Antibody-drug conjugates

Abstract

Antibody-drug conjugate compounds that include linkers, and methods of using such compounds, are provided.

Description

[Technical Field]

[0001] Provided herein are novel protein (e.g., antibody) drug conjugates comprising hydrophilic solubilizing groups and / or linkers comprising hydrophilic solubilizing groups, as well as methods of treating diseases, disorders, and conditions comprising administering protein drug conjugates comprising hydrophilic solubilizing groups and / or linkers thereof. [Background technology]

[0002] Antibody-drug conjugates (ADCs) are antibodies operably linked to biologically active small molecules (also called toxins or payloads). ADCs selectively deliver potent payloads to target-expressing cells, potentially reducing off-target side effects and / or toxicity and improving the therapeutic index. The lipophilicity of many payloads (i.e., drugs) can adversely affect the properties of ADCs, resulting in inefficient delivery of the payload to target cells. The low bioavailability of lipophilic payloads can narrow the therapeutic window of ADC therapy. Furthermore, the hydrophobicity of the payload can pose a challenge to its conjugation to antibodies, a reaction that proceeds under aqueous conditions. Therefore, there is a continuing need for the development of hydrophilic linkers for protein conjugates (e.g., ADCs) that enable the conjugation of lipophilic payloads, improved modulation of biological targets, increased bioavailability, and improved therapeutic windows.

[0003] Monoclonal antibody (mAb) therapy is gaining increasing acceptance as both adjuvant and first-line cancer therapy. The success of mAb therapies to treat colon cancer with AVASTIN® (anti-VEGF), non-Hodgkin's lymphoma with RITUXAN® (rituximab, anti-CD20), and breast adenocarcinoma with HERCEPTIN® (anti-Her2) demonstrates that unconjugated antibodies improve patient survival without significantly increasing the incidence of toxicity.

[0004] Monoclonal antibodies can be conjugated to therapeutic agents to form antibody-drug conjugates. For example, the HERCEPTIN® antibody described above is conjugated with a mertansine payload to form the ADC KADCYLA®. ADCs can exhibit higher efficacy compared to unconjugated antibodies. The conjugation between the antibody and the drug can be direct or indirect via a linker. The linker can be cleavable or non-cleavable. One factor considered important for developing efficient and well-tolerated ADCs is the composition and stability of the linker. For certain ADCs, the linker ideally provides serum stability while selectively releasing the drug within target cells.

[0005] Linking of the linker to the mAb can be achieved in a variety of ways, including via surface lysines, reductive coupling with oxidized carbohydrates, and cysteine ​​residues released from reduced interchain disulfide bonds. Various ADC conjugation systems have been described in the literature, including hydrazone-, disulfide-, and peptide-based linkages. Some hydrazone- and disulfide-based linkers can be unstable in the circulation, resulting in unwanted release of the drug outside of the target tissue. This premature drug release may result in systemic or organ-specific toxicity and / or less than ideal therapeutic efficacy. While peptide-based linker strategies can provide more stable linkers, the associated increased hydrophobicity of some linkers can lead to aggregation, particularly of highly hydrophobic drugs. Such aggregation can lead to a number of adverse effects, including ADC precipitation, administration difficulties, and nonspecific uptake of the ADC into non-target tissues, potentially affecting non-target toxicity and reducing efficacy.

[0006] Exatecan is a drug that is a structural analog of camptothecin and has antitumor activity. See Abou-Alfa et al., "Randomized Phase III Study of Exatecan and Gemcitabine Compared with Gemcitabine Alone in Untreated Advanced Pancreatic Cancer," Journal of Clinical Oncology, 24(27): 4441-7, September 20, 2006. Monomethyl auristatin E (MMAE) is a synthetic antitumor drug. Due to its toxicity, it cannot be used as a medicine by itself. MMAE is actually desmethyl auristatin E, i.e., the N-terminal amino group has only one methyl substituent, rather than two as in auristatin E itself. See Dosio et al., "Immunotoxins and Anticancer Drug Conjugate Assemblies: The Role of the Linkage between Components," Toxins. 3(12): 848-883, 2011.

[0007] In the field of small molecule therapeutics, strategies have been developed to provide prodrugs of active chemicals. Such prodrugs are administered in a relatively inactive (or significantly less active) form. Once administered, the prodrug is metabolized in vivo to the active compound. Such prodrug strategies can improve the selectivity of the drug for its intended target and reduce side effects.

[0008] Thus, there remains a need for targeted delivery of toxins to eliminate target cells while reducing toxicity to non-target cells. Furthermore, some antibodies, such as Patritumab, exhibit visually observable aggregation during rapid buffer exchange. The aggregation temperature (Tagg) detected by dynamic light scattering (DLS) and the tendency for self-association detected by AC-SINS analysis are both inferior to those of well-performing mAbs. Patritumab's tendency to aggregate leads to the aggregation of the corresponding ADC. Summary of the Invention [Problem to be solved by the invention]

[0009] Thus, there is an unmet medical need to produce ADCs with linker systems that provide a high level of linker serum stability and increased solubility, thereby allowing efficient conjugation of hydrophobic drugs and achieving intracellular delivery of the drugs. [Means for solving the problem]

[0010] Provided herein are ADCs having linker systems that offer a high level of linker serum stability and increased solubility, thereby enabling efficient conjugation of hydrophobic drugs and achieving intracellular delivery of the drugs.

[0011] In one embodiment, the ADC is a compound of formula (I): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, wherein the variables are as defined herein. In one embodiment, the ADC comprises one or more hydrophilic residues.

[0012] In another embodiment, the ADC is a compound of formula (Ia): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, wherein the variables are as defined herein.

[0013] In one embodiment, the ADC is a compound of formula (Ib): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, wherein the variables are as defined herein.

[0014] In another embodiment, the present specification describes a method of treating a disease, disorder, or condition in a patient in need thereof, comprising administering to the patient a compound described herein. Further provided is the use of a compound described herein for treating a disease, disorder, or condition described herein. Further provided is the use of a compound described herein for use in the manufacture of a medicament for treating a disease, disorder, or condition described herein. In some embodiments, the compound is an antibody-drug conjugate. In another embodiment, the present specification describes a method for preparing an antibody-drug conjugate, comprising contacting a binding agent with a linker-payload compound described herein under conditions suitable for forming a bond between the binding agent and the linker-payload compound. [Brief explanation of the drawings]

[0015] [Figure 1]The human plasma stability profile of ADC2-B is shown. The ADC2-B_Tab curve shows the ratio of the antibody concentration at different times to its concentration at TO (the so-called total antibody curve). This includes the non-payload-conjugated antibody and payload-conjugated antibody captured by the antigen (HER2) in the plasma. The ADC2-B_ADC curve shows the ratio of the drug-conjugated antibody ADC2-B concentration at different times to its concentration at TO (the so-called ADC curve). The ADC2-B_Payload curve shows the comparison of the payload release rate at different times to its concentration at TO (the so-called payload release curve). The Dxd release rate of ADC2-B is less than 5% at 168 h. ADC2-B does not undergo the maleimide ring-opening process. [Figure 2] The human plasma stability profile of ADC2-40 is shown. ADC2-40_Tab is the total antibody curve. This includes the unpayloaded and payload-bound antibody captured by the antigen (HER2) in plasma. ADC2-40_ADC is the ADC curve. ADC2-40_payload curve is the payload release curve. ADC2-40 has undergone a maleimide ring-opening process (ring opening) and has a P5 modification. The payload release curve shows less free payload release (<2%) than ADC2-B, indicating good linker stability. Furthermore, as the maleimide ring opens, the ADC curve closely matches the Tab curve, indicating fewer deconjugation events occur. [Figure 3] Figure 1 shows the human plasma stability profile of ADC2-32. The ADC2-32_Tab curve is the whole antibody curve. The ADC2-32_ADC curve is the ADC curve. The ADC2-32_payload curve is the payload release curve. ADC2-32 does not have ring opening but does have a P5 modification. ADC2-32 exhibits a lower payload release rate (<2%) at T168h than ADC2-B. [Figure 4]Figure 2 shows the human plasma stability profile of ADC2-42. The ADC2-42_Tab curve is the whole antibody curve. The ADC2-42_ADC curve is the ADC curve. The ADC2-42_payload curve is the payload release curve. ADC2-42 has ring-opening and P5 modification. ADC2-42 is similar to ADC2-40, with the ADC and Tab curves nearly identical and a lower payload release rate (e.g., T168h) than ADC2-B. Figures 2 and 4 illustrate that the antibody and ADC curves are nearly identical, respectively, indicating that the human plasma stability of ADC2-40 and ADC2-42 is consistent with that of the corresponding antibodies. [Figure 5] Figure 5 shows the human plasma stability profiles of ADC2-B, ADC2-32, ADC2-40, and ADC2-42, along with the corresponding payload release curves. ADC2-B does not have ring opening. ADC2-32 does not have ring opening but contains the corresponding P5 modification. ADC2-40 and ADC2-42 are ring-opened and contain the corresponding P5 modification. Figure 5 shows that in human plasma, the free payload release of the maleimide ring-opened ADCs is lower than that of the unopened ADCs. Under the P5 modification, ADC2-32 releases less payload than ADC2-B. [Figure 6] Figure 1 shows rat PK profiles based on concentration curves of whole antibodies (including antibodies with and without payload conjugates) for ADC2-B, ADC2-32, ADC2-40, and ADC2-42. ADC2-B and ADC2-32 are not ring-opened but contain the corresponding P5 modifications. ADC2-40_Tab and ADC2-42_Tab are ring-opened and contain the corresponding P5 modifications. The rat PK profiles based on antibody concentration are similar for both open and non-open ADCs. [Figure 7]Figure 1 shows rat PK profiles based on ADC (payload-bound antibody) concentration curves for ADC2-B, ADC2-32, ADC2-40, and ADC2-42. ADC2-B and ADC2-32 are not ring-opened but contain the corresponding P5 modifications. ADC2-40 and ADC2-42 are ring-opened and contain the corresponding P5 modifications. The rat PK profiles based on ADC concentration show that the elimination rate of the ring-opened ADCs is slower than that of the non-ring-opened ADCs. [Figure 8] 1 shows the rat PK profile of ADC2-B. The ADC2-B_ADC curve shows the concentration of ADC (payload-bound antibody). The ADC2-B_Tab total antibody curve shows the concentration of total antibody, including non-payload-bound antibody and payload-bound antibody. ADC2-B is not open-circuited. [Figure 9] Figure 9 shows the rat PK profile of ADC2-40. The ADC2-40_ADC curve shows the concentration of ADC (payload-conjugated antibody). ADC2-40_Tab shows the total antibody curve, which shows the concentration of all antibodies, including non-payload-conjugated and payload-conjugated antibodies. ADC2-40 is open-chain but contains the corresponding P5 modification. Figure 9 shows that the curves for ADC2-40_ADC and ADC2-40_Tab are nearly identical, indicating that the PK of the open-chain ADC is driven by the PK of the corresponding antibody. [Figure 10] Figure 1 shows the rat PK profile of ADC2-32. The ADC2-32_ADC curve shows the concentration of ADC (payload-bound antibody). The ADC2-32_Tab total antibody curve shows the concentration of total antibody, including non-payload-bound antibody and payload-bound antibody. ADC2-32 is not ring-opened but contains the corresponding P5 modification. [Figure 11] Figure 11 shows the rat PK profile of ADC2-42. The ADC2-42_ADC curve shows the concentration of ADC (payload-bound antibody). The ADC2-42_Tab total antibody curve shows the concentration of total antibody, including non-payload-bound antibody and payload-bound antibody. ADC2-42 is open-chain and contains the corresponding P5 modification. Figure 11 shows that the curves for ADC2-42_ADC and ADC2-42_Tab are nearly identical, indicating that the PK of the open-chain ADC is driven by the PK of the corresponding antibody. [Figure 12] Figure 1 shows the percent MMAE release of ADC2-A, ADC2-2, ADC2-4, ADC2-11, and ADC2-9-1 in a human plasma stability study. ADC2-A does not contain a P3 modification, while ADC2-2, ADC2-4, ADC2-11, and ADC2-9-1 contain a P3 modification. [Figure 13] Figures 12 and 13 show the percent MMAE release of ADC2-A, ADC2-2, ADC2-4, ADC2-11, and ADC2-9-1 in a mouse plasma stability study. ADC2-A does not contain a P3 modification, while ADC2-2, ADC2-4, ADC2-11, and ADC2-9-1 contain a P3 modification. Figures 12 and 13 show that the P3 modification of MMAE ADCs can reduce cleavage of the VC-PAB linker by the Ces1C enzyme in a mouse system, i.e., MMAE ADCs with at least the P3 modification (e.g., ADC2-2, ADC2-4, ADC2-11, and ADC2-9-1) exhibit reduced early free MMAE release, thereby reducing early free MMAE release in a mouse system. [Figure 14] Figure 14 shows the mouse PK profiles of ADC2-A, ADC2-2, ADC2-4, ADC2-11, and ADC2-9-1. ADC2-A does not contain a P3 modification, while ADC2-2, ADC2-4, ADC2-11, and ADC2-9-1 contain a P3 modification. Figure 14 shows that P3 modification of MMAE ADCs (e.g., ADC2-2, ADC2-4, ADC2-11, and ADC2-9-1) can reduce premature free MMAE release in a mouse system compared to MMAE ADCs without P3 modification (e.g., ADC2-A). In other words, Figure 14 shows that MMAE ADCs without P3 modification (i.e., ADC2-A) have more premature MMAE release compared to MMAE ADCs with P3 modification (e.g., ADC2-2, ADC2-4, ADC2-11, and ADC2-9-1). [Figure 15] 1 shows the hydrophobic interaction chromatography (HIC) profile of ADC 2-6-2 (HIC DAR 8.0). [Figure 16]Figure 1 shows the size exclusion chromatography (SEC) profile of ADC 2-6-2 with purity >98%. [Figure 17] The ADC 2-9-2 HIC profile (HIC DAR 8.0) is shown. [Figure 18] Figure 1 shows the ADC 2-9-2 SEC profile (>99% purity). [Figure 19] The ADC2-36-2 HIC profile (HIC DAR 7.8) is shown. [Figure 20] SEC profile of ADC2-36-2 with purity >99% is shown. [Figure 21] Different cathepsin B-mediated rhodamine release rates are shown. The order of rates is as follows: NAcCys-15~NAcCys-13 (reference)>NAcCys-14>NAcCys-17. [Figure 22] Inhibition of Ovcar3 cell line is shown. [Figure 23] Inhibition of A549 cell line is shown. [Figure 24] 1 shows data for detecting NCI-N87 cell viability. [Figure 25] 1 shows data for detecting NCI-N87 cell viability. [Figure 26] 1 shows data for detecting NCI-N87 cell viability. [Figure 27] 1 shows data for detecting NCI-N87 cell viability. [Figure 28] 1 shows data for detecting NCI-N87 cell viability. [Figure 29] 1 shows data for detecting NCI-N87 cell viability. [Figure 30] 1 shows data for detecting NCI-N87 cell viability. [Figure 31] 1 shows data for detecting NCI-N87 cell viability. [Figure 32] 1 shows data for detecting NCI-N87 cell viability. [Figure 33] Figure 1 shows the % payload release of the ADC in human plasma. [Figure 34]1 shows free MMAE release in mouse PK studies. [Figure 35] 1 shows the ADC DAR changes of ADC2-B, ADC2-32, and ADC2-42 in human plasma. [Figure 36] 1 shows the ADC DAR changes of ADC2-B and ADC2-42 in human plasma. [Figure 37] 1 shows the ADC DAR changes of ADC2-B, ADC2-32, ADC2-40, and ADC2-42 in rat PK. [Figure 38] 1 shows a comparison of HCC1569 cell killing between P5-modified ADCs (i.e., ADC3-2, ADC3-3, ADC3-4, ADC3-6) and ADC3-1 in part A, and between P5-modified ADCs (i.e., ADC3-5, ADC3-7, ADC3-8, ADC3-9, ADC3-10) and ADC3-1 in part B. [Figure 39] 1 shows a comparison of HCC1569 cell killing between P3-modified ADCs (i.e., ADC3-12 and ADC3-15) and ADC3-1 in part A, and between P3-modified ADCs (i.e., ADC3-11 and ADC3-14) and ADC3-1 in part B. [Figure 40] Figure 1 shows the human plasma stability profiles of ADC3-1, ADC3-2, ADC3-3, ADC3-4, ADC3-5, ADC3-6, ADC3-7, ADC3-8, ADC3-9, and ADC3-10, as measured by the corresponding antibody concentration. ADC3-1_TAB, ADC3-2_TAB, ADC3-3_TAB, ADC3-4_TAB, ADC3-5_TAB, ADC3-6_TAB, ADC3-7_TAB, ADC3-8_TAB, ADC3-9_TAB, and ADC3-10_TAB are full antibody curves, showing the ratio of antibody concentration to its TO concentration at different times, including non-payload-bound and payload-bound antibodies, respectively. This figure shows that the full antibody curves of the corresponding P5-modified open-ring ADCs and the ADC curves of the P5-modified non-open-ring ADCs are nearly identical, indicating that the human plasma stability of the P5-modified open-ring ADCs and the P5-modified non-open-ring ADCs, as measured by the concentrations of the corresponding antibodies, is consistent. [Figure 41]Figure 1 shows the human plasma stability profiles of ADC3-1, ADC3-2, ADC3-3, ADC3-4, ADC3-5, ADC3-6, ADC3-7, ADC3-8, ADC3-9, and ADC3-10, as measured by the concentration of the corresponding ADC. ADC3-1_ADC, ADC3-2_ADC, ADC3-3_ADC, ADC3-4_ADC, ADC3-5_ADC, ADC3-6_ADC, ADC3-7_ADC, ADC3-8_ADC, ADC3-9_ADC, and ADC3-10_ADC are ADC curves, which show the ratio of the concentration of the corresponding drug-conjugated antibody at different time points to its concentration at time TO. This figure shows that the P5-modified ring-opened ADCs (ADC3-4, ADC3-5, ADC3-6, ADC3-7, and ADC3-8) have better stability than the P5-modified non-ring-opened ADCs. However, the stability of the P5-modified ring-opened ADCs is similar. The stability of the P5-modified non-ring-opened ADCs is also similar. [Figure 42] 1 shows the human plasma stability profiles, as measured by free payload release, of ADC3-1, ADC3-2, ADC3-3, ADC3-4, ADC3-5, ADC3-6, ADC3-7, ADC3-8, ADC3-9, and ADC3-10. The ADC3-1_ADC_payload, ADC3-2_ADC_payload, ADC3-3_ADC_payload, ADC3-4_ADC_payload, ADC3-5_ADC_payload, ADC3-6_ADC_payload, ADC3-7_ADC_payload, ADC3-8_ADC_payload, ADC3-9_ADC_payload, and ADC3-10_ADC_payload curves are payload release curves, which show the rate of payload release at different time points compared to its concentration at time TO. This figure shows that the free payload concentration in human plasma for all ADCs was less than 3% after 168 hours, indicating that the ADCs have an acceptable plasma stability. The open-ring ADCs release less payload than the non-open-ring ADCs. [Figure 43]Figure 1 shows the human plasma stability profiles of ADC3-1, ADC3-11, ADC3-12, ADC3-13, ADC3-14, and ADC3-15, as measured by the concentration of the corresponding antibody. The ADC3-1_TAB, ADC3-11_TAB, ADC3-12_TAB, ADC3-13_TAB, ADC3-14_TAB, and ADC3-15_TAB curves are whole antibody curves, each showing the ratio of the antibody concentration at different times to its TO concentration, including both non-payload-bound and payload-bound antibodies. This figure shows that the whole antibody curves for the corresponding P3-modified ADCs closely match each other, indicating that the human plasma stability of the P3-modified ADCs, as measured by the concentration of the corresponding antibody, is similar. [Figure 44] Figure 1 shows the human plasma stability profiles of ADC3-1, ADC3-11, ADC3-12, ADC3-13, ADC3-14, and ADC3-15, as measured by the concentration of the corresponding ADC. Curves ADC3-1_ADC, ADC3-11_ADC, ADC3-12_ADC, ADC3-13_ADC, ADC3-14_ADC, and ADC3-15_ADC ​​are ADC curves. This figure shows that the P3-modified ring-opened ADCs (ADC3-11, ADC3-12, and ADC3-13) have better stability than the P3-modified non-ring-opened ADCs. However, the stability of the P3-modified ring-opened ADCs is similar. The stability of the P3-modified non-ring-opened ADCs is also similar. [Figure 45]Figure 1 shows the human plasma stability profiles of ADC3-1, ADC3-11, ADC3-12, ADC3-13, ADC3-14, and ADC3-15 as measured by free payload release. The ADC3-1_ADC_payload, ADC3-11_ADC_payload, ADC3-12_ADC_payload, ADC3-13_ADC_payload, ADC3-14_ADC_payload, and ADC3-15_ADC_payload curves are payload release curves, which show the concentration of free payload released by the corresponding ADC. This figure shows that the free payload concentration in human plasma for all ADCs after 168 hours was less than 3%, indicating that the human plasma stability of the ADCs is within an acceptable range. Open-ring ADCs release less payload than non-open-ring ADCs. [Figure 46] The results of studies of ADC3-1, ADC3-3, and ADC3-4 in xenograft tumor models are shown, and the P5-modified non-open-ring ADC (ADC3-3) and P5-modified open-ring ADC (ADC3-4) show efficacy similar to that of ADC3-1. [Figure 47] The results of studies on ADC3-1, ADC3-3, and ADC3-4 in xenograft tumor models are shown, and the P5-modified non-open-ring ADC (ADC3-3) and the P5-modified open-ring ADC (ADC3-4) show efficacy similar to that of ADC3-1. [Figure 48] The results of a study of ADC3-1 and ADC3-7 in the A375 xenograft tumor model are shown, and the P5-modified open-ring ADC (ADC3-7) exhibits efficacy similar to that of ADC3-1. [Figure 49] The results of a study of ADC3-1 and ADC3-7 in the A375 xenograft tumor model are shown, and the P5-modified open-ring ADC (ADC3-7) exhibits efficacy similar to that of ADC3-1. [Figure 50] The results of a study of ADC3-1, ADC3-12, and ADC3-15 in the A375 xenograft tumor model are shown, and the P3-modified non-open-ring ADC (ADC3-15) and the P5-modified open-ring ADC (ADC3-12) show efficacy similar to that of ADC3-1. [Figure 51]This paper presents the results of a study of ADC3-1, ADC3-12, and ADC3-15 in an A375 xenograft tumor model, showing that the P3-modified non-open-ring ADC (ADC3-15) and the P5-modified open-ring ADC (ADC3-12) exhibited superior efficacy compared to ADC3-1. In particular, 1 / 8 CR and 2 / 8 PR were observed for ADC3-12. [Figure 52]

[0033] Figure 1 shows the results of a study of ADC3-1, ADC3-3, and ADC3-4 in the A375 xenograft tumor model. The results demonstrate that the P5-modified non-open-ring ADC (ADC3-3) and the P5-modified open-ring ADC (ADC3-4) exhibit similar pharmacokinetic profiles to the reference ADC3-1. [Figure 53] The results of pharmacokinetic studies on ADC3-1, ADC3-7, ADC3-12, and ADC3-15 in mice are shown, demonstrating that the P5-modified ring-opened ADC (ADC3-7), the P5-modified ring-opened ADC (ADC3-12), and the P3-modified non-ring-opened ADC (ADC3-15) exhibit pharmacokinetic profiles similar to those of ADC3-1, indicating that the efficacy of the ADC is positively correlated with the pharmacokinetic profile of the ADC. [Figure 54] 1 shows that ADC2-62-1 and ADC2-62-2 have comparable cell-killing activity compared to a reference ADC (ADC2-A with DAR4) in JIMT-1 (moderate expression level of HER2). [Figure 55] ADC2-62-2 with DAR8 shows better cell killing activity in Capan-1 (low expression level of HER2) compared to the reference ADC (ADC2-A with DAR4). [Figure 56] ADC2-63-1 (DAR4) has equivalent cell-killing activity to the reference ADC (ADC2-A with DAR4) in JIMT-1 (moderate expression level of HER2), and ADC2-63-2 (DAR8) has better cell-killing activity than the reference ADC in JIMT-1 (moderate expression level of HER2). [Figure 57]ADC2-63-1 (DAR4) has equivalent cell-killing activity to the reference ADC (ADC2-A with DAR4) in Capan-1 (low HER2 expression level), and ADC2-63-2 (DAR8) has better cell-killing activity than the reference ADC in Capan-1 (low HER2 expression level). [Figure 58] ADC2-64-1 with DAR4 has cell-killing activity equivalent to that of the reference ADC (ADC2-A with DAR4) in JIMT-1 (a group with a moderate expression level of HER2), and ADC2-64-2 with DAR8 has cell-killing activity better than that of the reference ADC (ADC2-A with DAR4) in JIMT-1 (a group with a moderate expression level of HER2). [Figure 59] ADC2-64-1 (DAR4) has cell-killing activity equivalent to that of the reference ADC (ADC2-A with DAR4) in Capan-1 (low HER2 expression level), and ADC2-64-2 (DAR8) has cell-killing activity better than that of the reference ADC (ADC2-A with DAR4) in Capan-1 (low HER2 expression level). [Figure 60] ADC2-65-1 with DAR4 has cell-killing activity equivalent to that of the reference ADC (ADC2-A with DAR4) in JIMT-1 (a tumor with a moderate expression level of HER2), and ADC2-65-2 with DAR8 has cell-killing activity better than that of the reference ADC (ADC2-A with DAR4) in JIMT-1 (a tumor with a moderate expression level of HER2). [Figure 61] ADC2-65-1 (DAR4) has cell-killing activity equivalent to that of the reference ADC (ADC2-A with DAR4) in Capan-1 (low HER2 expression level), and ADC2-65-1 (DAR8) has cell-killing activity better than that of the reference ADC (ADC2-A with DAR4) in Capan-1 (low HER2 expression level). [Figure 62] ADC2-62-2 HIC profile (HIC DAR 7.7) is shown. [Figure 63]SEC profile of ADC2-62-2 with purity >99% is shown. [Figure 64] ADC2-63-2 HIC profile (HIC DAR 8.0) is shown. [Figure 65] SEC profile of ADC2-63-2 with purity >99% is shown. [Figure 66] ADC2-64-2 HIC profile (HIC DAR 7.7) is shown. [Figure 67] SEC profile of ADC2-64-2 with purity >99% is shown. [Figure 68] ADC2-65-2 HIC profile (HIC DAR 7.5) is shown. [Figure 69] SEC profile of ADC2-65-2 with purity >99% is shown. [Figure 70] Examples 66 and 69 exhibit stronger cell killing activity than Dxd in A375 and Calu-6 cancer cell lines, while Examples 68 and 72 are less potent than Dxd in A375 and Calu-6 cancer cell lines. [Figure 71] Examples 66 and 69 exhibit stronger cell killing activity than Dxd in A375 and Calu-6 cancer cell lines, while Examples 68 and 72 are less potent than Dxd in A375 and Calu-6 cancer cell lines. [Figure 72] It shows that ADC-C2 has stronger killing efficacy than ADC-C1 in MDA-MB-453, and ADC-C3 and ADC-C5 exhibit better cell killing activity than ADC-C1 in A375. [Figure 73] It shows that ADC-C2 has stronger killing efficacy than ADC-C1 in MDA-MB-453, and ADC-C3 and ADC-C5 exhibit better cell killing activity than ADC-C1 in A375. [Figure 74] It shows that ADC-C2 has stronger killing efficacy than ADC-C1 in MDA-MB-453, and ADC-C3 and ADC-C5 exhibit better cell killing activity than ADC-C1 in A375. [Figure 75] This shows that ADC-C3 has higher bystander cell killing activity than ADC-C1 in the MDA-MB-453 and Calu-6-nanoLuc systems, and ADC-C3 and ADC-C5 exhibit better bystander killing efficacy than ADC-C1 in the A375 and Calu-6-nanoLuc systems. [Figure 76] This shows that ADC-C3 has higher bystander cell killing activity than ADC-C1 in the MDA-MB-453 and Calu-6-nanoLuc systems, and ADC-C3 and ADC-C5 exhibit better bystander killing efficacy than ADC-C1 in the A375 and Calu-6-nanoLuc systems. [Figure 77] This shows that ADC-C3 has higher bystander cell killing activity than ADC-C1 in the MDA-MB-453 and Calu-6-nanoLuc systems, and ADC-C3 and ADC-C5 exhibit better bystander killing efficacy than ADC-C1 in the A375 and Calu-6-nanoLuc systems. [Figure 78] The sequences of the antibodies used herein are shown. [Figure 79] 1 shows the in vivo efficacy of ADC-C1, ADC-C3, and ADC-C5 compared to A-375 melanoma xenografts grown subcutaneously in BALB / c nude mice. [Figure 80] 1 shows the in vivo efficacy of ADC-C1 and ADC-C2 in A-375 melanoma xenografts grown subcutaneously in BALB / c nude mice. DETAILED DESCRIPTION OF THE INVENTION

[0016] Provided herein are compounds, compositions, ADCs, and methods useful for the treatment of various human cancers, including, but not limited to, colorectal cancer, gastric cancer, breast cancer, non-small cell lung cancer (NSCLC), ovarian cancer, head and neck cancer, pancreatic cancer, and cervical cancer. In one embodiment, provided herein are compounds, compositions, ADCs, and methods useful for the treatment of various human cancers.

[0017] In this disclosure, it is understood that the disclosure is not limited to the particular methods and / or experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described. All patents, applications, and non-patent publications mentioned herein are incorporated by reference in their entirety.

[0018] 5.1.Definition When referring to the compounds provided herein, the following terms have the following meanings unless otherwise explained. 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. In the event that there are multiple definitions for a term provided herein, those definitions prevail unless otherwise explained. As used in this specification and the appended claims, the indefinite articles "a" and "an" and the definite article "the" include plural as well as singular referents unless the context clearly dictates otherwise.

[0019] As used herein, unless otherwise stated, the terms "about" and "approximately" when used in connection with the amount or weight percent of a component of a composition refer to an amount or weight percent that would be recognized by one of ordinary skill in the art as providing a pharmacological effect equivalent to that obtained with the specified amount or weight percent. In certain embodiments, the terms "about" and "approximately," when used in this context, encompass amounts or weight percents that are within 30%, 20%, 15%, 10%, or 5% of the specified amount or weight percent.

[0020] As used herein, unless otherwise explained, the terms "about" and "approximately," when used in conjunction with a value or range of values ​​provided to characterize a particular solid form, such as a particular temperature or temperature range such as those describing melting, dehydration, desolvation, or glass transition temperature; mass change such as mass change with temperature or humidity; solvent or water content expressed by mass or percentage, or the like; or peak position in analysis such as by IR or Raman spectroscopy or XRPD, indicates that the value or range of values ​​may deviate to an extent that would be considered reasonable by one of ordinary skill in the art while still being representative of the solid form. Techniques used to characterize crystalline forms and amorphous solids include, but are not limited to, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), single crystal X-ray diffraction, vibrational spectroscopy (e.g., infrared (IR) and Raman spectroscopy), solid-state and solution nuclear magnetic resonance (NMR) spectroscopy, optical microscopy, hot-stage optical microscopy, scanning electron microscopy (SEM), electron crystallographic and quantitative analysis, particle size analysis (PSA), surface area analysis, solubility studies, and dissolution studies. In certain embodiments, the terms "about" and "approximately," as used in this context, indicate that the indicated value or range of values ​​may vary within 30%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1.5%, 1%, 0.5%, or 0.25% of the recited value or range of values. For example, in some embodiments, the values ​​of the XRPD peak positions can vary by ±0.2 degrees 2θ (or ±0.2 degrees 2θ) while still representing a particular XRPD peak.

[0021] An "alkyl" group is a saturated, partially saturated, or unsaturated, straight-chain or branched, acyclic hydrocarbon having 1 to 10 carbon atoms, typically 1 to 8 carbon atoms, or in some embodiments, 1 to 6, 1 to 4, or 2 to 6 carbon atoms. Representative alkyl groups include -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, and n-hexyl; saturated branched alkyls include -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, allyl, CH=CH(CH), -CH=C(CH), -C(CH)=CH, -C(CH)=CH(CH), C(CHCH)=CH, C≡CH, -C≡C(CH), -C≡C(CHCH), -CHC≡CH, -CHC≡C(CH), and CHC≡C(CHCH), etc. The alkyl group can be substituted or unsubstituted. When alkyl groups described herein are referred to as "substituted," they may be substituted with any one or more substituents, such as those found in the exemplary compounds and embodiments disclosed herein, as well as halogen (chlorine, iodine, bromine, or fluorine), hydroxyl, alkoxyl, alkoxyalkyl, amino, alkylamino, carboxyl, nitro, cyano, thiol, thioether, imine, imide, amidine, guanidine, enamine, aminocarbonyl, acylamino, phosphonate, phosphine, thiocarbonyl, sulfonyl, sulfone, sulfonamide, ketone, aldehyde, ester, urea, carbamate, oxime, hydroxylamine, alkoxylamine, arylalkoxylamine, N-oxide, hydrazine, hydrazide, hydrazone, azide, isocyanate, isothiocyanate, cyanate, thiocyanate, B(OH), or O(alkyl)aminocarbonyl, etc.

[0022] "Alkenyl" refers to a straight-chain or branched acyclic hydrocarbon having 2 to 10 carbon atoms, typically 2 to 8 carbon atoms, and containing at least one carbon-carbon double bond. Representative straight-chain and branched (C2C8) alkenyl groups include -vinyl, -allyl, -1-butenyl, -2-butenyl, -isobutenyl, -1-pentenyl, 2-pentenyl, -3-methyl-1-butenyl, -2-methyl-2-butenyl, -2,3-dimethyl-2-butenyl, -1-hexenyl, 2-hexenyl, -3-hexenyl, -1-heptenyl, -2-heptenyl, -3-heptenyl, -1-octenyl, -2-octenyl, 3-octenyl, and the like. The double bond of an alkenyl group can be unconjugated or conjugated to another unsaturated group. An alkenyl group can be unsubstituted or substituted.

[0023] A "cycloalkyl" group is a saturated or partially saturated cyclic alkyl group having 3 to 10 carbon atoms, having a single ring or multiple fused or bridged rings, and optionally substituted with 1 to 3 alkyl groups. In some embodiments, the cycloalkyl group has 3 to 8 ring members; in other embodiments, the number of ring carbon atoms ranges from 3 to 5, 3 to 6, or 3 to 7. Such cycloalkyl groups include, for example, monocyclic structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, and 2-methylcyclooctyl, or polycyclic or bridged ring structures such as adamantyl. Examples of unsaturated cycloalkyl groups include cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl. The cycloalkyl group can be substituted or unsubstituted. Examples of such substituted cycloalkyl groups include cyclohexanone.

[0024] An "aryl" group is an aromatic carbocyclic group of 6 to 14 carbon atoms having a single ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl or anthryl). In some embodiments, an aryl group has 6 to 14 carbons, and in other embodiments, the ring portion of an aryl group contains 6 to 12, or even 6 to 10, carbon atoms. Specific aryl groups include phenyl, biphenyl, naphthyl, and the like. Aryl groups can be substituted or unsubstituted. The phrase "aryl group" also includes groups containing fused rings, such as fused aromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, and the like).

[0025] A "heteroaryl" group is an aryl ring system having 1 to 4 heteroatoms as ring atoms in the heteroaromatic ring system, with the remaining atoms being carbon atoms. In some embodiments, heteroaryl groups contain 5 to 6 ring atoms, and in other embodiments, 6 to 9 or even 6 to 10 atoms in the ring portion of the group. Suitable heteroatoms include oxygen, sulfur, and nitrogen. In certain embodiments, heteroaryl ring systems are monocyclic or bicyclic. Non-limiting examples include, but are not limited to, the following groups: For example, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyrrolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, benzothienyl, furanyl, benzofuranyl (e.g., isobenzofuran-1,3-diimine), indolyl, azaindolyl (e.g., pyrrolopyridinyl or 1H-pyrrolo[2,3-b]pyridinyl), indazolyl, benzimidazolyl (e.g., 1H-benzo[d]imidazolyl, aryl), imidazopyridinyl (e.g., azabenzimidazolyl, 3H-imidazo[4,5-b]pyridinyl or 1H-imidazo[4,5-b]pyridinyl), pyrazolopyridinyl, triazolopyridinyl, benzotriazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, isoxazolopyridinyl, thionaphthyl, purinyl, xanpurinyl, adenyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl.

[0026] A "heterocyclyl group" is an aromatic (also called heteroaryl) or non-aromatic cycloalkyl in which 1 to 4 ring carbon atoms are independently replaced with heteroatoms from the group consisting of O, S, and N. In some embodiments, heterocyclyl groups contain 3 to 10 ring members, while other such groups have 3 to 5, 3 to 6, or 3 to 8 ring members. Heterocyclyl groups can also be attached to other groups at any ring atom (i.e., any carbon atom or heteroatom of the heterocycle). Heterocyclyl groups can be substituted or unsubstituted. Heterocyclyl groups include unsaturated, partially saturated, and saturated ring systems such as imidazolyl, imidazolinyl, and imidazolidinyl. The term heterocyclyl group includes fused ring species, including those containing fused aromatic and non-aromatic groups, such as benzotriazolyl, 2,3-dihydrobenzo[1,4]dioxinyl, and benzo[1,3]dioxolyl. The phrase also includes bridged polycyclic ring systems containing heteroatoms, including, but not limited to, quinuclidinyl.Representative examples of heterocyclyl include aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, thiazolidinyl, tetrahydrothienyl, tetrahydrofuranyl, dioxolyl, furanyl, thienyl, pyrrolyl, pyrrolinyl, imidazolyl, imidazolinyl, pyrazolyl, pyrazolinyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, thiazolinyl, isothiazolyl, thiadiazolyl, oxadiazolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, Tetrahydropyranyl (e.g., tetrahydro-2H-pyranyl), tetrahydrothiopyranyl, oxathiane, dioxy, dithianyl, pyranyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl, dihydropyridinyl, dihydrodithiazinyl, dihydrodisulfinyl, homopiperazinyl, quinuclidinyl, indolyl, indolinyl, isoindolyl, azaindolyl (pyrrolopyridinyl), indazolyl, indolizinyl, benzotriazolyl, benzimidazolyl, benzofuranyl, benzothie nyl, benzothiazolyl, benzoxadiazolyl, benzoxazinyl, benzodithiazinyl, benzoxathiazinyl, benzothiazinyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[1,3]dioxolyl, pyrazolopyridinyl, imidazopyridinyl (azabenzimidazolyl such as 1H-imidazo[4,5-b]pyridinyl or 1H-imidazo[4,5-b]pyridin-2(3H)-onyl), triazolopyridinyl, isoxazolopyridinyl, purinyl, xanpurinyl, adenyl, These include, but are not limited to, guaninyl, quinolinyl, isoquinolinyl, quinolidinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl, pteridinyl, thionaphthyl, dihydrobenzothiazinyl, dihydrobenzofuranyl, dihydroindolyl, dihydrobenzimidazolyl, tetrahydrobenzotriazolyl, tetrahydropyrrolopyridinyl, tetrahydropyrazolopyridinyl, tetrahydroimidazopyridinyl, tetrahydrotriazolopyridinyl, and tetrahydroquinolinyl.Representative substituted heterocyclyl groups may be mono- or multiply substituted, for example, but not limited to, pyridyl or morpholinyl, and may be 2-, 3-, 4-, 5-, or 6-substituted, or disubstituted with a variety of substituents (e.g., those listed below).

[0027] A "cycloalkylalkyl" group is a radical of the formula: -alkyl-cycloalkyl, where alkyl and cycloalkyl are as defined above. Substituted cycloalkylalkyl groups may be substituted on the alkyl, cycloalkyl, or both the alkyl and cycloalkyl portions of the group. Representative cycloalkylalkyl groups include, but are not limited to, cyclopentylmethyl, cyclopentylethyl, cyclohexylmethyl, cyclohexylethyl, and cyclohexylpropyl. Representative substituted cycloalkylalkyls may be mono- or multiply substituted.

[0028] An "aralkyl" group is a radical of the formula: -alkyl-aryl, where alkyl and aryl are as defined above. Substituted aralkyl groups may be substituted on the alkyl, the aryl, or both the alkyl and aryl portions of the group. Representative aralkyl groups include, but are not limited to, benzyl and phenethyl, and fused (cycloalkylaryl)alkyl groups (e.g., 4-ethyl-indanyl).

[0029] A "heterocyclylalkyl" group is a radical of the formula: -alkyl-heterocyclyl, where alkyl and heterocyclyl are as defined above. Substituted heterocyclylalkyl groups may be substituted on the alkyl, heterocyclyl, or both the alkyl and heterocyclyl portions of the group. Representative heterocycloalkyl groups include, but are not limited to, 4-ethylmorpholinyl, 4-propylmorpholinyl, furan-2-ylmethyl, furan-3-ylmethyl, pyridin-3-ylmethyl, (tetrahydro-2H-pyran-4-yl)methyl, (tetrahydro-2H-pyran-4-yl)ethyl, tetrahydrofuran-2-ylmethyl, tetrahydrofuran-2-ylethyl, and indol-2-ylpropyl.

[0030] "Halogen" is chlorine, iodine, bromine or fluorine. A "hydroxyalkyl" group is an alkyl group as defined above that is substituted with one or more hydroxy groups. An "alkoxy" group is O(alkyl), where alkyl is as defined above. An "alkoxyalkyl" group is (alkyl)O(alkyl), where alkyl is as defined above. An "amine" group is a group of the formula: NH2.

[0031] A “hydroxylamine” group has the formula: N(R # )OH or NHOH groups, where R # is a substituted or unsubstituted alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, or heterocyclylalkyl as defined herein. An "alkoxyamine" group is a group of the formula: -N(R # )O-alkyl or -NHO-alkyl groups, where R # is as defined above. An “aralkoxyamine” group has the formula: N(R # ) O-aryl or NHOaryl groups, where R #is as defined above. An "alkylamine" group is a radical of the formula: NH alkyl or -N(alkyl) 2 , where each alkyl is independently as defined above.

[0032] An "aminocarbonyl" group is a group of the formula: -C(=O)N(R # )2, -C(=O)NH(R#) or -C(=O)NH2, where each R # is as defined above. An "acylamino" group is a group of the formula: NHC(=O)(R # ) or -N(alkyl)C(=O)(R # ) where each alkyl and R # are independently as defined above. An "O(alkyl)aminocarbonyl" group is a group of the formula: O(alkyl)C(=O)N(R#), -O(alkyl)C(=O)NH(R # ) or —O(alkyl)C(═O)NH2, where each R # are independently as defined above. An "N-oxide" group is a group of the formula: -N + -O - is the basis of

[0033] A "carboxy" group is a radical of the formula: C(=O)OH. A "ketone" group is a group of the formula: C(=O)(R # ), where R # is as defined above. An "aldehyde" group is a radical of the formula: --CH(.dbd.O). An "ester" group is an ester of the formula: C(=O)O(R # ) or -OC(=O)(R # ), where R # is as defined above.

[0034] A "urea" group is a group of the formula: -N(alkyl)C(=O)N(R # )2, -N(alkyl)C(=O)NH(R #), -N(alkyl)C(=O)NH2, -NHC(=O)N(R # )2, -NHC(=O)NH(R # ) or -NHC(=O)NH2 # where each alkyl and R # are independently as defined above. An "imine" group is a group of the formula: -N=C(R # )2 or -C(R # )=N(R # ), where each R # are independently as defined above. An "imido" group is a group of the formula: -C(=O)N(R # )C(=O)(R # ) or -N((C=O)(R # ))2 groups, where each R # are independently as defined above.

[0035] A "carbamate" group is a group of the formula: -OC(=O)N(R # )2, -OC(=O)NH(R # ), -N(R # )C(=O)O(R # ) or -NHC(=O)O(R # ), where each R # are independently as defined above. An "amidine" group is a group of the formula: -C(=N(R # ))N(R # )2, -C(=N(R # ))NH(R # ), -C(=N(R # ))NH2, -C(=NH)N(R # )2, -C(=NH)NH(R # ), -C(=NH)NH2, -N=C(R # )N(R # )2, -N=C(R # )NH(R # ), -N=C(R # )NH2, -N(R # )C(R # )=N(R # ), -NHC(R #)=N(R # ), -N(R # )C(R # )=NH or -NHC(R # )=NH group, where each R # are independently as defined above.

[0036] A "guanidine" group has the formula: -N(R # )C(=N(R # ))N(R # )2, -NHC(=N(R # ))N(R # )2, -N(R # )C(=NH)N(R # )2, -N(R # )C(=N(R # ))NH(R # ), -N(R # )C(=N(R # ))NH2, -NHC(=NH)N(R # )2, -NHC(=N(R # ))NH(R # ), -NHC(=N(R # ))NH2, -NHC(=NH)NH(R # ), -NHC(=NH)NH2, -N=C(N(R # )2)2, -N=C(NH(R # ))2 or -N=C(NH2)2, where each R # are independently as defined above.

[0037] An "enamine" group is a group of the formula: -N(R # )C(R # )=C(R # )2, -NHC(R # )=C(R # )2, -C(N(R # )2)=C(R # )2, -C(NH(R # ))=C(R # )2, -C(NH2)=C(R # )2, -C(R # )=C(R # )(N(R # )2), -C(R #)=C(R # )(NH(R # )) or -C(R # )=C(R # )(NH), where each R # are independently as defined above. An "oxime" group is a group of the formula: -C(=NO(R # ))(R # ), -C(=NOH)(R # ), -CH(=NO(R # )) or —CH(═NOH), where each R # are independently as defined above.

[0038] A "hydrazide" group has the formula: -C(=O)N(R # )N(R # )2, -C(=O)NHN(R#)2, -C(=O)N(R # )NH(R # ), -C(=O)N(R # )NH2, -C(=O)NHNH(R # )2 or —C(═O)NHNH2, where each R # are independently as defined above. A “hydrazine” group has the formula: —N(R # )N(R # )2, -NHN(R # )2, -N(R#)NH(R # ), -N(R # )NH2, -NHNH(R # )2 or -NHNH2, where each R # are independently as defined above.

[0039] A "hydrazone" group has the formula: -C(=NN(R # )2)(R # )2, -C(=N-NNH(R # ))(R # )2, -C(=N-NH2)(R # )2, -N(R # )(N=C(R # )2) or -NH(N=C(R #)2), where each R # are independently as defined above. An "azido" group is a radical of the formula: -N3. An "isocyanate" group is a group of the formula: N=C=O. An "isothiocyanate" group is a group of the formula: N=C=S. A "cyanate" group is a radical of the formula: OCN. A "thiocyanate" group is a radical of the formula: SCN.

[0040] A "thioether" group has the formula: -S(R # ), where R # is as defined above. A "thiocarbonyl" group is a group of the formula: -C(=S)(R # ), where R # is as defined above. A "sulfinyl" group is a group of the formula: -S(=O)(R # ), where R # is as defined above. A "sulfone" group is a group of the formula: -S(=O)(R # ), where R # is as defined above. A "sulfonylamino" group is a group of the formula: -NHSO(R # ) or a group of -N(alkyl)SO2(R#), where each alkyl and R# are as defined above. A "sulfonamide" group is a group of the formula: -S(=O)N(R # )2 or -S(=O)2NH(R # ) or —S(═O)2NH2, where each R # are independently as defined above.

[0041] A "phosphonate" group has the formula: -P(=O)(O(R # ))2, -P(=O)(OH)2, -OP(=O)(O(R # ))(R # ) or -OP(=O)(OH)(R #), where each R # are independently as defined above. A "phosphine" group has the formula: -P(R # )2 groups, where each R # are independently as defined above.

[0042] When a group described herein (excluding alkyl groups) is referred to as "substituted," it can be substituted with any suitable substituent. Illustrative substituents include those found in the exemplary compounds and embodiments disclosed herein, as well as halogen (chlorine, iodine, bromine, or fluorine), alkyl, hydroxyl, alkoxy, alkoxyalkyl, amine, alkylamine, carboxyl, nitro, cyano, thiol, thioether, imine, imide, amidine, guanidine, enamine, aminocarbonyl, acylamino, phosphonate, phosphine, thiocarbonyl, sulfinyl, sulfone, sulfonamide, ketone, aldehyde, ester, urea, carbamate, oxime, hydroxylamine, alkoxyamine, aralkoxyamine, N-oxide, hydrazine, hydrazide, hydrazone, azide, isocyanate, isothiocyanate, cyanate, thiocyanate, oxygen (=O), B(OH), O(alkyl)aminocarbonyl, and cycloalkyl groups. and heterocyclic groups, which may be monocyclic or fused or non-fused polycyclic (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl); or heterocyclic groups, which may be monocyclic or fused or non-fused polycyclic (e.g., pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, or thiazinyl); monocyclic or fused or non-fused polycyclic aryl or heteroaryl (e.g., phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridinyl, quinolinyl, isoquinolinyl, acridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, benzimidazolyl, benzothienyl, or benzofuranyl); aryloxy, aralkoxy, heterocyclyloxy, and heterocyclylalkoxy.

[0043] As used herein, the term "one or more pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic acids or bases, including inorganic acids and bases and organic acids and bases. As used herein, unless otherwise explained, the term "clathrate" means a compound or salt thereof in the form of a crystal lattice containing spaces (e.g., channels) in which a guest molecule (e.g., solvent or water) is trapped, or in which the compound is a guest molecule.

[0044] As used herein, unless otherwise stated, the term "solvate" means a compound or a salt thereof that further includes a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. In one embodiment, the solvate is a hydrate. As used herein, unless otherwise explained, the term "hydrate" means a compound or a salt thereof that further includes a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces.

[0045] As used herein, the term "prodrug" refers to a derivative of a compound that can be hydrolyzed, oxidized, or otherwise reacted under biological conditions (in vitro or in vivo) to provide an active compound, particularly a compound. Examples of prodrugs include, but are not limited to, derivatives and metabolites of a compound containing a biohydrolyzable moiety, such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureides, and biohydrolyzable phosphate analogs. In certain embodiments, prodrugs of compounds with a carboxyl functional group are lower alkyl esters of the carboxylic acid. Carboxylic acid esters can be conveniently formed by esterifying any carboxylic acid moiety present on the molecule. Prodrugs can typically be prepared using well-known methods, such as those described by Burger's Medicinal Chemistry and Drug Discovery, 6th Edition (Donald J. Abraham, ed., 2001, Wiley) and Design and Application of Prodrugs (H. Bundgaard, ed., 1985, Harwood Academic Publishers GmbH).

[0046] Unless otherwise specified, the term "stereoisomer" or "stereoisomerically pure" means one stereoisomer of a compound that is substantially free of other stereoisomers of the compound. For example, a stereoisomerically pure compound having one chiral center is substantially free of the opposite enantiomer of the compound. A stereoisomerically pure compound having two chiral centers is substantially free of other diastereomers of the compound. A typical stereoisomerically pure compound contains greater than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of the other stereoisomers of the compound, greater than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of the other stereoisomers of the compound, greater than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of the other stereoisomers of the compound, or greater than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of the other stereoisomers of the compound. Compounds may have chiral centers and may occur as racemates, individual enantiomers or diastereomers, and mixtures thereof. All such isomeric forms, including mixtures thereof, are encompassed by the embodiments disclosed herein. The embodiments disclosed herein encompass the use of stereomerically pure forms of such compounds, as well as the use of mixtures of these forms. For example, mixtures containing equal or unequal amounts of the enantiomers of a particular compound can be used in the methods and compositions disclosed herein. These isomers can be asymmetrically synthesized or resolved using standard techniques, such as chiral columns or chiral resolving agents.See, for example, Jacques, J. et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen, SH et al., Tetrahedron 33:2725 (1977); Eliel, EL, Stereochemistry of Carbon Compounds (McGrawHill, NY, 1962); and Wilen, SH, Tables of Resolving Agents and Optical Resolutions, p. 268 (E.L. Eliel, ed., University of Notre Dame Press, Notre Dame, IN, 1972).

[0047] It should also be noted that the compounds may include E and Z isomers, or mixtures thereof, as well as cis and trans isomers, or mixtures thereof. In certain embodiments, the compounds are isolated as either cis or trans isomers. In other embodiments, the compounds are mixtures of cis and trans isomers.

[0048] "Tautomers" refer to isomeric forms of a compound that are in equilibrium with each other. The concentration of isomeric forms can vary depending on the environment in which the compound is placed, for example, whether the compound is a solid or in an organic or aqueous solution. For example, in aqueous solution, pyrazole can exhibit the following isomeric forms, called tautomers of each other: [ka]

[0049] As will be readily understood by one of ordinary skill in the art, various functional groups and other structures may exhibit tautomerism, and all tautomers of the compounds are within the scope of the invention. It should also be noted that the compounds may contain unnatural proportions of atomic isotopes at one or more atoms. For example, the compounds may contain tritium ( 3 H), iodine-125(125 I), sulfur 35( 35 S) or carbon-14 ( 14 C) or may be radiolabeled with deuterium ( 2 H), carbon-13( 13 C) or nitrogen-15( 15 The compound may be isotopically enriched with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 112, 113, 120, 121, 132, 133, 144, 150, 165, 170, 171, 182, 191, 192, 193, 194, 195, 196, 197, 198, 19

[0050] It should be noted that if there is a discrepancy between the illustrated structure and the name of the structure, the illustrated structure should bear more weight. The term "effective amount" with respect to a compound means an amount that is capable of completely or partially alleviating the symptoms, or slowing or stopping the further progression or worsening of the symptoms. It will be apparent to those skilled in the art that the effective amount of the compositions disclosed herein may vary depending on the severity of the therapeutic indication.

[0051] As used herein, "alkynyl" refers to a monovalent hydrocarbon moiety containing at least two carbon atoms and one or more carbon-carbon triple bonds. Alkynyl groups may be optionally substituted and may be linear, branched, or cyclic. Alkynyl groups include groups having 2 to 20 carbon atoms, i.e., C 2-20 Alkynyl, a group having 2 to 12 carbon atoms, i.e., C 2-12 Alkynyl, a group having 2 to 8 carbon atoms, i.e., C 2-8 Alkynyl, a group having 2 to 6 carbon atoms, i.e., C 2-6 Alkynyl and groups having 2 to 4 carbon atoms, i.e., C 2-4 Examples of alkynyl moieties include, but are not limited to, ethynyl, propynyl, and butynyl.

[0052] As used herein, "haloalkyl" refers to an alkyl group, as defined above, which contains at least one substituent selected from a halogen, such as fluorine (F), chlorine (Cl), bromine (Br), or iodine (I). Examples of haloalkyl groups include, but are not limited to, -CF, -CHCF, -CClF, and -CCl. As used herein, "haloalkoxy" refers to an alkoxy group, as defined above, which contains at least one substituent selected from a halogen, such as F, Cl, Br, or I.

[0053] As used herein, "arylalkyl" refers to a monovalent moiety that is a radical of an alkyl compound, where the alkyl compound is substituted with an aromatic substituent, i.e., the aromatic compound contains a single bond to the alkyl group, and the group is located on the alkyl group. The arylalkyl group is attached to the depicted chemical structure via the alkyl group. Arylalkyl groups can be represented by structures such as B-CH2-, B-CH2-CH2-, B-CH2-CH2-CH2-, B-CH2-CH2-CH2-, B-CH(CH3)-CH2-CH2-, B-CH2-CH(CH3)-CH2-, etc., where B is an aromatic moiety such as phenyl. Arylalkyl groups are optionally substituted, i.e., the aryl and / or alkyl may be substituted as disclosed herein. Examples of arylalkyl groups include, but are not limited to, benzyl.

[0054] As used herein, "alkylaryl" refers to a monovalent moiety that is a radical of an aryl compound, where the aryl compound is substituted with an alkyl substituent, i.e., the aryl compound contains a single bond to an alkyl group, and the group is located on the aryl group. The alkylaryl group is attached to the depicted chemical structure via the aryl group. The alkylaryl group can be represented by structures such as -B-CH, -B-CH-CH, -B-CH-CH-CH, -B-CH-CH-CH, -B-CH(CH)-CH-CH, -B-CH-CH(CH)-CH, and the like, where B is an aromatic moiety such as phenyl. The alkylaryl group is optionally substituted, i.e., the aryl and / or alkyl may be substituted as disclosed herein. Examples of alkylaryl groups include, but are not limited to, toluyl.

[0055] As used herein, "aryloxy" refers to a monovalent moiety that is a radical of an aromatic compound where the ring atoms are carbon atoms and the ring is substituted with an oxygen group, i.e., the aromatic compound contains a single bond to the oxygen atom and the group is on the oxygen atom, e.g., C6H5-O- of phenoxy. The aryloxy substituent is attached to the compound it substitutes through this oxygen atom. The aryloxy group may be substituted. Aryloxy groups include groups having 6 to 20 ring carbon atoms, i.e., C 6-20 Aryloxy groups, groups having 6 to 15 ring carbon atoms, i.e., C 6-15 Aryloxy groups and groups having 6 to 10 ring carbon atoms, i.e., C 6-10 Examples of aryloxy moieties include, but are not limited to, phenoxy, naphthoxy, and anthroxy.

[0056] As used herein, the term "residue" refers to the chemical moiety within a compound that remains after a chemical reaction. For example, the term "amino acid residue" or "N-alkyl amino acid residue" refers to the product after amide or peptide coupling of an amino acid or an N-alkyl amino acid with a suitable coupling partner, e.g., after amide or peptide coupling of an amino acid or an N-alkyl amino acid, a water molecule is released, thereby resulting in a product incorporating the amino acid residue or N-alkyl amino acid residue.

[0057] As used herein, "sugar" or "glycosyl" or "sugar residue" refers to a carbohydrate moiety that may include a 3-carbon (triose) unit, a 4-carbon (tetrose) unit, a 5-carbon (pentose) unit, a 6-carbon (hexose) unit, a 7-carbon (heptose) unit, or a combination thereof, and may be a monosaccharide, disaccharide, trisaccharide, tetrasaccharide, pentasaccharide, oligosaccharide, or any other polysaccharide. In some cases, a "sugar" or "glycosyl" or "sugar residue" includes a furanose (e.g., ribofuranose, fructofuranose) or a pyranose (e.g., glucopyranose, galactopyranose) or a combination thereof. In some cases, a "sugar" or "glycosyl" or "sugar residue" includes an aldose or a ketose, or a combination thereof. Non-limiting examples of monosaccharides include ribose, deoxyribose, xylose, arabinose, glucose, mannose, galactose, and fructose. Non-limiting examples of disaccharides include sucrose, maltose, lactose, lactulose, and trehalose. Other "sugars" or "glycosyl" or "sugar residues" include polysaccharides and / or oligosaccharides, including, but not limited to, amylose, amylopectin, glycogen, inulin, and cellulose. In some cases, the "sugar" or "glycosyl" or "sugar residue" is an amino sugar. In some cases, the "sugar" or "glycosyl" or "sugar residue" is a glucosamine residue (1-amino-1-deoxy-D-glucitol), which is attached to the rest of the molecule via its amino group, thereby forming an amide bond (i.e., gluconamide) with the rest of the molecule.

[0058] As used herein, "inorganic acid residues" refers to orthophosphoric and pyrophosphoric acid, phosphoric acid and sulfuric acid residues.

[0059] As used herein, "organic acid residue" refers to a residue of an alkanecarboxylic acid, an amino acid, or an oligopeptide. In one embodiment, the alkanecarboxylic acid is formic acid, acetic acid, propanoic acid, butanoic acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, tridecanoic acid, tetradecanoic acid, pentadecanoic acid, hexadecanoic acid, heptadecanoic acid, stearic acid, nonadecanoic acid, or eicosanoic acid. In one embodiment, the alkanecarboxylic acid is formic acid, acetic acid, propanoic acid, or butanoic acid.

[0060] Certain groups, moieties, substituents, and atoms are represented by wavy lines crossing one or more bonds to indicate to which atom the group, moiety, substituent, or atom is bonded, for example, phenyl substituted with propyl. [ka] is shown to have the following structure: [ka] .

[0061] As used herein, diagrams showing substituents attached to cyclic groups (e.g., aromatic, heteroaromatic, fused ring, and saturated or unsaturated cycloalkyl or heterocycloalkyl) via a bond between ring atoms are intended to indicate that the cyclic group may be substituted with a substituent at any ring position in a cyclic group or on any ring in a fused ring group in accordance with techniques described herein or known in the art to which this disclosure pertains, unless otherwise stated. As used herein, "binding agent" refers to any molecule, e.g., an antibody, that can specifically bind to a given binding partner (e.g., an antigen).

[0062] As used herein, the term "amino acid" refers to an organic compound containing an amine (-NH) and a carboxyl (-COOH) functional group, as well as a side chain (R group) specific to each amino acid. Amino acids may be proteinogenic or non-proteinogenic. "Proteogenic" means that the amino acid is one of the 20 naturally occurring amino acids found in proteins. Proteinogenic amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. "Non-proteinogenic" means that the amino acid is not naturally found in proteins or is not produced directly by cellular machinery (e.g., is a product of post-translational modification). Non-limiting examples of non-proteinogenic amino acids include gamma-aminobutyric acid (GABA), taurine (2-aminoethanesulfonic acid), theanine (L-gamma-glutamylacetamide), hydroxyproline, beta-alanine, ornithine, and citrulline.

[0063] As used herein, "peptide" is defined in its broadest sense and in various grammatical forms to refer to a compound of two or more subunit amino acids, amino acid analogs, or other peptidomimetics. The subunits may be linked to each other by peptide bonds or other bonds, such as ester bonds, ether bonds, etc. As used herein, the term "amino acid" refers to natural and / or unnatural, proteinogenic, non-proteinogenic, or synthetic amino acids, including glycine and its D or L optical isomers, as well as amino acid analogs and peptidomimetics. When peptide chains are short, e.g., two, three, or more amino acids, they are generally referred to as oligopeptides. When peptide chains are long, the peptides are typically referred to as polypeptides or proteins. The definition encompasses full-length proteins, analogs, variants, and fragments thereof. The term also includes post-expression modifications of the polypeptide, such as glycosylation, acetylation, phosphorylation, etc. Furthermore, certain peptides can be obtained as acidic or basic salts or in neutral form due to the presence of ionizable amino and carboxyl groups in the molecule. Peptides can be obtained directly from a source organism, or can be produced recombinantly or synthetically.

[0064] The amino acid sequence of an antibody can be numbered using any known scheme, including the following: Kabat et al. ("Kabat" numbering scheme); Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948 ("Chothia" numbering scheme); MacCallum et al., 1996, J. Mol. Biol. 262:732-745 ("Contact" numbering scheme); Lefranc et al., Dev. Comp. Immunol., 2003, 27:55-77 ("IMGT" numbering scheme); and Honegge and Pluckthun, J. Mol. Biol., 2001, 309:657-70 ("AHo" numbering scheme). Unless otherwise stated, the numbering scheme used herein is the Kabat numbering scheme. However, the choice of numbering scheme does not imply differences in sequences that do not differ, and one skilled in the art can easily ascertain the sequence positions by examining the amino acid sequences of one or more antibodies. Unless otherwise stated, the "EU numbering scheme" is generally used when referring to residues in antibody heavy chain constant regions (e.g., as reported in Kabat et al., supra).

[0065] As used herein, the term "anti-HER2 antibody" refers to an antibody that selectively binds to the HER2 receptor, such as trastuzumab. In one embodiment, trastuzumab can be produced and used as described in US Pat. No. 6,407,213 and US Pat. No. 5,821,337, the entire disclosures of which are incorporated herein by reference. As used herein, the term "anti-HER3 antibody" refers to an antibody that selectively binds to the HER3 receptor, such as patritumab. In one embodiment, patritumab can be made and used as described in US Patritumab 2013, the entire disclosure of which is incorporated herein by reference.

[0066] As used herein, the term "anti-PTK7 antibody" refers to an antibody that selectively binds to the PTK7 receptor, e.g., cofetuzumab. In one embodiment, cofetuzumab can be made and used as described in US9777070, the entire disclosure of which is incorporated herein by reference.

[0067] The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. A "tumor" comprises one or more cancerous cells. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More specific examples of such cancers include squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), lung cancer (including small cell lung cancer, non-small cell lung cancer ("NSCLC"), lung adenocarcinoma, and lung squamous cell carcinoma), peritoneal cancer, hepatocellular carcinoma, gastric cancer (including digestive cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatocellular carcinoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, salivary gland cancer, kidney cancer, renal cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, and head and neck cancer.

[0068] As used herein, the term "cell killing activity" refers to activity that reduces or decreases cell viability of a test cell line. In the following claims and in the preceding description of the invention, unless the context otherwise requires to indicate the express language or necessary meaning, the words "comprise" or variations such as "comprising" are used in the inclusive sense, i.e., to specify the presence of stated features in various embodiments of the invention but do not exclude the presence of further features or the addition of further features.

[0069] 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. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods, devices, and materials are now described.

[0070] 5.2. Complex In some other examples, the specification describes an ADC, or a pharmaceutically acceptable salt thereof, comprising a protein linked to at least one payload moiety and to at least one hydrophilic moiety via a covalent linker, wherein the covalent linker is directly or indirectly attached to each of the protein, payload moiety, and hydrophilic moiety. In some embodiments, the protein is an antibody or antigen-binding fragment thereof.

[0071] As shown herein, in some examples, the binding agent is directly attached to a covalent linker, such as a linker described herein. This means that the binding agent is only one binding site away from the covalent linker described herein. In some of these examples, the covalent linker is also directly attached to the payload moiety. This means that the covalent linker is only one binding site away from the payload (e.g., but not limited to, Dxd, MMAE or a stereoisomer thereof, or any payload described herein). In some of these examples, the covalent linker is also directly attached to the hydrophilic moiety. This means that the covalent linker is only one binding site away from the hydrophilic residue (e.g., a hydrophilic residue described herein). In some of these examples, the covalent linker is a covalent linker described herein.

[0072] In other instances, the binding agent is indirectly attached to the covalent linker, meaning that the binding agent is separated from the covalent linker by more than one binding site. This also means that the binder is bound to the covalent linker via another moiety. For example, the binder may be bound to a maleimide group bound to a polyethylene glycol group bound to the covalent linker. In some of these examples, the covalent linker is also indirectly bound to the payload moiety. This means that the covalent linker is separated from the payload (e.g., but not limited to, Dxd, MMAE or a stereoisomer thereof, or any payload described herein) by more than one binding site. This also means that the covalent linker is bound to the payload via another moiety. For example, the covalent linker can be attached to a dipeptide such as, but not limited to, Val-Ala or Val-Cit, which may be attached to a PAB, and the PAB may be attached to the payload. In some of these examples, the covalent linker is also indirectly bound to the hydrophilic moiety. This means that the covalent linker is separated from the hydrophilic moiety (e.g., a hydrophilic residue described herein) by more than one binding site. This also means that the covalent linker is bound to the hydrophilic moiety via another moiety.

[0073] In some cases, the hydrophilic residue comprises a terminal hydrophilic group. In some cases, the hydrophilic residue comprises at least one sugar residue. In some cases, the hydrophilic residue comprises one sugar residue. In some cases, the hydrophilic residue comprises one terminal sugar residue. In further cases, the hydrophilic residue comprises more than one sugar residue. In some cases, the hydrophilic residue comprises more than one terminal sugar residue.

[0074] In another embodiment, the payload provided herein is a chromophore functional group, and with respect to the functional group, the compound provided herein can be used to detect, monitor, or study the interaction of a cell-binding molecule with a target cell. The chromophore functional group is a functional group capable of absorbing specific light, such as ultraviolet light, fluorescence, infrared light, near-infrared light, or visible light. The pigment cell functional group is a functional group selected from the class or subclass of yellow pigment cells, red pigment cells, iridocytes, white pigment cells, melanocytes, and blue pigment cells, the class or subclass of fluorophore molecules, which are fluorescent compounds that re-emit light under light irradiation, the class or subclass of phototransduction molecules, the class or subclass of photophore molecules, the class or subclass of luminescent molecules, and the class or subclass of fluorescein compounds.

[0075] 5.2.1. Aspect 1 The present specification describes a compound according to formula (I): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof;

[0076] wherein BA is a binding agent selected from a humanized, chimeric, or human antibody or antigen-binding antibody fragment thereof, L is a covalent linker, PA is a payload residue, and subscript x is 1 to 30. In some cases, x is 1 to 4. In some cases, x is about 1. In some cases, x is about 2. In some cases, x is about 3. In some cases, x is about 4.

[0077] In one example, the BA is an antibody. In one example, the antibody is a humanized, chimeric, or human antibody, or an antigen-binding antibody fragment of an antibody. In one example, the antibody is a humanized, chimeric, or human anti-HER2 or anti-HER3 antibody, or an antigen-binding antibody fragment of an anti-HER2 or anti-HER3 antibody. In one example, the antibody is a monoclonal antibody. In one example, the BA is an antibody. In one example, the antibody is a humanized, chimeric, or human antibody, or an antigen-binding antibody fragment of cofetuzumab, patritumab, or trastuzumab.

[0078] In certain embodiments, the antibodies described herein bind to one or more receptors selected from the group consisting of CD7, CD19, CD22, CD27, CD30, CD33, CD37, CD70, CD74, CD79b, CD138, CD142, CA6, placental cadherin, CEA, CEACAM5, C4.4a, DLL3, EGFR, EGFRVIII, ENPP3, EphA2, EphrinA, FLOR1, FGFR2, GCC, HER2, HER3, cKIT, LIV1, LY6E, MSLN, MUC16, NaPi2b, Nectin4, gpNMB, PSMA, SLITRK6, STEAP1, TROP2, 5T4, SSEA4, GloboH, Gb5, STn, and Tn. In certain embodiments, the antibodies described herein bind to one or more receptors selected from the group consisting of B7H3, MUC1, FGFR2b, CLL1, CCR7, GPC1, and GPC3. In certain embodiments, the antibodies described herein bind to the CEA receptor. In certain embodiments, the antibodies described herein are bispecific antibodies.

[0079] 5.2.2. Mode 2 The present specification describes a compound according to formula (Ia), [ka] or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof;

[0080] RG 1 is a reactive group residue, and RG 2 is any reactive group residue, and SP 1 and SP 2is independently in each occurrence an optional spacer group residue, HG is a hydrophilic residue, PAB is an optional self-immolative unit, the subscript p is 0 or 1, and the subscript x is 1 to 30.

[0081] In some embodiments, x is 1 to 15. In some embodiments, x is 2 to 10. In some embodiments, x is 3 to 9. In one embodiment, x is about 3. In one embodiment, x is about 4. In one embodiment, x is about 5. In one embodiment, x is about 6. In one embodiment, x is about 7. In one embodiment, x is about 8. In one embodiment, x is about 9.

[0082] In some embodiments, the compound of formula (Ia) is a compound having a P3 modification, AA 2 contains the formula (W), and [ka] AA 3 -valine-alanine-, -valine-citrulline- or [ka] is a dipeptide residue of R 6 is —CH3 or —(CH2)3—NHC(═O)NH2.

[0083] In one embodiment [ka] teeth [ka] is.

[0084] In some embodiments, PAB represents —NH—CH—O—, formula (Y1), or formula (Y2), [ka] [ka] represents the bond that connects the PAB to the adjacent group in the formula.

[0085] In some embodiments, RG 1 teeth [ka] , -(succinimide-3-yl-N)-, [ka] is.

[0086] In some embodiments, RG 1 teeth [ka] and EWG is an electron withdrawing group, e.g., -CN, -NO2, halogen, -CF3, -C(=O)OR 1 and -C(=O)R 1 and R 1 is substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted heteroaryl.

[0087] In some embodiments, RG 1 teeth [ka] is.

[0088] In some embodiments, RG 1 teeth [ka] and EWG is an electron withdrawing group, e.g., -CN, -NO2, halogen, -CF3, -C(=O)OR 1 and -C(=O)R 1 and R 1is substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted heteroaryl.

[0089] In some embodiments, the ring-opened compound of Formula (Ia) is RG 1 In some embodiments, RG is a compound of formula (Ia), wherein RG is an open-ring heterocycle. 1 teeth [ka] is.

[0090] In some embodiments, RG 2 is a bond, —C(═O)—NH—, or —NHC(═O)—. In some embodiments, SP 1 is -(CH2) n1 -C(=O)-, -(CH2CH2O) n2 -CH2CH2-C(=O)-, -CH[-(CH2) n3 -COOH]-C(=O)-, -CH2-C(=O)-NH-(CH2) n4 -C(=O)-, -CH2-C(=O)-NH-(CH2) n3 -C(=O)-NH-(CH2) n4 -C(=O)- or -C(=O)-(CH2) n5 -C(=O)-, and n1, n2, n3, n4 and n5 each independently represent an integer of 1 to 8.

[0091] In some embodiments, SP 2 is -(CH2) n6 -, and n6 represents an integer of 1 to 8.

[0092] In some embodiments, HG is [ka] [ka] each n7 is independently 1 to 15, each n8 is independently 0 or 1, each n9 is independently 1 or 2, each n10 is independently an integer of 4 to 16, for example, 4, 8, or 12, each n11 is independently an integer of 0 to 5, n12 is an integer of 0 to 3, d is 0 to 3, and R 2 is H or Me, and R 3 -OH, -NH2, -NHCH2-CH2-(PEG) x -OH or -NHCH2-CH2-(PEG) x -OMe and R 4 is OH or NH2, and X, Y and Z are each independently -CH2-, -NH-, -S- or -O-.

[0093] In some embodiments, HG is [ka] each n7 is independently 1 to 15, each n8 is independently 0 or 1, each n9 is independently 1 or 2, each n10 is independently an integer of 4 to 16, for example, 4, 8, or 12, d is 0 to 3, and R 2 is H or Me, and R 3 -OH, -NH2, -NHCH2-CH2-(PEG) x -OH or -NHCH2-CH2-(PEG) x -OMe and R 4 is OH or NH2.

[0094] In some embodiments, HG is [ka] each n8 is independently 0 or 1; R 1 is H or Me.

[0095] In some embodiments, HG is [ka] wherein each n11 is independently an integer of 0 to 5; n12 is an integer of 0 to 3, and each X, Y, and Z is independently -CH2-, -NH-, -S-, or -O-.

[0096] In some embodiments, HG is —NHSO2NH2, —SO3H, —SO2NH2, —PO3H2, and RG 2 is a bond. In some embodiments, each PA independently represents one chromophore functional group. In some embodiments, each pigment cell functional group is a functional group independently selected from the following classes or subclasses: yellow pigment cells, red pigment cells, iridescent pigment cells, white pigment cells, melanin pigment cells, and blue pigment cells; fluorophore molecules, which are fluorescent compounds that re-emit light under light irradiation; phototransduction molecules, photophore molecules, luminescent molecules, and fluorescein compounds.

[0097] In some embodiments, each PA is selected from the group consisting of monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), monomethyl auristatin D (MMAD), mertansine (mertansine DM1 / DM4), paclitaxel, docetaxel, epothilone B, epothilone A, CYT997, auristatin tyramine phosphate, auristatin aminoquinolines, halocombstatins, calicheamicins, and the like. n) θ, 7-ethyl-10-hydroxycamptothecin (SN-38), pyrrolobenzodiazepines (PBDs), pancratistatin, cyclic phosphates, cribrostatin-6, kitastatin, turbostatin 1-4, halocombstatins, eribulin, hemiasterlin, PNU, and cilastatins.

[0098] In some embodiments, each PA is independently represented by formula (D1): [ka] R 4 , R 5a and R 5b are each independently hydrogen, a sugar residue, a substituted or unsubstituted inorganic or organic acid residue, a substituted or unsubstituted C 1-8 alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted non-aromatic heterocyclyl, substituted or unsubstituted cycloalkylalkyl, or substituted or unsubstituted heterocyclylalkyl;

[0099] R 5a and R 5b together with the atom to which they are attached form a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted non-aromatic heterocyclyl. In some embodiments, R 4 is hydrogen, [ka] and

[0100] In the formula, R 5a and R 5b are each independently H, CH3 or CF3, or R 5a and R 5b together with the atom to which they are attached form a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted non-aromatic heterocyclyl.

[0101] In some embodiments, R 4 is hydrogen, [ka] and In the formula, R 5a and R 5b are each independently H, CH3 or CF3, or R 5a and R 5b together with the atom to which they are attached form a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted non-aromatic heterocyclyl.

[0102] In some embodiments, each PA independently comprises: [ka] It is expressed as:

[0103] In some embodiments, each PA is independently represented by formula (D2): [ka] wherein ring B is a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heterocyclyl, or a substituted or unsubstituted heteroaryl.

[0104] In some embodiments, each PA independently comprises: [ka] is. In some embodiments, each PA is independently represented by formula (D3): [ka] In the formula, S 2 is an enzymatically hydrolyzable hydrophilic group.

[0105] In some embodiments, S 2 The group is hydrogen or represents one of the following formulae: [ka]

[0106] In some embodiments, each PA is independently represented by formula (E1): [ka] In the formula, R 7 and R 8 are each independently hydrogen, halogen, or alkyl.

[0107] In one embodiment, R 7 and R 8 is hydrogen. In one embodiment, R 7 and R 8 is methyl. In one embodiment, R 7 is methyl and R 8 is F. In one embodiment, R 7 and R 8 The carbon to which is attached is in the S configuration. In one embodiment, R 7 and R 8 The carbon to which is attached is in the R configuration.

[0108] In some embodiments, each PA is independently represented by the formula: [ka]

[0109] In some embodiments, each PA is independently Dxd or is independently represented by the formula: [ka]

[0110] In some embodiments, each PA is independently represented by the formula: [ka]

[0111] In some embodiments, AA 2 is glycine, or [ka] are the amino acid residues of

[0112] 5.2.3. Aspect 3 The present specification describes compounds of formula (Ia), or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof,

[0113] During the ceremony, A.A. 2 contains the formula (W), and [ka] AA 3 is -glycine-glycine-phenylalanine-glycine- or [ka] is a tetrapeptide residue of

[0114] In some embodiments, x is 1 to 15. In some embodiments, x is 2 to 10. In some embodiments, x is 3 to 9. In one embodiment, x is about 3. In one embodiment, x is about 4. In one embodiment, x is about 5. In one embodiment, x is about 6. In one embodiment, x is about 7. In one embodiment, x is about 8. In one embodiment, x is about 9. In one embodiment, BA, RG 1 , SP 1 , SP 2 , R.G. 2 , HG, PAB, p and PA are as provided herein.

[0115] 5.2.4. Aspect 4 The present specification provides a compound according to formula (Ib): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof;

[0116] During the ceremony, A.A. 2 contains the formula (W), and [ka] AA 1 is -valine-alanine-, -valine-citrulline- or [ka] where R 6 is —CH3 or —(CH2)3—NHC(═O)NH2.

[0117] In one embodiment, [ka] teeth [ka] is.

[0118] In some embodiments, x is 1 to 15. In some embodiments, x is 2 to 10. In some embodiments, x is 3 to 9. In one embodiment, x is about 3. In one embodiment, x is about 4. In one embodiment, x is about 5. In one embodiment, x is about 6. In one embodiment, x is about 7. In one embodiment, x is about 8. In one embodiment, x is about 9.

[0119] In some embodiments, the ring-opened compound of Formula (Ib) is RG 1 is an open-ring heterocycle. In some embodiments, the compound of formula (Ib) with ring-opening is 1 In some embodiments, RG is a compound of formula (Ib) wherein RG is an open-ring heterocycle. 1 teeth, [ka] is.

[0120] In one embodiment, BA, RG 1 , SP 1 , SP 2 , R.G. 2 , HG, PAB, p and PA are as provided herein.

[0121] 5.2.5. Aspect 5 The present specification describes a compound of formula (Ib), or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof,

[0122] During the ceremony, A.A. 2 contains the formula (W), and [ka]

[0123] AA 1 is -glycine-glycine-phenylalanine-glycine- or [ka] is a tetrapeptide residue of

[0124] In one embodiment, BA, RG 1 , SP 1 , SP 2 , R.G. 2 , HG, PAB, p, x and PA are as provided herein.

[0125] 5.2.6. Aspect 6 The present specification provides a compound according to formula (Ic): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof;

[0126] During the ceremony, A.A. 3 is -valine-alanine-, -valine-citrulline- or [ka] where R 6 is —CH3 or —(CH2)3—NHC(═O)NH2.

[0127] In some embodiments, x is 1 to 15. In some embodiments, x is 2 to 10. In some embodiments, x is 3 to 9. In one embodiment, x is about 3. In one embodiment, x is about 4. In one embodiment, x is about 5. In one embodiment, x is about 6. In one embodiment, x is about 7. In one embodiment, x is about 8. In one embodiment, x is about 9.

[0128] In some embodiments, the ring-opened compound of Formula (Ic) is RG 1 is an open-ring heterocycle. In some embodiments, the compound of formula (Ic) with ring-opening is 1 In some embodiments, RG is a compound of formula (Ic), wherein RG is an open heterocycle. 1 teeth, [ka] is.

[0129] In one embodiment, BA, RG 1 , SP 1 , PAB, p, x and PA are as provided herein.

[0130] 5.2.7. Aspect 7 The present specification describes a compound of formula (Ic), or a pharmaceutically acceptable salt, solvate, stereoisomer, or derivative thereof,

[0131] During the ceremony, A.A. 3is -glycine-glycine-phenylalanine-glycine- or [ka] is a tetrapeptide residue of

[0132] In one embodiment, BA, RG 1 , SP 1 , PAB, p, x and PA are as provided herein.

[0133] 5.2.8. Aspect 8 In some embodiments, the compound is selected from the group consisting of the compounds in Table 3. In some embodiments, the compound is selected from the group consisting of ADC2-62-1, ADC2-62-2, ADC2-63-1, ADC2-63-2, ADC2-64-1, ADC2-64-2, ADC2-65-1, and ADC2-65-2.

[0134] 5.3. Linker with Payload (Platform) In some embodiments, the compounds described herein comprise a reactive linker attached to at least one payload moiety. In some embodiments, the present specification describes a compound, or a pharmaceutically acceptable solvate, stereoisomer, or derivative thereof, comprising a payload unit attached to at least one hydrophilic residue via a covalent linker, wherein the covalent linker is directly or indirectly attached to each payload unit and to the hydrophilic residue. In one embodiment, the hydrophilic residue comprises a terminal hydrophilic group. In one embodiment, the hydrophilic residue comprises a sugar residue.

[0135] 5.3.1. Aspect 9 In some embodiments, the present disclosure provides a compound having formula (II): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, wherein L is a covalent linker and PA is a payload residue.

[0136] 5.3.2. Aspect 10 The present specification provides a compound according to formula (IIa): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof;

[0137] During the ceremony, A.A. 2 contains the formula (W), and [ka]

[0138] AA 3 -valine-alanine-, -valine-citrulline- or [ka] is a dipeptide residue of R 6 is —CH3 or —(CH2)3—NHC(═O)NH2.

[0139] In one embodiment, [ka] teeth [ka] is.

[0140] In one embodiment, RG 1 teeth, [ka] Succinimide-N-, [ka] is.

[0141] In one embodiment, RG 1 teeth [ka] and EWG is an electron withdrawing group, e.g., -CN, -NO2, halogen, -CF3, -C(=O)OR 1 and -C(=O)R 1 and R 1 is substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted heteroaryl.

[0142] In one embodiment, RG 1 teeth, [ka] is.

[0143] In one embodiment, RG 1 teeth [ka] and EWG is an electron withdrawing group, e.g., -CN, -NO2, halogen, -CF3, -C(=O)OR 1 and -C(=O)R 1 and R 1 is substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted heteroaryl.

[0144] In some embodiments, the ring-opened compound of formula (II) is RG 1 is an open-ring heterocycle. In some embodiments, the compound of formula (II) with ring-opening is 1 In some embodiments, RG is a compound of formula (II) wherein RG is an open heterocycle.1 teeth [ka] and EWG is an electron withdrawing group, e.g., -CN, -NO2, halogen, -CF3, -C(=O)OR 1 and -C(=O)R 1 and R 1 is substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted heteroaryl.

[0145] In some embodiments, the ring-opened compound of Formula (IIa) is RG 1 is an open-ring heterocycle. In some embodiments, the compound of formula (IIa) with ring-opening is 1 In some embodiments, RG is a compound of formula (IIa) wherein RG is an open heterocycle. 1 teeth, [ka] is. In one embodiment, RG 1 , SP 1 , SP 2 , R.G. 2 , HG, PAB, p and PA are as provided herein.

[0146] 5.3.3. Aspect 11 The present specification describes a compound according to formula (IIa), or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, During the ceremony, A.A. 2 contains the formula (W), and [ka] AA 3 is -glycine-glycine-phenylalanine-glycine- or [ka] is a tetrapeptide residue of In one embodiment, RG 1 , SP 1 , SP 2 , R.G. 2 , HG, PAB, p and PA are as provided herein.

[0147] 5.3.4. Aspect 12 The present specification provides a compound according to formula (IIb): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof;

[0148] During the ceremony, A.A. 2 contains the formula (W), and [ka]

[0149] AA 1 is -valine-alanine-, -valine-citrulline- or [ka] where R 6 is —CH3 or —(CH2)3—NHC(═O)NH2.

[0150] In one embodiment, [ka] teeth [ka] is.

[0151] In some embodiments, the ring-opened compound of Formula (IIb) is RG 1In some embodiments, RG is a compound of formula (IIb), wherein RG is an open heterocycle. 1 teeth, [ka] is.

[0152] In one embodiment, RG 1 , SP 1 , SP 2 , R.G. 2 , HG, PAB, p and PA are as provided herein.

[0153] 5.3.5. Aspect 13 The present specification describes a compound according to formula (IIb), or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, During the ceremony, A.A. 2 contains the formula (W), and [ka]

[0154] AA 1 is -glycine-glycine-phenylalanine-glycine- or [ka] is a tetrapeptide residue of In one embodiment, RG 1 , SP 1 , SP 2 , R.G. 2 , HG, PAB, p and PA are as provided herein.

[0155] 5.3.6. Aspect 14 The present specification provides a compound according to formula (IIc): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof;

[0156] During the ceremony, A.A. 3 is -valine-alanine-, -valine-citrulline- or [ka] where R 6 is —CH3 or —(CH2)3—NHC(═O)NH2.

[0157] In some embodiments, the ring-opened compound of Formula (IIc) is RG 1 In some embodiments, RG is a compound of formula (IIc) wherein RG is an open heterocycle. 1 teeth, [ka] is. In one embodiment, RG 1 , SP 1 , PAB, p and PA are as provided herein.

[0158] 5.3.7. Aspect 15 The present specification describes a compound of formula (IIc), or a pharmaceutically acceptable salt, solvate, stereoisomer, or derivative thereof, In the formula, RG 1 is the reactive group residue, and SP 1 is an optional spacer group residue, PAB is an optional self-immolative unit, the subscript p is 0 or 1, and PA is a payload residue;

[0159] During the ceremony, A.A. 3 is -glycine-glycine-phenylalanine-glycine- or [ka] is a tetrapeptide residue of In one embodiment, RG 1 , SP1 , PAB, p and PA are as provided herein.

[0160] 5.3.8. Aspect 16 In some embodiments, the compound is selected from the group consisting of the compounds in Table 1.

[0161] 5.3.9. Aspect 17 The present specification describes a ligand-drug conjugate comprising the structure of formula (E1) or a pharmaceutically acceptable salt or solvate thereof: [ka] During the ceremony, R 7 and R 8 are each independently hydrogen, halogen, or alkyl.

[0162] In some embodiments, the ligand-drug conjugate comprises the structure of formula (E2): [ka] During the ceremony, BA is a binding agent selected from a humanized, chimeric, or human antibody or an antigen-binding antibody fragment of an antibody; L is a covalent linker as described herein, and x is 1 to 10 and may be an integer or a decimal number.

[0163] In some embodiments, the antibody is patritumab, cofetuzumab, or trastuzumab. In one embodiment, the antibody is patritumab. In one embodiment, R 7 and R 8 is hydrogen. In one embodiment, R 7 and R 8 is methyl. In one embodiment, R 7 is methyl and R 8 is F. In one embodiment, R7 and R 8 The carbon to which is attached is in the S configuration. In one embodiment, R 7 and R 8 The carbon to which is attached is in the R configuration. In some embodiments, the Ligand-Drug conjugate is selected from Table 24. In one embodiment, the antibody is patritumab.

[0164] 5.3.10. Aspect 18 The present specification provides a compound having the formula (E3): [ka] or a pharmaceutically acceptable solvate, stereoisomer or derivative thereof, wherein L is a covalent linker as described herein; R 7 and R 8 are each independently hydrogen, halogen, or alkyl.

[0165] In one embodiment, R 7 and R 8 is hydrogen. In one embodiment, R 7 and R 8 is methyl. In one embodiment, R 7 is methyl and R 8 is F. In one embodiment, R 7 and R 8 The carbon to which is attached is in the S configuration. In one embodiment, R 7 and R 8 The carbon to which is attached is in the R configuration. In one embodiment, the compound is selected from Table 25.

[0166] Payload The present specification provides a compound or a pharmaceutically acceptable solvate, stereoisomer, or derivative thereof, said compound having the formula (D4): [ka] In the formula, R 4 , R 5a and R 5b are each independently hydrogen, a sugar residue, a substituted or unsubstituted inorganic or organic acid residue, a substituted or unsubstituted C 1-8 alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted non-aromatic heterocyclyl, substituted or unsubstituted cycloalkylalkyl, or substituted or unsubstituted heterocyclylalkyl; R 5a and R 5b together with the atom to which they are attached form a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted non-aromatic heterocyclyl.

[0167] In one embodiment, the compound has one of the following structures: [ka]

[0168] The present specification provides a compound or a pharmaceutically acceptable solvate, stereoisomer, or derivative thereof, said compound having the formula (D5): [ka] wherein ring B is a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted aryl, a substituted or unsubstituted heterocyclyl, or a substituted or unsubstituted heteroaryl.

[0169] In one embodiment, the compound has one of the following structures: [ka]

[0170] The present specification provides a compound or a pharmaceutically acceptable solvate, stereoisomer, or derivative thereof, said compound having the formula (D6): [ka] In the formula, S 2 is an enzymatically hydrolyzable hydrophilic group. In one embodiment, S 2 The groups are as described herein.

[0171] The present specification provides a compound or a pharmaceutically acceptable solvate, stereoisomer or derivative thereof, said compound having the formula (E4): [ka] or a pharmaceutically acceptable solvate, stereoisomer or derivative thereof, In the formula, R 7 and R 8 are each independently hydrogen, halogen, or alkyl.

[0172] In one embodiment, R 7 and R 8 is hydrogen. In one embodiment, R 7 and R 8 is methyl. In one embodiment, R 7 is methyl and R 8 is F. In one embodiment, R 7 and R 8 The carbon to which is attached is in the S configuration. In one embodiment, R 7 and R 8 The carbon to which is attached is in the R configuration. In one embodiment, the compound is selected from Table 26.

[0173] 5.5. Method or process for preparing the complex In some embodiments, the present disclosure describes a method for preparing an antibody-drug conjugate, comprising contacting a binder with a linker-payload compound under conditions suitable for forming a bond between the binder and the linker-payload compound. Also provided are methods for preparing a compound of Formula (I), Formula (Ia), Formula (Ib), or Formula (Ic) under conditions suitable for forming a bond between the binder and the linker-payload compound.

[0174] In certain embodiments, an antibody is reacted with or treated with a reactive linker-payload to form an antibody-payload complex. The reaction can be carried out under conditions deemed appropriate by one of ordinary skill in the art. In certain embodiments, the antibody is contacted with the reactive linker-payload compound under conditions suitable for forming a bond between the antibody and the linker-payload compound. Suitable reaction conditions are well known to those of ordinary skill in the art.

[0175] Examples of such reactions are provided in the examples below. In some embodiments, the present specification describes a method for preparing a conjugate, comprising treating or contacting a compound with a binder under coupling conditions, wherein the compound comprises a reactive linker attached to at least one payload moiety, and the compound that reacts with the binder is a compound of Formula (II), Formula (IIa), Formula (IIb), or Formula (IIc), or a pharmaceutically acceptable salt, solvate, stereoisomer, or derivative thereof.

[0176] 5.6. Pharmaceutical Compositions The present disclosure provides pharmaceutical compositions comprising a compound described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0177] 5.7. How to use In some embodiments, the present specification describes a method of treating a disease, disorder, or condition in a patient in need thereof, comprising administering to the patient a compound or pharmaceutical composition described herein. In some embodiments, the compound administered is an antibody-drug conjugate described herein.

[0178] In some embodiments, the present specification describes a method for treating or preventing a disease, disorder, or condition selected from the group consisting of a proliferative disorder, a neurodegenerative disorder, an immunological disorder, an autoimmune disease, an inflammatory disorder, a skin disorder, a metabolic disorder, a cardiovascular disease, and a gastrointestinal disorder, comprising administering to a subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. In some embodiments, the compound administered is an antibody-drug conjugate described herein.

[0179] In some embodiments, the present specification describes a method of treating a proliferative disease, metabolic disease, inflammation, or neurodegenerative disease in a subject, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. In some embodiments, the present specification describes a method of treating a proliferative disease in a subject, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. In some embodiments, the compound administered is an antibody-drug conjugate described herein.

[0180] In some embodiments, the present specification describes a method of treating a metabolic disorder in a subject, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. In some embodiments, the compound administered is an antibody-drug conjugate described herein.

[0181] In some embodiments, the present specification describes a method of treating inflammation in a subject, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. In some embodiments, the compound administered is an antibody-drug conjugate described herein. In some embodiments, the present specification describes a method of treating a neurodegenerative disease in a subject, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition described herein. In some embodiments, the compound administered is an antibody-drug conjugate described herein.

[0182] Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described. All patents, applications, and non-patent publications mentioned herein are incorporated by reference in their entirety. [Example]

[0183] Reagents and solvents were obtained from commercial sources such as Sinopharm Chemical Reagent Co. (SCRC), Sigma-Aldrich, Alfa, or other suppliers unless otherwise stated.

[0184] As used herein, the symbols and conventions used in these procedures, schemes, and examples (whether or not a particular abbreviation is specifically defined) are consistent with the symbols and conventions used in the contemporary scientific literature (e.g., Journal of the American Chemical Society or Journal of Biological Chemistry). In particular, but not by way of limitation, the following abbreviations may be used in the examples and throughout this specification: For the sake of brevity, several abbreviations are used herein. As an example, single-letter abbreviations are used to represent amino acid residues. The amino acids and their corresponding three-letter and one-letter abbreviations are as follows:

[0185] [Table 1]

[0186] [Table 2-1]

[0187] [Table 2-2]

[0188] Example 1 [ka]

[0189] Step 1 (3aS,4S,6aR)-6-((S)-4-(((benzyloxy)carbonyl)amino)-5-methoxy-5-oxopentanamido)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carboxylic acid (1b) Wet Pd / C (50 mg, 10% purity) was added to a mixture of ((3aR,4R,6R,6aR)-6-azido-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methanol 1a (400 mg, 1.75 mmol, see NUCLEOSIDES, NUCLEOTIDES AND NUCLEIC ACIDS 2018, 37, 79-88) and TEA (353.2 mg, 3.49 mmol) in THF (4 mL). The reaction mixture was purged with a H2 balloon three times and allowed to react under H2 balloon at room temperature for 2 h. After completion of the reaction, the mixture was filtered through diatomaceous earth and washed with methanol. The filtrate was concentrated in vacuo to give 1b (500 mg, crude).

[0190] Step 2 Methyl N 2 -((benzyloxy)carbonyl)-N 5 -((3aR,6R,6aR)-6-(hydroxymethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-L-glutamic acid (1d) TEA (511 mg, 5.06 mmol) was added to a mixture of 1b (500 mg, crude) and 5-(2,5-dioxopyrrolidin-1-yl)1-methyl ((benzyloxy)carbonyl)-L-glutamate 1c (550 mg, 1.40 mmol) in THF (5 mL). The reaction mixture was allowed to react overnight at room temperature. After completion of the reaction, the mixture was concentrated under vacuum. The residue was purified using preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 1d (300 mg, Yield 45.9%). 22 H 30 N2O9 exact mass [M+H] + Calculated value: 467.2, measured value: 467.4.

[0191] Step 3 (3aS,4S,6aR)-6-((S)-4-(((benzyloxy)carbonyl)amino)-5-methoxy-5-oxopentanamido)-2,2-dimethyltetrahydrofuran[3,4-d][1,3]dioxole-4-carboxylic acid (1) TEMPO (18 mg, 0.12 mmol) and PhI(OAc) (285 mg, 0.88 mmol) were added to a mixture of 1d (275 mg, 0.59 mmol) in acetonitrile / HO (4:1, 6 mL). The reaction mixture was reacted at 40 °C for 6 hours. After the reaction was completed, the mixture was concentrated under vacuum. The residue was purified using preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 1 (115 mg, 40% yield). 22 H 28 N2O 10 Accurate mass [M+H] + Calculated value 481.17, measured value 481.4

[0192] Example 2 [ka]

[0193] Step 1 N2-((benzyloxy)carbonyl)-N5-((3aR,4R,6aS)-6-carbamoyl-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-L-glutamic acid methyl ester (2a) PyBOP (433 mg, 0.83 mmol), HOBt (113 mg, 0.83 mmol), and DIEA (161 mg, 1.25 mmol) were added to a mixture of 1 (200 mg, 0.42 mmol) and NHCl (45 mg, 0.83 mmol) in DMF (2 mL). The reaction mixture was heated to 50 °C overnight and monitored by LCMS. The mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 2a (112 mg, 56% yield). C22H29N3O9 Exact Mass [M+H] + Calculated value: 480.19, measured value: 480.4.

[0194] Step 2 N2-((benzyloxy)carbonyl)-N5-((3aR,6S,6aS)-6-carbamoyl-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-L-glutamine (2b) 1M aqueous LiOH (0.36 mL, 0.36 mmol) was added to a solution of 2a (160 mg, 0.33 mmol) in THF / MeOH (4:1, 2.5 mL). The reaction mixture was allowed to react at room temperature for 1 hour. The mixture was acidified to pH=6 with 1N HCl, filtered, and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 2b (85 mg, 65% yield). C21H27N3O9 Exact Mass [M+H] + Calculated value: 466.17, measured value: 466.4.

[0195] Step 3 Benzyl ((S)-5-(((3aR,6S,6aS)-6-carbamoyl-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)amino)-1-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)carbamate (2d) HATU (83 mg, 0.22 mmol) and DIEA (71 mg, 0.55 mmol) were added to a solution of 2b (85 mg, 0.18 mmol) in DMF (1 mL). The reaction was allowed to proceed at room temperature for 10 min, followed by the addition of (S)-2-((S)-2-amino-3-methylbutanamido)-N-(4-(hydroxymethyl)phenyl)-5-ureidopentanamide 2c (69 mg, 0.18 mmol, commercially available) and an additional 1 h at the same temperature. The reaction was quenched with 1 mL of water, filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min) to give 2d (85 mg, 65% yield). C39H54N8O12 Accurate mass [M+H] + Calculated value: 827.39, measured value: 827.6.

[0196] Step 4 Benzyl ((S)-5-(((3aR,6S,6aS)-6-carbamoyl-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)amino)-1-(((S)-3-methyl-1-(((S)-1-(4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)-2-oxo-6-ureidohexan-3-yl)amino)-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)carbamate (2e) DIEA (40 mg, 0.31 mmol) was added to a mixture of 2d (85 mg, 0.10 mmol) and PNP (47 mg, 0.15 mmol). The reaction mixture was reacted at room temperature for 4 hours and monitored by LCMS. The mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 2e (70.2 mg, Yield 70%). C47H58N8O16 Exact Mass [M+H] + Calculated value: 991.40, measured value: 992.7.

[0197] Step 5 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutane- 2-yl)(methyl)carbamic acid 4-((5S,8S,11S)-5-(3-(((3aR,6S,6aS)-6-carbamoyl-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)amino)-3-oxopropyl)-8-isopropyl-3,6,9-trioxo-1-phenyl-11-(3-ureidopropyl)-2-oxa-4,7,10-triazadodecan-12-amide) benzyl ester (2g) HOBt (4.4 mg, 0.3 mmol) and DIEA (17 mg, 0.13 mmol) were added to a mixture of 2e (65 mg, 0.07 mmol) and 2f (47 mg, 0.07 mmol, commercially available) in DMF (1.5 mL). The reaction mixture was allowed to react at room temperature for 24 hours. The mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 2g (68 mg, 66% yield). C79H119N13O20 Exact Mass [M+H] + Calculated 1570.87, Found 1593.2 [M + Na] + .

[0198] Step 6 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)- 3-Methyl-1-oxobutan-2-yl)(methyl)carbamic acid 4-((S)-2-((S)-2-((S)-2-amino-5-(3-(((3aR,6S,6aS)-6-carbamoyl-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxolyl-4-yl)amino)-5-oxopentanamido)-3-methylbutanamido)-5-ureidopentanamido) benzyl ester (2h) Wet Pd / C (5 mg, 10% purity) was added to a solution of 2g (23 mg, 0.01 mmol) in MeOH (1 mL). The reaction mixture was purged with a H2 balloon three times and allowed to react at room temperature under a H2 balloon for 2 h. The reaction mixture was filtered through a syringe filter, and the filtrate was concentrated in vacuo to give 2h (20 mg, crude). C71H113N13O18 Exact Mass [M+H] + Calculated value: 1436.83, measured value: 1438.2.

[0199] Step 7 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutane- 2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamic acid 4-((S)-2-((S)-2-((S)-2-amino-5-(((3R,4S,5S)-5-carbamoyl-3,4-dihydroxytetrahydrofuran-2-yl)amino)-5-oxopentanamido)-3-methylbutanamido)-5-ureidopentanamido) benzyl ester (2i) A mixture of 2h (20 mg, crude) in TFA / HO (1:1, 500 μL) was stirred at room temperature for 2 h. The mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 2i (11 mg, 55% yield). C68H109N13O18 Exact Mass [M+H] + Calculated value: 1396.80, measured value: 1398.1.

[0200] Step 8 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxopropyl 4-((S)-2-((S)-2-((S)-5-(((3R,4S,5S)-5-carbamoyl-3,4-dihydroxytetrahydrofuran-2-yl)amino)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)-5-oxopentanamido)-3-methylbutanamido)-5-ureidopentanamido) benzyl ester (2) DIEA (3.1 mg, 0.02 mmol) was added to a mixture of 2i (11 mg, 0.01 mmol) and 2j (2.4 mg, 0.01 mmol) in DMF (200 μL). The reaction mixture was allowed to react at room temperature for 2 hours. The reaction mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 2i (11 mg, 55% yield). C78H120N14O21 Exact Mass [M+H] + Calculated value 1589.88, Found value 796.0 [M+2H] 2+ .

[0201] Example 3 [ka]

[0202] Step 1 (3aS,4S,6R,6aR)-6-Azido-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carboxylic acid (3a) 3a was synthesized according to the synthetic procedure in Step 3 of Example 1.

[0203] Step 2 (3aS,4S,6R,6aR)-6-Azido-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carboxylate methyl ester (3b) K2CO3 (1.024 g, 7.41 mmol) and MeI (956 mg, 6.74 mmol) were added to a solution of 3a (1.45 g, 6.74 mmol) in DMF (10 mL). The reaction mixture was allowed to react at room temperature for 4 hours. Water (20 mL) was added to the mixture, and the mixture was extracted with EtOAc (20 mL*3). After separation, the combined organic layer was washed with brine (20 mL*3), dried over Na2SO4, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel flash chromatography (A-petroleum ether; B-EtOAc) to give 3b (820 mg, 50% yield).

[0204] Step 3 (3aS,4S,6aR)-6-amino-2,2-dimethyltetrahydrofuran[3,4-d][1,3]dioxole-4-carboxylate methyl ester (3) Wet Pd / C (40 mg, 10% yield) was added to a solution of 3b (400 mg, 1.75 mmol) in MeOH (5 mL). The reaction mixture was purged with a balloon of H2 gas and stirred at room temperature for 2 h under a balloon of H2 gas. After completion of the reaction, the mixture was filtered through diatomaceous earth and washed with MeOH. Finally, it was concentrated under vacuum to give 3 (300 mg, 79% yield). C19H15NO5 Exact Mass [M+H] + Calculated value 218.10, actual value 218.2.

[0205] Example 4 [ka]

[0206] [ka]

[0207] Step 1 (S)-tert-Butyl 4-(((benzyloxy)carbonyl)amino)-5-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (4c) HATU (338 mg, 0.89 mmol) and DIEA (345 mg, 2.67 mmol) were added to a mixture of 4a (300 mg, 0.89 mmol) in DMF (5 mL). The reaction mixture was allowed to react at room temperature for 10 min. 2c (100 mg, 0.89 mmol) was then added and allowed to stand at the same temperature for an additional 2 h. The reaction was quenched with 10 mL of water, filtered, and the filter cake was triturated with EtOAc at room temperature. After re-filtration and vacuum drying of the filter cake, 4c (520 mg, 84% yield) was obtained.

[0208] Step 2 (S)-4-(((benzyloxy)carbonyl)amino)-5-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoic acid (4d) A mixture of 4c (300 mg, 0.43 mmol) in TFA / DCM (1:1, 4 mL) was stirred at room temperature for 2 h. The mixture was concentrated under vacuum to give a residue, which was then dissolved in MeOH (3 mL), cooled to 0 °C, and MeONa (200 mg) was added to the methanol solution and stirred at 0 °C for 30 min. The mixture was acidified to pH = 3-4 with 1 M HCl. Most of the precipitate was filtered, and the filter cake was washed with water. Finally, the filter cake was dried under vacuum to give 4d (140 mg, 51% yield). C31H42N6O9 Exact Mass [M+H] + Calculated value: 643.30, measured value: 643.5.

[0209] Step 3 (3aS,4S,6aR)-6-((S)-4-(((benzyloxy)carbonyl)amino)-5-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carboxylate methyl ester (4e) HATU (99 mg, 0.22 mmol) and DIEA (84 mg, 0.65 mmol) were added to a mixture of 4d (140 mg, 0.22 mmol) and 3 (47.32 mg, 0.22 mmol) in DMF (1.2 mL). The mixture was allowed to react at room temperature for 1 hour. The mixture was filtered and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 4e (92 mg, 49% yield). C40H55N7O13 Exact Mass [M+H] + Calculated 842.39, Found 842.6 [M + H] + .

[0210] Step 4 (3aS,4S,6R,6aR)-6-((S)-4-(((benzyloxy)carbonyl)amino)-5-(((S)-3-methyl-1-(((S)-1-((4-(((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-1-oxobutan-2-yl)amino)-5-oxopentanamido)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carboxylate methyl (4f) 4f was synthesized according to the synthetic procedure in Step 4 of Example 2 (37 mg, 88% yield). C47H58N8O17 Exact Mass [M+H] + Calculated value: 1007.39, measured value: 1007.6.

[0211] Step 5 (3aS,4S,6aR)-6-((S)-4-(((benzyloxy)carbonyl)amino)-5-(((S)-1-(((S)-1-((4-((5S,8S,11S,12R)-11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl) -2-oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carboxylate methyl (4h) 4h was synthesized according to the synthetic procedure in Step 5 of Example 2 (40 mg, 68% yield). C80H120N12O21 Exact Mass [M+H] + Calculated value: 1586.87, measured value: 1587.37.

[0212] Step 6 (3aS,4S,6aR)-6-((S)-4-amino-5-(((S)-1-(((S)-1-((4-((5S,8S,11S,12R)-11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2-oxoethyl )-5,8-Diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carboxylate methyl (4i) 4i was synthesized according to the synthetic procedure in Step 6 of Example 2 (18 mg, crude). C72H114N12O19 Exact Mass [M+H] + Calculated value: 1451.83, measured value: 1453.2.

[0213] Step 7 (2S,3S,4R)-5-((S)-4-amino-5-(((S)-1-(((S)-1-((4-((5S,8S,11S,12R)-11-((S)-tert-butyl)-12-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2 -oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)-3,4-dihydroxytetrahydrofuran-2-carboxylate methyl (4j) 4j was synthesized according to the synthetic procedure in Step 7 of Example 2 (10 mg, 70% yield). C69H110N12O19 Exact Mass [M+H] + Calculated value: 1411.80, measured value: 1413.2.

[0214] Step 8 (2S,3S,4R)-5-((S)-4-amino-5-(((S)-1-(((S)-1-((4-((5S,8S,11S,12R)-11-((S)-tert-butyl)-12-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)- 2-Oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)-3,4-dihydroxytetrahydrofuran-2-carboxylic acid (4k) 1M aqueous LiOH (0.2 mL, 0.2 mmol) was added to a solution of 4j (10 mg, 0.007 mmol) in MeOH (0.5 mL). The reaction mixture was allowed to react at room temperature for 30 minutes. The mixture was acidified to pH=6 with 1N HCl, filtered, and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 4k (5.6 mg, 57% yield). C68H108N12O19 Exact Mass [M+H] + Calculated value: 1397.79, measured value: 1399.2.

[0215] Step 9 (2S,3S,4R)-5-((S)-5-(((S)-1-(((S)-1-((4-((5S,8S,11S,12R)-11-((S)-tert-butyl)-12-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2-oxoethyl)-5,8-diisopropyl-4,1 0-Dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-4-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)-5-oxopentanamido)-3,4-dihydroxytetrahydrofuran-2-carboxylic acid (4) 4 was synthesized according to the synthetic procedure in Step 7 of Example 2 (4.3 mg, 67% yield). C78H118N13O22 Exact Mass [M+H] + Calculated value: 1590.86, measured value: 1591.8.

[0216] Example 5 [ka]

[0217] Step 1 (3aS,4S,6R)-6-((S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(benzyloxy)-5-oxopentanamido)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carboxylate methyl ester (5b) HATU (580 mg, 1.52 mmol) and DIEA (591 mg, 4.57 mmol) were added to a mixture of (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(benzyloxy)-5-oxopentanoic acid 5a (700 mg, 1.52 mmol) and 3 (320 mg, 1.47 mmol). The reaction mixture was allowed to react at room temperature for 2 hours. The reaction mixture was quenched with 20 mL of water and extracted with EtOAc (30 mL*3). After separation, the combined organics were washed with brine (20 mL*3), dried over Na2SO4, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel flash chromatography (A-petroleum ether, B-EtOAc) to give 5b (742.7 mg, 74% yield). C36H38N2O10 exact mass [M+H] + Calculated value 659.25, actual value 659.5.

[0218] Step 2 N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-((3aR,6S,6aS)-6-(methoxycarbonyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-L-glutamine (5c) Wet Pd / C (70 mg, 10% purity) was added to a solution of 5b (739 mg, 1.12 mmol) in MeOH (10 mL). The reaction mixture was purged with a balloon of H gas three times and then stirred under a balloon of H gas at room temperature for 2 h. The mixture was washed with diatomaceous earth, filtered with MeOH, and the filtrate was concentrated in vacuo to give 5c (600 mg, 94% yield). C29H32N2O10 exact mass [M+H] + Calculated value: 569.21, measured value: 569.5.

[0219] Step 3 N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-((3R,4S,5S)-3,4-dihydroxy-5-(methoxycarbonyl)tetrahydrofuran-2-yl)-L-glutamine (5) 5 was synthesized according to the synthetic procedure in Step 7 of Example 2 (35 mg, 47% yield). C26H28N2O10 Exact Mass [M+H] + Calculated value: 529.17, measured value: 529.4.

[0220] Example 6 [ka]

[0221] Step 1 (1S,2R)-2-((2R,3R)-3-((S)-1-((5S,8S,11S,12R)-1-(4-((S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)phenyl)-11-((S)-tert-butyl)-5,8-diisopropyl-12-methoxy-4,10-dimethyl-3,6,9-trioxo-2-oxa-4,7,10-triazatetradecan-14-oyl)pyrrolidin-2-yl)-3-methoxy-2-methylpropanamido)-1-phenylpropane hydrogen sulfate (6b) 6b was synthesized according to the synthetic procedure in Step 5 of Example 2 (50 mg, 50% yield). C73H104N10O17S Exact Mass [MH] - Calculated value: 1423.73, actual value: 1425.0.

[0222] Step 2 (1S,2R)-2-((2R,3R)-3-((S)-1-((5S,8S,11S,12R)-1-(4-((S)-2-((S)-2-amino-3-methylbutanamido)-5-ureidopentanamido)phenyl)-11-((S)-tert-butyl)-5,8-diisopropyl-12-methoxy-4,10-dimethyl-3,6,9-trioxo-2-oxa-4,7,10-triazatetradecan-14-oyl)pyrrolidin-2-yl)-3-methoxy-2-methylpropanamido)-1-phenylpropane hydrogen sulfate (6c) 6b (50 mg, 0.04 mmol) was dissolved in 20% piperidine (1 mL, DMF solution) and stirred at room temperature for 10 min. The mixture was acidified with 1N HCl to pH 5-6, then filtered and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5um 19*250mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min) to give 6c (34 mg, 81% yield). C58H94N10O15S Exact Mass [MH] - Calculated value: 1201.66, measured value: 1201.8.

[0223] Step 3 (2S,3S,4R)-5-((S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(((S)-1-(((S)-1-((4-((5S,8S,11S,12R)-11-((S)-tert-butyl)-5,8-diisopropyl-12-(2-((S)-2-((1R,2R)-1-methoxy-2-methyl-3-oxo-3-(((1S,2R)-1-phenyl-1-(sulfoxy)propane -2-yl)amino)propyl)pyrrolidin-1-yl)-2-oxoethyl)-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)-3,4-dihydroxytetrahydrofuran-2-carboxylate methyl (6d) HATU (15 mg, 0.04 mmol) and DIEA (15 mg, 0.119 mmol) were added to a solution of 5 (21 mg, 0.04 mmol) in DMF (2 mL). The reaction mixture was allowed to react at room temperature for 10 minutes. 6c (33.5 mg, 0.028 mmol) was then added and the mixture was left at the same temperature for another hour. The mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 6c (34 mg, 81% yield). C84H120N12O24S Exact Mass [MH] - Calculated value: 1711.83, measured value: 1713.2.

[0224] Step 4 (2S,3S,4R)-5-((S)-4-amino-5-(((S)-1-(((S)-1-((4-((5S,8S,11S,12R)-11-((S)-tert-butyl)-5,8-diisopropyl-12-(2-((S)-2-((1R,2R)-1-methoxy-2-methyl-3-oxo-3-(((1S,2R)-1-phenyl-1-(sulfoxy)propan-2-yl)amino)propan (I)pyrrolidin-1-yl)-2-oxoethyl)-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)-3,4-dihydroxytetrahydrofuran-2-carboxylic acid (6e) 2M LiOH aqueous solution (20 μL, 0.04 mmol) was added to a solution of 6d (25 mg, 0.01 mmol) in THF (0.2 mL). The reaction mixture was allowed to react at room temperature for 2 hours. The mixture was acidified to pH=6 with 1N HCl, filtered, and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 6e (85 mg, 48% yield). C68H108N12O22S exact mass [MH]- Calculated value: 1475.74, actual value: 1475.9.

[0225] Step 5 (2S,3S,4R)-5-((S)-5-(((S)-1-(((S)-1-((4-((5S,8S,11S,12R)-11-((S)-tert-butyl)-5,8-diisopropyl-12-(2-((S)-2-((1R,2R)-1-methoxy-2-methyl-3-oxo-3-(((1S,2R)-1-phenyl-1-(sulfooxy)propan-2-yl)amino)propyl)pyrrolidin-1-yl)-2-oxoethyl)-4, 10-Dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-4-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)-5-oxopentanamido)-3,4-dihydroxytetrahydrofuran-2-carboxylic acid (6) 6 was synthesized according to the synthetic procedure in Step 8 of Example 2 (3.6 mg, 80% yield). C78H119N13O25S exact mass [MH] - Calculated value: 1668.82, measured value: 1670.7.

[0226] Example 7 [ka]

[0227] Step 1 (2S,3S,4R)-5-((S)-5-(((S)-1-(((S)-1-((4-((5S,8S,11S,12R)-11-((S)-tert-butyl)-5,8-diisopropyl-12-(2-((S)-2-((1R,2R)-1-methoxy-2-methyl-3-oxo-3-(((1S,2R)-1-phenyl-1-(sulfooxy)propan-2-yl)amino)propyl)pyrrolidin-1-yl)-2-oxoethyl)-4,10-dimethyl methyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-4-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamido)-5-oxopentanamido)-3,4-dihydroxytetrahydrofuran-2-carboxylic acid (7) Compound 7 was synthesized according to the synthetic procedure in Step 8 of Example 2 (4.5 mg, 57% yield). MS(ESI) m / z: 1684.8 [MH] -

[0228] Example 8 [ka]

[0229] Step 1 2-((3aS,4S,6R,6aR)-6-((((benzyloxy)carbonyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)acetic acid (8b) 8a (740 mg, 1.95 mmol, synthesized according to the method described in Journal of Carbohydrate Chemistry 2000, 19, 653-657) was dissolved in a mixture of MeOH (4 mL) and HO (1 mL), and then NaOH (187 mg, 23.95 mmol) was added. The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, the mixture was concentrated under reduced pressure to remove most of the MeOH. HO (5 mL) was then added, and the mixture was adjusted to pH 5-6 with aqueous HCl (1 M) at 0 °C. The mixture was extracted with EA (20 mL * 3). The combined organic layer was dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give 8b (560 mg, 79% yield) as a colorless oil. MS(ESI) m / z: 364.2 [MH] -

[0230] Step 2 2,5-Dioxopyrrolidin-1-yl 2-((3aS,4S,6R,6aR)-6-((((benzyloxy)carbonyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)acetate (8c) 8b was dissolved in DCM (5 mL), and then DCC (365 mg, 1.77 mmol) and HOSu (204 mg, 1.77 mmol) were added. The resulting mixture was stirred at 25° C. for 2 hours. After the reaction was completed, the reaction mixture was filtered and concentrated to obtain a yellow solid in the form of crude product, which was purified by flash column chromatography (eluted with PE / EA). 8c (490 mg, 72% yield) was obtained as a pale yellow solid. MS(ESI) m / z: 485.4 [M+Na] +

[0231] Step 3 (2-((2S,3R,4S,5R)-5-((((benzyloxy)carbonyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)acetyl)glycylglycylglycylglycine (8e) 8c (490 mg, 1.06 mmol) was dissolved in THF (8 mL), and then 8d (381.70 mg, 1.06 mmol, commercially available) and saturated aqueous NaHCO3 (1 mL) were added. The resulting mixture was stirred at 25 °C for 2 h. After the reaction was completed, the mixture was adjusted to pH = 1 with aqueous HCl (1 M) at 0 °C and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give the final product 8e (100 mg, 19% yield). MS(ESI) m / z: 594.4 [M+H] +

[0232] Step 4 Methyl(2-((2S,3R,4S,5R)-5-((((benzyloxy)carbonyl)amino)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)acetyl)glycylglycylglycylglycine (8f) 8e (90 mg, 0.16 mmol) was dissolved in DMF (2 mL), and then CHCl (35 mg, 0.24 mmol) and NaHCO (27 mg, 0.33 mmol) were added sequentially. The resulting mixture was stirred at 25 °C for 16 h. After the reaction was completed, the reaction mixture was adjusted to pH = 6 with aqueous HCl (1 M) at 0 °C and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min). After lyophilization, 8f (85 mg, 83% yield) was obtained as a clear oil. MS(ESI) m / z: 590.4 [M+Na] +

[0233] Step 5 Methyl(2-((2S,3R,4S,5R)-5-(aminomethyl)-3,4-dihydroxytetrahydrofuran-2-yl)acetyl)glycylglycylglycylglycine (8g) 8f (85 mg, 0.15 mmol) is dissolved in MeOH (2 mL), and then Pd / C (wet, 10 mg, 10%) is added. The resulting mixture is stirred at 25° C. for 2 hours. After the reaction is complete, the reaction mixture is filtered through diatomaceous earth. The resulting filtrate is concentrated under reduced pressure to give a clear oil (65 mg) in the form of a crude product, which is used directly in the next step. MS(ESI) m / z: 434.3 [M+H] +

[0234] Step 6 Benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((2R,3S,4R,5S)-3,4-dihydroxy-5-(2-((2-((2-((2-((2-methoxy-2-oxoethyl)amino)-2-oxoethyl)amino)-2-oxoethyl)amino)-2-oxoethyl)amino)-2-oxoethyl)tetrahydrofuran-2-yl)methyl)-L-glutamate (8i) 8g (65 mg, 0.15 mmol) was dissolved in THF (2 mL), and then 8h (100 mg, 0.18 mmol, commercially available) and saturated NaHCO3 (0.5 mL) were added. The resulting mixture was stirred at 25 °C for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give 8i (132 mg, crude), which was used directly in the next step. MS(ESI) m / z: 875.5 [M+H] +

[0235] Step 7 N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((2R,3S,4R,5S)-3,4-dihydroxy-5-(2-((2-((2-((2-((2-methoxy-2-oxoethyl)amino)-2-oxoethyl)amino)-2-oxoethyl)amino)-2-oxoethyl)amino)-2-oxoethyl)tetrahydrofuran-2-yl)methyl)-L-glutamine (8) Compound 8 was synthesized according to the synthetic procedure in Step 2 of Example 5 (70 mg, 59% yield). MS(ESI) m / z: 785.5 [M+H] +

[0236] Example 9 [ka]

[0237] Step 1 4-((S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)benzyl ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate (9a) 9a was synthesized according to the synthetic procedure in Step 5 of Example 2 (60 mg, 32% yield). MS(ESI) m / z: 1347.1 [M+H] +

[0238] Step 2 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamic acid 4-((S)-2-((S)-2-amino-3-methylbutanamido)-5-ureidopentanamido)benzyl ester (9b) 9b was synthesized according to the synthetic procedure in Step 2 of Example 6 (31 mg, 70% yield). MS(ESI) m / z: 1124.7 [M+H]+

[0239] Step 3 (2-((2S,3R,4S,5R)-5-(((S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(((S)-1-(((S)-1-((4-(((((S)-1-((((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl- 1-Oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)acetyl)glycylglycylglycylglycine methyl ester (9d) 9d was synthesized according to the synthetic procedure in Step 3 of Example 6 (19 mg, 51% yield). MS(ESI) m / z: 946.1 [M+2H] 2+

[0240] Step 4 (2-((2S,3R,4S,5R)-5-(((S)-4-amino-5-(((S)-1-(((S)-1-((4-(((((S)-1-((((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl )(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)acetyl)glycylglycylglycylglycine (9e) 9d (19 mg, 0.01 mmol) was dissolved in a mixed solvent of CHCN and HO (v:v=1:1, 1 mL), and then piperidine (1.12 g, 2.936 mmol) was added. The resulting mixture was stirred at 25 °C for 2 h. The reaction mixture was then adjusted to pH=6 with HCl (1 M) at 0 °C and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 9e (14 mg, Yield: 87%). MS(ESI) m / z: 828.0 [M+2H] 2+

[0241] Step 5 (2-((2S,3R,4S,5R)-5-(((S)-4-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)-5-(((S)-1-(((S)-1-((4-(((((S)-1-((((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy -5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamoyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)acetyl)glycylglycylglycylglycine (9) Compound 9 was synthesized according to the synthetic procedure in Step 8 of Example 2 (13 mg, 83% yield). MS(ESI) m / z: 924.5 [M+2H] 2+

[0242] Example 10 [ka]

[0243] Step 1 ((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-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)carbamic acid 4-((R)-2-((R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)benzyl ester (10b) 6a (100 mg, 0.13 mmol) and 10a (69 mg, 0.13 mmol) were dissolved in DMF (1 mL), and then DIEA (51 mg, 0.39 mmol) was added. The resulting mixture was stirred at 25 °C for 4 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give the title compound 10b (200 mg, crude). MS(ESI) m / z: 1063.6 [M+H] +

[0244] Step 2 ((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-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)carbamic acid 4-((R)-2-((R)-2-amino-3-methylbutanamido)-5-ureidopentanamido)benzyl ester (10c) 10c was synthesized according to the synthetic procedure in Step 2 of Example 6 (52 mg, 66% yield).

[0245] Step 3 (2-((2S,3R,4S,5R)-5-(((S)-4-(((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(((R)-1-(((R)-1-((4-((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)carbamoyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)acetyl)glycylglycylglycylglycine methyl ester (10d) 10d was synthesized according to the synthetic procedure in Step 3 of Example 6 (28 mg, 28% yield). MS(ESI) m / z: 804.0 [M+2H]2+

[0246] Step 4 (2-((2S,3R,4S,5R)-5-(((S)-4-amino-5-(((R)-1-(((R)-1-((4-((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)carbamoyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)acetyl)glycylglycylglycylglycine (10e) 10e was synthesized according to the synthetic procedure in Step 4 of Example 9 (16 mg, 68% yield). MS(ESI) m / z: 1372.9 [M+H] +

[0247] Step 5 (2-((2S,3R,4S,5R)-5-(((S)-4-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)-5-(((R)-1-(((R)-1-((4-((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pi Iso[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanamido)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)acetyl)glycylglycylglycylglycine (10) Compound 10 was synthesized according to the synthetic procedure in Step 8 of Example 2 (7.5 mg, 34% yield). MS(ESI) m / z: 1565.5 [M+H] +

[0248] Example 11 [ka]

[0249] Step 1 N5-((3aR,6S,6aS)-6-((2,5,8,11-tetraoxatridecan-13-yl)carbamoyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-N2-((benzyloxy)carbonyl)-L-glutamic acid methyl ester (11b) 1 (300 mg, 0.62 mmol, synthesized according to the synthetic procedure of Example 1) was dissolved in DMF (4 mL), and then HATU (309 mg, 0.81 mmol) and DIEA (242 mg, 1.87 mmol) were added. The mixture was stirred at 25° C. for 15 minutes, followed by the addition of 11a (259 mg, 1.25 mmol). The resulting mixture was stirred at 25° C. for an additional 5 hours. After completion of the reaction, the reaction mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 11b (164 mg, Yield: 45%). MS(ESI) m / z: 670.6 [M+H] +

[0250] Step 2 N5-((3aR,6S,6aS)-6-((2,5,8,11-tetraoxatridecan-13-yl)carbamoyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-N2-((benzyloxy)carbonyl)-L-glutamine (11c) 11b (116 mg, 0.17 mmol) was dissolved in a mixture of THF and MeOH (v:v = 1:1, 0.6 mL), and then LiOH·HO (13 mg, 0.31 mmol) in HO (0.3 mL) was added. The resulting mixture was stirred at 40 °C for 1.5 h. After the reaction was completed, the reaction mixture was adjusted to pH 5 with aqueous HCl (1 M) and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min) to give 11c (62 mg, Yield: 55%). MS(ESI) m / z: 656.5 [M+H] + and 654.3 [MH] -

[0251] Step 3 Benzyl ((S)-5-(((3aR,6S,6aS)-6-((2,5,8,11-tetraoxatridecan-13-yl)carbamoyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)amino)-1-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)carbamate (11d) 11c (58 mg, 0.09 mmol) was dissolved in DMF (2 mL), followed by the addition of HATU (44 mg, 0.11 mmol) and DIEA (34 mg, 0.27 mmol). The resulting mixture was stirred at 25 °C for 15 minutes, followed by the addition of 2c (synthesized according to the procedure disclosed in patent WO2020 / 14541A2). The mixture was stirred at 25 °C for an additional 2 hours. After the reaction was completed, the mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to obtain 11d (58 mg, Yield: 65%). MS(ESI) m / z: 1017.8 [M+H] +

[0252] Step 4 Benzyl ((S)-5-(((3aR,6S,6aS)-6-((2,5,8,11-tetraoxatridecan-13-yl)carbamoyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)amino)-1-(((S)-3-methyl-1-(((S)-1-((4-(((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-1-oxobutan-2-yl)amino)-1,5-dioxopentan-2-yl)carbamate (11e) 11d (20 mg, 0.02 mmol) and p-nitrophenol carbonate (7 mg, 0.02 mmol) were dissolved in DMF (0.2 mL), and then DIEA (8 mg, 0.06 mmol) was added. The resulting mixture was stirred at 25 °C for 4 h. After the reaction was completed, HO (1 mL) was added to quench the reaction. The reaction mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 11e (18 mg, Yield: 78%). MS(ESI) m / z: 1017.8 [M+H] +

[0253] Step 5 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamic acid 4-((5S,8S,11S)-5-(3-(((3aR,6S,6aS)-6-((2,5,8,11-tetraoxatridecan-13-yl)carbamoyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)amino)-3-oxopropyl)-8-isopropyl-3,6,9-trioxo-1-phenyl-11-(3-ureidopropyl)-2-oxa-4,7,10-triazadodecan-12-amide)benzyl ester (11f) 11f was synthesized according to the synthetic procedure in Step 5 of Example 2 (31 mg, 68% yield). MS(ESI) m / z: 881.5 [M+2H] 2+

[0254] Step 6 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutane -2-yl)(methyl)carbamic acid 4-((S)-2-((S)-2-((S)-5-(((3aR,6S,6aS)-6-((2,5,8,11-tetraoxatridecan-13-yl)carbamoyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)amino)-2-amino-5-oxopentanamido)-3-methylbutanamido)-5-ureidopentanamido) benzyl ester (11g) 11g was synthesized according to the synthetic procedure in Step 6 of Example 2 (25 mg, crude). MS(ESI) m / z: 814.6 [M+2H] 2+

[0255] Step 7 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl -1-Oxobutan-2-yl)(methyl)carbamic acid 4-((S)-2-((S)-2-((S)-5-(((3R,4S,5S)-5-((2,5,8,11-tetraoxatridecan-13-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)amino)-2-amino-5-oxopentanamido)-3-methylbutanamido)-5-ureidopentanamido) benzyl ester (11h) 11h was synthesized according to the synthetic procedure in Step 7 of Example 2 (6.5 mg, 27% yield). MS(ESI) m / z: 1588.4 [M+H] +

[0256] Step 8 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl) Carbamic acid 4-((S)-2-((S)-2-((S)-5-(((3R,4S,5S)-5-((2,5,8,11-tetraoxatridecan-13-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)amino)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)-5-oxopentanamido)-3-methylbutanamido)-5-ureidopentanamido) benzyl ester (11) 11 was synthesized according to the synthetic procedure in Step 8 of Example 2 (3.5 mg, 48% yield). MS(ESI) m / z: 1781.0 [M+H] +

[0257] Example 12 [ka]

[0258] Step 1 ((S)-tert-Butyl 1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate (12a) Di-tert-butyl carbonate (60 μL, 0.15 mmol) was added to a solution of MMAE (100 mg, 0.14 mmol) in EtOH (1 mL). The mixture was stirred overnight. After completion of the reaction, the reaction mixture was concentrated under reduced pressure and purified by flash column chromatography (eluent: DCM / MeOH, 100 / 0 to 20 / 1 (v / v)) to give 12a (115 mg, quantitative). MS(ESI) m / z: 818.8 [M+H] +

[0259] Step 2 (1S,2R)-2-((2R,3R)-3-((S)-1-((6S,9S,12S,13R)-12-((S)-tert-butyl)-6,9-diisopropyl-13-methoxy-2,2,5,11-tetramethyl-4,7,10-trioxo-3-oxa-5,8,11-triazapentadecan-15-oyl)pyrrolidin-2-yl)-3-methoxy-2-methylpropanamido)-1-phenylpropane hydrogen sulfate (12b) At 0 °C, EtN (48 μL, 0.34 mmol) and chlorosulfonic acid (14 μL, 0.22 mmol) are added to a solution of 12a (70 mg, 0.08 mmol) in DCM (1 mL). The resulting mixture is stirred at this temperature for 1 h. After completion of the reaction, HO (1 mL) is added to quench the reaction. The mixture is extracted with DCM (1 mL × 4). The combined organic layers are dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give the crude product as a brown oil, which is purified by flash column chromatography (eluent: DCM / MeOH, 100 / 0 to 10 / 1 (v / v)) to give 12b (47 mg, 65% yield) as a white solid. MS(ESI) m / z: 896.6 [MH] -

[0260] Step 3 (1S,2R)-2-((2R,3R)-3-((S)-1-((3R,4S,5S)-4-((S)-N,3-dimethyl-2-((S)-3-methyl-2-(methylamino)butanamido)butanamido)-3-methoxy-5-methylheptanoyl)pyrrolidin-2-yl)-3-methoxy-2-methylpropanamido)-1-phenylpropane hydrogen sulfate (12c) 12b (40 mg, 0.04 mmol) was dissolved in DCM (0.8 mL), and then TFA (0.2 mL) was added. The resulting mixture was stirred at 0 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure to give crude 12c (50 mg), which was used directly in the next step. MS(ESI) m / z: 796.7 [MH] -

[0261] Step 4 (1S,2R)-2-((2R,3R)-3-((S)-1-((5S,8S,11S,12R)-11-((S)-tert-butyl)-1-(4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)-3-methylbutanamido)-5-ureidopentanamido)phenyl)-5,8-diisopropyl-12-methoxy-4,10-dimethyl-3,6,9-trioxo-2-oxa-4,7,10-triazatetradecan-14-oyl)pyrrolidin-2-yl)-3-methoxy-2-methylpropanamido)-1-phenylpropane hydrogen sulfate (12) 12c (27 mg, 0.03 mmol) and 12d (37 mg, 0.05 mmol, commercially available) were dissolved in DMF (0.5 mL), and then HOBt (0.9 mg, 0.01 mmol) and DIEA (56 μL, 0.34 mmol) were added. The mixture was stirred at 25 °C overnight. After the reaction was completed, the mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 12 (5.4 mg, 13% yield) as a white solid. MS(ESI) m / z: 1395.0 [M−H]-

[0262] Example 13 [ka]

[0263] Step 1 (3'-amino-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-6'-yl)ethyl carbamate (13b) Ethyl chloroformate (284 mg, 2.6 mmol) was added dropwise to a solution of 2-(6-amino-3-imino-3H-xanthen-9-yl)benzoic acid hydrochloride 13a (800 mg, 2.18 mmol) and DIEA (705 mg, 5.4 mmol) in DMF (5 mL) at 0 °C. After the addition was complete, the solution was stirred at room temperature for 15 h. The reaction was quenched by adding ice water (5 mL). The solution was extracted with EtOAc (20 mL * 3), washed with water (10 mL * 2) and brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and purified by flash column chromatography (petroleum ether / EtOAc = 100 / 0 to 70 / 30) to give the title compound 13b (321 mg, 36.3% yield). MS(ESI) m / z: 403.0 [M+H] +

[0264] Step 2 (3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-3',6'-diyl)biscarbamic acid 4-((14S,17S)-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-14-isopropyl-12,15-dioxo-17-(3-ureidopropyl)-3,6,9-trioxa-13,16-diazaoctadecane-18-amide) benzyl ester ethyl ester (13) To a solution of 13b (90 mg, 0.22 mmol) and 2,6-lutidine (72 mg, 0.67 mmol) in DCM (15 mL) was added triphosgene (33 mg, 0.11 mmol) portionwise under a nitrogen atmosphere at 0 °C and stirred at room temperature for 3 h. A solution of intermediate 13c (148 mg, 0.22 mmol) in DCM (3 mL) was added dropwise at 0 °C and stirred at room temperature for 15 h. The reaction was quenched by adding water (5 mL). The organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by flash column chromatography to give the title compound 13 (30 mg, 12.5% ​​yield). MS(ESI) m / z: 1091.4 [M+H] +

[0265] Example 14 [ka]

[0266] Step 1 (S)-Benzyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-((tert-butoxycarbonyl)amino)propanoate (14b) (S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-((tert-butoxycarbonyl)amino)propanoic acid 14a (10.0 g, 23.45 mmol) and benzyl bromide (16.0 g, 93.79 mmol) were dissolved in DMF (118 mL). Sodium bicarbonate (3.94 g, 46.90 mmol) was added to the solution and stirred at room temperature for 15 hours. EtOAc (1 L) was added and washed with water (200 mL*3) and brine (200 mL). The organic phase was dried over anhydrous sodium sulfate, concentrated under vacuum, and purified by flash column chromatography to give the title compound 14b (7 g, 57.8% yield). MS(ESI) m / z: 517.3 [M+H] +

[0267] Step 2 (S)-Benzyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-aminopropanoate (14c) To a solution of 14b (7.0 g, 13.55 mmol) in DCM (20 mL) was added TFA (10 mL) and stirred at room temperature for 2 h. The solution was concentrated, the pH of the solution was adjusted to 8 with saturated sodium bicarbonate, extracted with EtOAc (200 mL*3), dried over anhydrous sodium sulfate, concentrated under vacuum, and purified by flash column chromatography to give the title compound 14c (4.5 g, 79.7% yield). MS(ESI) m / z: 417.2 [M+H] +

[0268] Step 3 (S)-Benzyl 2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(sulfamoylamino)propanoate (14d) To a solution of 14c (4.0 g, 9.60 mmol) and TEA (1.5 g, 14.41 mmol) in DCM (100 mL) was added chlorosulfonamide (1.2 g, 10.57 mmol) portionwise at 0 °C and stirred at room temperature for 15 h. The reaction was quenched by adding water (100 mL), extracted with EtOAc (200 mL*3), dried over anhydrous sodium sulfate, concentrated in vacuo, and purified by flash column chromatography to give the title compound 14d (1.2 g, 25.2% yield). MS(ESI) m / z: 496.2 [M+H] +

[0269] Step 4 (S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-(sulfamoylamino)propanoic acid (14e) To a solution of 14d (1.1 g, 2.22 mmol) in MeOH (20 mL) and THF (20 mL) was added wet Pd / C (10%, 1.1 g) and stirred at room temperature under H2 (15 psi) for 1 h. The solution was filtered through diatomaceous earth and concentrated in vacuo to give the title compound 14e (600 mg, 66.7% yield). MS(ESI) m / z: 406.1 [M+H] +

[0270] Step 5 ((S)-1-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-1-oxo-3-(sulfamoylamino)propan-2-yl)carbamate (9H-fluoren-9-yl)methyl (14g) To a solution of 14e (600 mg, 1.48 mmol) and 14f (562 mg, 1.48 mmol) in DMF (10 mL) was added DIEA (383 mg, 2.96 mmol). HATU (732 mg, 1.92 mmol) was added to the solution at 0 °C and stirred at room temperature for 3 h. The reaction solution was purified by flash column chromatography to give the title compound 14g (600 mg, 52.9% yield). MS(ESI) m / z: 767.3 [M+H] +

[0271] Step 6 (3-Oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-3',6'-diyl)biscarbamic acid 4-((5S,8S,11S)-1-(9H-fluoren-9-yl)-8-isopropyl-3,6,9-trioxo-5-((sulfamoylamino)methyl)-11-(3-ureidopropyl)-2-oxa-4,7,10-triazadodecan-12-amide) benzyl ester ethyl ester (14h) 14h was synthesized according to the synthetic procedure described in Step 2 of Example 13 (50 mg, 8.0% yield). MS(ESI) m / z: 1195.4 [M+H] +

[0272] Step 7 (3-Oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-3',6'-diyl)biscarbamic acid 4-((S)-2-((S)-2-((S)-2-amino-3-(sulfamoylamino)propanamido)-3-methylbutanamido)-5-ureidopentanamido) benzyl ester ethyl ester (14i) To a solution of 14h (50 mg, 0.04 mmol) in DMF (3 mL), DBU (26 mg, 0.17 mmol) was added and stirred at room temperature for 2 h. The reaction solution was purified by flash column chromatography to give the title compound 14i (20 mg, 49.1% yield). MS(ESI) m / z: 973.4 [M+H] +

[0273] Step 8 (3-Oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-3',6'-diyl)biscarbamic acid 4-((2S,5S,8S)-21-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-5-isopropyl-4,7,10-trioxo-8-((sulfamoylamino)methyl)-2-(3-ureidopropyl)-13,16,19-trioxa-3,6,9-triazahencosamide) benzyl ester ethyl ester (14) To a solution of 14i (20 mg, 0.02 mmol), 14j (7 mg, 1.40 mmol), and DIEA (5.3 mg, 0.04 mmol) in DMF (3 mL), HATU (10 mg, 0.03 mmol) was added at 0° C. and stirred at room temperature for 3 h. The reaction solution was purified by preparative HPLC to give the title compound 14 (8.3 mg, 32.1% yield). MS(ESI) m / z: 1256.3 [M+H] +

[0274] Example 15 [ka]

[0275] Step 1 (tert-Butoxycarbonyl)(sulfo)-D-alanine (15b) To a solution of sulfo-D-alanine 15a (1.2 g, 7.1 mmol) and di-tert-butyl dicarbonate (1.9 g, 8.52 mmol) in DMF (10 mL), DIEA (1.4 g, 14.2 mmol) was added and stirred at room temperature overnight. The reaction solution was concentrated in vacuo to give the title compound 15b (2.0 g, crude) as a colorless oil. MS(ESI) m / z: 267.7 [MH] -

[0276] Step 2 (R)-2-((tert-butoxycarbonyl)amino)-3-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropane-1-sulfonic acid (15c) To a solution of 15b (100 mg, 0.37 mmol) and 14f (99 mg, 0.26 mmol) in DMF (3 mL), DIEA (1.5 g, 11.9 mmol) and BOP (246 mg, 0.56 mmol) were added and stirred in an ice bath for 0.5 h. The reaction solution was purified by preparative HPLC (water / MeCN = 100 / 0 to 80 / 20, 30 min, detection wavelength: 254 / 220 nm) to give the title compound 15c (180 mg, 77.1% yield) as a pale yellow solid. MS(ESI) m / z: 629.1 [MH] -

[0277] Step 3 (R)-2-Amino-3-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropane-1-sulfonic acid (15d) To a solution of 15c (210 mg, 0.33 mmol) in DCM (3 mL), add TFA (1 mL), stir at room temperature for 1 h, and concentrate. The residue is dissolved in 1,4-dioxane (2 mL), and a solution of NaOH (67 mg, 1.67 mmol) in water (1 mL) is added and stirred at room temperature for 1 h. The pH of the reaction solution is adjusted to 6 with 1 N HCl solution. The crude product is purified by preparative HPLC (water / MeCN = 100 / 0 to 60 / 40, 30 min, detection wavelength: 254 / 220 nm) to give the title compound 15d (160 mg, 91.4% yield) as a pale yellow solid. MS(ESI) m / z: 531.2 [M+H] +

[0278] Step 4 (R)-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-14-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)-12-oxo-3,6,9-trioxa-13-azapentadecane-15-sulfonic acid (15e) To a solution of 15d (180 mg, 0.34 mmol) and 14j (153 mg, 0.51 mmol) in DMF (5 mL), DIEA (88 mg, 0.68 mmol) and BOP (225 mg, 0.51 mmol) were added in an ice bath and stirred at 0 °C for 0.5 h. The reaction solution was purified by preparative HPLC (water / MeCN = 100 / 0 to 80 / 20, 30 min, detection wavelength: 254 / 220 nm) to give the title compound 15e (130 mg, 47.0% yield) as a pale yellow solid. MS(ESI) m / z: 814.2 [M+H] +

[0279] Step 5 (14R)-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-14-(((2S)-1-(((2S)-1-((4-((((3'-((ethoxycarbonyl)amino)-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-6'-yl)carbamoyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)-12-oxo-3,6,9-trioxa-13-azapentadecane-15-sulfonic acid (15) To a solution of 13b (64 mg, 0.16 mmol) and 2,6-lutidine (51 mg, 0.48 mmol) in DCM (10 mL) was added triphosgene (24 mg, 0.08 mmol) in an ice bath and stirred at room temperature for 3 h. To this was added a solution of 15e (130 mg, 0.16 mmol) in DCM (2 mL) and stirred at room temperature overnight. The reaction was quenched by adding ice water (0.2 mL). The reaction solution was concentrated. The residue was purified by flash column chromatography (DCM / MeOH = 6 / 1). The crude product was purified by preparative HPLC (column: XBridge Prep OBD C18 column, 30*150 mm, water (0.05% TFA) / MeCN=62 / 38~42 / 58, 7 min, detection wavelength: 254 / 220 nm) to give the title compound 15 (6.0 mg, 3.0% yield) as an orange solid. MS(ESI) m / z: 1242.3 [M+H] +

[0280] Example 16 [ka]

[0281] Step 1 (R)-2-Amino-3-methoxy-3-oxopropane-1-sulfonic acid hydrochloride (16a) To a solution of sulfo-D-alanine 15a (15 g, 88.7 mmol) in MeOH (500 mL) was added thionyl chloride (211 g, 1.77 mol) dropwise over 30 min at 0 °C. The reaction solution was stirred at room temperature overnight. The solution was concentrated in vacuo to give the title compound 16a (18 g, crude) as a white solid. MS(ESI) m / z: 184.0 [M-HCl+H] +

[0282] Step 2 (R)-2-(((benzyloxy)carbonyl)amino)-3-methoxy-3-oxopropane-1-sulfonic acid (16b) To a solution of 16a (18 g, 60.9 mmol) in THF (75 mL) was added a solution of sodium bicarbonate (12.8 g, 152.2 mmol) in water (25 mL). Benzyl chloroformate (12.5 g, 73.0 mmol) was added dropwise at 0 °C and stirred overnight at room temperature. The reaction solution was extracted with EtOAc (300 mL * 3), and the organic phase was washed with brine (200 mL * 3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give the title compound 16b (15 g, 77.6% yield) as a white solid. MS(ESI) m / z: 335.2 [M+NH3] +

[0283] Step 3 ((Benzyloxy)carbonyl)(sulfamoyl)-D-alanine methyl ester (16c) To a solution of 16b (14.0 g, 44.12 mmol) and triphenylphosphine (17.4 g, 66.2 mmol) in DCM (500 mL) was added thionyl chloride (8.9 g, 75.0 mmol) dropwise in an ice bath and stirred at room temperature for 3 h. The reaction solution was concentrated. The residue was dissolved in 0.5 M ammonia in THF (150 mL) and stirred at room temperature overnight. The reaction solution was concentrated. The residue was dissolved in MeOH (20 mL) and filtered. The filtrate was purified by preparative HPLC (XBridge Prep OBD C18 column, 30*150 mm, water (0.5% ammonium bicarbonate) / acetonitrile = 100 / 0 to 70 / 30, 60 min, detection wavelength: 254 / 220 nm) to give the title compound 16c (6.0 g, 43.0% yield) as a white solid. MS(ESI) m / z: 318.0 [M+H] +

[0284] Step 4 ((Benzyloxy)carbonyl)(sulfamoyl)-D-alanine (16d) Intermediate 16c (6.0 g, 19.0 mmol) and trimethyltin hydroxide (13.7 g, 75.9 mmol) were dissolved in 1,2-dichloroethane (50 mL) and stirred at 80 °C for 3 h. The reaction solution was filtered and purified by preparative HPLC (XBridge Prep OBD C18 column, 30 × 150 mm, water (0.5% ammonium bicarbonate) / acetonitrile = 100 / 0 ~ 80 / 20, 60 min, detection wavelength: 254 / 220 nm) to give the title compound 16d (1.8 g, 31.3% yield) as a pale yellow solid. MS(ESI) m / z: 325.0 [M+Na] +

[0285] Step 5 Benzyl ((R)-1-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-1-oxo-3-sulfamoylpropan-2-yl)carbamate (16e) To a solution of 16d (1.8 g, 5.9 mmol) and 14f (2.3 g, 5.9 mmol) in DMF (20 mL), DIEA (1.5 g, 11.9 mmol) and HATU (2.7 g, 7.2 mmol) were added and stirred at room temperature for 3 h. The reaction solution was purified by preparative HPLC (XBridge Prep OBD C18 column, 30 x 150 mm, water (0.5% ammonium bicarbonate) / acetonitrile = 100 / 0 to 20 / 80, 60 min, detection wavelength: 254 / 220 nm) to give the title compound 16e (500 mg, 12.8% yield) as a pale yellow solid. MS(ESI) m / z: 664.3 [M+H] +

[0286] Step 6 (S)-2-((S)-2-((R)-2-amino-3-sulfamoylpropanamido)-3-methylbutanamido)-N-(4-(hydroxymethyl)phenyl)-5-ureidopentanamide (16f) Intermediate 16e (400.0 mg, 0.6 mmol) was dissolved in TFA (5 mL), stirred at 50 °C for 6 h, and concentrated. The residue was dissolved in 1,4-dioxane (5 mL), and a solution of lithium hydroxide (72.2 mg, 3.0 mmol) in water (2.5 mL) was added and stirred at room temperature for 1 h. The reaction mixture was concentrated and purified by preparative HPLC (XBridge Prep OBD C18 column, 30*150 mm, water (10 M ammonium bicarbonate) / acetonitrile = 80 / 20 to 50 / 50, 8 min, detection wavelength: 254 / 220 nm) to give the title compound 16f (150 mg, 47.2% yield) as a white solid. MS(ESI) m / z: 530.3 [M+H] +

[0287] Step 7 (S)-2-((14R,17S)-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-17-isopropyl-12,15-dioxo-14-(sulfamoylmethyl)-3,6,9-trioxa-13,16-diazaoctadecane-18-amido)-N-(4-(hydroxymethyl)phenyl)-5-ureidopentanamide (16g) To a solution of 16f (150 mg, 0.28 mmol) and 14j (93.9 mg, 0.31 mmol) in DMF (5 mL), DIEA (73.2 mg, 0.57 mmol) and HATU (129.2 mg, 0.34 mmol) were added and stirred at room temperature for 2 h. The reaction solution was purified by preparative HPLC (Xselect CSH OBD column, 30*150 mm, 5 μm column, water (0.1% FA) / acetonitrile = 95 / 5 to 68 / 32, 10 min, detection wavelength: 254 / 220 nm) to give the title compound 16g (110 mg, 48.3% yield) as a white solid. MS(ESI) m / z: 813.4 [M+H] +

[0288] Step 8 (3-Oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-3',6'-diyl)biscarbamic acid 4-((2S,5S,8R)-21-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-5-isopropyl-4,7,10-trioxo-8-(sulfamoylmethyl)-2-(3-ureidopropyl)-13,16,19-trioxa-3,6,9-triazahencosamide) benzyl ester ethyl ester (16) To a solution of 13b (65.3 mg, 0.16 mmol) and 2,6-lutidine (52.2 mg, 0.49 mmol) in DCM (20 mL) was added triphosgene (24.1 mg, 0.08 mmol) in an ice bath and stirred at room temperature for 4 h. 16g (110.0 mg, 0.14 mmol) was added to the solution and stirred at room temperature overnight. Water (0.2 mL) was added to quench the reaction and the mixture was concentrated. The residue was purified by flash column chromatography (DCM / MeOH = 5 / 1). The crude product was purified by preparative HPLC (column: XBridge Prep OBD C18 column, 19*250 mm, 5 μm, water (0.1% FA) / acetonitrile = 80 / 20 to 50 / 50, 8 min, detection wavelength: 254 / 220 nm) to give the title compound 16 (5.9 mg, yield 3.4%) as a pale pink solid. MS(ESI) m / z: 1241.3 [M+H] +

[0289] Example 17 [ka]

[0290] Step 1 (R)-2-((tert-butoxycarbonyl)amino)-3-(diethoxyphosphoryl)propanoic acid methyl ester (17c) (R)-2-((tert-butoxycarbonyl)amino)-3-iodopropanoate 17a (10 g, 30.38 mmol, commercially available) and triethyl phosphite (100 mL, 601.84 mmol) were mixed and stirred overnight at 140° C. The reaction solution was concentrated and purified by flash column chromatography (petroleum ether / ethyl acetate, gradient elution) to give the title compound 17c (7.5 g, 72.8% yield). MS(ESI) m / z: 340.3 [M+H] +

[0291] Step 2 (R)-2-((tert-butoxycarbonyl)amino)-3-(diethoxyphosphoryl)propanoic acid (17d) To a solution of 17c (7.5 g, 22.1 mmol) in THF (40 mL) and water (20 mL), lithium hydroxide monohydrate (2.1 g, 88.53 mmol) was added and stirred overnight at room temperature. The pH of the reaction solution was adjusted to 6 with 1 M HCl, concentrated, and purified by reverse-phase column chromatography (column: C18, water (0.1% FA) / acetonitrile, gradient elution, flow rate: 20 mL / min). The desired fraction was lyophilized to give the title compound 17d (2.8 g, 38.9% yield). MS(ESI) m / z: 326.1 [M+H] +

[0292] Step 3 ((R)-tert-butyl 3-(diethoxyphosphoryl)-1-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-1-oxopropan-2-yl)carbamate (17e) To a solution of 17d (2.8 g, 8.61 mmol) in DMF (40 mL), HATU (3.9 g, 10.33 mmol) and DIEA (2.2 g, 17.22 mmol) were added and stirred at room temperature for 10 min. 14f (3.6 g, 9.47 mmol, commercially available) was added to the solution and stirred at room temperature for 2 h. The reaction solution was purified by reverse-phase column chromatography (column: C18, water (0.5% ammonium bicarbonate) / acetonitrile, gradient elution, flow rate: 20 mL / min). The desired fraction was lyophilized to give the title compound 17e (4.5 g, 76.1% yield). MS(ESI) m / z: 686.3 [M+H] +

[0293] Step 4 Diethyl ((R)-2-amino-3-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)phosphonate (17f) To a solution of 17e (4.5 g, 6.55 mmol) in DCM (20 mL), TFA (10 mL) was added and stirred at room temperature for 1 h. The reaction solution was concentrated and dissolved in 1,4-dioxane (30 mL) and water (10 mL). Lithium hydroxide monohydrate (1.4 g, 32.65 mmol) was added in an ice bath and stirred at room temperature for 2 h. The reaction solution was purified by reverse-phase column chromatography (column: C18, water (0.5% ammonium bicarbonate) / acetonitrile, gradient elution, flow rate: 20 mL / min). The desired fraction was lyophilized to give the title compound 17f (3.3 g, 85.9% yield). MS(ESI) m / z: 587.8 [M+H] +

[0294] Step 5 ((R)-2-amino-3-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)phosphonic acid (17g) To a solution of 17f (3.0 g, 5.11 mmol) in N-methyl-2-pyrrolidinone (15 mL), sodium bromide (2.2 g, 21.48 mmol) was added, and trimethylsilyl trifluoromethanesulfonate (9.1 g, 40.91 mmol) was added dropwise in an ice bath, followed by stirring at 60 °C overnight. The reaction solution was purified by reverse-phase column chromatography (column: C18, water (0.05% TFA) / acetonitrile, gradient elution, flow rate: 20 mL / min). The desired fraction was lyophilized to give the title compound 17f (1.5 g, 55.3% yield). MS(ESI) m / z: 531.3 [M+H] +

[0295] Step 6 ((R)-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-14-(((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)-12-oxo-3,6,9-trioxa-13-azapentadecan-15-yl)phosphonic acid (17i) To a solution of 17h (500 mg, 0.94 mmol) and 17g (413 mg, 1.04 mmol, commercially available) in DMF (10 mL) was added DIEA (365 mg, 2.83 mmol) in an ice bath and stirred at room temperature for 3 h. The reaction solution was purified by reverse-phase column chromatography (column: C18, water (0.05% TFA) / acetonitrile, gradient elution, flow rate: 20 mL / min). The desired fraction was lyophilized to give the title compound 17i (200 mg, 26.1% yield). MS(ESI) m / z: 814.2 [M+H] +

[0296] Step 7 ((14R)-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-14-(((2S)-1-(((2S)-1-((4-((((3'-((ethoxycarbonyl)amino)-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-6'-yl)carbamoyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)-12-oxo-3,6,9-trioxa-13-azapentadecan-15-yl)phosphonic acid (17) To a solution of 13b (119 mg, 0.30 mmol) in DCM (6 mL), 2,6-lutidine (95 mg, 0.89 mmol) was added, and triphosgene (44 mg, 0.15 mmol) was added dropwise in an ice bath. The mixture was stirred at room temperature for 4 h. A solution of 17i (200 mg, 0.25 mmol) in N-methyl-2-pyrrolidinone was added dropwise to the reaction solution in an ice bath and stirred at room temperature overnight. The reaction solution was purified by flash column chromatography (DCM / MeOH, gradient elution). The crude product was purified by preparative HPLC (column: Sunfire Prep C18 OBD 5 μm 19*250 mm, mobile phase: water (0.05% TFA) / acetonitrile, gradient elution, flow rate: 20 mL / min). The desired fraction was lyophilized to give the title compound 17 (10 mg, 3.2% yield). MS(ESI) m / z: 1242.5 [M+H] +

[0297] Example 18 [ka]

[0298] Step 1 Benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-L-glutamate (18b) To a solution of benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-((3aR,6R,6aR)-6-(hydroxymethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-L-glutamate 18a (100 mg, 0.16 mmol, available from WuXi AppTec) in THF (0.5 mL) was added water (1 mL) and TFA (1 mL) at 0 °C and stirred at room temperature for 2 h. Toluene (3 mL) was added and concentrated in vacuo to give the title compound 18b (95 mg, crude, 100% yield), which was used directly without further purification. MS(ESI) m / z: 591.5 [M+H]+

[0299] Step 2 N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-L-glutamine (18) To a solution of 18b (94 mg, 0.16 mmol, crude) in methanol (2 mL) was added 10% Pd / C (17 mg) under a nitrogen atmosphere. The mixture was stirred at room temperature under a hydrogen atmosphere (hydrogen gas balloon) for 1.5 h. The solution was filtered through diatomaceous earth and concentrated to give the title compound 18 (80 mg, 100% yield). MS(ESI) m / z: 499.2 [MH] -

[0300] Example 19 [ka]

[0301] Step 1 (((3aR,4R,6R,6aR)-6-azido-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)(tert-butyl)dimethylsilane (19a) TBSCl (770.4 mg, 5.11 mmol) and imidazole (632.7 mg, 9.29 mmol) are added to a mixture of 1a (1.00 g, 4.65 mmol) in DCM (10 mL). The mixture is reacted at room temperature overnight and monitored by TLC. The reaction is quenched with 50 mL of saturated NaHCO3 and extracted with DCM (50 mL*2). After separation, the combined organic layer is washed with brine (30 mL), dried over Na2SO4, filtered, and the filtrate is concentrated in vacuo to give a residue. The residue is purified by silica gel column chromatography (A-petroleum ether, B-EtOAc) to give 19a (1.40 g, 91% yield).

[0302] Step 2 (3aR,4R,6R,6aR)-6-(((tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-amine (19b) Wet Pd / C (150 mg, 10% purity) was added to a mixture of 19a (1.40 g, 4.25 mmol) in MeOH (20 mL). The reaction mixture was purged with a H2 balloon three times and allowed to react at room temperature for 1 h under a H2 balloon. After completion of the reaction, the mixture was filtered through diatomaceous earth and washed with MeOH. The filtrate was concentrated under vacuum and purified by silica gel column chromatography (A - DCM, B - MeOH) to give 19b (740.0 mg, 57.3% yield). MS(ESI) m / z: 326.4 [M+Na] +

[0303] Step 3 (3aS,6R,6aR)-6-((S)-4-(((benzyloxy)carbonyl)amino)-5-methoxy-5-oxopentanamido)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carboxylic acid (19c) To a mixture of 1 (255.0 mg, 0.531 mmol) in pyridine (1 mL) was added EDCI (101.7 mg, 0.531 mmol). The resulting yellow mixture was allowed to react at room temperature for 10 min. 19b (162.7 mg, 0.54 mmol) was added and the reaction was continued at the same temperature for another 4 h, monitored by LCMS. The mixture was concentrated in vacuo to remove most of the pyridine, then diluted with DMF (1 mL) and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 19c (112.0 mg, 27% yield). MS(ESI) m / z: 766.6 [M+H] +

[0304] Step 4 N2-((benzyloxy)carbonyl)-N5-((3aR,6S,6aS)-6-(((3aR,6R,6aR)-6-(hydroxymethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)carbamoyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-L-glutamic acid methyl ester (19d) To a mixture of 19c (112.0 mg, 0.15 mmol) in THF (2 mL) was added TBAF (1 M in THF, 161 μL, 0.161 mmol). The mixture was allowed to react at room temperature for 0.5 h. After the mixture was complete, the reaction was quenched with 10 mL of saturated NaHCO3 and extracted with EtOAc (10 mL*2). After separation, the combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered, and the filtrate was concentrated in vacuo to give 19d (95 mg, crude). MS(ESI) m / z: 674.5 [M+Na] +

[0305] Step 5 N2-((benzyloxy)carbonyl)-N5-((2R,3R,5S)-5-(((2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-L-glutamic acid methyl ester (19e) The mixture of 19d (112 mg, crude) and TFA / H2O (4:1, 1 mL) was reacted at room temperature for 0.5 h and monitored by LCMS. The mixture was concentrated in vacuo to give 19e (83.0 mg, crude). MS(ESI) m / z: 594.4[M+Na] +

[0306] Step 6 N2-((benzyloxy)carbonyl)-N5-((3R,4S,5S)-5-(((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-L-glutamine (19) To a mixture of 19e (83.0 mg, crude) in MeOH (1 mL) was added aqueous lithium hydroxide solution (1N, 0.3 mL). The mixture was reacted at room temperature for 0.5 h. The mixture was acidified to pH = 6, filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 19 (55 mg, Yield: 67.9%). MS(ESI) m / z: 580.4[M+Na] +

[0307] Example 20 [ka]

[0308] Step 1 Benzyl (((3aR,4R,6S,6aS)-6-(2-amino-2-oxoethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)carbamate (20a) To a suspension of 8b (2.0 g, 5.47 mmol) in DMF (20 mL) was added PyBOP (3.42 g, 6.57 mmol), HOBt (887.60 mg, 6.57 mmol), and DIPEA (2.12 g, 16.42 mmol). The resulting brown suspension was reacted at 50 °C overnight. The mixture was quenched with water (20 mL) and extracted with EtOAc (30 mL*3). After separation, the combined organics were washed with brine (30 mL*3), dried over Na2SO4, filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was purified by silica gel column chromatography (A-DCM; B-MeOH) to give 20a (4.3 g, crude). MS(ESI) m / z: 365.4 [M+H] +

[0309] Step 2 2-((3aS,4S,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)acetamide (20b) Compound 20b was synthesized according to the synthetic procedure in Step 2 of Example 19 (1.9 g, crude). MS(ESI) m / z: 231.4 [M+H] +

[0310] Step 3 Benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((3aR,4R,6S,6aS)-6-(2-amino-2-oxoethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)-L-glutamate (20d) To a mixture of 20b (900.0 mg, crude) and 20c [(1.74 g, 3.13 mmol), commercially available] in THF (20 mL) was added saturated NaHCO3 (5 mL). The mixture was allowed to react at room temperature for 1 h. After completion of the reaction, the mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL*2). The combined organics were washed with brine (30 mL), dried over Na2SO4, filtered, and the filtrate was concentrated in vacuo to give 20d (2.1 g, crude). MS(ESI) m / z: 672.6 [M+H] +

[0311] Step 4 N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((3aR,4R,6S,6aS)-6-(2-amino-2-oxoethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)-L-glutamine (20e) Compound 20e was synthesized according to the synthetic procedure in Step 2 of Example 19 (1.87 g, crude). MS(ESI) m / z: 582.5 [M+H] +

[0312] Step 5 N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-L-glutamine (20) Compound 20 was synthesized according to the synthetic procedure in Step 5 of Example 19 (630.0 g, 42.5% yield). MS(ESI) m / z: 542.5 [M+H] +

[0313] Example 21 [ka]

[0314] Step 1 2-((3aS,4S,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)acetic acid (21a) Compound 21a was synthesized according to the synthetic procedure in Step 2 of Example 19 (480 mg, 75% yield). MS(ESI) m / z: 232.4 [M+H] +

[0315] Step 2 2-((3aS,4S,6R,6aR)-6-(((S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(benzyloxy)-5-oxopentanamido)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)acetic acid (21b) Compound 21b was synthesized according to the synthetic procedure in Step 3 of Example 20 (1.5 g, crude). MS(ESI) m / z 673.4 [M+H] +

[0316] Step 3 Benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((3aR,4R,6S,6aS)-6-(2-((((3aR,4R,6S,6aS)-6-(2-amino-2-oxoethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)amino)-2-oxoethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)-L-glutamate (21c) To a mixture of 21b (410 mg, 0.609 mmol) in DMF (6 mL) was added HATU (231.73 mg, 0.609 mmol) and DIPEA (236.3 mg, 1.828 mmol). The resulting yellow mixture was allowed to react at room temperature for 10 min, and then 20b (168.4 mg, 0.731 mmol) was added. The mixture was allowed to react at room temperature for another 1 h. After completion of the reaction, the mixture was quenched with 20 mL of water and extracted with EtOAc (30 mL*2). After separation, the combined organic layer was washed with brine (30 mL*3), dried over Na2SO4, filtered, and the filtrate was concentrated under vacuum to give a residue. The residue was purified by C18 column flash chromatography [mobile phase: A-water (0.1% formic acid): B-acetonitrile] to give 21c (430.0 mg, 66.7% yield). MS(ESI) m / z: 885.4 [M+H] +

[0317] Step 4 N2-(((9H-Fluoren-9-yl)methoxy)carbonyl)-N5-(((3aR,4R,6S,6aS)-6-(2-((((3aR,4R,6S,6aS)-6-(2-amino-2-oxoethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)amino)-2-oxoethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)-L-glutamine (21d) Compound 21d was synthesized according to the synthetic procedure in Step 2 of Example 19 (377.5 g, crude). MS(ESI) m / z: 795.4 [M+H]+

[0318] Step 5 N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((2R,3S,4R,5S)-5-(2-((((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-L-glutamine (21) Compound 21 was synthesized according to the synthetic procedure in Step 5 of Example 19 (181.0 mg, 53.1% yield). MS(ESI) m / z: 715.5 [M+H] +

[0319] Example 22 [ka]

[0320] Step 1 (R)-2,2-Dimethyl-1,3-dioxolane-4-carboxylic acid (22b) 22a (2 g, 12.49 mmol) was dissolved in a mixed solvent of THF and HO (v / v = 1:1, 40 mL), and then LiOH (449 mg, 18.73 mmol) was added. The resulting mixture was stirred at 25 °C for 1 h. After the reaction was completed, the reaction mixture was extracted with EA (30 mL). The remaining aqueous layer was acidified to pH = 3 with aqueous HCl (1 M) and further extracted with EA (30 mL * 3). The combined organic layer was dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give 22b (1.51 g, 82.8% yield) as a clear oil, which was used directly in the next step without purification. 1H NMR: δ 11.15(br s, 1H), 4.63(dd, J = 7.6, 4.8Hz, 1H), 4.30(dd, J = 8.8, 7.6Hz, 1H), 4.19(dd, J = 8.8, 4.8Hz, 1H), 1.53(s, 3H), 1.42(s, 3H).

[0321] Step 2 (R)-Benzyl 2,2-dimethyl-1,3-dioxolane-4-carboxylate (22c) 22b (3 g, 20.53 mmol) was dissolved in DMF (21 mL), and then BnBr (5.27 g, 30.79 mmol) and KCO (4.26 g, 30.79 mmol) were added. The resulting mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction mixture was diluted with HO (30 mL) and extracted with EA (40 mL*3). The combined organic layers were dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give a brown oil in the form of crude product, which was purified by flash column chromatography (PE / EA = 100 / 0 to 80 / 20) to give 22c (2.52 g, 52.0% yield) as a clear oil. 1H NMR: δ 7.38-7.31(m, 5H), 5.21(m, 2H), 4.62(dd, J = 7.2, 5.2Hz, 1H), 4.30(dd, J = 8.8, 7.2Hz, 1H), 4.19(dd, J = 8.8, 5.2Hz, 1H), 1.49(s, 3H), 1.40(s, 3H).

[0322] Step 3 (R)-Benzyl 2,3-dihydroxypropanoate (22d) 22c (1 g, 4.23 mmol) was dissolved in DCM (13 mL), and then a mixture of TFA and HO (v / v = 9:1, 13 mL) was added at 0 °C. The resulting mixture was stirred at 0 °C for 1 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure and purified by flash column chromatography (PE / EA = 95 / 5 to 20 / 80) to give 22d (557 mg, 67.1% yield) as a colorless oil. 1H NMR: δ 7.40-7.31(m, 5H), 5.25(d, J = 2.8Hz, 2H), 4.31(t, J = 3.6Hz, 1H), 3.93-3.85(m, 2H), 2.89(br s, 2H).

[0323] Step 4 (R)-Benzyl 3-((tert-butyldiphenylsilyl)oxy)-2-hydroxypropanoate (22e) 22d (557 mg, 2.84 mmol) was dissolved in anhydrous DMF (8 mL), and then imidazole (290 mg, 4.26 mmol) and TBDPSCl (858 mg, 3.12 mmol) were added. The resulting mixture was stirred at 25 °C for 4 h. After the reaction was completed, the reaction mixture was diluted with EA (50 mL) and washed with brine (25 mL*2) and water (25 mL*3). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give a crude yellow oil, which was purified by flash column chromatography (PE / EA = 100 / 0 to 75 / 25) to give 22e (705 mg, 57.1% yield) as a clear oil. MS(ESI) m / z: 457.6 [M+Na] +

[0324] Step 5 (S)-11-benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2-oxa-4,7,10,13,16-pentaazaheptadecan-17-yl acetate (22 g) 22f (commercially available, 5 g, 8.12 mmol) was dissolved in anhydrous DMF (26 mL), followed by the addition of Cu(OAc) (561 mg, 3.09 mmol), Pb(OAc) (4.11 g, 9.26 mmol), and HOAc (1.11 g, 18.44 mmol). The resulting mixture was stirred at 60 °C for 70 min under N atmosphere. After completion of the reaction, the reaction mixture was diluted with DCM (160 mL) and washed with brine (3 x 40 mL) and HO (2 x 40 mL). The organic layer was dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give a brown oil in the form of crude product, which was purified by flash column chromatography (DCM / MeOH = 100 / 0 to 90 / 10) to give 22g (4.19 g, 81.8% yield) as a white solid. MS(ESI) m / z: 652.5 [M+Na] +

[0325] Step 6 Benzyl (11S,19R)-11-benzyl-19-(((tert-butyldiphenylsilyl)oxy)methyl)-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2,18-dioxa-4,7,10,13,16-pentaazaicosan-20-oate (22h) 22g (150 mg, 0.24 mmol), 22e (207 mg, 0.48 mmol), and 4 Å molecular sieves (200 mg) were added to anhydrous THF (3 mL). The resulting mixture was stirred at 25 °C for 30 min, followed by the addition of Sc(OTf) (117 mg, 0.24 mmol). The mixture was stirred at 25 °C for another 16 h. After the reaction was complete, the reaction mixture was filtered and concentrated under reduced pressure to obtain the crude product in the form of a yellow oil, which was purified by flash column chromatography (DCM / MeOH = 100 / 0 to 90 / 10) to give 22h (174 mg, 72.6% yield) as a white solid. MS(ESI) m / z: 1026.5 [M+Na] +

[0326] Step 7 (11S,19R)-11-benzyl-19-(((tert-butyldiphenylsilyl)oxy)methyl)-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2,18-dioxa-4,7,10,13,16-pentaazaicosan-20-enoic acid (22i) 22h (150 mg, 0.15 mmol) was dissolved in a mixture of THF (2.5 mL) and HO (2.5 mL), and then Pd / C (wet, 10%, 45 mg) was added. The mixture was stirred under H (15 psi) at 25 °C for 4 h. After the reaction was complete, the mixture was filtered and concentrated under reduced pressure to give 22i (136 mg, quantitative) as a white solid, which was used directly in the next step without further purification. MS(ESI) m / z: 936.6 [M+Na] +

[0327] Step 8 ((6R,14S)-14-benzyl-6-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)carbamoyl)-2,2-dimethyl-10,13,16,19-tetraoxo-3,3-diphenyl-4,7-dioxa-9,12,15,18-tetraaza-3-silaeicosan-20-yl)carbamate (9H-fluoren-9-yl)methyl (22k) 22i (85 mg, 0.09 mmol), HATU (42 mg, 0.11 mmol), and DIEA (36 mg, 0.28 mmol) were dissolved in DMF (2 mL). The mixture was stirred at 25 °C for 15 min, followed by the addition of exatecan mesylate (22j, commercially available, 59 mg, 0.11 mmol). The mixture was stirred at 25 °C for 30 min. After the reaction was complete, the mixture was filtered and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 22k (80 mg, Yield: 64.6%) as a pale yellow solid. MS(ESI) m / z: 1355.0 [M+Na] +

[0328] Step 9 (R)-2-(((S)-13-amino-7-benzyl-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)-3-hydroxypropanamide (22l) 22k (78 mg, 0.06 mmol) was dissolved in anhydrous THF (6 mL), and then TBAF (1 M THF solution, 70 μL, 0.07 mmol) was added. The resulting mixture was stirred at 25 °C for 1 h, and EtNH (60 μL, 0.58 mmol) was added. The mixture was stirred at 25 °C for another 1 h. After the reaction was completed, the reaction mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 22l (26 mg, Yield: 51.0%) as a white solid. MS(ESI) m / z: 871.6 [M+H] +

[0329] Step 10 N-((2R,10S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-2-(hydroxymethyl)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide (22) 22l (24 mg, 0.028 mmol) and 22m (9.4 mg, 0.030 mmol) were dissolved in DMF (0.5 mL), and TEA (2.5 mg, 0.025 mmol) was added. The resulting mixture was stirred at 25 °C for 30 min. After the reaction was completed, the reaction mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min) to give 22 (16.5 mg, Yield: 51.2%) as a white solid. MS(ESI) m / z: 1064.7 [M+H] +

[0330] Example 23 [ka] Compound 23 was synthesized according to the synthetic procedure of Example 22. MS(ESI) m / z: 1064.7 [M+H] +

[0331] Example 24 [ka]

[0332] Step 1 (S)-3-((11-benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2-oxa-4,7,10,13,16-pentaazaheptadecan-17-yl)oxy)bicyclo[1.1.1]pentane-1-carboxylate methyl ester (24b) To a mixture of 22g (50 mg, 0.08 mmol), methyl 3-hydroxybicyclo[1.1.1]pentane-1-carboxylate 24a (23 mg, 0.16 mmol), and dry 4 Å molecular sieves (140 mg) in anhydrous THF (1 mL) was added scandium triflate (47 mg, 0.1 mmol) and stirred overnight at room temperature. The solution was filtered through diatomaceous earth, concentrated, and purified by flash column chromatography (DCM / MeOH = 10 / 1) to give the title compound 24b (41 mg, 72.5% yield) as a white solid. MS(ESI) m / z: 734.5 [M+Na] +

[0333] Step 2 (S)-3-((13-amino-7-benzyl-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecyl)oxy)bicyclo[1.1.1]pentane-1-carboxylic acid (24c) To a solution of 24b (41 mg, 0.06 mmol) in methanol (1 mL) and water (0.2 mL), potassium carbonate (83 mg, 0.6 mmol) was added and stirred at room temperature for 2 h. The reaction solution was extracted with hexane (2 mL*3), and the pH of the aqueous phase was adjusted to 6 with 2 N HCl. The aqueous solution was concentrated and lyophilized to give the title compound 24c (230 mg, crude), which was used directly without further purification. MS(ESI) m / z: 474.3 [MH] -

[0334] Step 3 (S)-3-((7-benzyl-20-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3,6,9,12,15-pentaoxo-2,5,8,11,14-pentaazaicosyl)oxy)bicyclo[1.1.1]pentane-1-carboxylic acid (24d) To a solution of 24c (230 mg, crude, theoretical 27 mg, 0.06 mmol) and 2j (35 mg, 0.11 mmol) in anhydrous DMF (1 mL) was added DIEA (47 μL, 0.28 mmol) and stirred at room temperature for 15 min. The reaction was quenched with AcOH (40 μL). The solution was purified by preparative HPLC (0.1% FA in water / MeCN) and lyophilized to give the title compound 24d (24 mg, 63.0% yield) as a white solid. MS(ESI) m / z: 667.4 [MH] -

[0335] Step 4 3-(((S)-7-Benzyl-20-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3,6,9,12,15-pentaoxo-2,5,8,11,14-pentaazaicosyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)bicyclo[1.1.1]pentane-1-carboxamide (24) To a solution of 24d (24 mg, 35.9 μmol) and NHS (6.2 mg, 53.8 μmol) in anhydrous DMF (0.3 mL), DCC (11 mg, 53.8 μmol) was added and stirred overnight at room temperature. The reaction solution was slowly added to a solution of exatecan mesylate (23 mg, 43.1 μmol) and DIEA (8 μL, 43.1 μmol) in anhydrous DMF (0.3 mL) and stirred overnight at room temperature. The reaction solution was filtered, purified by preparative HPLC (water / MeCN with 0.1% FA), and lyophilized to give the title compound 24 (2.6 mg, 10.7% yield) as a white solid. MS(ESI) m / z: 1086.9 [M+H] +

[0336] Example 25 [ka]

[0337] Step 1 Benzyl (1R,3s)-benzyl 3-(((S)-11-benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2-oxa-4,7,10,13,16-pentaazaheptadecan-17-yl)oxy)cyclobutane-1-carboxylate (25c) To a solution of 25a (100 mg, 0.159 mmol) and 25b (31.76 mg, 0.318 mmol) in THF (1.5 mL) was added 4 Å molecular sieves. The mixture was stirred at room temperature for 10 min, then Sc(OTf) (78.16 mg, 0.159 mmol) was added and allowed to react for another 16 h at room temperature. The suspended mixture was filtered through a pad of diatomaceous earth, the filter cake was washed with THF (30 mL), and the filtrate was quenched by adding saturated NaHCO (30 mL) and extracted with EtOAc (30 mL * 2). After separation, the combined organic layer was washed with brine (50 mL), dried over NaSO, filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was purified by silica gel column chromatography (A - DCM, B - MeOH) to give 25c (115 mg, 93.3% yield). MS(ESI) m / z: 798.5 [M+Na] +

[0338] Step 2 (1R,3S)-3-(((S)-11-benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2-oxa-4,7,10,13,16-pentaazaheptadecan-17-yl)oxy)cyclobutane-1-carboxylic acid (25d) To a solution of 25c (115 mg, 0.183 mmol) in MeOH (2 mL) was added wet Pd / C (25 mg). The black suspension was purged with a H2 balloon three times and then reacted under a H2 balloon at room temperature for 2 h. After completion of the reaction, the black suspension was filtered through a pad of diatomaceous earth, the filter cake was washed with MeOH, and the combined organic layers were concentrated in vacuo to give 25d (90 mg, 63.5% yield). MS(ESI) m / z: 708.5 [M+Na] +

[0339] Step 3 (9H-fluoren-9-yl)methyl ((S)-7-benzyl-1-((1R,3R)-3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)carbamoyl)cyclobutoxy)-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecan-13-yl)carbamate (25e) To a mixture of 25d (80 mg, 0.117 mmol) and TSTU (35.15 mg, 0.117 mmol) in DMF (2 mL) was added DIPEA (30.18 mg, 0.233 mmol). The mixture was reacted at room temperature for 10 minutes, and the acid was converted to the activated ester according to LCMS. 10a (68.1 mg, 0.128 mmol) was added and reacted at the same temperature for another 5 hours. The reaction mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 25e (55 mg, Yield: 42.6%). MS(ESI) m / z: 1125.7 [M+Na] +

[0340] Step 4 (1R,3R)-3-(((S)-13-amino-7-benzyl-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)cyclobutane-1-carboxamide (25f) To a solution of 25e (55 mg, 0.050 mmol) in DMF (1 mL) was added piperidine (85.15 mg, 1.0 mmol). The mixture was allowed to react at room temperature for 10 minutes. The reaction mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 25f (35 mg, 80% yield). MS(ESI) m / z: 903.7 [M+Na] +

[0341] Step 5 (1R,3R)-3-(((S)-7-benzyl-20-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3,6,9,12,15-pentaoxo-2,5,8,11,14-pentaazaicosyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)cyclobutane-1-carboxamide (25) Compound 25 was synthesized according to the synthetic procedure in Step 8 of Example 2 (12.5 mg, 68.2% yield).

[0342] Example 26 [ka]

[0343] Step 1 (1S,3r)-3-(((S)-11-benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2-oxa-4,7,10,13,16-pentaazaheptadecan-17-yl)oxy)cyclobutane-1-carboxylate methyl ester (26b) Compound 26b was synthesized according to the synthetic procedure in Step 1 of Example 25 (105 mg, 94.5% yield). MS(ESI) m / z: 722.5 [M+Na] +

[0344] Step 2 (1S,3R)-3-(((S)-13-amino-7-benzyl-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecyl)oxy)cyclobutene-1-carboxylic acid (26c) To a solution of 26b (105 mg, 0.15 mmol) in MeOH (1 mL) and HO (0.2 mL) was added KCO (414.76 mg, 3.0 mmol). The resulting white suspension was allowed to react at room temperature for 3 h. The white suspension was acidified to pH = 6 with 1 N HCl, and the mixture was freeze-dried to give 26c (485 mg, crude). MS(ESI) m / z: 486.4 [M+Na] +

[0345] Step 3 (1S,3r)-3-(((S)-7-benzyl-20-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3,6,9,12,15-pentaoxo-2,5,8,11,14-pentaazaicosyl)oxy)cyclobutane-1-carboxylic acid (26d) Compound 26d was synthesized according to the synthetic procedure in Step 8 of Example 2 (23 mg, 24% yield). MS(ESI) m / z: 655.4 [MH] -

[0346] Step 4 (1S,3S)-3-(((S)-7-benzyl-20-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3,6,9,12,15-pentaoxo-2,5,8,11,14-pentaazaicosyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)cyclobutane-1-carboxamide (26) Compound 26 was synthesized according to the synthetic procedure in Step 3 of Example 25 (20.5 mg, 54.5% yield). MS(ESI) m / z: 1074.9 [M+H] +

[0347] Example 27 [ka]

[0348] Step 1 Acetic acid (5S,8S)-1-(9H-fluoren-9-yl)-5-isopropyl-3,6,9-trioxo-8-(3-ureidopropyl)-2-oxa-4,7,10-triazaundecan-11-yl ester (27b) 27b was synthesized according to the synthetic procedure in Step 5 of Example 22 (585 mg, 51.8% yield). MS(ESI) m / z: 590.6 [M+H] +

[0349] Step 2 Benzyl (5S,8S)-1-(9H-fluoren-9-yl)-5-isopropyl-3,6,9-trioxo-8-(3-ureidopropyl)-2,12-dioxa-4,7,10-triazatetradecane-14-oate (27d) To a solution of 27b (90 mg, 0.16 mmol) and benzyl 2-hydroxyacetate (132 mg, 0.79 mmol) in anhydrous THF (5 mL) was added dry 4 Å molecular sieves (600 mg) and stirred at room temperature for 30 min. Scandium triflate (94 mg, 0.19 mmol) was added to the solution and stirred at room temperature for 6 h. The solution was filtered through diatomaceous earth, concentrated, and purified by flash column chromatography (DCM / MeOH = 10 / 1) to give 27d (82 mg, 76.1% yield) as a white solid. MS(ESI) m / z: 696.7 [M+Na] +

[0350] Step 3 (5S,8S)-1-(9H-fluoren-9-yl)-5-isopropyl-3,6,9-trioxo-8-(3-ureidopropyl)-2,12-dioxa-4,7,10-triazatetradecan-14-oic acid (27e) To a solution of 27d (151 mg, 0.22 mmol) in MeOH (3 mL), 10% Pd / C (35 mg) was added, and the suspension was purged with a H gas balloon three times and then stirred under H pressure at room temperature for 90 min. After the reaction was complete, the Pd / C was filtered, and the methanol was removed under reduced pressure to give 27e (125 mg, 96% yield). MS(ESI) m / z: 606.4 [M+Na] +

[0351] Step 4 (S)-2-((S)-2-amino-3-methylbutanamido)-N-((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-2-oxoethoxy)methyl)-5-ureidopentanamide (27f) To a mixture of 27e (50 mg, 0.086 mmol) and TSTU (25.85 mg, 0.086 mmol) in DMF (2 mL) was added DIPEA (33.216 mg, 0.257 mmol). The mixture was allowed to react at room temperature for 10 minutes, and the acid was converted to the activated ester according to LCMS. 10a (45.67 mg, 0.086 mmol) was added and the reaction was continued at the same temperature for another 5 hours. After the reaction was complete, piperidine (72.30 mg, 0.849 mmol) was added and the mixture was allowed to stand at room temperature for another 10 minutes. The mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 27f (43 mg, Yield: 65.0%). MS(ESI) m / z: 779.6 [M+H] +

[0352] Step 5 (3S,4R,5R)-methyl 5-((7S,10S,13S)-13-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-10-isopropyl-1,6,9,12-tetraoxo-7-(3-ureidopropyl)-3-oxa-5,8,11-triazahexadecan-16-amido)-3,4-dihydroxytetrahydrofuran-2-carboxylate (27g) HATU (30.21 mg, 0.079 mmol) and DIPEA (30.80 mg, 0.238 mmol) were added to a solution of 5 (42 mg, 0.079 mmol) in DMF (2 mL). The reaction mixture was allowed to react at room temperature for 10 minutes. 27f (30.94 mg, 0.040 mmol) was then added and the mixture was left at the same temperature for another hour. The mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 27g (21 mg, 41% yield). MS(ESI) m / z: 1311.8 [M+Na] +

[0353] Step 6 (3S,4R,5R)-5-((7S,10S,13S)-13-amino-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-10-isopropyl-1,6,9,12-tetraoxo-7-(3-ureidopropyl)-3-oxa-5,8,11-triazahexadecan-16-amido)-3,4-dihydroxytetrahydrofuran-2-carboxylic acid (27h) To a solution of 27g (21.0 mg, 0.016 mmol) in THF (2 mL), add aqueous lithium hydroxide (1 N, 0.977 mL, 0.977 mmol). Stir the mixture at room temperature for 2 h. After the reaction is complete, the mixture is acidified to pH = 6 with 1 N HCl, filtered, and the filtrate is purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min) to obtain 27g (11.1 mg, Yield: 64.7%). MS(ESI) m / z: 1053.7 [M+H] +

[0354] Step 7 (3S,4R,5R)-5-((7S,10S,13S)-13-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-10-isopropyl-1,6,9,12-tetraoxo-7-(3-ureidopropyl)-3-oxa-5,8,11-triazahexadecan-16-amido)-3,4-dihydroxytetrahydrofuran-2-carboxylic acid (27) Compound 27 was synthesized according to the synthetic procedure in Step 8 of Example 2 (5.7 mg, 48% yield).

[0355] Example 28 [ka]

[0356] Step 1 (S)-11-Benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2-oxa-4,7,10,13,16-pentaazaheptadecan-17-yl acetate (28b) To a solution of 28a (3.0 g, 4.873 mmol) in DMF (26 mL) was added catalytic amounts of Cu(OAc) (336.33 mmol), Pb(OAc) (2.46 g, 5.55 mmol), and HOAc (632.62 μL, 11.062 mmol). The resulting blue mixture was purged with N three times and stirred at 60 °C for 70 min. After completion of the reaction, the mixture was cooled to room temperature, quenched with water (100 mL), and extracted with EtOAc (100 mL). After separation, the combined organic layer was washed with brine (150 mL), dried over NaSO, filtered, and the filtrate was concentrated in vacuo to give a residue. Purification by silica gel column rapid chromatography using 10% methanol in dichloromethane as an eluent gave 28b (3.0 g, 97.8% yield). MS(ESI) m / z: 652.5 [M+Na] +

[0357] Step 2 (S)-11-Benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2,18-dioxa-4,7,10,13,16-pentaazaicosan-20-oate (28d) Compound 28c was synthesized according to the synthetic procedure in Step 1 of Example 25 (995 mg, 77.4% yield). MS(ESI) m / z: 758.5 ​​[M+Na] +

[0358] Step 3 (S)-11-Benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2,18-dioxa-4,7,10,13,16-pentaazaicosan-20-enoic acid (28e) To a solution of 28e (995 mg, 1.352 mmol) in MeOH (5 mL) was added 10% Pd / C (100 mg), the suspension was purged with a H gas balloon three times, and then stirred under H pressure at room temperature for 90 min. After the reaction was complete, the Pd / C was filtered and the methanol was removed under reduced pressure to give 28e (770 mg, 88.2% yield). MS(ESI) m / z: 668.5 [M+Na]+

[0359] Step 4 (9H-fluoren-9-yl)methyl ((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)carbamate (28f) HATU (453.447 mg, 1.193 mmol) and DIPEA (462.38 mg, 3.578 mmol) are added to a solution of 28e (770 mg, 1.193 mmol) in DMF (10 mL). The reaction mixture is allowed to react at room temperature for 10 min. Then 10a (639.3 mg, 1.204 mmol) is added and left at the same temperature for another 1 h. The mixture is diluted with EtOAc (50 mL) and washed with brine (30 mL*3). The organic layer is dried over Na2SO4, filtered, and the filtrate is concentrated in vacuo to give 28f (1.27 g, crude). MS(ESI) m / z: 1085.7 [M+Na] +

[0360] Step 5 (S)-2-(2-(2-aminoacetamido)acetamido)-N-(2-((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-2-oxoethoxy)methyl)amino)-2-oxoethyl)-3-phenylpropanamide (28) To a mixture of 28f (1.27 g, crude) in THF (20 mL) was added EtN (1.74 g, 23.855 mmol). The mixture was allowed to react at room temperature for 2 hours. After completion of the reaction, the reaction mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 28 (296 mg, Yield: 29.5%). MS(ESI) m / z: 841.6 [M+H] +

[0361] Example 29 [ka]

[0362] Step 1 2-((3aS,4S,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)acetic acid (29b) 29a (purchased from WuXi Apptec, 450 mg, 1.23 mmol) was dissolved in THF (9 mL) and MeOH (9 mL), and then Pd / C (wet, 10%, 110 mg) was added. The resulting mixture was stirred at 25 °C for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give 29b (284 mg, quantitative) as a clear oil, which was used directly in the next step without purification. MS(ESI) m / z: 230.2 [MH] -

[0363] Step 2 2-((3aS,4S,6R,6aR)-6-(((S)-4-(((benzyloxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanamido)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)acetic acid (29d) 29d (520 mg, 76.8%) was synthesized from 29b and 29c (commercially available) according to the synthesis method in Step 2 of Example 1. MS(ESI) m / z: 573.5 [M+Na] +

[0364] Step 3 N5-(((3aR,4R,6S,6aS)-6-(2-(benzyloxy)-2-oxoethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)-N2-((benzyloxy)carbonyl)-L-glutamic acid tert-butyl ester (29e) 29e was synthesized according to the synthetic procedure in Step 3 of Example 22 (573 mg, 80.5% yield). MS(ESI) m / z: 663.5 [M+Na] +

[0365] Step 4 N5-(((2R,3S,4R,5S)-5-(2-(benzyloxy)-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N2-((benzyloxy)carbonyl)-L-glutamine (29f) 29e (150 mg, 0.23 mmol) was dissolved in DCM (2 mL), and then a mixed solvent of TFA / HO (v / v=9:1, 2 mL) was added at 0 °C. The mixture was first stirred at 0 °C for 1.5 h, and then stirred at 20 °C for another 1 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain a pale yellow oil in the form of crude product, which was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to obtain 29f (27 mg, Yield: 21.2%) as a white solid. MS(ESI) m / z: 545.5 [M+H] +

[0366] Step 5 Benzyl 2-((2S,3R,4S,5R)-5-((6S,15S)-15-benzyl-6-(((benzyloxy)carbonyl)amino)-24-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-3,7,10,13,16,19,24-heptaoxo-22-oxa-2,8,11,14,17,20-hexaazatetraconyl)-3,4-dihydroxytetrahydrofuran-2-yl)acetate (29g) 29g was synthesized according to the synthetic procedure in Step 8 of Example 22 (26 mg, 41.4% yield). MS(ESI) m / z: 1389.7 [M+Na] +

[0367] Step 6 2-((2S,3R,4S,5R)-5-((6S,15S)-6-amino-15-benzyl-24-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-3,7,10,13,16,19,24-heptaoxo-22-oxa-2,8,11,14,17,20-hexaazatetraconyl)-3,4-dihydroxytetrahydrofuran-2-yl)acetic acid (29h) 29h was synthesized according to the synthetic procedure in Step 7 of Example 22 (22 mg, crude). MS(ESI) m / z: 1143.7 [M+H] +

[0368] Step 7 2-((2S,3R,4S,5R)-5-((6S,15S)-6-amino-15-benzyl-24-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-3,7,10,13,16,19,24-heptaoxo-22-oxa-2,8,11,14,17,20-hexaazatetraconyl)-3,4-dihydroxytetrahydrofuran-2-yl)acetic acid (29) 29 was synthesized according to the synthetic procedure in Step 10 of Example 22 (4.5 mg, 77.0% yield). MS(ESI) m / z: 1358.8 [M+Na] +

[0369] Example 30 [ka]

[0370] Step 1 (S)-2-amino-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)pentanediamide (30a) To a mixture of 20 (20 mg, 0.037 mmol) and HATU (14.05 mg, 0.037 mmol) in DMF (1 mL) was added DIPEA (14.33 mg, 0.111 mmol). The mixture was allowed to react at room temperature for 10 min. 28 (40.0 mg, 0.048 mmol) was added and allowed to react at the same temperature for an additional 15 min. After the reaction was complete, piperidine (139.34 mg, 0.366 mmol) was added and allowed to stand at room temperature for an additional 10 min. The mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min) to give 30a (30.0 mg, Yield: 71.7%). MS(ESI) m / z: 1164.8 [M+Na] +

[0371] Step 2 (S)-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)pentanediamide (30) Compound 30 was synthesized according to the synthetic procedure in Step 8 of Example 2 (7.5 mg, 42.8% yield). MS(ESI) m / z: 1335.9 [M+H] +

[0372] Example 31 [ka]

[0373] Step 1 2-((3aS,4S,6R,6aR)-6-(hydroxymethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)acetic acid (31b) 31b was synthesized according to the synthetic procedure in Step 2 of Example 22 (231 mg, crude, 87.1% yield).

[0374] Step 2 N2-(((9H-Fluoren-9-yl)methoxy)carbonyl)-N6-(2-((3aS,4S,6R,6aR)-6-(hydroxymethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)acetyl)-L-lysine tert-butyl ester (31d) 31d was synthesized according to the synthetic procedure in Step 8 of Example 22 (393 mg, 64.9% yield). MS(ESI) m / z: 639.6 [M+H] +

[0375] Step 3 N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N6-(2-((2S,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)acetyl)-L-lysine (31e) 31e was synthesized according to the synthetic procedure in Step 4 of Example 29 (27 mg, 63.5% yield). MS(ESI) m / z: 543.5 [M+H] +

[0376] Step 4 (9H-fluoren-9-yl)methyl ((10S,19S)-10-benzyl-26-((2S,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15,18,25-heptaoxo-3-oxa-5,8,11,14,17,24-hexaazahexacosan-19-yl)carbamate (31f) 31f was synthesized according to the synthetic procedure in Step 8 of Example 22 (40 mg, 51.3% yield). MS(ESI) m / z: 1387.9 [M+Na] +

[0377] Step 5 (S)-2-Amino-N-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-6-(2-((2S,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)acetamido)hexanamide (31g) 31f (20 mg, 0.015 mmol) was dissolved in DMF (1 mL), and then EtNH (15 μL, 0.15 mmol) was added. The resulting mixture was stirred at 25 °C for 2 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give 31g (19 mg, quantitative) as a gray solid, which was used directly in the next step without purification. MS(ESI) m / z: 1143.8 [M+H] +

[0378] Step 6 (S)-N-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentane Tetraoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-6-(2-((2S,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)acetamido)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)hexanamide (31) 31 was synthesized according to the synthetic procedure in Step 8 of Example 22 (13.9 mg, 70.4% yield). MS(ESI) m / z: 1358.9 [M+Na] +

[0379] Example 32 [ka]

[0380] Step 1 (9H-Fluoren-9-yl)methyl ((10S,19S)-10-benzyl-22-(((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)amino)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15,18,22-heptaoxo-3-oxa-5,8,11,14,17-pentaazadocos-19-yl)carbamate (32a) To a solution of 18 (40 mg, 0.08 mmol), 28 (60 mg, 0.07 mmol), and HATU (30 mg, 0.08 mmol) in anhydrous DMF (1 mL) was added DIEA (36 μL, 0.21 mmol) and stirred at room temperature for 1.5 h. The reaction solution was filtered, purified by preparative HPLC (water / MeCN with 0.1% FA), and lyophilized to give the title compound 32a (52 mg, 55.1% yield) as a beige solid. MS(ESI) m / z: 1345.8 [M+Na] +

[0381] Step 2 (2S)-2-Amino-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-N5-((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pentanediamide (32b) To a solution of 32a (52 mg, 0.04 mmol) in DMF (1 mL), EtNH (81 μL, 0.79 mmol) was added and stirred at room temperature for 1 h. The solution was extracted with petroleum ether (3 mL * 4), and the DMF phase was concentrated to give the title compound 32b (44.4 mg, 102.6% yield) as a beige solid. MS(ESI) m / z: 1101.8 [M+H] +

[0382] Step 3 (2S)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12 ,15-Pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-N5-((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)pentanediamide (32) To a solution of 32b (25 mg, 22.7 μmol), 6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoic acid 32c (5.3 mg, 25.0 μmol), and HATU (9.5 mg, 25.0 μmol) in anhydrous DMF (1 mL) was added DIEA (8 μL, 45.4 μmol) and stirred at room temperature for 20 min. The solution was filtered, purified by preparative HPLC (0.1% FA in water / MeCN), and lyophilized to give the title compound 32 (10 mg, 34.0% yield) as a white solid. MS(ESI) m / z: 1316.8 [M+Na] +

[0383] Example 33 [ka]

[0384] Step 33-1 Benzyl((10S,19S)-10-benzyl-22-(((3R,4S,5S)-5-(((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)amino)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15,18,22-heptaoxo-3-oxa-5,8,11,14,17-pentaazadocos-19-yl)carbamate (33a) To a mixture of 19 (26 mg, 0.047 mmol) and HATU (17.73 mg, 0.047 mmol) in DMF (1 mL) was added DIPEA (18.08 mg, 0.14 mmol). The mixture was allowed to react at room temperature for 10 min. 28 (39.21 mg, 0.047 mmol) was added and allowed to react at the same temperature for another 15 min. After the reaction was complete, the mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min) to give 33a (35.0 mg, Yield: 54.4%). MS(ESI) m / z: 1403.9 [M+Na] +

[0385] Step 33-2 (2S)-2-amino-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9, 12,15-Pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-N5-((3R,4S,5S)-5-(((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)pentanediamide (33b) To a solution of 33a (35 mg, 0.025 mmol) in MeOH (1 mL), 10% Pd / C (5.3 mg) was added, and the suspension was purged with a H gas balloon three times and then stirred under H pressure at room temperature for 4 h. After the reaction was complete, the Pd / C was filtered, and the methanol was removed under reduced pressure to give 33b (28.3 mg, 89.6% yield). MS(ESI) m / z: 1246.8 [M+H] +

[0386] Step 33-3 (2S)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11, 14-tetraazahexadecan-16-yl)-N5-((3R,4S,5S)-5-(((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)pentanediamide (33) Compound 33 was synthesized according to the synthetic procedure in Step 8 of Example 2 (11.8 mg, 36.1% yield). MS(ESI) m / z: 1462.9 [M+Na] +

[0387] Example 34 [ka]

[0388] Step 1 N-methyl-N-(N-methyl-N-(N-methyl-N-(2-(methyl-12-azanyl)acetyl)glycyl)glycyl)glycinate methyl ester (34b) To a solution of compound 34a (200 mg, 0.44 mmol) in MeOH (6 mL) was added wet Pd / C (20 mg, 10 wt%). The mixture was stirred at room temperature under an atmosphere of H2 (15 psi) for 2 hours. The mixture was filtered through a pad of diatomaceous earth and concentrated to give the crude product. Compound 34b (180 mg, crude) was obtained as a colorless oil. MS(ESI) m / z: 317.4 [M+H] + .

[0389] Step 2 Methyl 1-((3 aS,6R,6 aR)-6-((S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(benzyloxy)-5-oxopentanamido)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-2,5,8,11-tetramethyl-1,4,7,10-tetraoxo-2,5,8,11-tetraazatridecan-13-oate (34d) To a solution of compound 34c (368 mg, 0.57 mmol) in pyridine (4 mL) was added 34b (180 mg, 0.63 mmol) and EDCI (130 mg, 0.63 mmol). The mixture was stirred at room temperature for 6 hours. The mixture was concentrated, and the residue was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 34d (120 mg, 22.3% yield) was obtained as a white solid. MS(ESI) m / z: 943.7 [M+H] + .

[0390] Step 3 Methyl 1-((3S,4R,5R)-5-((S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(benzyloxy)-5-oxopentanamido)-3,4-dihydroxytetrahydrofuran-2-yl)-2,5,8,11-tetramethyl-1,4,7,10-tetraoxo-2,5,8,11-tetraazatridecan-13-oate (34e) To a solution of compound 34d (120 mg, 0.13 mmol) in THF (1 mL) was added THF-HO (v:v=4:1, 5 mL). The mixture was stirred at room temperature for 4 hours. The mixture was concentrated to give the crude product, which was used in the next step without further purification. Compound 34e (115 mg, crude) was obtained as a white solid. MS(ESI) m / z: 903.7 [M+H] + .

[0391] Step 4 N2-(((9H-Fluoren-9-yl)methoxy)carbonyl)-N5-((3R,4S,5S)-3,4-dihydroxy-5-(methyl(5,8,11-trimethyl-3,6,9,12-tetraoxo-2-oxa-5,8,11-triazatridecan-13-yl)carbamoyl)tetrahydrofuran-2-yl)-L-glutamine (34f) To a solution of compound 34e (115 mg, 0.13 mmol) in MeOH (4 mL) was added wet Pd / C (15 mg, 10%). The mixture was stirred under H2 (15 psi) at room temperature for 5 h. The mixture was filtered through a pad of diatomaceous earth and concentrated. The crude was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 34f (74 mg, 64.3% yield) was obtained as a white solid. MS(ESI) m / z: 813.6 [M+H] + .

[0392] Step 5 N5-((3R,4S,5S)-5-((2-((2-((2-((2-amino-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-L-glutamine (34g) A solution of compound 34f (74 mg, 0.09 mmol) in NH-MeOH (6 mL, 7 M) was stirred in a sealed tube at 40 °C for 16 h. The mixture was concentrated, dissolved in HO (10 mL), and washed with EA (4 * 10 mL). The aqueous solution was lyophilized to give the crude product. Compound 34g (68 mg, crude) was obtained as a white solid. MS(ESI) m / z: 576.5 [M+H] + .

[0393] Step 6 N2-(((9H-Fluoren-9-yl)methoxy)carbonyl)-N5-((3R,4S,5S)-5-((2-((2-((2-((2-amino-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-L-glutamine(34h) To a solution of compound 34g (68 mg, 0.12 mmol) in CHCN (3 mL) and HO (1.5 mL) was added Fmoc-OSu (30.8 mg, 0.09 mmol) and NaCO (18.4 mg, 0.17 mmol). The mixture was stirred at room temperature for 2 hours. The mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 34h (29 mg, 30.2% yield) was obtained as a white solid. MS(ESI) m / z: 798.6 [M+H] + .

[0394] Step 7 ((10S,19S)-22-(((3R,4S,5S)-5-((2-((2-((2-((2-amino-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)amino)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro- 9-Hydroxy-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)amino)-1,6,9,12,15,18,22-heptaoxo-3-oxa-5,8,11,14,17-pentaazadocos-19-yl)carbamate (9H-fluoren-9-yl)methyl (34j) To a solution of compound 34i (29 mg, 0.036 mmol) in DMF (2 mL) was added 34h (32.1 mg, 0.038 mmol), HATU (20.7 mg, 0.054 mmol), and DIEA (7.1 mg, 0.054 mmol). The mixture was stirred at room temperature for 20 minutes. The mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 34j (32 mg, 54.0%) was obtained as a white solid. MS(ESI) m / z: 1644.0 [M+Na] + .

[0395] Step 8 (S)-2-Amino-N5-((3R,4S,5S)-5-((2-((2-((2-((2-amino-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-N1-((S)-10-benzyl-1-(((1S,9S)-9 -Ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)pentanediamide (34k) To a solution of compound 34j (32 mg, 0.02 mmol) in DMF (2 mL) was added EtNH (28.9 mg, 0.395 mmol). The mixture was stirred at room temperature for 1 h. The mixture was concentrated and coevaporated with toluene (3*2 mL). The crude was used in the next step without further purification. Compound 34k (34 mg, crude) was obtained as an off-white solid. MS(ESI) m / z: 1400.1 [M+H] + .

[0396] Step 9 (S)-N5-((3R,4S,5S)-5-((2-((2-((2-((2-amino-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)amino)-2-oxoethyl)(methyl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)pentanediamide (34) To a solution of compound 34i (6.3 mg, 0.03 mmol) in DMF (2 mL) was added HATU (11.3 mg, 0.03 mmol) and DIEA (3.8 mg, 0.03 mmol). The mixture was stirred at room temperature for 15 minutes. Compound 34k (27.6 mg, 0.02 mmol) was added to the mixture. The mixture was stirred at room temperature for 15 minutes. The mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 34 (21 mg, 66.8% yield) was obtained as a white solid. MS(ESI) m / z: 1614.1 [M+Na] + .

[0397] Example 35 [ka]

[0398] Step 1 N-(2-amino-2-oxoethyl)-2-(N,5,8,11,14,17,20,23,26,29-decamethyl-4,7,10,13,16,19,22,25,28-nonaoxo-2,5,8,11,14,17,20,23,26,29-decaazahentriacontan-31-amido)-N-methylacetamide (35b) To a solution of compound 35a (200 mg, 0.196 mmol) in MeOH (4 mL) was added wet Pd / C (20 mg, 10 wt%). The mixture was stirred at room temperature under an atmosphere of H2 (15 psi) for 2 hours. The mixture was filtered through a pad of diatomaceous earth and concentrated to give the crude product. Compound 35b (185 mg, crude) was obtained as a colorless oil.

[0399] Step 2 Benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-((3aR,6S,6aS)-6-((35-amino-3,6,9,12,15,18,21,24,27,30,33-undecamethyl-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxo-3,6,9,12,15,18,21,24,27,30,33-undecaazapentatriacontyl)(methyl)carbamoyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-L-glutamate (35d) To a solution of compound 35c (134 mg, 0.208 mmol) in pyridine (4 mL) was added 35b (185 mg, 0.208 mmol) and EDCI (47 mg, 0.01 mmol). The mixture was stirred at room temperature for 6 hours. The mixture was concentrated, and the residue was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 35d (305 mg, 98.1% yield) was obtained as a white solid. MS(ESI) m / z: 1535.0 [M+Na] + .

[0400] Step 3 Benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-((3R,4S,5S)-5-((35-amino-3,6,9,12,15,18,21,24,27,30,33-undecamethyl-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxo-3,6,9,12,15,18,21,24,27,30,33-undecaazapentatriacontyl)(methyl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-L-glutamate (35e) To a solution of compound 35d (305 mg, 0.202 mmol) in THF (2 mL) was added THF-HO (v:v=4:1, 10 mL). The mixture was stirred at room temperature for 4 hours. The mixture was concentrated to give the crude product, which was used in the next step without further purification. Compound 35e (295 mg, crude) was obtained as a white solid. MS(ESI) m / z: 1495.0 [M+Na] + .

[0401] Step 4 N2-(((9H-Fluoren-9-yl)methoxy)carbonyl)-N5-((3R,4S,5S)-5-((35-amino-3,6,9,12,15,18,21,24,27,30,33-undecamethyl-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxo-3,6,9,12,15,18,21,24,27,30,33-undecaazapentatriacontyl)(methyl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-L-glutamine (35f) To a solution of compound 35e (295 mg, 0.2 mmol) in MeOH (6 mL) was added wet Pd / C (30 mg, 10%). The mixture was stirred under H2 (15 psi) at room temperature for 5 h. The mixture was filtered through a pad of diatomaceous earth and concentrated. The crude was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 35f (162 mg, 58.5% yield) was obtained as a white solid. MS(ESI) m / z: 1404.9 [M+Na] + .

[0402] Step 5 N5-((3R,4S,5S)-5-((35-amino-3,6,9,12,15,18,21,24,27,30,33-undecamethyl-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxo-3,6,9,12,15,18,21,24,27,30,33-undecaazapentatriacontyl)(methyl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-L-glutamine (35g) A solution of compound 35f (162 mg, 0.117 mmol) in NH-MeOH (6 mL, 7 M) was stirred in a sealed tube at 40 °C for 16 h. The mixture was concentrated, dissolved in HO (15 mL), and washed with EA (4 * 10 mL). The aqueous solution was lyophilized to give the crude product. Compound 35g (142 mg, crude) was obtained as a white solid. MS(ESI) m / z: 1145.9 [M+H] + .

[0403] Step 6 N2-(((9H-Fluoren-9-yl)methoxy)carbonyl)-N5-((3R,4S,5S)-5-((35-amino-3,6,9,12,15,18,21,24,27,30,33-undecamethyl-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxo-3,6,9,12,15,18,21,24,27,30,33-undecaazapentatriacontyl)(methyl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-L-glutamine (35h) To a solution of compound 35g (126 mg, 0.11 mmol) in CHCN (3 mL) and HO (1.5 mL) was added FmocOSu (40.9 mg, 0.121 mmol) and NaCO (23.3 mg, 0.22 mmol). The mixture was stirred at room temperature for 2 hours. The mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 35h (62 mg, 41.1% yield) was obtained as a white solid. MS(ESI) m / z: 1388.9 [M+Na] + .

[0404] Step 7 ((10S,19S)-22-(((3R,4S,5S)-5-((35-amino-3,6,9,12,15,18,21,24,27,30,33-undecamethyl-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxo-3,6,9,12,15,18,21,24,27,30,33-undecaazapentatriacontyl)(methyl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)amino)-10-benzyl-1 -(((1S,9S)-9-Ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15,18,22-heptaoxo-3-oxa-5,8,11,14,17-pentaazadocos-19-yl)carbamate (9H-fluoren-9-yl)methyl (35j) To a solution of compound 35i (20 mg, 0.024 mmol) in DMF (2 mL) was added 35h (35.8 mg, 0.026 mmol), HATU (13.6 mg, 0.04 mmol), and DIEA (6.2 mg, 0.05 mmol). The mixture was stirred at room temperature for 30 minutes. The mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 35j (33 mg, 63.5% yield) was obtained as a white solid. MS(ESI) m / z: 2189.7 [M+H] + .

[0405] Step 8 (S)-2-amino-N5-((3R,4S,5S)-5-((35-amino-3,6,9,12,15,18,21,24,27,30,33-undecamethyl-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxo-3,6,9,12,15,18,21,24,27,30,33-undecaazapentatriacontyl)(methyl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-N1-(( S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)pentanediamide (35k) To a solution of compound 35j (33 mg, 0.015 mmol) in DMF (2 mL) was added EtNH (22 mg, 0.30 mmol). The mixture was stirred at room temperature for 45 min. The mixture was concentrated and coevaporated with toluene (3*2 mL). The crude was used in the next step without further purification. Compound 35k (35 mg, crude) was obtained as an off-white solid. MS(ESI) m / z: 1968.1 [M+H] + .

[0406] Step 9 (S)-N5-((3R,4S,5S)-5-((35-amino-3,6,9,12,15,18,21,24,27,30,33-undecamethyl-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxo-3,6,9,12,15,18,21,24,27,30,33-undecaazapentatriacontyl)(methyl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)-N1-((S)-10-benzyl-1-(((1S,9S)-9- Ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)pentanediamide (35) To a solution of compound 35i (4.8 mg, 0.023 mmol) in DMF (2 mL) was added HATU (8.7 mg, 0.023 mmol) and DIEA (2.96 mg, 0.023 mmol). The mixture was stirred at room temperature for 15 minutes. Compound 35k (30 mg, 0.015 mmol) was added to the mixture. The mixture was stirred at room temperature for 15 minutes. The mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 35 (19.8 mg, Yield: 60.09%) was obtained as a white solid. MS(ESI) m / z: 2161.4 [M+H] + .

[0407] Example 36 [ka]

[0408] Step 1 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamic acid 4- ((5S,8S,11S)-5-(3-(((3R,4S,5S)-5-(((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)amino)-3-oxopropyl)-8-isopropyl-3,6,9-trioxo-1-phenyl-11-(3-ureidopropyl)-2-oxa-4,7,10-triazadodecan-12-amide)benzyl ester (36a) Compound 36a was synthesized according to the synthetic procedure in Step 1 of Example 33 (34.7 mg, 36.5% yield). MS(ESI) m / z: 1664.4 [M+H] +

[0409] Step 2 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl (methyl)carbamic acid 4-((2S)-2-((2S)-2-((2S)-2-amino-5-(((3R,4S,5S)-5-(((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)amino)-5-oxopentanamido)-3-methylbutanamido)-5-ureidopentanamido) benzyl ester (36b) Compound 36b was synthesized according to the synthetic procedure in Step 2 of Example 33, 19.2 mg, 60.2% yield. MS(ESI) m / z: 1529.2 [M+H] +

[0410] Step 3 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamic acid 4-((2S )-2-((2S)-2-((2S)-5-(((3R,4S,5S)-5-(((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)amino)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)-5-oxopentanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl ester (36) Compound 36 was synthesized according to the synthetic procedure in Step 8 of Example 2 (10.7 mg, 49.5% yield). MS(ESI) m / z: 1722.9 [M+H] +

[0411] Example 37 [ka]

[0412] Step 1 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamic acid 4-((5S,8S,11S)-5-(3-(((3R,4S,5S)-5-(((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl) Tetrahydrofuran-2-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)amino)-3-oxopropyl)-8-isopropyl-3,6,9-trioxo-1-phenyl-11-(3-ureidopropyl)-2-oxa-4,7,10-triazadodecan-12-amido)benzyl ester (37a) Compound 37a was synthesized according to the synthetic procedure in Step 1 of Example 33 (15.0 mg, 24.0% yield). MS(ESI) m / z: 1381.8 [M+H] +

[0413] Step 2 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl (methyl)carbamic acid 4-((2S)-2-((2S)-2-((2S)-2-amino-5-(((3R,4S,5S)-5-(((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)amino)-5-oxopentanamido)-3-methylbutanamido)-5-ureidopentanamido) benzyl ester (37b) Compound 37b was synthesized according to the synthetic procedure in Step 2 of Example 33, 12.5 mg, 90.9% yield. MS(ESI) m / z: 1268.9 [M+Na] +

[0414] Step 3 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamic acid 4-((2S )-2-((2S)-2-((2S)-5-(((3R,4S,5S)-5-(((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)carbamoyl)-3,4-dihydroxytetrahydrofuran-2-yl)amino)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)-5-oxopentanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl ester (37) Compound 37 was synthesized according to the synthetic procedure in Step 8 of Example 2 (7.3 mg, 50.6% yield). MS(ESI) m / z: 1441.0 [M+H] +

[0415] Example 38 [ka]

[0416] Step 1 (S)-N4-((2R,3R,4R,5S,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-2-amino-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)succinimide (38b) To a mixture of 38a (30 mg, 0.044 mmol) and TSTU (13.21 mg, 0.044 mmol) in DMF (1 mL) was added DIPEA (17.01 mg, 0.132 mmol). The mixture was allowed to react at room temperature for 10 min, and the acid was converted to the activated ester by LCMS. 28 (29.52 mg, 0.035 mmol) was added and the reaction was continued at the same temperature for another 1 h. After the reaction was complete, piperidine NH2NH2·HO (200 μL, 80%) was added, and the mixture was allowed to stand at room temperature for another 10 min. The mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min) to give 38b (6.9 mg, 17.2% yield). MS(ESI) m / z: 1158.9 [M+H] +

[0417] Step 2 (S)-N4-((2R,3R,4R,5S,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)succinimide (38) Compound 38 was synthesized according to the synthetic procedure in Step 8 of Example 2 (4.5 mg, 55.9% yield). MS(ESI) m / z: 1373.8 [M+Na] +

[0418] Example 39 [ka]

[0419] Step 1 2,5-Dioxopyrrolidin-1-yl 2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetate (39b) To a solution of compound 39a (600 mg, 3.87 mmol) in DMF (8 mL) was added DCC (958 mg, 4.64 mmol) and HOSu (534 mg, 4.64 mmol). The mixture was stirred at room temperature for 4 h. The mixture was filtered to remove the solids, and the filtrate was concentrated to give the crude product. Compound 39b (855 mg, crude) was obtained as an off-white solid. MS(ESI) m / z: 275.2 [M+Na] + .

[0420] Step 2 3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanoic acid (39d) To a solution of compound 39b (855 mg, crude) in DMSO (8 mL) was added 3-aminopropanoic acid (379 mg, 4.26 mmol). The mixture was stirred at 40 °C for 4 h. The mixture was diluted with H2O (20 mL) and washed with MTBE (3 * 20 mL). The aqueous solution was extracted with EA (6 * 100 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered and concentrated. A solution of compound 39d (875 mg, theoretical) in DMSO was obtained, which was used directly in the next step without further purification. MS(ESI) m / z: 227.2 [M+H] + .

[0421] Step 3 2,5-Dioxopyrrolidin-1-yl 3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanoate To a solution of compound 39e (875 mg, theoretical) in DCM (10 mL) was added DCC (958 mg, 4.64 mmol) and HOSu (534 mg, 4.64 mmol). The mixture was stirred at room temperature for 3 hours. The mixture was filtered and concentrated. The mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 39e (195 mg, 15.6% yield) was obtained as a colorless gum. MS(ESI) m / z: 346.3 [M+Na] + .

[0422] Step 4 (S)-2-(2-(2-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamide)acetamido)acetamido)-N-(2-((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-2-oxoethoxy)methyl)amino)-2-oxoethyl)-3-phenylpropanamide (39) To a solution of compound 39f (25 mg, 0.03 mmol) in DMF (0.8 mL) was added 39e (12.5 mg, 0.04 mmol) and DIEA (2.2 mg, 0.03 mmol). The mixture was stirred at room temperature for 2 hours. The pH was adjusted to 6 with 0.1% TFA solution. The mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 39 (8.0 mg, 25.6% yield) was obtained as a white solid. MS(ESI) m / z: 1071.7 [M+Na] + .

[0423] Example 40 [ka]

[0424] Step 1 (2S)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaerythritol N-oxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-N5-((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamido)pentanediamide (40) To a solution of 32b (20 mg, 18.2 μmol) in anhydrous DMF (1 mL) was added 7a (8.8 mg, 27.2 μmol) and stirred at room temperature for 1.5 h. The solution was filtered, purified by preparative HPLC (water / MeCN with 0.1% FA), and lyophilized to give the title compound 40 (9.5 mg, 39.9% yield) as a white solid. MS(ESI) m / z: 1331.8 [M+Na] +

[0425] Example 41 [ka] (S)-2-(2-(2-(2-(4-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)phenyl)acetamido)acetamido)acetamido)-N-(2-((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-2-oxoethoxy)methyl)amino)-2-oxoethyl)-3-phenylpropanamide (41) 28 (synthesized according to the synthetic procedure of Example 28, 5.0 mg, 0.022 mmol), 41a (commercially available, 18 mg, 0.022 mmol), and HATU (9.0 mg, 0.024 mmol) were dissolved in DMF (0.5 mL), and DIEA (11.5 μL, 0.065 mmol) was added. The resulting mixture was stirred at 25 °C for 15 min. After the reaction was completed, the reaction mixture was acidified to pH = 6 with HO containing 0.1% TFA and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min) to give 41 (11 mg, Yield: 48.2%) as a white solid. MS(ESI) m / z: 1076.5 [M+Na] +

[0426] Example 42 [ka]

[0427] Step 1 ((10S,19S)-10-benzyl-19-(3-(((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)amino)-3-oxopropyl)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15,18,21-heptaoxo-3-oxa-5,8,11,14,17,20-hexaazatricos-23-yl)carbamate (9H-fluoren-9-yl)methyl (42b) To a solution of 32b (24 mg, 21.8 μmol), Fmoc-glycine 42a (7.5 mg, 24.0 μmol), and HATU (9.1 mg, 24.0 μmol) in anhydrous DMF (1.5 mL) was added DIEA (7 μL, 43.6 μmol) and stirred at room temperature for 25 min. The solution was filtered, purified by preparative HPLC (0.1% FA in water / MeCN), and lyophilized to give the title compound 42b (16.5 mg, 54.3% yield) as a white solid. MS(ESI) m / z: 1416.8 [M+Na] +

[0428] Step 2 (2S)-2-(3-aminopropanamido)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-N5-((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pentanediamide (42c) To a solution of 42b (16.5 mg, 11.8 μmol) in anhydrous DMF (1 mL) was added EtNH (24 μL, 0.24 mmol) and stirred at room temperature for 30 min. The solution was concentrated in vacuo. The residue was triturated with MTBE (5 mL), filtered, and washed with MTBE (1 mL*3). The filter cake was collected to give the title compound 42c (15 mg, 108.4% yield) as a light brown solid, which was used directly in the next step without further purification. MS(ESI) m / z: 1172.8 [M+H] +

[0429] Step 3 (2S)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo -3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-N5-((3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-(3-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamide)propanamide)pentanediamide (42) The title compound 42 (9.1 mg, 55.2%) was obtained according to the procedure described in Step 1 of Example 40. MS(ESI) m / z: 1402.8 [M+Na] +

[0430] Example 43 [ka]

[0431] Step 1 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl) ethyl)carbamic acid 4-((5S,8S,11S)-5-(3-((((2S,3R,4S,5R)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-3-oxopropyl)-1-(9H-fluoren-9-yl)-8-isopropyl-3,6,9-trioxo-11-(3-ureidopropyl)-2-oxa-4,7,10-triazadodecan-12-amide) benzyl ester (43a) To a mixture of 20 (25 mg, 0.046 mmol) and HATU (17.55 mg, 0.046 mmol) in DMF (1 mL) was added DIPEA (17.90 mg, 0.138 mmol). The mixture was allowed to react at room temperature for 10 minutes. 9b (51.86 mg, 0.046 mmol) was added and the mixture was allowed to react at the same temperature for another 15 minutes. After the reaction was complete, the mixture was filtered and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min) to give 43a (55.9 mg, Yield: 73.5%). MS(ESI) m / z: 824.6 [M+2H] 2+

[0432] Step 2 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino) -3-Methyl-1-oxobutan-2-yl)(methyl)carbamic acid 4-((S)-2-((S)-2-((S)-2-amino-5-((((2S,3R,4S,5R)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-5-oxopentanamido)-3-methylbutanamido)-5-ureidopentanamido) benzyl ester (43b) To a mixture of 43a (55.9 mg, 0.034 mmol) in DMF (1 mL) was added EtN (49.65 mg, 0.479 mmol). The mixture was allowed to react at room temperature for 30 min. After completion of the reaction, the mixture was concentrated under reduced pressure to give 43b (59 mg, crude). MS(ESI) m / z: 713.5 [M+2H] 2+

[0433] Step 3 ((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamic acid 4-((21S,24S,27S) -21-(3-((((2S,3R,4S,5R)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-3-oxopropyl)-1-((1R,8S,9s)-bicyclo[6.1.0]non-4-yn-9-yl)-24-isopropyl-3,19,22,25-tetraoxo-27-(3-ureidopropyl)-2,7,10,13,16-pentaoxa-4,20,23,26-tetraazaoctacosan-28-amide) benzyl ester (43) To a mixture of 43c (10.0 mg, 0.023 mmol) and HATU (8.61 mg, 0.023 mmol) in DMF (0.3 mL) was added DIPEA (8.78 mg, 0.068 mmol). The mixture was allowed to react at room temperature for 10 minutes. 43b (32.27 mg, crude) was added and allowed to react at the same temperature for another 15 minutes. After the reaction was complete, the mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min) to give 43 (22.2 mg, Yield: 53.0%). MS(ESI) m / z: 1849.2 [M+H] +

[0434] Example 44 [ka]

[0435] Step 1 (9H-fluoren-9-yl)methyl ((2R,10S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-2-(hydroxymethyl)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)carbamate (44a) 22k (125 mg, 0.094 mmol) was dissolved in THF (3 mL), and then TBAF (1 M THF stock solution, 113 μL, 0.11 mmol) was added. The resulting mixture was stirred at 25 °C for 1 h. EtNH (97 μL, 0.94 mmol) was added, and the mixture was stirred at 25 °C for an additional 1 h. After completion of the reaction, the reaction mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 44a (38 mg, 46.4% yield) as a white solid. MS(ESI) m / z: 871.7 [M+H] +

[0436] Step 2 ((6S,15S,23R)-1-((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)-15-benzyl-24-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,1 2H-Benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)amino)-23-(hydroxymethyl)-3,7,10,13,16,19,24-heptaoxo-22-oxa-2,8,11,14,17,20-hexaazatetracosan-6-yl)carbamate (9H-fluoren-9-yl)methyl (44b) 44b was synthesized according to the synthetic procedure in Step 8 of Example 22 (14.5 mg, 50.3% yield). MS(ESI) m / z: 1416.9 [M+Na] +

[0437] Step 3 (S)-2-Amino-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((2R,10S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-2-(hydroxymethyl)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)pentanediamide (44c) 44c was synthesized according to the synthetic procedure in Step 9 of Example 22 (15.2 mg, crude). MS(ESI) m / z: 1173.8 [M+H] +

[0438] Step 4 (S)-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((2R,10S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-2-(hydroxymethyl)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-2-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamido)pentanediamide (44c) 44 was synthesized according to the synthetic procedure in Step 10 of Example 22 (5.3 mg, 36.9% yield). MS(ESI) m / z: 1402.9 [M+Na] +

[0439] Example 45 [ka]

[0440] Step 1 3-(((5S,14S)-5-(3-((((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-3-oxopropyl)-14-benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15,18-hexaoxo-2-oxa-4,7,10,13,16,19-hexaazaicos-20-yl)oxy)bicyclo[1.1.1]pentane-1-carboxylic acid (45a) To a solution of 20 (68 mg, 0.12 mmol) and TSTU (37 mg, 0.12 mmol) in anhydrous DMF (1 mL) was added DIEA (56 μL, 0.34 mmol) and stirred at room temperature for 10 min. 24c (53 mg, 0.11 mmol) was added and stirred at room temperature for 1 h. The solution was filtered, purified by preparative HPLC (water / MeCN with 0.1% FA), and lyophilized to give the title compound 45a (52 mg, 46.3% yield) as a white solid. MS(ESI) m / z: 997.7 [MH] -

[0441] Step 2 ((7S,16S)-21-((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)-7-benzyl-1-((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12 H-Benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)bicyclo[1.1.1]pentan-1-yl)oxy)-3,6,9,12,15,19-hexaoxo-2,5,8,11,14,20-hexaazahenicocin-16-yl)carbamate (9H-fluoren-9-yl)methyl (45b) To a solution of 45a (52 mg, 52.0 μmol), exatecan mesylate (29 mg, 54.7 μmol), and HATU (21 mg, 54.7 μmol) in anhydrous DMF (2 mL) was added DIEA (17 μL, 0.1 mmol) and stirred at room temperature for 30 min. The solution was filtered, purified by preparative HPLC (0.1% FA in water / MeCN), and lyophilized to give the title compound 45b (62 mg, 84.2% yield) as an off-white solid. MS(ESI) m / z: 1438.9 [M+Na] +

[0442] Step 3 (S)-2-Amino-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-7-benzyl-1-((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)carbamoyl)bicyclo[1.1.1]pentan-1-yl)oxy)-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecan-13-yl)pentanediamide (45c) To a solution of 45b (60 mg, 42.4 μmol) in anhydrous DMF (1 mL) was added EtNH (88 μL, 0.85 mmol) and stirred at room temperature for 2 h. The solution was concentrated in vacuo. The residue was triturated with MTBE (5 mL), filtered, and washed with MTBE (1 mL*3). The filter cake was collected to give the title compound 45c (56 mg, 110.6% yield) as a light brown solid, which was used directly in the next step without further purification. MS(ESI) m / z: 1194.9 [M+H] +

[0443] Step 4 (S)-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-7-benzyl-1-((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pi Lano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)bicyclo[1.1.1]pentan-1-yl)oxy)-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecan-13-yl)-2-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamido)pentanediamide (45) The title compound 45 (13 mg, 44.1%) was obtained according to the procedure described in Step 1 of Example 40. MS(ESI) m / z: 1424.8 [M+Na] +

[0444] Example 46 [ka] 46 (5.6 mg, 44.1% yield) is synthesized using a synthetic procedure similar to that of Example 44 (see Example 21 for the synthesis of intermediate 21). MS(ESI) m / z: 1575.9 [M+Na] +

[0445] Example 47 [ka]

[0446] Step 1 3-(((5S,14S)-5-(3-((((2R,3S,4R,5S)-5-(2-((((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-3-oxopropyl)-14-benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15,18-hexaoxo-2-oxa-4,7,10,13,16,19-hexaazaicos-20-yl)oxy)bicyclo[1.1.1]pentane-1-carboxylic acid (38b) To a mixture of 21 (95.0 mg, 0.133 mmol) and TSTU (40.02 mg, 0.133 mmol) in DMF (2 mL) was added DIPEA (51.54 mg, 0.399 mmol). The mixture was reacted at room temperature for 10 minutes, and the acid was converted to the activated ester according to LCMS. 47a (385 mg, crude) was added and reacted at the same temperature for another hour. The reaction was complete. The mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 47b (66.5 mg, Yield: 42.9%). MS(ESI) m / z: 1170.8 [MH] -

[0447] Step 2 ((7S,16S)-21-((2R,3S,4R,5S)-5-(2-((((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)-7-benzyl-1-((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)carbamoyl)bicyclo[1.1.1]pentan-1-yl)oxy)-3,6,9,12,15,19-hexaoxo-2,5,8,11,14,20-hexaazahenicocin-16-yl)carbamate (9H-fluoren-9-yl)methyl (47c) Compound 47c was synthesized according to the synthetic procedure in Step 4 of Example 28, 29.5 mg, 90.5% yield. MS(ESI) m / z: 1613.0 [M+Na] +

[0448] Step 3 (S)-2-amino-N5-(((2R,3S,4R,5S)-5-(2-((((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-7-benzyl-1-((3-(((1S,9S)-9-ethyl-5-fluoro -9-Hydroxy-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)carbamoyl)bicyclo[1.1.1]pentan-1-yl)oxy)-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecan-13-yl)pentanediamide (47d) Compound 47d was synthesized according to the synthetic procedure in Step 2 of Example 43 (29.6 g, crude). MS(ESI) m / z: 1368.1 [M+H] +

[0449] Step 4 (S)-N5-(((2R,3S,4R,5S)-5-(2-((((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-7-benzyl-1-((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)carbamoyl)bicyclo[1.1.1]pentan-1-yl)oxy)-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecan-13-yl)-2-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamido)pentanediamide (47) Compound 47 was synthesized according to the synthetic procedure in Step 8 of Example 2 (19.9 mg, 58.3% yield). MS(ESI) m / z: 1598.9 [M+Na] +

[0450] Example 48 [ka]

[0451] Step 1 (S)-2-amino-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)pentanediamide (30a) Compound 30a was synthesized according to the synthetic procedure in Step 1 of Example 30 (20 mg, 36.8% yield). MS(ESI) m / z: 1142.8 [M+H+]

[0452] Step 2 (S)-2-amino-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)pentanediamide (48) To a mixture of 48a (5 mg, 0.021 mmol) and HATU (8 mg, 0.021 mmol) in DMF (1 mL) was added DIPEA (5 mg, 0.035 mmol). The mixture was allowed to react at room temperature for 10 minutes. 30a (20 mg, 0.018 mmol) was added and the mixture was allowed to react at the same temperature for another 15 minutes. The mixture was filtered, and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 48 (4 mg, Yield: 16.9%). MS(ESI) m / z: 1372.8 [M+Na] +

[0453] Example 49 [ka]

[0454] Step 1 2,5-Dioxopyrrolidin-1-yl 2-((3aS,4S,6R,6aR)-2,2-dimethyl-6-((((Z)-2-oxo-2-phenylethylidene)amino)methyl)tetrahydrofuro[3,4-d][1,3]dioxol-4-yl)acetate (49b) 49f was synthesized according to the synthetic procedure in Step 1 of Example 39b (215 mg, theoretical).

[0455] Step 2 14-((3aS,4S,6R,6aR)-6-((((benzyloxy)carbonyl)amino)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-3,6,9,12-tetramethyl-4,7,10,13-tetraoxo-3,6,9,12-tetraazatetradecanoic acid (49d) To a solution of compound 49c (156 mg, 0.37 mmol) in DMF (3 mL) was added 49b (215.16 mg, 0.47 mmol) and DIEA (289 mg, 2.24 mmol). The mixture was stirred at room temperature for 16 hours. The mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 49d (115 mg, 47.3% yield) was obtained as a white solid. MS(ESI) m / z: 650.6 [M+H] + .

[0456] Step 3 Benzyl (((3aR,4R,6S,6aS)-6-(14-amino-3,6,9,12-tetramethyl-2,5,8,11,14-pentaoxo-3,6,9,12-tetraazatetradecanyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)carbamate (49e) To a solution of compound 49d (105 mg, 0.16 mmol) in DMF (1 mL) was added PyBOP (127 mg, 0.24 mmol), HOBt (33 mg, 0.24 mmol), DIEA (105 mg, 0.81 mmol), and NH3-MeOH (230 μL, 7 M). The mixture was stirred at room temperature for 1 h. The mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 49e (87 mg, 83.0% yield) was obtained as a white solid. MS(ESI) m / z: 649.6 [M+H] + .

[0457] Step 4 N-(2-amino-2-oxoethyl)-2-(2-(2-(2-((3aS,4S,6R,6aR)-6-(aminomethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-N-methylacetamide)-N-methylacetamide)-N-methylacetamide)-N-methylacetamide (49f) 49f was synthesized according to the synthetic procedure in Step 1 of Example 34b (85.0 mg, crude). MS(ESI) m / z: 515.6 [M+H] + .

[0458] Step 5 Benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((3aR,4R,6S,6aS)-6-(14-amino-3,6,9,12-tetramethyl-2,5,8,11,14-pentaoxo-3,6,9,12-tetraazatetradecanyl)-2,2 dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)-L-glutamate (49h) To a solution of compound 49g (83.5 mg, 0.18 mmol) in DMF (2 mL), HATU (94.2 mg, 0.25 mmol) and DIEA (64 mg, 0.5 mmol) were added. The mixture was stirred at room temperature for 15 minutes. Compound 49f (85.0 mg, 0.165 mmol) was added to the mixture and stirred at room temperature for 2 hours. The mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min). Compound 49h (83.0 mg, 52.5% yield) was obtained as a white solid. MS(ESI) m / z: 956.8 [M+H] + .

[0459] Step 6 Benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((2R,3S,4R,5S)-5-(14-amino-3,6,9,12-tetramethyl-2,5,8,11,14-pentaoxo-3,6,9,12-tetraazatetradecanyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-L-glutamate (49i) 49i was synthesized according to the synthetic procedure in Step 3 of Example 34e (79.0 mg, crude). MS(ESI) m / z: 916.7 [M+H] + .

[0460] Step 7 N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((2R,3S,4R,5S)-5-(14-amino-3,6,9,12-tetramethyl-2,5,8,11,14-pentaoxo-3,6,9,12-tetraazatetradecanyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-L-glutamine (49j) 49j was synthesized according to the synthetic procedure in Step 4 of Example 34f (42.0 mg, 59.2% yield). MS(ESI) m / z: 826.7 [M+H] + .

[0461] Step 8 ((6S,15S)-1-((2R,3S,4R,5S)-5-(14-amino-3,6,9,12-tetramethyl-2,5,8,11,14-pentaoxo-3,6,9,12-tetraazatetradecanyl)-3,4-dihydroxytetrahydrofuran-2-yl)-15-benzyl-24-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-di oxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)amino)-3,7,10,13,16,19,24-heptaoxo-22-oxa-2,8,11,14,17,20-hexaazatetracosan-6-yl)carbamate (9H-fluoren-9-yl)methyl (49l) 49l was synthesized according to the synthetic procedure in Step 7 of Example 34j (55.0 mg, theoretical). MS(ESI) m / z: 1672.0 [M+Na] + .

[0462] Step 9 (S)-2-amino-N5-(((2R,3S,4R,5S)-5-(14-amino-3,6,9,12-tetramethyl-2,5,8,11,14-pentaoxo-3,6,9,12-tetraazatetradecanyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro- 9-Hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)pentanediamide (49m) 49m was synthesized according to the synthetic procedure in Step 8 of Example 34k (30.28 mg, theoretical). MS(ESI) m / z: 1428.1 [M+H] +

[0463] Step 10 (S)-N5-(((2R,3S,4R,5S)-5-(14-amino-3,6,9,12-tetramethyl-2,5,8,11,14-pentaoxo-3,6,9,12-tetraazatetradecanyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-2-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamido)pentanediamide (49) 49 was synthesized according to the synthetic procedure in Step 4 of Example 39 (11.2 mg, 31.5% yield). MS(ESI) m / z: 1658.0 [M+Na] +

[0464] Example 50 [ka]

[0465] Step 1 (2S,3R,4R,5S,6R)-2-(aminomethyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (50b) To a mixture of 50a (200 mg, 0.896 mmol) in MeOH / HO (V / V = 1:1, 4 mL) was added wet Pd / C (35 mg, 10% purity). The black suspension was purged with a H gas balloon three times and stirred at room temperature for 2 h. LCMS showed complete consumption of 50a. The black suspension was filtered through a pad of diatomaceous earth, and the filtrate was concentrated to give 50b (173 mg, crude). MS(ESI) m / z: 194.2 [M+H] +

[0466] Step 2 Benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((2S,3R,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)methyl)-L-glutamate (50c) To a mixture of 50b (173 mg, crude) and 20c (431.43 mg, 0.775 mmol) in MeOH / HO (8 mL) was added saturated NaHCO (3 mL) and stirred at room temperature for 2 h. LCMS showed complete consumption of 50b. The mixture was concentrated in vacuo to remove most of the MeOH, and the aqueous phase was acidified to pH = 6 with 1 N HCl, filtered, and the filtrate was purified by preparative HPLC (FA) column: XBridge Prep C18 OBD 5 um 19*250 mm, mobile phase: A-water (0.1% formic acid): B-acetonitrile, flow rate: 20 mL / min, to give 50c (35 mg, 7.1% yield). MS(ESI) m / z: 635.5 [M+H] +

[0467] Step 3 N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((2S,3R,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)methyl)-L-glutamine (50d) Compound 50d was synthesized according to the synthetic procedure in Step 2 of Example 19 (30 mg, crude). MS(ESI) m / z: 545.5 [M+H] +

[0468] Step 4 ((6S,15S)-15-benzyl-24-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-3,7,10, 13,16,19,24-Heptaoxo-1-((2S,3R,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-22-oxa-2,8,11,14,17,20-hexaazatetracosan-6-yl)carbamate (9H-fluoren-9-yl)methyl (50e) Compound 50e was synthesized according to the synthetic procedure in Step 1 of Example 38, 32.5 mg, 43.1% yield. MS(ESI) m / z: 1389.9 [M+Na] +

[0469] Step 5 (S)-2-Amino-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-N5-(((2S,3R,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)methyl)pentanediamide (50f) Compound 50f was synthesized according to the synthetic procedure in Step 3 of Example 42 (29.5 mg, crude). MS(ESI) m / z: 1145.8 [M+H] +

[0470] Step 6 (S)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxo (2S,3R,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)methyl)pentanediamide (50) Compound 50 was synthesized according to the synthetic procedure in Step 8 of Example 2 (9.5 mg, 34.2% yield). MS(ESI) m / z: 1354.4 [M+H] +

[0471] Example 51 [ka]

[0472] Step 1 (S)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaox So-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)-N5-(((2S,3R,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)methyl)pentanediamide (51) Compound 51 was synthesized according to the synthetic procedure in Step 8 of Example 2 (24.2 mg, 34.5% yield). MS(ESI) m / z: 1339.2 [M+H] +

[0473] Example 52 [ka]

[0474] Step 1 (2S,3R,4R,5S,6R)-2-(aminomethyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (52b) To a solution of compound 52a (200 mg, 0.89 mmol) in MeOH (3 mL) was added HCOONH4 (226 mg, 3.58 mmol) and Pd / C (20 mg, 10%). The mixture was stirred at 68 °C for 1 h. H2O (10 mL) was added to the mixture, which was then filtered. The filtrate was concentrated to give the crude product. Compound 52b (210 mg, crude) was obtained as a white solid. MS(ESI) m / z: 194.3 [M+H] + .

[0475] Step 2 Benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((3aR,4R,6S,6aS)-2,2-dimethyl-6-(2-oxo-2-((((2S,3R,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)methyl)amino)ethyl)tetrahydrofuran-[3,4-d][1,3]dioxol-4-yl)methyl)-L-glutamate (52d) 52d was synthesized according to the synthetic procedure in Step 1 of Example 45a (152 mg, 48.2% yield). MS(ESI) m / z: 848.7 [M+H] + .

[0476] Step 3 (2S,3S,4R,5R,6S)-6-((2-((3aR,4R,6S,6aS)-6-(((S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(benzyloxy)-5-oxopentanamido)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)acetamido)methyl)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (52e) To a solution of compound 52d (152 mg, 0.18 mmol) in THF (4 mL) and HO (2 mL) was added TEMPO (2.8 mg, 0.018 mmol), KBr (2.1 mg, 0.018 mmol), and NaHCO (151 mg, 1.79 mmol). NaClO (953 mg, 0.90 mmol, 7%) was added dropwise to the mixture at 0 °C. The mixture was stirred at room temperature for 1 h. The mixture was adjusted to pH 5. The mixture was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min). Compound 52e (115 mg, 74.7% yield) was obtained as a white solid. MS(ESI) m / z: 862.6 [M+Na] + .

[0477] Step 4 Benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((3aR,4R,6S,6aS)-6-(2-((((2S,3R,4R,5S,6S)-6-carbamoyl-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl)amino)-2-oxoethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)-L-glutamate (52f) 52f was synthesized according to the synthetic procedure in Step 3 of Example 49e (52 mg, 45.2% yield). MS(ESI) m / z: 861.7 [M+H] + .

[0478] Step 5 N2-(((9H-Fluoren-9-yl)methoxy)carbonyl)-N5-(((3aR,4R,6S,6aS)-6-(2-((((2S,3R,4R,5S,6S)-6-carbamoyl-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl)amino)-2-oxoethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)-L-glutamine (52g) 52g was synthesized according to the synthetic procedure in Step 7 of Example 49j (45 mg, crude). MS(ESI) m / z: 771.6 [M+H] + .

[0479] Step 6 N2-(((9H-Fluoren-9-yl)methoxy)carbonyl)-N5-(((3aR,4R,6S,6aS)-6-(2-((((2S,3R,4R,5S,6S)-6-carbamoyl-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl)amino)-2-oxoethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methyl)-L-glutamine (52h) 52h was synthesized according to the synthetic procedure in Step 6 of Example 49i (32 mg, 71.7% yield). MS(ESI) m / z: 731.5 [M+H] + .

[0480] Step 7 N2-(((9H-Fluoren-9-yl)methoxy)carbonyl)-N5-(((2R,3S,4R,5S)-5-(2-((((2S,3R,4R,5S,6S)-6-carbamoyl-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl)amino)-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-L-glutamine (52i) 52i was synthesized according to the synthetic procedure in Step 8 of Example 49l (24 mg, 35.3% yield). MS(ESI) m / z: 1575.9 [M+Na] + .

[0481] Step 8 ((6S,15S)-15-benzyl-1-((2R,3S,4R,5S)-5-(2-((((2S,3R,4R,5S,6S)-6-carbamoyl-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl)amino)-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)-24-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl (9H-fluoren-9-yl)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)amino)-3,7,10,13,16,19,24-heptaoxo-22-oxa-2,8,11,14,17,20-hexaazatetracosan-6-yl)carbamate (52j) 52j was synthesized according to the synthetic procedure in Step 9 of Example 49m (20.6 mg, theoretical). MS(ESI) m / z: 1354.8 [M+Na] + .

[0482] Step 9 (S)-2-Amino-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxo (2R,3S,4R,5S)-5-(2-((((2S,3R,4R,5S,6S)-6-carbamoyl-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl)amino)-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)pentanediamide (52) 52 was synthesized according to the synthetic procedure in Step 8 of Example 34k (7.8 mg, 32.7% yield). MS(ESI) m / z: 1561.9 [M+Na] +

[0483] Example 53 [ka]

[0484] Step 1 (2S,3S,4R,5R,6S)-6-(((S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(benzyloxy)-5-oxopentanamido)methyl)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (53a) To a mixture of 50c (200 mg, 0.32 mmol) in THF (1 mL) and water (1 mL) was added sodium bicarbonate (158.84 mg, 1.89 mmol). To the mixture was added TEMPO (9.85 mg, 0.06 mmol) and potassium bromide (11.25 mg, 0.09 mmol). The mixture was cooled to 0 °C in an ice bath, and sodium hypochlorite solution (aqueous, 3%-6% chlorine) (1.03 g, 0.69 mmol) was added dropwise. After the addition, the reaction mixture was concentrated in vacuo (without heating) to remove organic volatiles. The aqueous layer was acidified with 1N HCl to pH 2, filtered, and the filtrate was purified by preparative HPLC (FA) column: XBridge Prep C18 OBD 5um 19*250mm, mobile phase: A-water (0.1% formic acid): B-acetonitrile, flow rate: 20mL / min to give 53a (137.7mg, yield 67%). MS(ESI) m / z: 649.8 [M+H] +

[0485] Step 2 Benzyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((2S,3R,4R,5S,6S)-6-carbamoyl-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl)-L-glutamate (53b) To a mixture of 53a (137.7 mg, 0.21 mmol), PyBOP (113.78 mg, 0.22 mmol), and HOBt (29.55 mg, 0.22 mmol) in DMF (1 mL) was added DIPEA (27.44 mg, 0.21 mmol) and NH₃·MeOH (7 M, 0.42 mmol, 60 μL). The resulting yellow mixture was allowed to react at room temperature for 1 h. LCMS showed complete consumption of 53b. The mixture was purified by preparative HPLC (FA) column: XBridge Prep C18 OBD 5 μm 19*250 mm, mobile phase: A-water (0.1% formic acid): B-acetonitrile, flow rate: 20 mL / min, to give 53b (61.7 mg, 45% yield). MS(ESI) m / z: 648.5 [M+H] +

[0486] Step 3 N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N5-(((2S,3R,4R,5S,6S)-6-carbamoyl-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl)-L-glutamine (53c) Compound 53c was synthesized according to the synthetic procedure in Step 2 of Example 19 (30 mg, crude). MS(ESI) m / z: 558.5 [M+H] +

[0487] Step 4 ((6S,15S)-15-benzyl-1-((2S,3R,4R,5S,6S)-6-carbamoyl-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)-24-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-3,7,10,13,16,19,24-heptaoxo-22-oxa-2,8,11,14,17,20-hexaazatetracosan-6-yl)carbamate (9H-fluoren-9-yl)methyl (53d) Compound 53d was synthesized according to the synthetic procedure in Step 1 of Example 38, 66.7 mg, 50.9% yield. MS(ESI) m / z: 1402.8 [M+Na] +

[0488] Step 5 (S)-2-Amino-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-N5-(((2S,3R,4R,5S,6S)-6-carbamoyl-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl)pentanediamide (53e) Compound 53e was synthesized according to the synthetic procedure in Step 3 of Example 42 (55.5 mg, crude). MS(ESI) m / z: 1158.9 [M+H] +

[0489] Step 6 (S)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3 -oxa-5,8,11,14-tetraazahexadecan-16-yl)-N5-(((2S,3R,4R,5S,6S)-6-carbamoyl-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl)-2-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamide)pentanediamide (53) To a mixture of 53f (7.91 mg, 0.035 mmol) and HATU (13.30 mg, 0.035 mmol) in DMF (2 mL) was added DIPEA (3.01 mg, 0.023 mmol). The mixture was allowed to react at room temperature for 5 minutes. 53e (27.7 mg, crude) was added and allowed to react at room temperature for 10 minutes. LCMS showed complete consumption of 53e. The mixture was purified by preparative HPLC (FA) column: XBridge Prep C18 OBD 5um 19*250 mm, mobile phase: A-water (0.1% formic acid): B-acetonitrile, flow rate: 20 mL / min to give 53 (23.9 mg, 50.0% yield). MS(ESI) m / z: 1367.1 [M+H] +

[0490] Example 54 [ka] (S)-N1-((S)-10-benzyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-N5-(((2S,3R,4R,5S,6S)-6-carbamoyl-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl)-2-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamido)pentanediamide (54) Compound 54 was synthesized according to the synthetic procedure in Step 6 of Example 53 (21.5 mg, 45.5% yield). MS(ESI) m / z: 1352.3 [M+H] +

[0491] Example 55 [ka] (S)-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-7-benzyl-1-((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)carbamoyl)bicyclo[1.1.1]pentan-1-yl)oxy)-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecan-13-yl)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamido)pentanediamide (55) 55 was synthesized according to the synthetic procedure in Step 9 of Example 34 (12.5 mg, 59.8% yield). MS(ESI) m / z: 1388.3 [M+H] +

[0492] Example 56 [ka] Benzyl (11S,19R)-11-benzyl-19-(((tert-butyldiphenylsilyl)oxy)methyl)-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2,18-dioxa-4,7,10,13,16-pentaazaicosan-20-oate (22h) See Example 22 for the synthesis of 22h. MS(ESI) m / z: 1026.5 [M+Na] +

[0493] Step 1 (2R,3S,4S,5R,6R)-2-(acetoxymethyl)-6-(((11S,19R)-11-benzyl-19-((benzyloxy)carbonyl)-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2,18-dioxa-4,7,10,13,16-pentaazaicosan-20-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (56a) 22h (127 mg, 0.17 mmol), 56b (62 mg, 0.15 mmol), and 4 Å molecular sieves (300 mg) were dissolved in anhydrous DCE (20 mL). The mixture was stirred at 25 °C for 1 h, followed by the addition of AgOTf (46 mg, 0.18 mmol) and NIS (41 mg, 0.18 mmol). The resulting mixture was stirred at 0 °C for 2 h and then at 25 °C for 16 h. After the reaction was complete, the mixture was filtered through diatomaceous earth, and the filtrate was washed with saturated aqueous NaSO (20 mL) and brine (20 mL). The organic layer was dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give the crude product as a brown oil, which was purified by flash column chromatography (DCM / MeOH = 100 / 0 to 85 / 15) to give 56c (32 mg, 21.1% yield) as a white solid. MS(ESI) m / z: 788.6 [M+Na] +

[0494] Step 2 (2R,3S,4S,5R,6R)-2-(acetoxymethyl)-6-(((11S,19R)-11-benzyl-19-((benzyloxy)carbonyl)-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2,18-dioxa-4,7,10,13,16-pentaazaicosan-20-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (56c) 22h (127 mg, 0.17 mmol), 56b (62 mg, 0.15 mmol), and 4 Å molecular sieves (300 mg) were dissolved in anhydrous DCE (20 mL). The mixture was stirred at 25 °C for 1 h, followed by the addition of AgOTf (46 mg, 0.18 mmol) and NIS (41 mg, 0.18 mmol). The resulting mixture was stirred at 0 °C for 2 h and then at 25 °C for 16 h. After the reaction was complete, the mixture was filtered through diatomaceous earth, and the filtrate was washed with saturated aqueous NaSO (20 mL) and brine (20 mL). The organic layer was dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give the crude product as a brown oil, which was purified by flash column chromatography (DCM / MeOH = 100 / 0 to 85 / 15) to give 56c (32 mg, 21.1% yield) as a white solid. MS(ESI) m / z: 1118.7 [M+Na] +

[0495] Step 3 (11S,19R)-11-benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-19-((((2R,3R,4S,5S,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)oxy)methyl)-2,18-dioxa-4,7,10,13,16-pentaazaicosan-20-enoic acid (56d) 56d was synthesized according to the synthetic procedure in Step 7 of Example 22 (30 mg, crude). MS(ESI) m / z: 1028.8 [M+Na] +

[0496] Step 4 (2R,3S,4S,5R,6R)-2-(acetoxymethyl)-6-(((11S,19R)-11-benzyl-19-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)carbamoyl)-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2,18-dioxa-4,7,10,13,16-pentaazaicos-20-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (56f) 56f was synthesized according to the synthetic procedure in Step 8 of Example 22 (33 mg, 77.7% yield). MS(ESI) m / z: 1446.8 [M+Na] +

[0497] Step 5 (R)-2-(((S)-13-amino-7-benzyl-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)-3-(((2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propanamide (56g) 56f (30 mg, 0.021 mmol) was dissolved in anhydrous MeOH (7 mL), and then aqueous MeONa solution (1 M in MeOH, 0.011 mmol) was added. The resulting mixture was stirred at 25 °C for 3 h. After the reaction was completed, the reaction mixture was neutralized with a cation exchange resin and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 um 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 56g (9 mg, 41.3% yield) as a white solid. MS(ESI) m / z: 1033.7 [M+H] +

[0498] Step 6 (R)-2-(((S)-7-benzyl-20-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3,6,9,12,15,19-hexaoxo-2,5,8,11,14,18-hexaazaicosyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)-3-(((2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propanamide (56) 56g (8 mg, 0.0077 mmol) was dissolved in DMF (0.5 mL), followed by the addition of 56h (13 mg, 0.039 mmol). The mixture was stirred at 25 °C for 15 min and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A - water (0.1% formic acid): B - acetonitrile, Flow rate: 20 mL / min) to give 56 (6.5 mg, Yield: 67.6%) as a white solid. MS(ESI) m / z: 1263.8 [M+Na] +

[0499] Example 57 [ka]

[0500] Step 1 ((6S,9S,12S)-1-((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)-18-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzyl) (9H-Fluoren-9-yl)methyl benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)amino)-9-isopropyl-3,7,10,13,18-pentaoxo-12-(3-ureidopropyl)-16-oxa-2,8,11,14-tetraazaoctadecan-6-yl)carbamate (57a) Compound 57a was synthesized according to the synthetic procedure in Step 1 of Example 38 (64.0 mg, 44.4% yield). MS(ESI) m / z: 1325.0 [M+Na] +

[0501] Step 2 (S)-2-Amino-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-1-(((S)-1-((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-2-oxoethoxy)methyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide (57b) Compound 57b was synthesized according to the synthetic procedure in Step 3 of Example 42 (60.0 mg, crude). MS(ESI) m / z: 1080.8 [M+H] +

[0502] Step 3 (S)-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-2-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamide)-N1-((S)-1-(((S)-1-(((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxybenzoyl)methyl ...benzoyl)methyl)-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-2-(3-(2-(2,5-dioxo-2,5-dihydro-1 hydroxy-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)amino)-2-oxoethoxy)methyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide (50) Compound 57 was synthesized according to the synthetic procedure in Step 8 of Example 2 (16.1 mg, 45.0% yield). MS(ESI) m / z: 1289.4 [M+H] +

[0503] Example 58 [ka]

[0504] Step 1 Acetic acid (5S,8S)-1-(9H-fluoren-9-yl)-5-isopropyl-8-methyl-3,6,9-trioxo-2-oxa-4,7,10-triazaundecan-11-yl ester (58b) 58b was synthesized according to the synthetic procedure in Step 5 of Example 22g (850 mg, 82.5% yield).

[0505] Step 2 Benzyl (5S,8S)-1-(9H-fluoren-9-yl)-5-isopropyl-8-methyl-3,6,9-trioxo-2,12-dioxa-4,7,10-triazatetradecan-14-oate (58d) 58d was synthesized according to the synthetic procedure in Step 6 of Example 22h (450 mg, 92% yield). MS(ESI) m / z: 610.5 [M+Na] + .

[0506] Step 3 (5S,8S)-1-(9H-Fluoren-9-yl)-5-isopropyl-8-methyl-3,6,9-trioxo-2,12-dioxa-4,7,10-triazatetradecan-14-oic acid (58e) 58e was synthesized according to the synthetic procedure in Step 4 of Example 34f (215 mg, theoretical). MS(ESI) m / z: 521.5 [M+Na] + .

[0507] Step 4 (9H-fluoren-9-yl)methyl ((S)-1-(((S)-1-((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-2-oxoethoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate (58g) 58g was synthesized according to the synthetic procedure in step 8 of example 22k (72 mg, theoretical). MS(ESI) m / z: 937.7 [M+Na] + .

[0508] Step 5 (S)-2-Amino-N-((S)-1-((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-2-oxoethoxy)methyl)amino)-1-oxopropan-2-yl)-3-methylbutanamide (58h) 58h was synthesized according to the synthetic procedure in Step 8 of Example 34k (54.5 mg, theoretical). MS(ESI) m / z: 693.6 [M+H] + .

[0509] Step 6 (9H-Fluoren-9-yl)methyl ((6S,9S,12S)-1-((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)-18-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-9-isopropyl-12-methyl-3,7,10,13,18-pentaoxo-16-oxa-2,8,11,14-tetraazaoctadecan-6-yl)carbamate (58j) 58j was synthesized according to the synthetic procedure in Step 7 of Example 34j (42 mg, 43.9% yield). MS(ESI) m / z: 1239.0 [M+Na] + .

[0510] Step 7 (S)-2-amino-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-1-(((S)-1-((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-2-oxoethoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide (58k) 58k was synthesized according to the synthetic procedure in Step 8 of Example 34k (34.3 mg, theoretical). MS(ESI) m / z: 994.9 [M+H] + .

[0511] Step 8 (S)-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-2-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamide)-N1-((S)-1-(((S)-1-(((2-(((1S,9S)-9-ethyl-5-fluoro- 9-Hydroxy-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)amino)-2-oxoethoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide (58) 58 was synthesized according to the synthetic procedure in Step 4 of Example 39 (22.1 mg, 53.1% yield). MS(ESI) m / z: 1225.2 [M+Na] + .

[0512] Example 59 [ka]

[0513] Step 1 Methyl 3-(((5S,8S)-1-(9H-fluoren-9-yl)-5-isopropyl-3,6,9-trioxo-8-(3-ureidopropyl)-2-oxa-4,7,10-triazaundecan-11-yl)oxy)bicyclo[1.1.1]pentane-1-carboxylate (59a) To a mixture of 27b (300 mg, 0.53 mmol), 24a (188 mg, 1.32 mmol), and dry 4 Å molecular sieves (1.2 g) in anhydrous THF (6 mL) was added scandium triflate (392 mg, 0.79 mmol). The resulting mixture was stirred overnight at room temperature. The solution was filtered through diatomaceous earth and quenched with saturated aqueous NaHCO3 (15 mL). The aqueous phase was extracted with ethyl acetate (20 mL*3). The combined organic layers were dried over anhydrous sodium sulfate. After filtration and evaporation, the residue was purified by flash column chromatography (eluent: DCM / MeOH, 100 / 0 to 10 / 1 (v / v)) to give 59a (250 mg, 72.8% yield). MS(ESI) m / z: 672.6 [M+Na] +

[0514] Step 2 Methyl 3-(((S)-2-((S)-2-amino-3-methylbutanamido)-5ureidopentanamido)methoxy)bicyclo[1.1.1]pentane-1-carboxylate (59b) To a solution of 59a (120 mg, 0.18 mmol) in DMF (3 mL) was added diethylamine (203 mg, 2.77 mmol). The reaction was stirred at room temperature for 1 h. The reaction solution was extracted with petroleum ether (3 mL*3). The aqueous solution was concentrated to give the title compound 59b (77 mg, crude), which was used directly without further purification. MS(ESI) m / z: 450.2 [M+Na] +

[0515] Step 3 Methyl 3-(((5S,8S,11S)-5-(3-((((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-3-oxopropyl)-1-(9H-fluoren-9-yl)-8-isopropyl-3,6,9,12-tetraoxo-11-(3-ureidopropyl)-2-oxa-4,7,10,13-tetraazatetradecan-14-yl)oxy)bicyclo[1.1.1]pentane-1-carboxylate (59c) To a solution of 20 (107 mg, 0.20 mmol) and TSTU (60 mg, 0.20 mmol) in anhydrous DMF (3 mL) was added DIEA (90 μL, 0.54 mmol). The resulting mixture was stirred at room temperature for 10 min. 59b (77 mg, 0.18 mmol) was added. The reaction was stirred at room temperature for 1 h. The solution was filtered, purified by preparative HPLC (water / MeCN with 0.1% FA), and lyophilized to give the title compound 59c (50 mg, 29.2% yield) as a white solid. MS(ESI) m / z: 973.8 [M+Na] +

[0516] Step 4 3-(((5S,8S,11S)-5-(3-((((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-3-oxopropyl)-1-(9H-fluoren-9-yl)-8-isopropyl-3,6,9,12-tetraoxo-11-(3-ureidopropyl)-2-oxa-4,7,10,13-tetraazatetradecan-14-yl)oxy)bicyclo[1.1.1]pentane-1-carboxylic acid (59d) To a solution of 59c (50 mg, 0.05 mmol) in THF (1 mL) and water (1 mL) was added lithium hydroxide (1 N aqueous solution) (0.75 mL, 0.75 mmol). The reaction was stirred at 0 °C for 5 h. The aqueous solution was concentrated to give the title compound 59d (46 mg, crude), which was used directly without further purification. MS(ESI) m / z: 959.7 [M+Na] +

[0517] Step 5 ((6S,9S,12S)-1-amino-17-((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)-6-((((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[a]phenyl]-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl) 9H-Fluoren-9-yl)methyl 2-[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)carbamoyl)bicyclo[1.1.1]pentan-1-yl)oxy)methyl)carbamoyl)-9-isopropyl-1,8,11,15-tetraoxo-2,7,10,16-tetraazaheptadecan-12-yl)carbamate (59e) To a solution of 59d (46 mg, 0.05 mmol), exatecan mesylate (29 mg, 0.05 mmol), and HATU (19.6 mg, 0.05 mmol) in anhydrous DMF (2 mL) was added DIEA (30 μL, 0.15 mmol). The reaction was stirred at room temperature for 1 min. The solution was filtered, purified by preparative HPLC (water / MeCN with 0.1% FA), and lyophilized to give the title compound 59e (40 mg, 60.2% yield) as a white solid. MS(ESI) m / z: 1377.9 [M+Na] +

[0518] Step 6 (S)-2-amino-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-1-((((S)-1-((((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)carbamoyl)bicyclo[1.1.1]pentan-1-yl)oxy)methyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide (59f) To a solution of 59e (40 mg, 0.03 mmol) in DMF (2 mL) was added diethylamine (202.8 mg, 2.77 mmol). The reaction was stirred at room temperature for 1 h. The reaction solution was extracted with petroleum ether (3 mL*3). The aqueous solution was concentrated to give the title compound 59f (31 mg, crude), which was used directly without further purification. MS(ESI) m / z: 1132.8 [M+H] +

[0519] Step 7 (S)-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-2-(3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)acetamido)propanamide)-N1-((S)-1-(((S)-1-(((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl 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)carbamoyl)bicyclo[1.1.1]pentan-1-yl)oxy)methyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide (59) To a solution of 59f (31 mg, 0.05 mmol), exatecan mesylate (6.8 mg, 0.03 mmol), and HATU (10.94 mg, 0.03 mmol) in anhydrous DMF (1 mL) was added DIEA (10 μL, 0.04 mmol). The reaction was stirred at room temperature for 15 min. The solution was filtered, purified by preparative HPLC (water / MeCN with 0.1% FA), and lyophilized to give the title compound 59 (12.6 mg, 34.3% yield) as a white solid. MS(ESI) m / z: 1362.8 [M+Na] +

[0520] Example 60 [ka]

[0521] Step 1 (S)-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoamide)-N1-((S)-1-(((S)-1-(((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy- 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)amino)-2-oxoethoxy)methyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide (67) Compound 60 was synthesized according to the synthetic procedure in Step 8 of Example 2 (18.0 mg, 33.9% yield). MS(ESI) m / z: 1274.3 [M+H] +

[0522] Example 61 [ka] Compound 61 was synthesized according to the synthetic procedure in Step 9 of Example 34 (56.4 mg, 79.1% yield). MS(ESI) m / z: 1209.9 [M+Na] + .

[0523] Example 62 [ka]

[0524] Example 62 Compound 62 was synthesized according to the synthetic procedure of Example 9 (18.8 mg, 45.1% yield). MS(ESI) m / z: 1918.0 [M+H] + .

[0525] Example 63 [ka]

[0526] Example 63 Compound 63 was synthesized according to the synthetic procedure of Example 9 (24.3 mg, 58.1% yield). MS(ESI) m / z: 2486.5 [M+H] + .

[0527] Example 64 [ka]

[0528] Example 64 Compound 64 was synthesized according to the synthetic procedure of Example 9 (9.8 mg, 35.2% yield). MS(ESI) m / z: 2771.1 [M+H] + .

[0529] Example 65 [ka] Compound 65 was synthesized according to the synthetic procedures of Examples 9 and 50 (22.8 mg, 41.8% yield). MS(ESI) m / z: 2502.9 [M+H] + .

[0530] Example 66 [ka]

[0531] Step 1 N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)-3-hydroxy-2,2-dimethylpropanamide (66) To a mixture of 66a (5 mg, 0.042 mmol) and HATU (16 mg, 0.042 mmol) in DMF (1 mL) was added DIPEA (21 μL, 16 mg, 0.13 mmol) and 66b (purchased from ShangHai HaoYuan MedChemExpress CO. LTD, 23 mg, 0.043 mmol). The resulting brown mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was purified by preparative HPLC (TFA) (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A - water (0.1% TFA): B - acetonitrile, Flow rate: 20 mL / min). The fractions were lyophilized to give 66 (15 mg, 65.5%) as a white powder. 1H NMR(400MHz, DMSO): δ 8.00(d, J = 8.4Hz, 1H), 7.79(d,J = 11.2Hz, 1H), 7.31(s, 1H), 6.52(s, 1H), 5.59-5.54(m, 1H), 5.42(s, 2H), 5.18(q, J = 19.2Hz, 2H), 4.87(t,J = 5.2Hz, 1H), 3.45(dd, J = 10.2, 4.8Hz, 1H), 3.41-3.28(m, 1H), 3.15(t, J = 5.6Hz, 2H), 2.40(s, 3H), 2.24-2.07(m, 2H), 1.92-1.80(m, 2H), 1.11(d, J = 7.6Hz, 6H), 0.87(t, J = 7.2Hz, 3H). MS(ESI) m / z: 536.4 [M+H] +

[0532] Example 67 [ka]

[0533] Step 1 Benzyl (S)-benzyl 11-benzyl-1-(9H-fluoren-9-yl)-20,20-dimethyl-3,6,9,12,15-pentaoxo-2,18-dioxa-4,7,10,13,16-pentaazahenicosan-21-oate (67c) To a solution of 67a (250 mg, 0.40 mmol) and 67b (83 mg, 0.40 mmol) in THF (5 mL) was added 4 Å molecular sieves. The mixture was stirred at room temperature for 10 min, then Sc(OTf) (195 mg, 0.40 mmol) was added and the mixture was allowed to react for another 16 h at room temperature. The suspended mixture was filtered through a pad of diatomaceous earth, and the filter cake was washed with THF (10 mL). The filtrate was then quenched with saturated NaHCO (10 mL) and extracted with EtOAc (30 mL). After separation, the combined organic layer was washed with brine (50 mL), dried over NaSO, filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was purified by silica gel column chromatography (A - DCM; B - MeOH, MeOH / DCM = 0 / 100 to 95 / 5) to give 67c (90 mg, 29.2% yield). MS(ESI) m / z: 800.5 [M+Na] +

[0534] Step 2 (S)-11-Benzyl-1-(9H-fluoren-9-yl)-20,20-dimethyl-3,6,9,12,15-pentaoxo-2,18-dioxa-4,7,10,13,16-pentaazahenicosan-21-oic acid (67d) To a solution of 67c (80 mg, 0.10 mmol) in MeOH (3 mL) was added wet Pd / C (20 mg). The black suspension was purged with a H2 balloon three times and then reacted under a H2 balloon at room temperature for 2 h. After completion of the reaction, the black suspension was filtered through a pad of diatomaceous earth, the filter cake was washed with MeOH, and the combined organic layers were concentrated in vacuo to give 67d (61 mg, 84.8% yield). MS(ESI) m / z: 710.4 [M+Na] +

[0535] Step 3 (9H-fluoren-9-yl)methyl ((S)-7-benzyl-17-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)amino)-16,16-dimethyl-2,5,8,11,17-pentaoxo-14-oxa-3,6,9,12-tetraazaheptadecanyl)carbamate (67f) To a mixture of 67d (60 mg, 0.087 mmol) and HATU (33 mg, 0.087 mmol) in DMF (2 mL) was added DIPEA (43 μL, 34 mg, 0.26 mmol). The mixture was allowed to react at room temperature for 10 minutes. 67e (46 mg, 0.087 mmol) was added and the mixture was allowed to react for another hour at the same temperature. After the reaction was complete, the mixture was filtered and the filtrate was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19*250 mm, Mobile phase: A-water (0.1% formic acid): B-acetonitrile, Flow rate: 20 mL / min) to give 67f (85 mg, Yield: 88.2%). MS(ESI) m / z: 1105.5 [M+H] +

[0536] Step 4 3-(((S)-13-amino-7-benzyl-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecyl)oxy)-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-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)-2,2-dimethylpropanamide (67g) To a soluti...

Claims

1. A compound of formula (I), 【Chemistry 1】 or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, wherein BA is a binding agent selected from a humanized, chimeric, or human antibody or an antigen-binding fragment of an antibody; L is a covalent linker; PA is a payload residue; and subscript x is 1-30.

2. A compound of formula (Ia), 【Chemistry 2】 or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, During the ceremony, RG １ is a reactive group residue, RG ２ is an arbitrary reactive group residue, and SP １ and SP ２ is independently in each occurrence an optional spacer group residue, HG is a hydrophilic residue, PAB is an optional self-immolative unit, and the subscript p is 0 or 1; A.A. ２ includes the formula (W), 【Transformation 3】 A.A. ３ is -valine-alanine-, -valine-citrulline- or 【Chemistry 4】 is a dipeptide residue of the formula ６ is -CH ３ or -(CH ２ ) ３ -NHC(=O)NH ２ or AA ３ is -glycine-glycine-phenylalanine-glycine- or 【Transformation 5】 2. The compound of claim 1, which is a tetrapeptide residue of

3. The compound is a compound of formula (Ib): 【Transformation 6】 or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, During the ceremony, RG １ is a reactive group residue, RG ２ is an arbitrary reactive group residue, and SP １ and SP ２ is independently in each occurrence an optional spacer group residue, HG is a hydrophilic residue, PAB is an optional self-immolative unit, and the subscript p is 0 or 1; A.A. ２ comprises the formula (W), and 【Transformation 7】 A.A. １ is -valine-alanine-, -valine-citrulline- or 【Transformation 8】 is a dipeptide residue of the formula ６ is -CH ３ Or -(CH ２ ) ３ -NHC(=O)NH ２ or A.A. １ is -glycine-glycine-phenylalanine-glycine- or 【Chemistry 9】 2. The compound of claim 1, which is a tetrapeptide residue of

4. The compound is a compound of formula (Ic): 【Chemistry 10】 or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope or prodrug thereof, where BA is a binder and RG １ is a reactive group residue, and SP １ is an optional spacer group residue, PAB is an optional self-immolative unit, the subscript p is 0 or 1, PA is a payload residue, and the subscript x is 1 to 30; A.A. ３ is -valine-alanine-, -valine-citrulline- or 【Chemistry 11】 is a dipeptide residue of the formula ６ is -CH ３ Or -(CH ２ )3-NHC(=O)NH ２ or In the formula, AA ３ is -glycine-glycine-phenylalanine-glycine- or 【Chemistry 12】 2. The compound of claim 1, which is a tetrapeptide residue of

5. The antibody may be a humanized, chimeric, or human antibody or antigen-binding fragment thereof, and may be any of CD7, CD19, CD22, CD27, CD30, CD33, CD37, CD70, CD74, CD79b, CD138, CD142, CA6, placental cadherin, CEA, CEACAM5, C4.4a, DLL3, EGFR, EGFR VIII, ENPP3, EphA2, Ephrin A, FLOR1, FGFR2, GCC, HER2, HER3, cKIT, , LIV1, LY6E, MSLN, MUC16, NaPi2b, Nectin4, gpNMB, PSMA, SLITRK6, STEAP1, TROP2, 5T4, SSEA4, GloboH, Gb5, STn, and Tn, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof.

6. 5. The compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, wherein the antibody is a humanized, chimeric, or human antibody or antigen-binding fragment thereof and binds to one or more receptors selected from the group consisting of B7H3, MUC1, FGFR2b, CLL1, CCR7, GPC1, and GPC3.

7. RG １ teeth, 【Chemistry 13】 【Chemistry 14】 3. The compound of claim 2, wherein:

8. RG １ teeth, 【Chemistry 15】 and EWG is -CN, halogen, -CF ３ , -C(=O)OR １ and -C(=O)R １ and R is an electron-withdrawing group selected from １ is substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted heteroaryl, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof.

9. RG １ teeth, 【Chemistry 16】 3. The compound of claim 2, wherein:

10. RG １ teeth, 【Chemistry 17】 and EWG is -CN, halogen, -CF ３ , -C(=O)OR １ and -C(=O)R １ and R is an electron-withdrawing group selected from １ is substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted heteroaryl, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof.

11. A compound of formula (II), [Chemistry 18] or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, wherein L is a covalent linker and PA is a payload residue.

12. The compound is a compound of formula (IIa): 【Chemistry 19】 or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, During the ceremony, RG １ is a reactive group residue, RG ２ is an arbitrary reactive group residue, and SP １ and SP ２ is independently in each occurrence an optional spacer group residue, HG is a hydrophilic residue, PAB is an optional self-immolative unit, and the subscript p is 0 or 1; A.A. ２ includes the formula (W), 【Chemistry 20】 A.A. ３ is -valine-alanine-, -valine-citrulline- or 【Chemistry 21】 is a dipeptide residue of the formula ６ is -CH ３ or -(CH ２ ) ３ -NHC(=O)NH ２ or A.A. ３ is -glycine-glycine-phenylalanine-glycine- or 【Chemistry 22】 12. The compound of claim 11, which is a tetrapeptide residue of:

13. The compound is a compound of formula (IIb): 【Chemistry 23】 or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, During the ceremony, RG １ is a reactive group residue, RG ２ is an arbitrary reactive group residue, and SP １ and SP ２ is independently in each occurrence an optional spacer group residue, HG is a hydrophilic residue, PAB is an optional self-immolative unit, and the subscript p is 0 or 1; A.A. ２ includes the formula (W), 【Chemistry 24】 A.A. １ is -valine-alanine-, -valine-citrulline- or 【Chemistry 25】 is a dipeptide residue of the formula ６ is -CH ３ Or -(CH ２ ) ３ -NHC(=O)NH ２ or A.A. １ is -glycine-glycine-phenylalanine-glycine- or 【Chemistry 26】 12. The compound of claim 11, which is a tetrapeptide residue of:

14. The compound is a compound of formula (IIc): 【Chemistry 27】 or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, In the formula, RG １ is a reactive group residue, and SP １ is an optional spacer group residue, PAB is an optional self-immolative unit, the subscript p is 0 or 1, PA is a payload residue, In the formula, AA ３ is -valine-alanine-, -valine-citrulline- or 【Chemistry 28】 is a dipeptide residue of the formula ６ is -CH ３ Or -(CH ２ ) ３ -NHC(=O)NH ２ or In the formula, AA ３ is -glycine-glycine-phenylalanine-glycine- or 【Chemistry 29】 12. The compound of claim 11, which is a tetrapeptide residue of:

15. RG １ teeth, 【Transformation 30】 【Chemistry 31】 15. The compound of any one of claims 12 to 14, wherein:

16. RG １ teeth, 【Chemistry 32】 and EWG is -CN, halogen, -CF ３ , -C(=O)OR １ and -C(=O)R １ and R is an electron withdrawing group selected from the group consisting of １ is substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted heteroaryl, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof.

17. RG １ teeth, 【Transformation 33】 15. The compound of any one of claims 12 to 14, wherein:

18. RG １ teeth, 【Transformation 34】 and EWG is -CN, halogen, -CF ３ , -C(=O)OR １ and -C(=O)R １ and R is an electron-withdrawing group selected from １ is substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted heteroaryl, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof.

19. SP １ is -(CH ２ ) ｎ１ -C(=O)-, -(CH ２ CH ２ O) ｎ２ -CH ２ CH ２ -C(=O)-, -CH[-(CH ２ ) ｎ３ -COOH] -C(=O)-, -CH ２ -C(=O)-NH-(CH ２ ) ｎ４ -C(=O)-, -CH ２ -C(=O)-NH-(CH ２ ) ｎ３ -C(=O)-NH-(CH ２ ) ｎ４ -C(=O)- or -C(=O)-(CH ２ ) ｎ５ The compound according to claim 2 or 12, wherein n1, n2, n3, n4, and n5 each independently represent an integer of 1 to 8, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof.

20. SP ２ is -(CH ２ ) ｎ６ - and n6 represents an integer of 1 to 8, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof.

21. RG ２ is a bond, -C(=O)-NH-, or -NHC(=O)-, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof.

22. HG is, 【Chemistry 35-1】 【Chemistry 35-2】 each n7 is independently 1 to 15, each n8 is independently 0 or 1, each n9 is independently 1 or 2, each n10 is independently an integer from 4 to 16, each n11 is independently an integer from 0 to 5, n12 is an integer from 0 to 3, d is 0 to 3, and R ２ is H or Me, and R ３ -OH, -NH ２ , -NHCH ２ -CH ２ -(PEG) ｘ -OH or -NHCH ２ -CH ２ -(PEG) ｘ -OMe, and R ４ is OH or NH ２ X, Y and Z are each independently —CH ２ 13. The compound of claim 2 or 12, wherein the group is -, -NH-, -S-, or -O-, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof.

23. HG is -NHSO ２ NH ２ , -SO ３ H, -SO ２ NH ２ , or -PO ３ H ２ and RG ２ 13. The compound of claim 2 or 12, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, wherein: is a bond.

24. PAB is -NH-CH ２ -O-, formula (Y1), or formula (Y2), 【Transformation 36】 【Chemistry 37】 13. The compound of claim 2 or 12, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, wherein: represents the bond connecting said PAB to the adjacent group in said formula.

25. each PA independently represents formula (D1); 【Transformation 38】 In the formula, R ４ , R ５ａ and R ５ｂ are each independently hydrogen, a sugar residue, a substituted or unsubstituted inorganic or organic acid residue, a substituted or unsubstituted C １－８ alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted non-aromatic heterocyclyl, substituted or unsubstituted cycloalkylalkyl, or substituted or unsubstituted heterocyclylalkyl; R ５ａ and R ５ｂ or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof.

26. each PA independently represents formula (D2); 【Chemistry 39】 12. The compound of claim 1 or 11, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, wherein ring B is substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or substituted or unsubstituted heteroaryl.

27. each PA independently represents formula (E1); 【Chemistry 40】 During the ceremony, R ７ and R ８ are each independently hydrogen, halogen, or alkyl, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof.

28. The compound of claim 1, 【Chemistry 41-1】 【Chemistry 41-2】 【Chemistry 41-3】 【Chemistry 41-4】 【Chemistry 41-5】 【Chemistry 41-6】 【Chemistry 41-7】 【Chemistry 41-8】 【Chemistry 41-9】 【Chemistry 41-10】 【Chemistry 41-11】 【Chemistry 41-12】 【Chemistry 41-13】 【Chemistry 41-14】 【Chemistry 41-15】 2. The compound of claim 1, wherein:

29. 10. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, and a pharmaceutically acceptable excipient.

30. A composition for treating a proliferative disease, metabolic disease, inflammation or neurodegenerative disease in a subject, comprising a compound of claim 1, or a pharmaceutically acceptable salt, tautomer, solvate, stereoisomer, enantiomer, isotope, or prodrug thereof, or a pharmaceutical composition of claim 29.

31. The compound of claim 30, 【Chemistry 42-1】 【Chemistry 42-2】 【Chemistry 42-3】 【Chemistry 42-4】 【Chemistry 42-5】 【Chemistry 42-6】 【Chemistry 42-7】 【Chemistry 42-8】 【Chemistry 42-9】 【Chemistry 42-10】 【Chemistry 42-11】 【Chemistry 42-12】 【Chemistry 42-13】 12. The compound of claim 11, wherein:

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