabolites: the anti-folates (e.g., methotrexate), fluoropyrimidines (e.g., fluorouracil and capecitabine), deoxynucleoside analogues and thiopurines. In another embodiment the chemoptheraputic reagent is an anti-microtubule agent such as vinca alkaloids (e.g., vincristine and vinblastine) and taxanes (e.g., paclitaxel and docetaxel). In another embodiment the chemotherapeutic reagent is a topoisomerase inhibitor or a cytotoxic antibiotic such as doxorubicin, mitoxantrone, bleomycin, actinomycin, and mitomycin. [00143] In embodiments, the prodrugs described herein increase the half-life of a small molecule active agent that is administered to a subject. In some embodiments, the half- life is increased by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% compared to a non-conjugated active agent. [00144] In embodiments, the prodrugs described herein increase the adsorption, distribution, metabolism and/or excretion of a small molecule active agent that is administered to a subject. In some embodiments, one or more is increased by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% compared to a non- conjugated active agent. EXAMPLES 43 PATENT TT3-003WO [00145] The following non-limiting examples are provided for illustrative purposes only in order to facilitate a more complete understanding of representative embodiments now contemplated. These examples are intended to be a mere subset of all possible contexts in which the components of the formulation may be combined. Thus, these examples should not be construed to limit any of the embodiments described in the present specification, including those pertaining to the type and amounts of components of the formulation and/or methods and uses thereof. Example 1 Chemical Synthesis of 1-oxidothiomorpholino (2-(((((1S,9S)-1-amino-9-ethyl-5- fluoro-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15- octahydrobenzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9- yl)oxy)carbonyl)oxy)ethyl)carbamate 2-((tert-butoxycarbonyl)amino)ethyl carbonochloridate (V4483-078) To a solution of tert-butyl (2-hydroxyethyl)carbamate (2.5 g x 2 batches, 15.53 mmol) in THF (40 mL) was dropwise added TEA (4.31 mL, 31.06 mmol) and triphosgene (1.4 g, 44 PATENT TT3-003WO 4.66 mmol , in 3 mL THF) at 0℃. The reaction mixture was stirred for 2h at 0℃. The solids was filtrated out. The filtrate was used directly in next step. (9H-fluoren-9-yl)methyl ((1S,9S)-9-(((2-((tert- butoxycarbonyl)amino)ethoxy)carbonyl)oxy)-9-ethyl-5-fluoro-4-methyl-10,13- dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2- b]quinolin-1-yl)carbamate (V4483-079) To a mixture of (9H-fluoren-9-yl)methyl ((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl- 10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]indolizino[1,2- b]quinolin-1-yl)carbamate (500 mg x 2 batches, 0.76 mmol), DMAP (929 mg, 7.4 mmol) and THF (10 mL) was dropwise added the filtrate from the above step. The reaction mixture was stirred overnight at room temperature. The resulting mixture was diluted with water (15mL), extracted with dichloromethane (30 mL x 2), dried with anhydrous sodium sulfate, and concentrated. The residue was purified by reversed phase column chromatography (water (0.05% FA)/MeCN) to afford the product as light-brown solid (1.08 g, crude). LCMS: Calculated Exact Mass = 844.3, Found [M+H]⁺ (ESI+) = 845.2. (9H-fluoren-9-yl)methyl ((1S,9S)-9-(((2-aminoethoxy)carbonyl)oxy)-9-ethyl-5- fluoro-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15- octahydrobenzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate hydrochloride (V4483-082) To a solution of (9H-fluoren-9-yl)methyl ((1S,9S)-9-(((2-((tert- butoxycarbonyl)amino)ethoxy)carbonyl)oxy)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo- 2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1- yl)carbamate (1.0 g, crude) in dioxane (40 mL) was added HCl (7.10 mL, 28.44 mmol, 4M in dioxane). The reaction mixture was stirred for 6h at room temperature. The resulting mixture was diluted with dichloromethane (50 mL) and methanol (50 mL), extracted with water (200 mL x 2). The aqueous phase was added saturated sodium bicarbonate aqueous solution (100 mL), then extracted with dichloromethane (300 mL x 45 PATENT TT3-003WO 2), dried with anhydrous sodium sulfate, and concentrated. This resulting in the product as brown solid (540 mg, crude). LCMS: Calculated Exact Mass = 744.3, Found [M+H]⁺ (ESI+) = 745.2. (9H-fluoren-9-yl)methyl ((1S,9S)-9-ethyl-5-fluoro-4-methyl-9-(((2-((((1- oxidothiomorpholino)oxy)carbonyl)amino)ethoxy)carbonyl)oxy)-10,13-dioxo- 2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2- b]quinolin-1-yl)carbamate (V4483-085) A mixture of (9H-fluoren-9-yl)methyl ((1S,9S)-9-(((2-aminoethoxy)carbonyl)oxy)-9-ethyl- 5-fluoro-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15- octahydrobenzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate hydrochloride (500 mg, crude) 4-nitrophenyl (1-oxidothiomorpholino) carbonate (213 mg, 0.71 mmol), DIEA (0.34 mL, 1.94 mmol) and DMF (5 mL) was stirred for 1h at room temperature. The reaction mixture was filtrated and purified by reversed phase column chromatography (water (0.05% FA)/MeCN) to afford the product as light-brown solid (380 mg, 65% yield). LCMS: Calculated Exact Mass = 905.3, Found [M+H]⁺ (ESI+) = 906.2. 1-oxidothiomorpholino (2-(((((1S,9S)-1-amino-9-ethyl-5-fluoro-4-methyl-10,13- dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]indolizino[1,2- b]quinolin-9-yl)oxy)carbonyl)oxy)ethyl)carbamate (V4483-087) To a solution of (9H-fluoren-9-yl)methyl ((1S,9S)-9-ethyl-5-fluoro-4-methyl-9-(((2-((((1- oxidothiomorpholino)oxy)carbonyl)amino)ethoxy)carbonyl)oxy)-10,13-dioxo- 2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1- yl)carbamate (330 mg, 0.36 mmol) in DCM (40 mL) was dropwise added DBU (67.0 mg, 0.44 mmol) at 0℃. The reaction mixture was stirred for 30 min at 0℃. The resulting mixture was adjusted pH to 5 with FA (50% in water). The aqueous phase was purified by reverse column chromatography (water (0.05% FA)/MeCN) twice to afford the product as white solid (83.0 mg, 33% yield). LCMS: Calculated Exact Mass = 683.2, Found [M+H]⁺ (ESI+) = 684.0. 46 PATENT TT3-003WO ¹H NMR (400 MHz, DMSO-d6) δ 8.18 (s, 1H), 7.76 (d, J = 11.0 Hz, 1H), 7.67 (t, J = 5.1 Hz, 1H), 7.02 (s, 1H), 5.70 – 5.37 (m, 4H), 4.43 – 4.34 (m, 1H), 4.21-4.06 (m, 2H), 3.28 – 2.87 (m, 12H), 2.38 (s, 3H), 2.24 – 1.97 (m, 4H), 0.89 (t, J = 7.4 Hz, 3H). Example 2 Endoperoxide–Fe(II) reaction initiates a series of reactions ending with release of native payload [00146] In this example, a drug (i.e., exatecan) is linked to a trioxolane molecule for administration as a prodrug. In the presence of Iron, the drug is released. X _{O Fe+2} O N Payload _{H2N Payload ad} Example 3 No TOP1 inhibition observed with either unreleasable Exatecan conjugate (DLX-1) or releasable exatecan-invention conjugate (CONJ-33) [00147] Topoisomerases are nuclear enzymes that play essential roles in DNA replication, transcription, chromosome segregation, and recombination. All cells have two major forms of topoisomerases: Type I, which makes single-stranded cuts in DNA, and Type II enzymes, which cut and pass double-stranded DNA. DNA topoisomerases are important targets of approved and experimental anti-cancer agents. The protocols 47 PATENT TT3-003WO used herein were used to assess pro-drug compounds for their ability to inhibit Type I. The vitro assay for topoisomerase I activity is based on relaxation of supercoiled DNA. [00148] The assay described herein shows Topoisomerases 1 (TOP1) activity when cells are exposed to an agent (i.e., Exatecan, Exatecan mesylate, Tatara chem-1 (DLX- 1) or Tatara chem-4 (Conj-33)). Normal TOP1 activity results in relaxed DNA; TOP1 inhibition results in presence of supercoiled plasmid DNA. [00149] The activity of human topoisomerase enzyme was determined prior to the testing of the compounds – 1 unit (U) was defined as the amount of enzyme required to give full relaxation of the supercoiled DNA substrate. Test compounds were first prepared in 100% DMSO to achieve a 50 mM stock concentration. The stock was then diluted in 100% DMSO to achieve a 6-point dilution, 3-fold from a top concentration of 90uM for test compounds and 30uM for exatecan control. 1 U of human topoisomerase was then incubated with 0.5 μg of supercoiled pBR322 DNA in a 30 μL reaction at 37^oC for 30 minutes under the following conditions: 20 mM Tris.HCl (pH 7.5), 200 mM NaCl, 0.25 mM EDTA, 5 % (v/v) glycerol and 5% DMSO (+/- test compound). Each reaction was then stopped by the addition of 30 μl chloroform/iso-amyl alcohol (24:1) and 30 μL Stop Dye (40% sucrose (w/v), 100 mM Tris.HCl (pH 7.5), 10 mM EDTA, 0.5 μg/ml bromophenol blue), before being loaded on a 1.0% TAE agarose gel. Samples were ran at 90V for 1.5 hours. Bands were visualized by ethidium bromide staining for 10 minutes, de-stained for 10 minutes in water and analysed by gel documentation equipment (Syngene, Cambridge, UK) and quantified using Syngene Gene Tools software. Raw gel data (fluorescent band volumes) was calculated as a % of the 100% control (fully relaxed DNA) and converted to % activity with addition of the test compound. Fluorescence exceeding 100% of signal were normalized to 100%. [00150] FIG.2A and 2B show the results of the topoisomerase assay. Structures of the control compounds (DLX-1, TRX-1 and TRX-2) are shown in FIG.2C. The top arrow shows relaxed DNA (“R DNA”) with normal TOP1 activity; the bottom arrow shows supercoiled plasmid DNA (“SC DNA”) with TOP1 inhibition. 48 PATENT TT3-003WO [00151] DLX-1 was modified chemically to prevent release (also verifies caging of trioxolane conjugates); CONJ-33 is a releasable conjugate and lack of supercoiled DNA shows little to no inhibition of TOP1. [00152] In the next study, IC₅₀ values were determined for several compounds. The experiment was designed to ensure that TOP1 inhibitors conjugated to the N-O cage described herein are inactivated and thus do not engage TOP1. [00153] using the following protocol. 40 µL of HCT116 (or other) cell suspension at appropriate cell concentration was plated in each well of a 384-well plate. Plates were covered and allowed to equilibrate for 30 min at room temperature, then transferred to a CO₂ incubator at 37^oC overnight. A 10mM test compound or 1mM reference compound stock solution was made and 45 µL transferred to a 384-well plate. 3-fold 10-point dilutions were made using a TECAN EVO200 liquid handler. Diluted compound plates were spun at room temperature at 1000 RPM for 1 min, and shaken on a plate shaker for 2 min. 40 nL of diluted compound was then added to the cell plate and returned to the CO2 incubator. After 3 – 5 days of treatment, depending on assay endpoint, 40 µL of cell-titer glow reagent was added to each well (cell plate and reagent equilibrated to room temperature) for 30 min, and luminescence detected using a plate reader (EnVision). Cell growth inhibition was calculated using the formula: IC50 = 100 x (ReadoutHC – ReadoutSample) / (ReadoutHC –ReadoutLC) with HC = 0.1% DMSO and LC = 1uM Exatecan. [00154] IC₅₀ Values are shown in Table 2 below. Table 3 IC₅₀ (μM) 49 PATENT TT3-003WO TRX-1 >90 >90 >90 TRX-2 23.7 27.8 25.8 [00155] easing its exatecan payload. Its IC₅₀ was reported as ND because there was insufficient TOP1 inhibition over the concentration range studied in the assay. CONJ-33 is, importantly, a releasable conjugate, but is sufficiently stable to prevent the release of the TOP1 inhibitor payload. Similarly, its IC₅₀ is also reported as ND due to insufficient TOP1 inhibition over the concentration range studied. Example 4 Antibodies and Preparation of Linker-Payloads [00156] In embodiments, the payload is an antibody or antibody fragment. For example, trastuzumab is anti-neoplastic therapeutic antibody that can be use according to aspects. Maleimides can be conjugated to antibodies as shown in FIG.6A (maximum DAR of 8). Aspects include a compound of conj-71, conj-72 and conj-73. O F F O N H N F OH [00157] s-tosy bearng n er payoads t at can be conjugated as s own n G.6 (maximum DAR of 4). Aspects include a compound of conj-74, conj-75 and conj-76. 50 _{PATENT TT3-003WO [00158] Table 2 shows the DAR for each compound (conj-73, conj-74, conj-75 and} conj-76) conjugated to trastuzumab. Table 3 A_{ntibody Conjugate ADC DAR Tr t z m b C n 73 ADC 1 73 3 [00159] Experimental procedures for synthesis of these compounds are described} below. Procedures for Conjugates 71-76 Procedures for preparation of conjugate 71 Project Summary The synthetic route is based on a procedure supplied by WuXi: NO O² PATENT TT3-003WO H₂N BocHN BocHN F F NO F O ₂ N _{B TEA} N N HO O _{Genera procedure or preparat on o compound 5-} HCl Cl Alloc Cl HN N _{5-1 5-2} To a solution of 2-chloro-N-(2-chloroethyl)ethanamine(hydrochloride) (10.0 g, 56.0 mmol, 1.00 eq) and DIEA (18.1 g, 140 mmol, 24.40 mL, 2.50 eq) in DCM (200 mL) was added allyl carbonochloridate (6.75 g, 56.0 mmol, 5.94 mL, 1.00 eq) at 0 °C. The mixture was stirred at 20 °C for 16 hrs. TLC (Petroleum ether: Ethylacetate = 10 : 1, R_f = 0.30) indicated Reactant 1 52 PATENT TT3-003WO was disappear, and one major new spot with lower polarity was detected. The mixture were washed with 10% citric acid solution (1000 mL) and NaCl solution (100 mL). The organic phase was dried over anhydrous Na₂SO₄, filtered and concentrated to give a residue. The residue was purified by flash silica gel chromatography (ISCO^®; 400 g SepaFlash^® Silica Flash Column, Eluent of 0~15% Ethylacetate/Petroleum ether gradient @ 45 mL/min). Compound allyl N,N- bis(2-chloroethyl)carbamate (11.8 g, 52.2 mmol, 93.2% yield) was obtained as a colorless oil. Spectrum: ¹H NMR HNMR-ET95292-33-P1C, DMSO-d₆400MHz J = 5.3, 10.5, 17.2 Hz, 1H), 5.38 - 5.10 (m, 2H), 4.56 (td, J = 1.5, 5.2 Hz, 2H), 3.78 - 3.67 (m, 4H), 3.66 - 3.55 (m, 4H). _{General procedure for preparation of compound 5-3} Alloc Cl N ^NH _{2OH. HCl Alloc} N OH - - A mixture of allyl N,N-bis(2-chloroethyl)carbamate (6.00 g, 26.5 mmol, 1.00 eq) and NH₂OH.HCl (5.53 g, 79.6 mmol, 3.00 eq) in Et₃N (60 mL) was degassed and purged with N₂ for three times, and then the mixture was stirred at 90 °C for 48 hrs under N₂ atmosphere. TLC Plate 1 (Petroleum ether : Ethyl acetate = 10 : 1, R_f = 0.00) and Plate 2 (Petroleum ether : Ethyl acetate = 0 : 1, R_f = 0.30) indicated some Reactant 1 reamined and one major new spot with larger polarity was detected. After cooling to 25 °C, the reaction mixture was filtered and the filter cake was washed with EtOAc (80 mL). The combined filtrate was concentrated to give a residue. The residue was purified by flash silica gel chromatography (ISCO^®; 40.0 g SepaFlash^® Silica Flash Column, Eluent of 0~100% Ethylacetate/Petroleum ether gradient @ 50 mL/min). Compound allyl 4-hydroxypiperazine-1-carboxylate (0.98 g, 5.26 mmol, 19.8% yield) was obtained as an off-white solid. _{General procedure for preparation of compound 5-4} 53 PATENT TT3-003WO NO O ₂ Alloc _{Cl O} 4-2a Alloc NO N O ₂ To a solution of ally -hydroxypiperazine-1-carboxylate (150 mg, 1.00 eq) and (4- nitrophenyl) carbonochloridate (162 mg, 9.98e-1.00 eq) in THF (3 mL) was added TEA (163 mg, 2.00 eq). The reaction mixture was stirred at 20 °C for 1 hr. LCMS showed reactant 1 was consumed completely and one main peak (no mass response) was detected. The reaction mixture (300 mg, crude) as a light yellow liquid was used to next step directly without work-up. _{General procedure for preparation of compound 1} H₂N BocHN F To a solution of (10S,23S)-23-amino-10-ethyl-18-fluoro-10-hydroxy-19-methyl-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1(24),2(14),6(11),12,15,17,19- heptaene-5,9-dione; methanesulfonic acid (6.40 g, 12.0 mmol, 1.00 eq) in THF (128 mL) were added TEA (2.44 g, 24.1 mmol, 3.35 mL, 2.00 eq) and Boc₂O (2.89 g, 13.2 mmol, 3.04 mL, 1.10 eq) at 0 °C. The mixture was stirred at 20 °C for 20 hrs. LC-MS showed some Reactant 1 remained and desired compound was detected. The mixture was filtered and the solid was washed with cold THF (150 mL). The solid was dried under reduced pressure to give tert-butyl 54 PATENT TT3-003WO N-[(10S,23S)-10-ethyl-18-fluoro-10-hydroxy-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 23-yl]carbamate (4.90 g, 9.00 mmol, 74.7% yield, 98.4% purity) was obtained as off-white solid. Spectrum: ¹H NMR HNMR-ET95292-4-P1C, DMSO-d₆400MHz - 7.49 (m, 2H), 7.25 (s, 1H), 6.50 (s, 1H), 5.48 - 5.34 (m, 2H), 5.20 - 5.06 (m, 2H), 4.81 (br d, J = 19.0 Hz, 1H), 3.23 - 3.15 (m, 1H), 3.06 - 2.95 (m, 1H), 2.26 (s, 3H), 2.20 (br dd, J = 5.1, 13.3 Hz, 1H), 2.04 - 1.94 (m, 1H), 1.86 (td, J = 7.0, 14.3 Hz, 2H), 1.75 (td, J = 3.3, 6.6 Hz, 1H), 1.47 (s, 9H), 0.87 (t, J = 7.3 Hz, 3H). LCMS LCMS-ET95292-4-P1C: m/z 536.3 (M+1) _{General procedure for preparation of compound 2} BocHN BocHN F O₂ To a solution of tert-butyl N-[(10S,23S)-10-ethyl-18-fluoro-10-hydroxy-19-methyl-5,9-dioxo-8- oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-23-yl]carbamate (2.00 g, 3.73 mmol, 1.00 eq) in DCM (20 mL) was added DMAP (1.14 g, 9.34 mmol, 2.50 eq) and a solution of (4-nitrophenyl) carbonochloridate (753 mg, 3.73 mmol, 1.00 eq) in DCM (20 mL) at 0 °C. The mixture was stirred at 20 °C for 1 hr. LCMS showed 7% of Reactant 1 was remaining and 55.3% peak with desired ms was detected. The reaction mixture (theoretical amount 2.6 g) as a yellow liquid, which was used to next step directly. 55 PATENT TT3-003WO _{General procedure for preparation of compound 3} BocHN BocHN F F bz To a s , y y y y-5,9- dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl] (4-nitrophenyl) carbonate (2.6 g, 3.74 mmol, 1.00 eq) in DCM (5 mL) was added DMAP (914 mg, 7.48 mmol, 2.00 eq) and a solution of benzyl N-(2- hydroxyethyl)carbamate (876 mg, 4.49 mmol, 1.20 eq) in DCM (5 mL) at 0 °C. The mixture was stirred at 20 °C for 1 hr. LC-MS showed 11.8% of Reactant 1 was remaining and 33.9% peak with desired mass was detected. The reaction mixture was concentrated to give a residue. The residue was purified by prep-HPLC (neutral condition, column: Waters Xbridge BEH C18 _{250*70mm* mobile phase: [H2O(10mM NH4HCO3)-ACN]; gradient: 40%-85% B over 18.0} min). Compound 2-(benzyloxycarbonylamino)ethyl [(10S,23S)-23-(tert-butoxycarbonylamino)- 10-ethyl-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] carbonate (1.40 g, 1.75 mmol, 46.8% yield, 94.5% purity) was obtained as an off-white solid. Spectrum: ¹H NMR HNMR-ET95757-6-P1P1, DMSO-d₆400MHz J = 5.5 Hz, 1H), 7.32 - 7.22 (m, 5H), 7.03 (s, 1H), 5.51 (s, 2H), 5.31 - 5.12 (m, 2H), 5.04 - 4.96 (m, 1H), 4.94 (s, 2H), 4.10 (br t, J = 5.4 Hz, 2H), 3.16 - 3.04 (m, 1H), 2.55 - 2.52 (m, 1H), 2.36 - 2.34 (m, 3H), 2.34 - 2.31 (m, 1H), 2.25 - 2.10 (m, 3H), 2.10 - 1.98 (m, 1H), 1.51 (s, 9H), 0.89 (t, J = 7.3 Hz, 3H). 56 PATENT TT3-003WO LCMS LCMS-ET95757-6-P1P1: m/z 757.4 (M+1) _{General procedure for preparation of compound 4} BocHN BocHN F F H₂ To a soluti on o -( enzyoxycar onyamno)e y [( , )- -(er- uoxycar onylamino)-10- ethyl-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] carbonate (500 mg, 1.00 eq) in DMF (20 mL) was added Pd/C (282 mg, 265 10.0% purity, 4.01e-1.00 eq) under N₂. The suspension was degassed and purged with H₂ for three times. The resulting mixture was stirred at 20 °C for 1 hr under H₂ (15 Psi). LC-MS showed 7.5% of Reactant 1 was remaining and 79.6% peak with desired mass was detected. The reaction mixture (400 mg, crude) was filtered and the filtrate as a yellow liquid was used to next step directly. _{General procedure for preparation of compound 5} BocHN BocHN lloc PATENT TT3-003WO To a solution of 2-aminoethyl [(10S,23S)-23-(tert-butoxycarbonylamino)-10-ethyl-18-fluoro-19- methyl-5,9-dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl] carbonate (400 mg, 1.00 eq) in DMF (20 mL) were added Et₃N (130 mg, 2.00 eq) and a solution of allyl 4-(4- nitrophenoxy)carbonyloxypiperazine-1-carboxylate (248 mg, 1.10 eq) in THF (3 mL) at 20 °C, the reaction mixture was stirred at 20 °C for 2 hrs. LCMS showed Reactant 1 was consumed completely and 72.6% peak with desired mass was detected. The reaction mixture was concentrated to give a residue. The residue was purified by Prep-HPLC (neutral condition; _{column: Waters Xbridge BEH C18250*70mm* mobile phase: [H2O(10mM NH4HCO3)-} ACN]; gradient: 40%-85% B over 18.0 min). allyl 4-[2-[[(10S,23S)-23-(tert- butoxycarbonylamino)-10-ethyl-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl]oxycarbonyloxy]ethylcarbamoyloxy]piperazine-1-carboxylate (200 mg, 34.7% yield, 93.0% purity) was obtained as a white solid. Spectrum: ¹H NMR HNMR-ET95757-23-P1C1, DMSO-d₆400MHz J = 10.9 Hz, 1H), 7.69 (br d, J = 8.9 Hz, 1H), 7.63 - 7.57 (m, 1H), 7.01 (s, 1H), 5.98 - 5.86 (m, 1H), 5.51 (s, 2H), 5.35 - 5.10 (m, 4H), 4.52 (d, J = 5.3 Hz, 2H), 4.21 - 4.06 (m, 2H), 3.97 - 3.74 (m, 2H), 3.28 - 3.20 (m, 2H), 3.20 - 3.03 (m, 4H), 2.70 - 2.65 (m, 1H), 2.65 - 2.57 (m, 1H), 2.37 (s, 3H), 2.34 - 2.30 (m, 1H), 2.27 - 2.13 (m, 3H), 2.11 - 2.01 (m, 1H), 1.51 (s, 9H), 0.93 - 0.86 (m, 3H) LCMS LCMS-ET95757-23-P1C1: m/z 835.2 (M+1) _{General procedure for preparation of compound 6} BocHN BocHN H PATENT TT3-003WO To a solution of allyl 4-[2-[[(10S,23S)-23-(tert-butoxycarbonylamino)-10-ethyl-18-fluoro-19- methyl-5,9-dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl]oxycarbonyloxy]ethylcarbamoyloxy]piperazine-1- carboxylate (100 mg, 1.00 eq) in THF (1 mL) was added Pd(PPh₃)₄ (27.7 mg, 24.0 0.20 eq) and 1,3-dimethylhexahydropyrimidine-2,4,6-trione (112 mg, 6.00 eq) .The mixture was stirred at 20 °C for 18 hrs. LCMS showed 1% of reactant 1 was remaining and 49% of peak with desired mass was detected. The reaction mixture was diluted with H₂O (5 mL) and washed with EtOAc (5 mL * 3). The aqueous phase was lyophilized to give a red amorphous solid. The residue was dissolved in DMF (5 mL), and the resulting solution was _{purified by prep-HPLC (column: WePure Biotech XP tC18150*40* mobile phase:} [H₂O(10mM NH₄HCO₃)-ACN]; gradient: 20%-50% B over 10 min). Compound [(10S,23S)-23- (tert-butoxycarbonylamino)-10-ethyl-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] 2-(piperazin-1-yloxycarbonylamino)ethyl carbonate (40.0 mg, 44.5% yield) was obtained as a red solid. _{General procedure for preparation of compound 7} BocHN O O BocHN F O F O To a solution of [(10S,23S)-23-(tert-butoxycarbonylamino)-10-ethyl-18-fluoro-19-methyl-5,9- dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl] 2-(piperazin-1-yloxycarbonylamino)ethyl carbonate (30.0 mg, 1.00 eq) in DMF (0.5 mL) were added TEA (12.1 mg, 3.00 eq) and (2,5-dioxopyrrolidin-1-yl) 6-(2,5-dioxopyrrol-1-yl)hexanoate (13.6 mg, 1.10 eq) and the resulting mixture was stirred at 20 °C for 1 hr. LCMS showed Reactant 1 was consumed completely and 13% peak with desired mass was detected. The reaction mixture was concentrated to give a residue. The residue was purified by perp-HPLC(TFA,column : WePure _{Biotech XP tC18150*40* mobile phase : [H2O(10mM NH4HCO3)-ACN]; gradient: 25%-} 55% B over 8.0 min). Compound [(10S,23S)-23-(tert-butoxycarbonylamino)-10-ethyl-18-fluoro- 19-methyl-5,9-dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 59 PATENT TT3-003WO 1,6(11),12,14,16,18,20(24)-heptaen-10-yl] 2-[[4-[6-(2,5-dioxopyrrol-1-yl)hexanoyl]piperazin-1- yl]oxycarbonylamino]ethyl carbonate (10.0 mg, 26.5% yield) was obtained as a yellow solid. _{General procedure for preparation of compound Target 1} o a sou on o [( , )- -(er- uoxycar onyamno)- -e y- - uoro- -me y- , - dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl] 2-[[4-[6-(2,5-dioxopyrrol-1-yl)hexanoyl]piperazin-1- yl]oxycarbonylamino]ethyl carbonate (7.00 mg, 1.00 eq) in EtOAc (0.035 mL) was added HCl/EtOAc (2 M, 0.035 mL, 9.44 eq). The mixture was stirred at 10 °C for 10 min. LCMS showed Reactant 1 was remaining and desired mass was detected. The reaction mixture was concentrated to give a residue. The residue was purified by prep-HPLC(TFA,column: WePure _{Biotech XP tC18100*30* mobile phase: [H2O(0.1% TFA)-ACN]; gradient: 5%-40% B over} 8.0 min). Compound [(10S,23S)-23-amino-10-ethyl-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] 2-[[4-[6-(2,5-dioxopyrrol-1-yl)hexanoyl]piperazin-1-yl]oxycarbonylamino]ethyl carbonate (5.00 mg, 68.7% yield, 97.6% purity, TFA) was a pale yellow solid. Spectrum: ¹H NMR HNMR-ET96034-91-P1P2, DMSO-d₆400MHz – 0.80 (m, 1H) 0.81 – 0.98 (m, 3H) 1.02 – 1.55 (m, 7H) 2.02 - 2.22 (m, 3H) 2.42 (br d, J = 8.38 Hz, 4H) 2.78 – 2.89 (m, 3H) 3.04 – 3.21 (m, 7H) 3.67 – 3.81 (m, 2H) 3.96 – 4.04 (m, 1H) 4.13 – 4.35 (m, 2H) 4.36 – 4.48 (m, 1H) 4.71 – 5.04 (m, 1H) 5.29 – 5.57 (m, 2H) 6.59 – 6.79 (m, 1H) 6.94 – 7.34 (m, 1H) 7.60 – 8.14 (m, 2H) 8.22 – 8.44 (m, 2H) LCMS LCMS-ET96034-91-P1P4: m/z 844.2 (M+1) SFC SFC-ET96034-91-P1F1: de% = 100 60 PATENT TT3-003WO Procedures for the preparation of conjugate 72 O₂N O Cl Boc N ^NH _{2OH. HCl} Boc N _{4-2a O Cl O2N O} Boc N oc _{General procedure for preparation of compound 6-2} Cl Boc N NH₂OH. HCl Boc PATENT TT3-003WO A mixture of tert-butyl N,N-bis(2-chloroethyl)carbamate (25.0 g, 103 mmol, 1.00 eq) and NH₂OH.HCl (21.5 g, 310 mmol, 3.00 eq) in TEA (250 mL) was degassed and purged with N₂ for three times, and then the mixture was stirred at 90 °C for 48 hrs under N₂ atmosphere. TLC (Petroleum ether: Ethylacetate = 0 : 1) indicated some Reactant 1 remained and one new spot (R_f = 0.40) formed. After cooling to 25 °C, the reaction mixture was filtered and the filter cake was washed with EtOAc (400 mL). The combined filtrate was concentrated to give a residue. The residue was purified by flash silica gel chromatography (ISCO^®; 120 g SepaFlash^® Silica Flash Column, Eluent of 0~68% Ethyl acetate/Petroleum ether gradient @ 100 mL/min). Plate 1 (Petroleum ether: Ethylacetate = 0 : 1, R_f = 0.40). Compound tert-butyl 4-hydroxypiperazine-1- carboxylate (3.00 g, 14.8 mmol, 14.4% yield) was obtained as an off-white solid. General procedure for preparation of compound 6-3 O_{2N O} Boc _{4-2a O2N} Boc To a solution of tert-butyl 4-hydroxypiperazine-1-carboxylate (200 mg, 1.00 eq) in THF (2 mL) was added DIEA (256 mg, 2.00 eq) and (4-nitrophenyl) carbonochloridate (199 mg, 1.00 eq) at 20 °C under N₂ atmosphere. The mixture was stirred at 20 °C for 1 hr under N₂ atmosphere. TLC (Petroleum ether : Ethyl acetate = 0 : 1, R_f = 0.90) indicated Reactant 1 was consumed completely and one new spot (R_f = 0.90) formed. No work up was carried out. Compound tert-butyl 4-(4-nitrophenoxy) carbonyloxypiperazine-1- carboxylate (360 mg, crude) as a yellow liquid in THF (2 mL) was used in next step directly. General procedure for preparation of compound 2 62 PATENT TT3-003WO FmocHN FmocHN F F oc To a , 9-methyl- 5,9-dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-23-yl]carbamate (400 mg, 1.00 eq), 4-(tert- butoxycarbonylamino)-3,3-dimethyl-butanoic acid (366 mg, 1.58 mmol, 2.60 eq) in DCM (6 mL) was added DMAP (14.9 mg, 0.20 eq), DIC (154 mg, 2.00 eq) at 20 °C. The mixture was stirred at 20 °C for 12 hrs. LC-MS showed Reactant 1 was consumed completely and ~64% of desired compound was detected. The reaction mixture was diluted with 10 mL H₂O and extracted with EtOAc (10 mL * 3). The combined organic phase was dried with anhydrous Na₂SO₄, the mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by flash silica gel chromatography (ISCO^®; 12.0 g SepaFlash^® Silica Flash Column, Eluent of 0~30% Ethyl acetate/Petroleum ethergradient @ 80 mL/min). (Petroleum ether/Ethyl acetate = 1/2, R_f = 0.61). Compound [(10S,23S)-10-ethyl-23-(9H-fluoren-9- ylmethoxycarbonylamino)-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] 4-(tert-butoxycarbonylamino) -3,3-dimethyl-butanoate (420 mg, 79.3% yield) was obtained as a yellow solid. _{General procedure for preparation of compound 3} 63 PATENT TT3-003WO FmocHN FmocHN F F H₂ A solution of [(10S,23S)-10-ethyl-23-(9H-fluoren-9-ylmethoxycarbonylamino)-18-fluoro-19- methyl-5,9-dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl] 4-(tert-butoxycarbonylamino)-3,3-dimethyl-butanoate (300 mg, 1.00 eq) in HCl/EtOAc (4 M, 4 mL, 46.5 eq) was stirred at 20 °C for 1 hr. LC- MS showed Reactant 1 was consumed completely and ~89% of desired compound was detected. The mixture was concentrated to give a residue. Compound [(10S,23S)-10-ethyl-23-(9H-fluoren- 9-ylmethoxycarbonylamino)-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] 4-amino-3,3-dimethyl-butanoate (260 mg, crude) was obtained as a yellow solid. General procedure for preparation of compound 4 FmocHN _{freshly prepared FmocHN} F F Boc To a solution of [(10S,23S)-10-ethyl-23-(9H-fluoren-9-ylmethoxycarbonylamino)-18-fluoro-19- methyl-5,9-dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl] 4-amino-3,3-dimethyl-butanoate (260 mg, 1.00 eq), DIPEA (87.2 mg, 2.00 eq) in THF (2 mL) was added tert-butyl 4- 64 PATENT TT3-003WO (4-nitrophenoxy)carbonyloxypiperazine-1-carboxylate (124 mg, 1.00 eq) in DMF (2 mL) at 0 °C, while stirring under a nitrogen atmosphere, the reaction mixture was stirred at 20 °C for 0.5 hr. LC-MS showed Reactant 1 was consumed completely and ~47% of desired compound was detected. The mixture was concentrated to give a residue. The residue was purified by prep- HPLC (neutral column: WePure Biotech XPt C18150 x 40 x mobile phase: [H₂O (10mM NH₄HCO₃)-ACN]; gradient: 65%-95% B over 8.0 min. Compound tert-butyl 4-[[4-[[(10S,23S)-10- ethyl-23-(9H-fluoren-9-ylmethoxycarbonylamino)-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl]oxy]-2,2-dimethyl-4-oxo-butyl]carbamoyloxy]piperazine-1-carboxylate (200 mg, 59.4% yield) was obtained as a yellow solid. General procedure for preparation of compound 5 FmocHN FmocHN F F NH A solution of tert-butyl 4-[[4-[[(10S,23S)-10-ethyl-23-(9H-fluoren-9-ylmethoxycarbonylamino)-18- fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl]oxy]-2,2-dimethyl-4-oxo-butyl]carbamoyloxy]piperazine-1-carboxylate (80.0 mg, 1.00 eq) in HCl/EtOAc (2 M, 4.00 mL) was stirred at 0 °C for 10 min. LC-MS showed ~45% of Reactant 1 was remained and ~42% of desired compound was detected. The reaction mixture was concentrated under vacuum. The crude product was used directly. Compound [(10S,23S)- 10-ethyl-23-(9H-fluoren-9-ylmethoxycarbonylamino)-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] 3,3-dimethyl-4-(piperazin-1-yloxycarbonylamino)butanoate (70.0 mg, crude) was obtained as a yellow solid. 65 PATENT TT3-003WO _{General procedure for preparation of compound 6} FmocHN FmocHN F O O O F N N O T - methyl-5,9-dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl] 3,3-dimethyl-4-(piperazin-1- yloxycarbonylamino)butanoate (70.0 mg, 1.00 eq) in DMF (1 mL) was added DIPEA (10.1 mg, 1.00 eq) and (2,5-dioxopyrrolidin-1-yl) 6-(2,5-dioxopyrrol-1- yl)hexanoate (24.0 mg, 1.00 eq) at 0 °C. The mixture was stirred at 20 °C for 0.5 hr. LC-MS showed Reactant 1 was consumed completely and ~30% of desired compound was detected. The mixture was concentrated to give a residue. The residue was purified by prep- HPLC (TFA column: WePure Biotech XPt C18150 x 40 mm x mobile phase: [H₂O (10mM NH₄HCO₃)-ACN]; gradient: 55%-85% B over 8.0 min). Compound [(10S,23S)-10-ethyl-23-(9H- fluoren-9-ylmethoxycarbonylamino)-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl]4-[[4-[6-(2,5-dioxopyrrol-1-yl)hexanoyl]piperazin-1-yl]oxycarbonylamino]-3,3-dimethyl- butanoate (6.00 mg, _{General procedure for preparation of compound Target 2} PATENT TT3-003WO yl]oxycarbonylamino]-3,3-dimethyl-butanoate (11.0 mg, 1.00 eq) and DBU (1.99 mg, 1.30 eq) in DCM (1 mL) was stirred at 0 °C for 0.5 hr. LC-MS showed Reactant 1 was consumed completely and ~36% of desired compound was detected. The reaction mixture was quenched by addition 1M HCl at 0 °C to pH = 5, and the mixture was concentrated under vacuum. The residue was purified by prep-HPLC (FA column: column: Phenomenex luna C18100 x 40 mm x mobile phase: [H₂O (0.2% FA)-ACN]; gradient: 12%-42% B over 10.0 min). Compound [(10S,23S)-23-amino-10-ethyl-18-fluoro-19-methyl-5,9- dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl] 4-[[4-[6-(2,5-dioxopyrrol-1-yl)hexanoyl]piperazin-1- yl]oxycarbonylamino]-3,3-dimethyl-butanoate (2.90 mg, 33.1% yield, 100% purity) was obtained as a white solid. HNMR-ET95337-37-P1B, QC-ET95337-37-P1Q, SFC-ET95337-37-P1A showed it was the pure desired compound. Spectrum: ¹H NMR HNMR-ET95337-37-P1B, DMSO-d₆400MHz d, J = 10.88 Hz, 2H), 6.96 (s, 1H), 6.44 (s, 1H), 5.42 - 5.51 (m, 1H), 5.24 - 5.33 (m, 1H), 4.09 - 4.55 (m, 2H), 3.67 - 4.01 (m, 2H), 3.53 (br dd, J = 17.39, 4.75 Hz, 1H), 3.13 - 3.26 (m, 6H), 2.89 - 3.08 (m, 4H), 2.38 (s, 6H), 2.06 - 2.20 (m, 4 H), 1.54 - 2.02 (m, 4H), 1.19 - 1.51 (m, 8H), 1.08 (s, 6H), 0.93 (br s, 3H) LCMS LCMS-ET95337-37-P1Q: m/z 870.4 (M+1) SFC SFC-ET95337-37-P1A: de% = 100% Procedures for the preparation of conjugate 73 67 PATENT TT3-003WO General procedure for preparation of compound 1 F Boc F N O H To a solution of tert-butyl 4-(4-nitrophenoxy)carbonyloxypiperazine-1-carboxylate (363 mg, 988 1.00 eq) in DMF (20 mL) were added Et₃N (200 mg, 2.00 eq) and a solution of (10S,23S)-23-amino-10-ethyl-18-fluoro-10-hydroxy-19-methyl-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)- heptaene-5,9-dione (430 mg, 1.00 eq) in THF (4 mL) at 20 °C, the reaction mixture was stirred at 20 °C for 30 min. LCMS showed Reactant 1 was consumed completely and 85% peak with desired mass was detected. The reaction mixture was concentrated to give a residue. 68 PATENT TT3-003WO The residue was purified by prep-HPLC (neutral condition, column: Waters Xbridge BEH C18 _{250*70* moblie phase: [H2O(10mM NH4HCO3)-ACN]; gradient: 30%-70% B over 18.0} min). Compound tert-butyl 4-[[(10S,23S)-10-ethyl-18-fluoro-10-hydroxy-19-methyl-5,9-dioxo-8- oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-23-yl]carbamoyloxy]piperazine-1-carboxylate (430 mg, 648 65.6% yield) was obtained as a yellow solid. Spectrum: ¹H NMR HNMR-ET96034-5-P1A3, DMSO 400MHz J = 7.32 Hz, 3H) 1.35 - 1.50 (m, 9H) 1.79 - 1.95(tt, J = 14.48, 7.04 Hz, 2H) 2 - 2.11(m, 1H) 2.27 - 2.33 (m, 1H) 2.41 - 2.47 (s, 3H) 2.55 (s, 2H) 2.69 - 2.86 (m, 2H) 3.05 – 3.07 (m, 3H) 3.23 – 3.28 (m, 3H) 3.79 – 3.96 (s, 2H) 5.12 - 5.21 (m, 1H) 5.24 - 5.37 (m, 2H) 5.5 (s, 2H) 6.52 - 6.57 (s, 1H) 7.26 – 7.34 (s, 1H) LCMS LCMS-ET96034-5-P1A4: m/z 664.4 (M+1) _{General procedure for preparation of compound 2} Boc F F N O HN O , , 8- oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-23-yl]carbamoyloxy]piperazine-1-carboxylate (30.0 mg, 1.00 eq) in EtOAc (0.1 mL) was added HCl/EtOAc (4 M, 0.1 mL), and the reaction mixture was stirred at 25 °C for 15 min under N₂. LCMS showed Reactant 1 was consumed completely and 96% peak with desired mass was detected. The reaction mixture was concentrated to give a residue. Compound piperazin-1-yl N-[(10S,23S)-10-ethyl-18-fluoro-10- 69 PATENT TT3-003WO hydroxy-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 23-yl]carbamate (26.1 mg, 96.2% yield, HCl) was obtained as a white solid and used into the next step without further purification. General procedure for preparation of compound Target 3 To a solution of piperazin-1-yl N-[(10S,23S)-10-ethyl-18-fluoro-10-hydroxy-19-methyl-5,9-dioxo- 8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-23-yl]carbamate (27.1 mg, 1.00 eq, HCl) in DMF (0.3 mL) and DCM (0.3 mL) was added TEA (13.7 mg, 3.00 eq) and (2,5- dioxopyrrolidin-1-yl) 6-(2,5-dioxopyrrol-1-yl)hexanoate (18.1 mg, 1.30 eq). The mixture was stirred at 20 °C for 10 min. LCMS showed 24% of Reactant1 was remaining and 24% peak with desired mass was detected. The residue was purified by prep-HPLC (neutral _{condition, column : WePure Biotech XP tC18150*40* mobile phase: [H2O(10mM} NH₄HCO₃)-ACN]; gradient : 21%-47% B over 10.0 min). Compound [4-[6-(2,5-dioxopyrrol-1- yl)hexanoyl]piperazin-1-yl] N-[(10S,23S)-10-ethyl-18-fluoro-10-hydroxy-19-methyl-5,9-dioxo-8- oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-23-yl]carbamate (5.60 mg, 16.0% yield, 98.0% purity) was obtained as a white solid. HNMR: ET96034-23-P1P1.LCMS: ET96034-23-P1P3.SFC: ET96034-23-P1P1. Spectrum: ¹H NMR HNMR-ET96034-23-P1P1, DMSO-d₆400MHz 70 PATENT TT3-003WO J = 7.32 Hz, 3H) 1.11 - 1.29 (m, 2H) 1.48 (dq, J = 14.20, 7.11 Hz, 4H) 1.78 - 1.97 (m, 2H) 1.98 - 2.20 (m, 1H) 2.24 - 2.31 (m, 2H) 2.39 (s, 3H) 2.58 - 2.65 (m, 1H) 2.68 - 2.84 (br dd, J = 7.50, 5.25 Hz, 2H) 2.93 - 3.03 (m, 1H) 3.07 - 3.17 ( m, 2H) 3.21- 3.27 (br d, J = 4.88 Hz, 2H) 3.28 - 3.30 (m, 2H) 3.38 (br s, 1H) 3.40 (br s, 1H) 3.79 - 3.95 (m, 1H) 4.12 - 4.37 (m, 1H) 5.14 - 5.23 (m, 1H) 5.25 - 5.38 (m, 2H) 5.43 (s, 2H) 6.52 (s, 1H) 6.96 - 7.04 (m, 1H) 7.32 (s, 1H) 7.79 (d, J = 10.88 Hz, 1H) 8.30 (br d, J = 8.88 Hz, 1H). LCMS LCMS-ET96034-23-P1P3: m/z 757.4 (M+1) SFC SFC-ET96034-23-P1P1: de% = 100% Procedures for the preparation of conjugate 74 O OH O BocHN O F General procedure for preparation of compound 2: 71 PATENT TT3-003WO O OH O BocHN O F S was stirred at 20 °C for 2 hrs. A solution of tert-butyl ((1S,9S)-9-ethyl-5-fluoro-4-methyl-10,13- dioxo-9-(((2-(((piperazin-1-yloxy)carbonyl)amino)ethoxy)carbonyl)oxy)-2,3,9,10,13,15- hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (29.0 mg, ture and the resulting mixture was stirred at 20 °C for 2 hrs. LCMS showed starting material consumed completely and 96.2% peak with desired mass was detected. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give crude product. The residue was dissolved in DMF (1 mL) and _{purified by prep- 2O(0.2%} FA)-ACN]; gradient: 40%-95% B over 8.0 min) to give tert-butyl ((1S,9S)-9-ethyl-5-fluoro-4- methyl-10,13-dioxo-9-(((2-((((4-(4-(3-tosyl-2-(tosylmethyl)propanoyl)benzoyl)piperazin-1- yl)oxy)carbonyl)amino)ethoxy)carbonyl)oxy)-2,3,9,10,13,15-hexahydro-1H,12H- benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamate (15.0 mg, 13.1% yield) as a white solid. General procedure for preparation of Target 4B: PATENT TT3-003WO To a solution of tert-butyl ((1S,9S)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-9-(((2-((((4-(4-(3-tosyl- 2-(tosylmethyl)propanoyl)benzoyl)piperazin-1-yl)oxy)carbonyl)amino)ethoxy)carbonyl)oxy)- 2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1- mixture was stirred at 20 °C for 2 hrs. LCMS showed some starting material remained and desired mass was detected. The reaction mixture was concentrated under reduced pressure under 20 °C to give a residue. The residue was dissolved in DMF (1 mL) and purified by prep-HPLC (FA 2O(0.2% FA)-ACN]; gradient: 25%-70% B over 8.0 min) to give Target 4B (5.60 mg, 51.8% yield, 93.9% purity, 0.1FA) as a white solid . ¹H NMR: ET97226-16-P1A8 (400 MHz, DMSO-d₆) - 0.11 (m, 3H) 1.25 - 1.42 (m, 4H) 1.44 - 1.47 (m, 1H) 1.53 - 1.56 (m, 3H) 1.56 - 1.59 (m, 1H) 1.62 (s, 4H) 1.86 - 1.99 (m, 2H) 2.27 (br s, 1H) 2.32 - 2.41 (m, 3H) 2.84 - 2.88 (m, 1H) 2.96 - 3.06 (m, 2H) 3.11 - 3.20 (m, 1H) 3.23 - 3.51 (m, 3H) 3.63 - 3.75 (m, 2H) 4.58 - 4.63 (m, 1H) 4.69 (br s, 2H) 4.77 - 4.82 (m, 1H) 5.06 - 5.13 (m, 1H) 5.26 (s, 1H) 6.18 - 6.22 (m, 1H) 6.23 - 6.31 (m, 1H) 6.47 (d, J = 7.88 Hz, 1H) 6.57 - 6.68 (m, 6H) 6.69 - 6.78 (m, 3H) 6.80 - 6.89 (m, 1H) 6.90 - 6.99 (m, 2H) 7.30 - 7.33 (m, 1H) LCMS LCMS- ET97226-16-P1A8: m/z 1133.3 (M+1) SFC SFC- ET97226-16-P1A8: de% = 100% 73 PATENT TT3-003WO Procedures for the preparation of conjugate 75 O Cl O O OH H₂N OH NH PATENT TT3-003WO General procedure for preparation of compound 5B-2 O ^O H , _{120 o 2}N H_{N HCl C 12 h} OH To a solution of 4,4-dimethylpyrrolidin-2-one (16.0 g, 141 mmol, 1.00 eq) in H₂O (40 mL) was added HCl (40.8 g, 414 mmol, 40 mL, 37.0% purity, 2.93 eq) at 20 °C. The mixture was stirred at 120 °C for 12 hrs. LC-MS showed ~2% of Reactant 1 was remained and ~56% of desired compound was detected. The reaction mixture was quenched by addition NaHCO₃ saturated solution at 0 °C to pH = 9. No purification was carried out, it was used in next step directly. Compound 4-amino-3,3-dimethyl-butanoic acid (18.0 g, crude) was obtained as a yellow liquid in H₂O (40 mL) and it was used in next step directly General procedure for preparation of compound 5B-3 O Cl O OH To a solution of 4-amino-3,3-dimethyl-butanoic acid (18.0 g, 137 mmol, 1.00 eq) in H₂O (250 mL) and MeOH (25 mL) was added NaHCO₃ (11.5 g, 137 mmol, 5.34 mL, 1.00 eq) and benzyl carbonochloridate (28.1 g, 165 mmol, 23.51 mL, 1.20 eq) at 20 °C. The mixture was stirred at 20 °C for 2 hrs. LC-MS showed the reaction goes completely. The reaction was filtered and the filtrate was concentrated under vacuum. The residue was purified by prep-HPLC (TFA _{condition,column: Welch Xtimate C18250*100mm# mobile phase: [H2O(0.1% TFA)-} ACN]; gradient: 20%-50% B over 20.0 min). Compound 4-(benzyloxycarbonylamino)-3,3- dimethyl-butanoic acid (10.0 g, 37.7 mmol, 27.5% yield) was obtained as a yellow oil. Spectrum: ¹H NMR HNMR-ET95369-8-P1A, DMSO-d₆400MHz - 7.43 (m, 6H), 5.03 (s, 2H), 2.96 (br t, J = 5.69 Hz, 2H), 2.10 (d, J = 2.00 Hz, 2H), 0.91 (br d, J = 2.88 Hz, 6H) 75 PATENT TT3-003WO _{General procedure for preparation of compound 2} BocHN BocHN F 5B-3 F Cbz To a soluti y , y y y y ,9-dioxo-8- oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-23-yl]carbamate (2.80 g, 5.23 mmol, 1.00 eq) in DCM (40 mL) was added 4-(benzyloxycarbonylamino)-3,3-dimethyl-butanoic acid (2.77 g, 10.5 mmol, 2.00 eq), DIC (1.32 g, 10.5 mmol, 1.62 mL, 2.00 eq) and DMAP (128 mg, 1.05 mmol, 0.20 eq) at 20 °C. The mixture was stirred at 20 °C for 2 hrs. LC-MS showed no Reactant 1 was remained and ~72% of desired compound was detected. The reaction mixture was diluted with 10 mL H₂O and extracted with DCM (10 mL * 3). The combined organic phase was dried with anhydrous Na₂SO₄, the mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by flash silica gel chromatography (ISCO^®; 20.0 g SepaFlash^® Silica Flash Column, Eluent of 0~30% Ethyl acetate/Petroleum ethergradient @ 100 mL/min). (Petroleum ether/Ethyl acetate = 1/1, R_f = 0.64 by Plate 1). Compound [(10S,23S)-23-(tert- butoxycarbonylamino)-10-ethyl-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] 4-(benzyloxycarbonylamino)-3,3-dimethyl-butanoate (3.00 g, 3.83 mmol, 73.3% yield) was obtained as a yellow solid. _{General procedure for preparation of compound 3} 76 PATENT TT3-003WO BocHN BocHN F F H₂ To a solution of [(10S,23S)-23-(tert-butoxycarbonylamino)-10-ethyl-18-fluoro-19-methyl-5,9- dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl] 4-(benzyloxycarbonylamino)-3,3-dimethyl-butanoate (1.30 g, 1.66 mmol, 1.00 eq) in MeOH (20 mL) and THF (20 mL) was added Pd/C (1.77 g, 1.66 mmol, 10.0% purity, 1.00 eq) at 20 °C under N₂ atmosphere. The mixture was stirred at 20 °C for 15 min at 15 psi under H₂ atmosphere. LC-MS showed no Reactant 1 was remaining and ~65% of desired compound was detected. The reaction was filtered and the filtrate was concentrated under vacuum. Compound [(10S,23S)-23-(tert-butoxycarbonylamino)-10-ethyl-18- fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] 4-amino-3,3-dimethyl-butanoate (1.00 g, crude) was obtained as a white solid. _{General procedure for preparation of compound 4} BocHN BocHN F lloc To a solution of [(10S,23S)-23-(tert-butoxycarbonylamino)-10-ethyl-18-fluoro-19-methyl-5,9- dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl] 4-amino-3,3-dimethyl-butanoate (700 mg, 1.08 mmol, 77 PATENT TT3-003WO 1.00 eq) in DMF (5 mL) was added DIPEA (279 mg, 2.00 eq) and allyl 4- (4-nitrophenoxy)carbonyloxypiperazine-1-carboxylate (379 mg, 1.08 mmol, 1.00 eq). The mixture was stirred at 20 °C for 1 hr. LC-MS showed Reactant 1 was consumed completely and 21% desired compound was detected. The mixture was concentrated to give a residue. The residue was purified by prep-HPLC (neutral condition; column: WePure Biotech XP tC18 _{150*40* mobile phase: [H2O(10mM NH4HCO3)-ACN]; gradient: 50%-80% B over 8.0 min).} Compound allyl 4-[[4-[[(10S,23S)-23-(tert-butoxycarbonylamino)-10-ethyl-18-fluoro-19-methyl- 5,9-dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl]oxy]-2,2-dimethyl-4-oxo- butyl]carbamoyloxy]piperazine-1-carboxylate (200 mg, 21.5% yield) was obtained as a white solid. _{General procedure for preparation of compound 5} BocHN BocHN F O F NH 4 5 To a solution of allyl 4-[[4-[[(10S,23S)-23-(tert-butoxycarbonylamino)-10-ethyl-18-fluoro-19- methyl-5,9-dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl]oxy]-2,2-dimethyl-4-oxo- butyl]carbamoyloxy]piperazine-1-carboxylate (200 mg, 1.00 eq) in THF (2 mL) was added Pd(PPh₃)₄ (53.7 mg, 0.20 eq) and 1,3-dimethylhexahydropyrimidine-2,4,6- trione (218 mg, 1.39 mmol, 6.00 eq). The mixture was stirred at 20 °C for 1 hr under N₂ atmosphere. LC-MS showed Reactant 1 was consumed completely and 32% desired compound was detected. The mixture was concentrated to give a residue. The residue was purified by prep- _{HPLC (neutral condition; column: WePure Biotech XP tC18 150*40* mobile phase:} [H₂O(10mM NH₄HCO₃)-ACN]; gradient: 35%-75% B over 8.0 min). Compound [(10S,23S)-23- (tert-butoxycarbonylamino)-10-ethyl-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- 78 PATENT TT3-003WO diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] 3,3-dimethyl-4-(piperazin-1-yloxycarbonylamino)butanoate (150 mg, 83.1% yield) was obtained as a yellow solid. _{General procedure for preparation of compound 6A} O S O To a solution of 4-[3-(p-tolylsulfonyl)-2-(p-tolylsulfonylmethyl)propanoyl]benzoic acid (99.2 mg, 2.20 eq) in DCM (3 mL) was added ethyl 2-ethoxy-2H-quinoline-1-carboxylate (24.5 mg, 1.10 eq) at 20 °C. The mixture was stirred at 20 °C for 1 hr. Then [(10S,23S)-23- (tert-butoxycarbonylamino)-10-ethyl-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] 3,3-dimethyl-4-(piperazin-1-yloxycarbonylamino)butanoate (70.0 mg, 1.00 eq) was added. The mixture was stirred at 20 °C for 1 hr. LC-MS showed the reaction complete. The mixture was concentrated to give a residue. The residue was purified by prep-HPLC (neutral _{condition,column: WePure Biotech XP tC18150*40* mobile phase: [H2O(10mM NH4HCO3)-} ACN]; gradient: 50%-85% B over 8.0 min). Compound [(10S,23S)-23-(tert-butoxycarbonylamino)- 10-ethyl-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] 3,3-dimethyl-4-[[4-[4-[3-(p-tolylsulfonyl)-2-(p- tolylsulfonylmethyl)propanoyl]benzoyl]piperazin-1-yl]oxycarbonylamino]butanoate (37.0 mg, 28.8 32.0% yield, 98.0% purity) was obtained as a yellow solid. _{General procedure for preparation of compound Target 5B} 79 PATENT TT3-003WO dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-10-yl] 3,3-dimethyl-4-[[4-[4-[3-(p-tolylsulfonyl)-2-(p- tolylsulfonylmethyl)propanoyl]benzoyl]piperazin-1-yl]oxycarbonylamino]butanoate (25.0 mg, 1.00 eq) in DCM (2 mL) was added HCl/EtOAc (4 M, 1.00 mL, 202 eq). The mixture was stirred at 20 °C for 12 hrs. LC-MS showed Reactant 1 was consumed completely and 83% desired compound was detected. The mixture was concentrated to give a residue. The residue was purified by prep-HPLC (FA condition,column: Phenomenex luna C18100*40mm* mobile phase: [H₂O(0.2% FA)-ACN]; gradient: 30%-60% B over 8.0 min,LCMS: ET95369-50- P1A2). Compound [(10S,23S)-23-amino-10-ethyl-18-fluoro-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 10-yl] 3,3-dimethyl-4-[[4-[4-[3-(p-tolylsulfonyl)-2-(p- tolylsulfonylmethyl)propanoyl]benzoyl]piperazin-1-yl]oxycarbonylamino]butanoate (5.05 mg, 19.0% yield, 90.1% purity, FA) was obtained as a white solid. Spectrum: 1H NMR HNMR-ET95369-50-P1B, CD₃CN 400MHz - 7.65 (m, 5H), 7.32 - 7.46 (m, 8H), 7.16 (br d, J = 5.75 Hz, 1H), 7.03 (s, 1H), 5.27 - 5.64 (m, 4H), 4.37 (br d, J = 4.13 Hz, 1H), 3.92 - 4.11 (m, 1H), 3.62 - 3.73 (m, 2H), 3.48 - 3.54 (m, 2H), 3.10 - 3.30 (m, 6H), 2.77 - 3.05 (m, 2H), 2.43 - 2.48 (m, 10H), 2.38 (s, 3H), 2.11 (dt, J = 5.00, 2.50 Hz, 4H), 1.77 (t, J = 2.44 Hz, 2H), 1.05 (d, J = 6.25 Hz, 6H), 0.98 (t, J = 7.38 Hz, 3H) LCMS LCMS-ET95369-50-P1Q4: m/z 1159.4 (M+1) SFC SFC-ET95369-50-P1A: de% = 100% Procedures for the preparation of conjugate 76 80 PATENT TT3-003WO G_{eneral procedure for preparation of compound 1} F Boc F N O H To a solution of (10S,23S)-23-amino-10-ethyl-18-fluoro-10-hydroxy-19-methyl-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)- heptaene-5,9-dione (2.00 g, 4.59 mmol, 1.00 eq) in DMF (80 mL) were added Et₃N (930 mg, 9.19 mmol, 1.28 mL, 2.00 eq) and a solution of tert-butyl 4-(4- nitrophenoxy)carbonyloxypiperazine-1-carboxylate (1.67 g, 4.55 mmol, 0.99 eq) in THF (16 mL) at 20 °C, the reaction mixture was stirred at 20 °C for 30 min. LCMS showed the reaction complete. The reaction mixture was concentrated to remove THF. The residue was purified by p_{rep-HPLC (neutral condition, column: Waters Xbridge BEH C18250*70mm* mobile} 81 PATENT TT3-003WO phase: [H₂O(10mM NH₄HCO₃)-ACN]; gradient: 30%-70% B over 18.0 min). Compound tert-butyl 4-[[(10S,23S)-10-ethyl-18-fluoro-10-hydroxy-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 23-yl]carbamoyloxy]piperazine-1-carboxylate (1.50 g, 2.26 mmol, 49.2% yield) was obtained as a yellow solid. _{General procedure for preparation of compound 2} Boc F F N O HN O To a solution of tert-butyl 4-[[(10S,23S)-10-ethyl-18-fluoro-10-hydroxy-19-methyl-5,9-dioxo-8- oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-23-yl]carbamoyloxy]piperazine-1-carboxylate (100 mg, 151 1.00 eq) 13.3 eq). The mixture was stirred at 25 °C for 15 min under N₂. LCMS showed 16% of Reactant 1 was remaining and 70% peak with desired mass was detected. The reaction mixture was concentrated to give a residue. The residue was purified by prep-HPLC (TFA, column: WePure Biotech XP tC18 _{100*30* mobile phase: [H2O(0.1% TFA)-ACN]; gradient: 10%-45% B over 8.0} min).LCMS(ET96034-89-P1A),HPLC(ET96034-89-P1A1).piperazin-1-yl N-[(10S,23S)-10-ethyl- 18-fluoro-10-hydroxy-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 23-yl]carbamate (50.0 mg, 49.0% yield, TFA) was obtained as a yellow solid. _{General procedure for preparation of compound Target 6B} 82 PATENT TT3-003WO To a solution of 4-[3-(p-tolylsulfonyl)-2-(p-tolylsulfonylmethyl)propanoyl]benzoic acid (25.9 mg, 1.00 eq) in DCM (3.5 mL) was added EEDQ (14.1 mg, 1.10 eq) and the reaction mixture was stirred at 20 °C for 1 hr under N₂. Then NMM (13.1 mg, 14.27 2.51 eq) and piperazin-1-yl N-[(10S,23S)-10-ethyl-18-fluoro-10-hydroxy-19-methyl-5,9- dioxo-8-oxa-4,15-diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa- 1,6(11),12,14,16,18,20(24)-heptaen-23-yl]carbamate (35.0 mg, 1.00 eq, TFA) was added, the reaction mixture was stirred at 20 °C for 1 hr under N₂. LCMS showed the reaction complete. The reaction mixture was concentrated to remove DCM. The reaction mixture was purified by prep-HPLC(neutral condition, column : WePure Biotech XP tC18150*40* mobile phase: [H₂O(10mM NH₄HCO₃)-ACN]; gradient: 33%-63% B over 8.0 min). Compound [4- [4-[3-(p-tolylsulfonyl)-2-(p-tolylsulfonylmethyl)propanoyl]benzoyl]piperazin-1-yl] N-[(10S,23S)-10- ethyl-18-fluoro-10-hydroxy-19-methyl-5,9-dioxo-8-oxa-4,15- diazahexacyclo[14.7.1.02,14.04,13.06,11.020,24]tetracosa-1,6(11),12,14,16,18,20(24)-heptaen- 23-yl]carbamate (8.30 mg, 15.1% yield, 98.5% purity) was obtained as a green solid. Spectrum: 1H NMR HNMR-ET96034-101-P1P1, DMSO-d₆400MHz J = 7.25 Hz, 3H) 1.84 (tt,J = 13.90,6.86 Hz, 2H) 2.12 - 2.23 (m, 1H) 2.25 – 2.30 (m, 1H) 2.39 (s, 3H) 2.44 (d, J = 3.13 Hz, 4H) 2.81 - 2.98 (m, 2H) 3.05 – 3.21 (m, 2H) 3.25 – 3.30 (m, 2H) 3.42 - 3.57 (m, 3H) 3.66- 3.76 (m, 2H) 3.79 - 3.89 (m, 2H) 3.96 – 4.04 (m, 1H) 4.30 – _{4.46 (m, 1H) 5.17 – 5.39 (m, 4H) 7.32 (s, 1H) 7.43 (dd, J = 8.13,4.25 Hz, 3H) 7.48 (s, 3H) 7.57} (dd, 2H) 7.80 (d, J = 10.88 Hz, 1H) 8.27 – 8.39 (m, 1H) LCMS LCMS-ET96034-101-P1P: m/z 1046.2 (M+1) 83 PATENT TT3-003WO SFC SFC-ET96034-101-P1P1: de% = 100% Procedures for conjugation Reagent preparation 1. DPBS: Gibco, Cat#14190136 2. TCEP: Sigma,Cat#646547 and dissolved in PBS 3. DMSO: Sigma, Cat#D2650 4. 30KDa cutoff membrane: cobetter, ULRC0300040P 5. 0.22 um membrane: Millipore, UFC40GV0S 6. Trastuzumab: Expressed in CHO cell, Purified by Protein A and exchange buffer to PBS. 7. Target: Dissolve the dry powder in DMSO to a concentration of 10 mg/ml. ADC-3: Trastuzumab+Target3 conjugation Reduction _{A 1.5 mL tube Put 52.8 uL DPBS in a 1.5 mL tube then take 127.7 uL trastuzumab with a concentration of 7.83 mg/mL into the 1.5 mL tube the pH was 7.4, followed by 19.44 uL 5 mmol/L TCEP. Finally, the concentration of antibody is 5.00 mg/mL in the solution the molar ratio of TCEP and antibody was 14:1. The reaction was carried out for 2 hours at 25 .} Conjugation After the reduction reaction, 8.4 uL Target3 with concentration of 10 mg/ml was added and _{vortexed. In this solution, the volume ratio of DMSO was 10% and the molar ratio of linker payload and antibody was 16:1. The reaction was carried out at 25 for 1 hour.} Purification 84 PATENT TT3-003WO _{After conjugation, the solution was purified by ultrafiltration using 30KDa cutoff membrane to} remove the free drug and exchange the buffer to 10 mM NaAc-HAc, pH 5.0. Finally, the solution was filtered by 0.22 um membrane, then QC analysis was carried out. DAR measurement Percentage Weighted peak area Drug Peak Total ADC-4: Trastuzumab+Target4B conjugation Reduction _{Put 222.5 uL DPBS in a 1.5 mL tube then take 510.9 uL trastuzumab with a concentration of 7.83 mg/ml into the 1.5 mL tube the pH was 8.0, followed by 66.67 uL 5 mmol/L TCEP. Finally, the concentration of antibody is 5.00 mg/mL in the solution the molar ratio of TCEP and antibody was 12:1. The reaction was carried out for 2 hours at 25 .} Conjugation 85 PATENT TT3-003WO After the reduction reaction, 125.9 uL Target4B with concentration of 10 mg/mL was added and _{vortexed. In this solution, the volume ratio of DMSO was 13.5% and the molar ratio of linker payload and antibody was 40:1. The reaction was carried out at 25 for 1 hour.} Purification _{After conjugation, the solution was purified by ultrafiltration using 30KDa cutoff membrane to} remove the free drug and exchange the buffer to 10 mM NaAc-HAc, pH5.0. Finally, the solution was filtered by 0.22 um membrane, then QC analysis was carried out. DAR measurement Percentage Weighted peak area Drug Total ADC-5: Trastuzumab+Target5B conjugation Reduction 86 PATENT TT3-003WO _{Put 222.5 uL DPBS in a 1.5 mL tube then take 510.9 uL trastuzumab with a concentration of 7.83 mg/mL into the 1.5 mL tube the pH was 8.0, followed by 66.67 uL 5 mmol/L TCEP. Finally, the concentration of antibody is 5.00 mg/mL in the solution the molar ratio of TCEP and antibody was 12:1. The reaction was carried out for 2 hours at 25 .} Conjugation After the reduction reaction, 128.8 uL Target5B with concentration of 10 mg/mL was added and _{vortexed. In this solution, the volume ratio of DMSO was 13.7% and the molar ratio of linker payload and antibody was 40:1. The reaction was carried out at 25 for 1 hour.} Purification _{After conjugation, the solution was purified by ultrafiltration using 30KDa cutoff membrane to} remove the free drug and exchange the buffer to 10 mM NaAc-HAc, pH5.0. Finally, the solution was filtered by 0.22 um membrane, then QC analysis was carried out. DAR measurement Weighted peak area Drug Percentage 87 PATENT TT3-003WO HHLL+4d 4 858236 18.203% 0.728 Reduction _{Put 222.5 uL DPBS in a 1.5 mL tube then take 510.9 ul trastuzumab with a concentration of 7.83 mg/mL into the 1.5 mL tube the pH was 8.0, followed by 66.67 uL 5 mmol/L TCEP.} Finally, the concentration of antibody is 5.00 mg/mL in the solution, the molar ratio of TCEP and antibody was 12 1. The reaction was carried out for 2 hours at 25 . Conjugation After the reduction reaction, 69.7 uL DMSO was added and vortexed, then followed by 69.7 uL Target6B with concentration of 10 mg/mL. In this solution, the volume ratio of DMSO was 10% and the molar ratio of linker payload and antibody was 24:1. The reaction was carried out at _{25 for 1 hour.} Purification _{After conjugation, the solution was purified by ultrafiltration using 30KDa cutoff membrane to} remove the free drug and exchange the buffer to 10 mM NaAc-HAc, pH5.0. Finally, the solution was filtered by 0.22um membrane, then QC analysis was carried out. DAR measurement Percentage Weighted peak area 88 PATENT TT3-003WO HL+1d+270 1 128467 4.090% 0.041 Pharmaceutical Compositions [00160] Embodiments include a pharmaceutical composition for the administration to a subject. The pharmaceutical composition disclosed herein may further include a pharmaceutically acceptable carrier, excipient, or diluent. As used herein, the term “pharmaceutically acceptable” means that the composition is sufficient to achieve the therapeutic effects without deleterious side effects, and may be readily determined depending on the type of the diseases, the patient’s age, body weight, health conditions, gender, and drug sensitivity, administration route, administration mode, administration frequency, duration of treatment, drugs used in combination or coincident with the composition disclosed herein, and other factors known in medicine. [00161] The pharmaceutical composition can also include a pharmaceutically acceptable carrier. For oral administration, the carrier can include, for example, a binder, a lubricant, a disintegrant, an excipient, a solubilizer, a dispersing agent, a stabilizer, a suspending agent, a colorant, and a flavorant. For injectable preparations, the carrier can include a buffering agent, a preserving agent, an analgesic, a solubilizer, an isotonic agent, and a stabilizer. For preparations for topical administration, the carrier can include a base, an excipient, a lubricant and a preserving agent. 89 PATENT TT3-003WO [00162] The disclosed compositions can be formulated into a variety of dosage forms in combination with the aforementioned pharmaceutically acceptable carriers. For example, for oral administration, the pharmaceutical composition can be formulated into tablets, troches, capsules, elixirs, suspensions, syrups or wafers. For injectable preparations, the pharmaceutical composition can be formulated into an ampule as a single dosage form or a multidose container. The pharmaceutical composition can also be formulated into solutions, suspensions, tablets, pills, capsules and long-acting preparations. [00163] The administration dose and frequency of the pharmaceutical composition disclosed herein are determined by the type of active ingredient, together with various factors such as the disease to be treated, administration route, patient’s age, gender, and body weight, and disease severity. [00164] The total effective dose of the compositions disclosed herein may be administered to a patient in a single dose or may be administered for a long period of time in multiple doses according to a fractionated treatment protocol. In the pharmaceutical composition disclosed herein, the content of active ingredient may vary depending on the disease severity. Preferably, the total daily dose of the peptide disclosed herein may be approximately 0.0001㎍ to 500 mg per 1 kg of body weight of a patient. However, the effective dose of the peptide is determined considering various factors including patient’s age, body weight, health conditions, gender, disease severity, diet, and secretion rate, in addition to administration route and treatment frequency of the pharmaceutical composition. In view of this, those skilled in the art may easily determine an effective dose suitable for the particular use of the pharmaceutical composition disclosed herein. The pharmaceutical composition disclosed herein is not particularly limited to the formulation, and administration route and mode, as long as it shows suitable effects. 90 PATENT TT3-003WO [00165] Moreover, the pharmaceutical composition may be administered alone or in combination or coincident with other pharmaceutical formulations showing prophylactic or therapeutic efficacy. [00166] Given the teachings and guidance provided herein, those skilled in the art will understand that a formulation described herein can be equally applicable to many types of biopharmaceuticals, including those exemplified, as well as others known in the art. Given the teachings and guidance provided herein, those skilled in the art also will understand that the selection of, for example, type(s) or and/or amount(s) of one or more excipients, surfactants and/or optional components can be made based on the chemical and functional compatibility with the biopharmaceutical to be formulated and/or the mode of administration as well as other chemical, functional, physiological and/or medical factors well known in the art. [00167] A pharmaceutical composition or cancer therapeutic is administered to an individual. An individual is typically a human being, but can be an animal, including, but not limited to, dogs, cats, birds, cattle, horses, sheep, goats, reptiles and other animals, whether domesticated or not. Typically, any individual who is a candidate for treatment is a candidate with some form of cancer, whether the cancer is benign or malignant, a tumor, solid or otherwise, a cancer cell not located in a tumor or some other form of cancer. Among the most common types of cancer include, but are not limited to, bladder cancer, breast cancer, colon and rectal cancer, endometrial cancer, kidney cancer, renal cancer, leukemia, lung cancer, melanoma, non-Hodgkins lymphoma, pancreatic cancer, prostate cancer, stomach cancer and thyroid cancer. Pre-operative evaluation typically includes routine history and physical examination in addition to thorough informed consent disclosing all relevant risks and benefits of the procedure. [00168] It is understood that a specific dose level for any particular patient will vary depending upon a variety of factors, including the activity of the specific active agent; the age, body weight, general health, sex and diet of the patient; the time of administration; the rate of excretion; possible drug combinations; the severity of the 91 PATENT TT3-003WO particular condition being treated; the area to be treated and the form of administration. One of ordinary skill in the art would appreciate the variability of such factors and would be able to establish specific dose levels using no more than routine experimentation. [00169] Pharmacokinetic parameters such as bioavailability, absorption rate constant, apparent volume of distribution, unbound fraction, total clearance, fraction excreted unchanged, first-pass metabolism, elimination rate constant, half-life, and mean residence time can be determined by methods well known in the art. [00170] If desired, other therapeutic agents can be employed in conjunction with those provided in the above-described compositions. The amount of active ingredients that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated, the nature of the disease, disorder, or condition, and the nature of the active ingredients. 92

Claims

_{PATENT TT3-003WO} CLAIMS What is claimed is: _{1. A compound of Formula (I):} D – L – X or a pharmaceutically acceptable salt thereof, wherein: D is an N-linked drug moiety or an O-linked drug moiety; L is a bond, a C1 – C12 alkylene, or a 3 – 12 membered heteroalkylene; X is -C(=O)O-R¹; and R¹ is an optionally substituted 4 – 12 membered heterocyclyl. _{2. The compound of claim 1 a pharmaceutically acceptable salt thereof, wherein the} compound is Formula (II): wherein: Y¹ is NH, O, S, or N linked to a C1-C10 straight or branched alkyl, Y² is NH, O, S, or N linked to a C1-C10 straight or branched alkyl, Z is O, or S, NH or N linked to a C1-C10 straight or branched alkyl, R¹, R², R³, R⁴, R⁵, R⁶ are independently H, C1-C9 straight or branched alkyl, W is O, S, NH or N linked to a C1-C10 straight or branched alkyl, and n is 0, 1, or 2. 93 PATENT TT3-003WO 3. The compound of claim 1 a pharmaceutically acceptable salt thereof, wherein the compound is Formula (III): wherein: wherein D is an N-linked Drug Moiety or an O-linked Drug Moiety; L is a bond, a C1 – C12 alkylene, or a 3 – 12 membered heteroalkylene; Y¹ – Y⁵ comprise a carboxycylic or heterocyclic ring, Y¹, Y², Y⁴, Y⁵ are CR¹⁰, R¹⁰ is H, a halide or C1-C10 straight or branched alkyl, and Y³ is CR¹⁰, O, S(O), SO2, CONH, SO2 -alkyl or a substituted alkyl. 4. The compound of claim 1 a pharmaceutically acceptable salt thereof, wherein the compound is Formula (IV): wherein: wherein D is an N-linked Drug Moiety or an O-linked Drug Moiety; L is a bond, a C1 – C12 alkylene, or a 3 – 12 membered heteroalkylene; Y¹, Y², Y⁴, Y⁵ are CR¹⁰, Y³ is O, CR¹⁰, S(O), SO2, CONH, SO2 -alkyl or a substituted alkyl, NS(O₂)N(CH₃)₂, 94 PATENT TT3-003WO R¹⁰ is H, a halide or C1-C10 straight or branched alkyl, CF3, OCF3, (CH3)OH, (CH3)NHCH2(OH)OC(CH3)3, (CH3)CO(CH3), (F)CO(OH), NHS(O)₂N(CH₃)₂, or CONHS(O)₂N(CH₃)₂, and optionally, wherein Y¹ and Y⁵ form a 5-membered ring. 5. A compound of Formula (I): D – L – X wherein D is a payload, X is a cage group and L is a linker selected from L1, L2, L3, L4, L5, L6, L7, L8, L9, L10, L11, L12, L13, L14, L15, L16, L17, L18, L19, L20, L21, L22, L23, L24, L25, L26, L27, L28 and L29: PATENT TT3-003WO 6. The compound of claim 5, wherein L is a linker and X is selected from C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, C30, C31, C32, C33, C34, C35 and C36: 96 PATENT TT3-003WO PATENT TT3-003WO 7. The compound of claim 5, wherein the compound is selected from Formula A, Formula B, Formula C and Formula D: 98 PATENT TT3-003WO F F H N OH 8. The compound of claim 1, wherein the drug moiety is an anti-neoplastic compound. 9. The compound of claim 5, wherein the payload is an anti-neoplastic compound. 10. The compound of claim 8 or 9, wherein the anti-neoplastic compound selected from exatecan, pimasertib, rucaparib, ASN007, Imatinib (Gleevec), Gefitinib (Iressa), Erlotinib (Tarceva), Sunitinib (sutent), Lapatinib (Tykerb), Nilotinib (Tasigna), Sorafenib (Nexavar), Temsirolimus (CCI-779), Everolimus (afinitor), Pazopanib (Votrient), Crizotinib (Xalkori), Ruxolitinib (jafaki), Vandetenib (Caprelsa) Axitinib (Inlyta), Bosutinib (Bosulif), Cabozantinib (Cometriq), Ponatinib (Iclusig), Regorafenib (Stivagra), Ibrutinib (Imbruvica), Trametinib (Mekinist), Perifosine, Bortezomib (Velcade), Carfilzomib (Kyprolis), Marizomib (NPI-0052), Batimastat (BB-94), Neovastat (AE-941), Prinomastat 99 PATENT TT3-003WO (AG-3340), Rebimastat (BMS-275291), Ganetespib, NVP-AUY922, Marimastat (BB- 2516), Obatoclax (GX15-070) and Navitoclax(ABT-263). 11. The compound of claim 1, wherein the drug moiety is an antibody or antibody fragment. 12. The compound of claim 5, wherein the payload is an antibody or antibody fragment. 13. The compound of claim 11 or 12, wherein the antibody is selected from Traztuzumab, Sacituzumab, Enfortumab, Mirvetuximab, Datopotamab, Samrotamab, Zolbetuximab, Sigvotagtug. 14. The compound of claim 1, wherein the drug moiety is an ERK inhibitor. 15. The compound of claim 5, wherein the payload is an ERK inhibitor. 16. The compound of claim 14 or 15, wherein the ERK inhibitor is selected from KO- 947 and Temuterkib®. 17. A method of treating an ailment, the method comprised of administering a therapeutic amount of the compound of claim 1, 2, 3, 4, 5, 6, 7, 8, 10, 11 or 12 to a subject. 18. The method of claim 17, wherein the ailment is associated with a cell or organism having an increased Fe^ll level. 19. The method of claim 17, wherein the ailment is cancer. 20. The method of claim 19, wherein the cancer is pancreatic cancer, lung cancer, colorectal cancer, thyroid cancer, testicle cancer, melanoma, bladder cancer, liver cancer, kidney cancer, myelodysplastic syndrome or leukemia. 100 PATENT TT3-003WO 21. The method of claim 19, further comprising administering one or more additional cancer therapies. 22. The method of claim 21, wherein the one or more additional cancer therapies is chemotherapy, radiation therapy, immunotherapy, surgery or hormone therapy. 23. The method of claim 21, wherein the one or more additional cancer therapies is a chemotherapeutic reagent is an alkylating agent. 24. The method of claim 21, wherein the one or more additional cancer therapies is a TOP1 inhibitor or a cytotoxic antibiotic such as doxorubicin, mitoxantrone, bleomycin, actinomycin and mitomycin. 25. The method of claim 21, wherein the one or more additional cancer therapies is selected from anti-CTLA-4 monoclonal antibodies, anti-PD-1 and PD-L1 monoclonal antibodies (e.g., immune checkpoint inhibitors), antibiotics/antineoplastics, proteasome inhibitors and VEGF/VEGFR inhibitors. 26. A method of treating an ailment associated with a cell or organism having an increased Fe^II level compared to a control, the method comprising administering an effective amount of the compound of claim 1, 2, 3, 4, 5, 6, 7, 8, 10, 11 or 12 to the subject. 101