Somatostatin subtype-2 receptor (SST2R)-targeted therapeutic agents and uses thereof

Abstract

Described herein are somatostatin subtype-2 receptor (SST2R)-targeted therapeutics that target tumor cells that express SST2R and their use in the treatment of cancer.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 387,235, filed December 13, 2022, and U.S. Provisional Patent Application No. 63 / 597,871, filed November 10, 2023, each of which is incorporated by reference herein in its entirety.

[0002] Described herein are somatostatin subtype-2 receptor (SST2R) drug conjugates and methods of using such drug conjugates as cancer therapeutics, diagnostics, or both. [Background technology]

[0003] Neoplasms are abnormal cell proliferations that cause significant medical burdens in humans, including morbidity and mortality. Neoplasms include benign or noncancerous neoplasms that do not exhibit malignant characteristics and are generally unlikely to be dangerous (e.g., adenomas). Malignant neoplasms exhibit characteristics such as genetic mutations, loss of normal function, rapid division, and the ability to metastasize (invade) to other tissues, resulting in neoplasms with uncertain or unknown behavior. Malignant neoplasms (i.e., cancerous solid tumors) are the leading cause of death in industrialized countries. Noncancerous neoplasms, including benign adenomas, can also cause significant morbidity and mortality. While standard treatments can achieve significant effects in tumor growth inhibition and even tumor elimination, the applied drugs only show limited selectivity for malignant tissue over healthy tissue, and their severe side effects limit their effectiveness and use. Specific targeting of neoplastic cells without affecting healthy tissues is the main hope for effective solid tumor therapy. Non-peptide SST2R ligands conjugated to suitable drug cargoes or payloads represent a novel class of small molecule drug conjugates (SMDCs) for selective cancer therapeutics or diagnostics. Summary of the Invention

[0004] Described herein are SST2R modulators that target the delivery of payloads to tumors that express SST2R and their use in tumor treatment. The present disclosure provides alternative and improved methods for tumor treatment. In some embodiments, the SMDCs disclosed herein provide an improved method for targeting tumor cells that express SST2R over conventional therapies with narrow therapeutic indices.

[0005] In one embodiment, a compound having the structure of formula (I):

[0006] [ka] During the ceremony, A is —N(H)— or —O—; R a is hydrogen or C1-C6 alkyl, R 2 is hydrogen or C1-C6 alkyl, R 6 is chloro or -C(=O)NH2, L is -L 1 -L 2 - and L 1 is an optional spacer, and L 2 is an optional linker, L 1 or L 2 At least one of R d is a payload moiety comprising a chemotherapeutic agent, or a pharmaceutically acceptable salt thereof.

[0007] In some embodiments, L 2 exists, and -(L 2a ) w -L 2b -or-L 2c - and Each L 2aare independently selected from natural or unnatural amino acids, and any free amine of the amino acid is optionally and independently replaced with —CH; L 2b is absent or -N(R 10 )(unsubstituted or substituted benzyl)-OC(=O)-, wherein substituted benzyl is -C(=O)NHR 12 or is substituted with a monosaccharide, Each R 10 are independently selected from hydrogen and C1-C6 alkyl; Each R 12 are independently hydrogen, C4-C 20 and polyethylene glycol, and unsubstituted or substituted C1-C6 alkyl, wherein the substituted C1-C6 alkyl is selected from -NHR 13 , -C(=O)NHR 13 , or -NHC(=O)R 13 is replaced by Each R 13 are independently hydrogen, C4-C 20 Polyethylene glycol and C4-C 20 polyethylene glycol-NH2; or R 13 exists and L 2a If there is at least one free carboxylic group of the amino acid of R 13 and L 2a The free carboxylic groups of the amino acids join together to form a ring, w is 1, 2, 3, 4, 5, or 6; Each L 2c is N-maleimidomethyl-cyclohexane-1-carbonyl (MCC) or -S-.

[0008] In some embodiments, L 1 exists and -X 2 -L 3 -L 4 - and X 2 is -C(=O)(CH2) p -, -(CH2) p-, -C(=O)CH(CH2SO3H)NHC(=O)-, or -(X 2a ) p - and each X 2a are independently selected from natural or unnatural amino acids, and any free amine of the amino acid is optionally and independently replaced with —CH; p is 0, 1, 2, 3, 4, 5, or 6; L 3 is absent, unsubstituted or substituted C1-C 10 Alkylene, unsubstituted or substituted C-C 10 Heteroalkylene, C4-C 20 Polyethylene glycol, or -(X 3 CH2CH2) t - and each X 3 are independently O and NR 10 is selected from each t is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; L 4 is absent or -L 4a -(CH2) u -L 4b -(CH2) u -L 4c - and L 4a -O-, -NR 10 -, -NR 10 C(=O)-, -C(=O)NR 10 - or -C(=O)-, L 4b is absent or is an unsubstituted or substituted N-containing 5-10 membered heterocycloalkylene, and any free amine of the N-containing 5-10 membered heterocycloalkylene is optionally and independently substituted with —CHCOH; L 4c -O-, -NR 10 -, -NR 10 C(=O)-, -C(=O)NR 10 -, -C(=O)NR 10 (CH2) u O(CH2) uC(=O)-, CH(CH2SO3H)C(=O)NR 10 (CH2) u O(CH2) u C(=O)-, -C(=O)-, -CH(=N)-, -CH(=N-NH)-, -CCH3(=N)-, -CCH3(=N-NH)-, -C(=O)-(C1-C6 alkylene)-, -C(=O)NR 10 -(C1-C6 alkylene)-, -NR 10 C(=O)-(C1-C6 alkylene)-, -NR 10 -(C1-C6 alkylene)- or C1-C6 alkylene-; each u is independently 0, 1, 2, 3, 4, 5, or 6; Each R 10 are independently selected from hydrogen and C1-C6 alkyl.

[0009] In some embodiments, R d teeth,

[0010] [ka] is.

[0011] In another aspect, described herein are methods for treating cancer, comprising administering an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, to a mammal having cancer.

[0012] In another aspect, described herein are methods for treating tumors, comprising administering to a mammal having a tumor an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0013] In some embodiments, the mammal has anal cancer, bladder cancer, intestinal cancer, brain cancer, breast cancer, colon cancer, colorectal cancer, endometrial cancer, esophageal cancer, gallbladder cancer, gastric cancer, heart cancer, kidney cancer, lung cancer, liver cancer, melanoma, uterine cancer, lymphoma, ovarian cancer, pancreatic cancer, prostate cancer, thymic cancer, pheochromocytoma, medullary thyroid cancer, head and neck cancer, or melanoma. In some embodiments, the mammal has an endocrine cancer. In some embodiments, the endocrine cancer comprises an adrenal tumor, a neuroendocrine tumor, a parathyroid tumor, a pituitary tumor, or a thyroid tumor. In some embodiments, the mammal has a neuroendocrine tumor. In some embodiments, the mammal has a somatostatin receptor-positive gastrointestinal pancreatic neuroendocrine tumor (GEP-NET).

[0014] In another aspect, described herein is a method for targeting the delivery of a chemotherapeutic agent to a tumor in a mammal, comprising administering to the mammal having the tumor a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0015] In another aspect, described herein is a method of killing tumors in a mammal that overexpresses the somatostatin subtype-2 receptor (SST2R), comprising administering to the mammal a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0016] In any of the embodiments disclosed herein, the mammal is a human.

[0017] Other objects, features, and advantages of the compounds, methods, and compositions described herein will become apparent from the following detailed description. It should be understood, however, that the detailed description and specific embodiments, while indicating specific embodiments, are given by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description. [Brief explanation of the drawings]

[0018] [Figure 1]Figure 1 shows time-activity curves for selected organ activity of In-Compound 1 in female Swiss nude mice bearing AR4J2 pancreatic tumors. Uptake of In-Compound 1 in tumor and normal tissues in tumor-bearing animals is shown (mean ± SD). [Figure 2] Figure 1 shows the uptake of In-Compound 1 alone and with excess In-Compound 1 at 2 hours post-dose in an AR42J-derived tumor-bearing xenograft mouse model. Organ activity in %ID / g of tissue at 2 hours post-dose and selective blockade of In-Compound 1 uptake in the presence of excess In-Compound 1 are shown. This study demonstrates that specific SST2R-mediated uptake in tumors is blocked by excess In-Compound 1 (mean ± SD). DETAILED DESCRIPTION OF THE INVENTION

[0019] Cancer is a disease in which some cells undergo genetic changes in the regulation of their growth and replication, leading to uncontrolled growth and spread. It is one of the leading causes of death worldwide. Common types of cancer include solid tumors (cancers typically originating in organs), carcinomas (cancers originating in the skin or tissues lining organs), sarcomas (cancers of connective tissue such as bone), leukemias (cancers of the bone marrow), and lymphomas and myelomas (cancers of the immune system). Neoplasms are abnormal proliferations of cells that result in solid tumors that can be benign (i.e., not exhibiting malignant characteristics and generally less likely to become dangerous, such as adenomas), malignant (i.e., exhibiting characteristics such as genetic mutations, loss of normal function, and rapid division, and capable of metastasizing (invading) other tissues), and of uncertain or unknown behavior. Cutting-edge treatment of neoplasms is achieved through a combination of surgery, chemotherapy, and radiation therapy. While surgery can be curative under some conditions, it often requires multiple interventions and the combination of radiation and chemotherapy. Chemotherapy has often proven to be a powerful weapon in the fight against cancer, but further optimization is needed. Chemotherapy is typically performed by the systemic administration of potent cytotoxic drugs, but these compounds lack tumor selectivity and therefore also kill healthy cells in the body. This nonspecific toxicity is the cause of the severe side effects commonly associated with chemotherapy. Radiation therapy is the use of high-energy radiation to kill cells. The radiation source can be external beam radiation (applied using an external source), internal radiation (placement of a radioactive substance near the target cells), or radiation therapy from the systemic administration of radioactive substances. Similar to chemotherapy, many radiation therapy options also lack the tumor cell-specific properties necessary to achieve the ultimate goal of targeted tumor therapy with drug molecules or radionuclides.

[0020] GPCRs are a large and diverse group of integral membrane receptors, and as a result, are expressed in all cell types in the body. GPCRs are generally poorly antigenic, making them difficult targets for antibody-based strategies. For many GPCRs, the majority of the protein population resides in intracellular compartments at any given time, reducing the total number of cell surface binding sites accessible to antibodies or peptides.

[0021] Many human tumors overexpress different GPCRs, often at significantly higher densities than other tissues. For example, gastrointestinal pancreatic (GEP) neuroendocrine tumors (NETs) overexpress somatostatin receptors, namely SSTR2, SSTR3, and SSTR5. Breast cancer also overexpresses SSTR2. Due to the complex GPCR overexpression profiles in neoplasms, simultaneously targeting multiple receptors could address issues such as heterogeneity, resistance, and phenotypic changes during disease progression that hinder many current treatment options.

[0022] The somatostatin receptor (SSTR) class consists of five members (SSTR1, SSTR2, SSTR3, SSTR4, and SSTR5) that are widely expressed in different tissues throughout the body, including nerves, pituitary glands, kidneys, lungs, and immune cells. Their natural ligand is the neuropeptide somatostatin (SST), which occurs in two active isoforms, SST-14 and SST-28. Both isoforms, in combination with their receptors, act as inhibitory hormones. An important physiological function of the SSTR / SST axis is, for example, the inhibition of growth hormone release. SSTRs, especially SSTR subtype 2, are found to be highly expressed in many neoplastic cells and tumor vasculature. Overexpression of SSTRs, especially SSTR2, has been found in various neuroendocrine tumors as well as other tumors, such as breast, ovarian, and lung cancers. Targeting SSTR2 for drug delivery has been achieved by using stabilized cyclic somatostatin analogs such as octreotate, octreotide, and lanreotide. For example, DOTA chelators such as octreotide (DOTA-TATE, DOTA-(Tyr)-2-methyl-2-propanol), and lanreotide (DOTA-(Tyr)-2-methyl-2-propanol) are used. 3 Covalent attachment of somatostatin to benzophenone-3 (also known as benzophenone-octreotate) enabled targeted delivery of the radionuclide to tumor cells expressing somatostatin receptors. 177 Lu DOTA-TATE therapy is a form of peptide receptor radionuclide therapy (PRRT) that targets somatostatin receptors and is a form of targeted drug delivery.

[0023] Most currently available drugs that target GPCRs act on receptors whose natural ligands are small molecules such as histamine, adrenaline, and neurotransmitters. Drugs that target GPCRs whose natural ligands are peptides or proteins are typically also peptides or proteins.

[0024] Peptides are inherently susceptible to proteolytic enzymes and peptidases present in most tissues and are rapidly degraded into multiple fragments that no longer have significant affinity for their intended receptors. While there are methods to stabilize peptides (e.g., incorporating peptidomimetic structures or using more stable D-amino acids in the peptide backbone), these modifications can result in loss of affinity and / or selectivity and can adversely affect physicochemical properties (e.g., poor solubility and tendency to aggregate). Additionally, peptides can trigger undesirable immunogenic responses that can complicate later stages of development by masking therapeutic efficacy and affecting safety evaluation.

[0025] When peptide ligands are linked to cytotoxic payloads, the resulting conjugates are often rapidly degraded in plasma, generating cytotoxic peptide fragments that can nonspecifically bind to both tumor and normal tissues. This rapid degradation of peptide-drug conjugates (PDCs) and antibody-drug conjugates (ADCs) reduces the amount of cytotoxic payload distributed to the target tumor, reducing therapeutic efficacy and potentially increasing toxicity. In addition, peptides are most likely excreted exclusively via the kidney, which may limit the application of PDCs. The significant renal uptake of some peptide-based therapeutics limits their routine use.

[0026] High-affinity small molecule ligands that bind to peptide GPCRs and protein GPCRs, including chemokine GPCRs, have been described. They are cell-permeable and can access receptor populations in the endoplasmic reticulum and endosomes. Due to their low molecular weight, non-peptide small molecules should have improved vascular permeability and tumor penetration compared to high-molecular-weight conjugates based on peptides and antibodies. In many cases, the binding affinity of small molecule non-peptide ligands exceeds that of FDA-approved antibodies by several orders of magnitude.

[0027] Provided herein are SST2R ligand drug conjugates, SMDCs. The conjugated drug cargo or payload moiety is attached to the small molecule SST2R ligand in a manner that does not affect the binding affinity of the ligand to SST2R. The conjugated drug cargo or payload moiety includes a chemotherapeutic agent and a radionuclide that are attached to the ligand using a spacer and / or linker moiety.

[0028] Solid tumors: benign and / or malignant neoplasms (cancer) In one embodiment, the compounds of formula (I) are used to treat benign and / or malignant neoplasms (solid tumors), the neoplasms comprising cells that overexpress cell surface GPCRs.

[0029] As used herein, the term "neoplasm" refers to an abnormal growth of cells that may grow in an uncontrolled manner and may have the capacity to metastasize (spread).

[0030] Neoplasms include solid tumors, adenomas, carcinomas, sarcomas, leukemias, and lymphomas at any stage of disease, with or without metastasis.

[0031] A solid tumor is an abnormal mass of tissue that usually does not contain cysts or liquid areas. Solid tumors can be benign (not cancer) or malignant (cancer). Different types of solid tumors are named by the type of cells that form them. Examples of solid tumors are sarcomas, carcinomas, and lymphomas. Leukemias (cancers of the blood) generally do not form solid tumors.

[0032] Solid tumors are typically cancers originating in organs such as the bladder, bowel, brain, breast, endometrium, heart, kidney, lung, liver, uterus, ovary, pancreas or other endocrine organs (thyroid), and prostate.

[0033] Adenomas are non-cancerous tumors. They begin in adenoid cells of epithelial tissue (thin layers of tissue that cover organs, glands, and other structures in the body). Adenomas can grow from many glandular organs, including the adrenal glands, pituitary gland, thyroid gland, prostate, and others. Over time, adenomas can become malignant, at which point they are called adenocarcinomas. While benign, they can compress other structures (mass effect), produce large amounts of hormones in an unregulated, feedback-independent manner (causing paraneoplastic syndromes), and cause serious health complications.

[0034] Adenomas typically occur in the colon (e.g., adenomatous polyps, which have a tendency to become malignant and lead to colon cancer), kidney (e.g., renal adenomas may be precursor lesions to renal cancer), adrenal glands (e.g., adrenal adenomas, such as pheochromocytomas, some of which secrete hormones such as cortisol, which causes Cushing's syndrome, aldosterone, which causes Conn's syndrome, or androgens, which cause hyperandrogenism), thyroid (e.g., thyroid adenomas), and pituitary gland (e.g., prolactinomas, etc.). They are found in the pituitary adenomas, parathyroid glands (e.g., adenomas of the parathyroid gland may inappropriately secrete large amounts of parathyroid hormone, thereby causing primary hyperparathyroidism), liver (e.g., hepatocellular adenomas), breast (e.g., fibroadenomas), appendix (e.g., cystic adenomas), bronchi (e.g., bronchial adenomas may cause carcinoid syndrome, a type of paraneoplastic syndrome), prostate (e.g., prostatic adenomas), sebaceous glands (e.g., parathyroid adenomas), and salivary glands.

[0035] Metastasis is the spread of malignant cells to new areas of the body, often via the lymphatic system or bloodstream. A metastatic tumor is a tumor that has spread from its primary site, or where it began, to various areas of the body. A metastatic tumor contains malignant cells that express cell surface GPCRs.

[0036] Tumors formed from disseminated cells are called secondary tumors. Tumors may spread to areas near the primary site, called regional metastasis, or to more distant parts of the body, called distant metastasis.

[0037] In some embodiments, the tumor being treated comprises tumor cells that express a GPCR, and the tumor is a primary or metastatic tumor. In some embodiments, the tumor being treated comprises tumor cells that express a GPCR, and the tumor is a primary or metastatic tumor of gastrointestinal origin, such as colorectal cancer, gastric cancer, small intestine cancer, or esophageal cancer. In some embodiments, the tumor being treated comprises tumor cells that express a GPCR, and the tumor is a primary or metastatic tumor of the pancreas. In some embodiments, the tumor being treated comprises tumor cells that express a GPCR, and the tumor is a primary or metastatic tumor of the lung, such as squamous cell carcinoma, adenosquamous carcinoma, or adenocarcinoma. In some embodiments, the tumor being treated comprises tumor cells that express a GPCR, and the tumor is a primary or metastatic neuroectodermal tumor, such as a pheochromocytoma or paraganglioma. In some embodiments, the tumor being treated comprises tumor cells that express a GPCR, and the tumor is a primary or metastatic bronchopulmonary tumor or a gastrointestinal neuroendocrine tumor. In some embodiments, the tumor being treated comprises tumor cells that express a GPCR, and the tumor is a primary or metastatic tumor of the rectum or colon.

[0038] In some embodiments, the compounds of formula (I) are used to treat sarcomas, such as leiomyosarcoma or rhabdomyosarcoma.

[0039] In some embodiments, the compounds of formula (I) are used to treat adenomas.

[0040] In another aspect, provided herein are methods for treating cancer in a mammal, comprising administering an SMDC disclosed herein to a mammal in need thereof. In some embodiments, the cancer comprises tumor cells that express one or more peptide hormone GPCRs. In some embodiments, the cancer comprises tumor cells that overexpress one or more GPCRs. In some embodiments, the cancer comprises a solid tumor. In some embodiments, the cancer comprises a sarcoma, carcinoma, or lymphoma. In some embodiments, the cancer comprises a neuroendocrine tumor. In some embodiments, the cancer comprises an insulinoma. In some embodiments, the cancer comprises a peptide hormone GPCR-positive (e.g., somatostatin receptor-positive) gastrointestinal pancreatic neuroendocrine tumor (GEP-NET).

[0041] In some embodiments, a compound of Formula (I) is administered to a patient with a tumor, hi some embodiments, the patient with a tumor has been diagnosed with a carcinoma, a sarcoma, a primary tumor, a metastatic tumor, a solid tumor, a non-solid tumor, a hematological tumor, a leukemia, or a lymphoma.

[0042] Carcinomas include, but are not limited to, esophageal carcinoma, hepatocellular carcinoma, basal cell carcinoma (a form of skin cancer), squamous cell carcinoma (various tissues), bladder cancer (including transitional cell carcinoma (malignant neoplasm of the bladder)), bronchial carcinoma, colon cancer, colorectal cancer, gastric cancer, lung cancer including small cell carcinoma and non-small cell carcinoma of the lung, adrenocortical carcinoma, thyroid cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, renal cell carcinoma, ductal carcinoma in situ or bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterine cancer, testicular cancer, osteogenic carcinoma, epithelial carcinoma, and nasopharyngeal carcinoma.

[0043] Sarcomas include, but are not limited to, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, chordoma, osteogenic sarcoma, osteosarcoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, lymphangioendothelial sarcoma, synovioma, mesothelioma, Ewing's sarcoma, leiomyosarcoma, rhabdomyosarcoma, and other soft tissue sarcomas.

[0044] Solid tumors include, but are not limited to, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, and retinoblastoma. Benign solid tumors include adenomas.

[0045] Leukemias include, but are not limited to, a) chronic myeloproliferative syndromes (neoplastic disorders of pluripotent hematopoietic stem cells), b) acute myeloid leukemia (neoplastic transformation of pluripotent hematopoietic stem cells or lineage-restricted hematopoietic cells, c) chronic lymphocytic leukemia (CLL, a clonal proliferation of immunologically immature and functionally incomplete small lymphocytes) including B-cell CLL, T-cell CLL, prolymphocytic leukemia, and hairy cell leukemia, and d) acute lymphoblastic leukemia (characterized by the accumulation of lymphoblasts). Lymphomas include, but are not limited to, B-cell lymphomas (e.g., Burkitt's lymphoma), Hodgkin's lymphoma, and the like.

[0046] Primary and metastatic tumors include, for example, lung cancer (including, but not limited to, lung adenocarcinoma, squamous cell carcinoma, large cell carcinoma, bronchioloalveolar carcinoma, non-small cell carcinoma, small cell carcinoma, and mesothelioma), breast cancer (including, but not limited to, ductal carcinoma, lobular carcinoma, inflammatory breast cancer, clear cell carcinoma, and mucinous carcinoma), colorectal cancer (including, but not limited to, colon carcinoma and rectal carcinoma), anal cancer, pancreatic cancer (including, but not limited to, pancreatic adenocarcinoma, pancreatic islet cell carcinoma, and neuroendocrine tumors), prostate cancer, ovarian cancer (including, but not limited to, serous tumors, endometrioid tumors, and mucinous cystic adenocarcinomas, ovarian epithelial carcinoma or surface epithelial-stromal tumors, including umbilical stromal tumors), liver and bile duct cancer (including, but not limited to, hepatocellular carcinoma, cholangiocarcinoma, and vascular carcinoma), cancers of the head and neck (including but not limited to squamous cell carcinoma), gastric cancer (including but not limited to gastric adenocarcinoma, gastrointestinal stromal tumor), thymic carcinoma, multiple myeloma, testicular cancer, germ cell tumors, neuroendocrine tumors, cervical cancer, carcinoids of the gastrointestinal tract, breast, and other organs, and signet ring cell carcinoma.

[0047] Small molecule SST2R ligands In one aspect, small molecule SST2R drug conjugates are described herein.

[0048] In some embodiments, the SMDC is a compound having the structure of Formula (A):

[0049] [ka] During the ceremony, A is —N(H)— or —O—; X 1 But -N(R a)-, -O-, -C(=O)-, -C(=O)N(R a )-, -S(=O)-, -(CH2)C(R a )=NO-(CH2)-, R a is hydrogen or C1-C6 alkyl, R b is hydrogen or C1-C6 alkyl, or R a and R b If both exist, R a and R b together with the intermediate atom to which they are attached form piperidine or pyrrolidine, q is 1, 2, or 3; R 1 is hydrogen, R 2 is hydrogen or C1-C6 alkyl, R 3 But hydrogen, -OR 8 , -N(R 8 )2, -CN, halogen, C1-C6 alkyl, or C1-C6 fluoroalkyl; or R 2 and R 3 together with the middle atom to which they are attached to form morpholine, Each R 4 and R 5 are independently hydrogen, halogen, C-C alkyl, C-C fluoroalkyl, substituted or unsubstituted C-C heteroalkyl, —CN, —N(R 8 )2, or -OR 8 and m is 1, 2, or 3; R c but,

[0050] [ka] and Each R 6 and R 7are independently hydrogen, halogen, C1-C4 alkyl, C1-C4 fluoroalkyl, C2-C4 alkenyl, C2-C4 alkynyl, substituted or unsubstituted C1-C6 heteroalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted C3-C6 cycloalkyl, -CN, -OR 8 , -CO2R 8 , -C(=O)N(R 8 )2, -N(R 8 )2, -NR 8 C(=O)R 9 , -NR 8 C(=O)OR 9 , -SR 8 , -S(=O)R 9 , -SO2R 9 , or -SO2N(R 8 )2, n is 1, 2, or 3; Each R 8 are independently hydrogen, C1-C4 alkyl, C1-C4 fluoroalkyl, substituted or unsubstituted C1-C4 heteroalkyl; Each R 9 are independently C1-C4 alkyl, C1-C4 fluoroalkyl, substituted or unsubstituted C1-C4 heteroalkyl; L is -L 1 -L 2 - and L 1 is an optional spacer, L 2 is an optional linker, L 1 Or L 2 At least one of R d is (i) a chemotherapeutic agent, or (ii) a payload moiety comprising a chelating moiety or a radionuclide conjugate thereof, or a pharmaceutically acceptable salt thereof.

[0051] In some embodiments, the SMDC is a compound having the structure of Formula (A):

[0052] [ka] During the ceremony, A is —N(H)— or —O—; X 1 But -N(R a )-, -O-, -C(=O)-, -C(=O)N(R a )-, -S(=O)-, -(CH2)C(R a )=NO-(CH2)-, R a is hydrogen or C1-C6 alkyl, R b is hydrogen or C1-C6 alkyl, or R a and R b If both exist, R a and R b together with the intermediate atom to which they are attached form piperidine or pyrrolidine, q is 1, 2, or 3; R 1 is hydrogen, R 2 is hydrogen or C1-C6 alkyl, R 3 But hydrogen, -OR 8 , -N(R 8 )2, -CN, halogen, C1-C6 alkyl, or C1-C6 fluoroalkyl; or R 2 and R 3 together with the middle atom to which they are attached to form morpholine, Each R 4 and R 5 are independently hydrogen, halogen, C-C alkyl, C-C fluoroalkyl, substituted or unsubstituted C-C heteroalkyl, —CN, —N(R 8 )2, or -OR 8 and m is 1, 2, or 3; R c but,

[0053] [ka] and Each R 6 and R 7 are independently hydrogen, halogen, C1-C4 alkyl, C1-C4 fluoroalkyl, C2-C4 alkenyl, C2-C4 alkynyl, substituted or unsubstituted C1-C6 heteroalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted C3-C6 cycloalkyl, -CN, -OR 8 , -CO2R 8 , -C(=O)N(R 8 )2, -N(R 8 )2, -NR 8 C(=O)R 9 , -NR 8 C(=O)OR 9 , -SR 8 , -S(=O)R 9 , -SO2R 9 , or -SO2N(R 8 )2, n is 1, 2, or 3; Each R 8 are independently hydrogen, C1-C4 alkyl, C1-C4 fluoroalkyl, substituted or unsubstituted C1-C4 heteroalkyl; Each R 9 are independently C1-C4 alkyl, C1-C4 fluoroalkyl, substituted or unsubstituted C1-C4 heteroalkyl; L is -L 1 -L 2 - and L 1 is an optional spacer, L 2 is an optional linker, L 1 Or L 2 At least one of R d is a payload moiety comprising a chelating moiety or a radionuclide conjugate thereof, or a pharmaceutically acceptable salt thereof.

[0054] In some embodiments, R 2is hydrogen and R 3 is hydrogen, -OH, or -OCH3, or R 2 and R 3 together with the middle atom to which they are attached to form morpholine.

[0055] In some embodiments, R 2 is hydrogen and R 3 is hydrogen.

[0056] In some embodiments, each R 4 and R 5 are independently hydrogen, F, Cl, Br, C1-C4 alkyl, C1-C4 fluoroalkyl, -CN, -N(R 8 )2, or -OR 8 is.

[0057] In some embodiments, each R 4 and R 5 are independently hydrogen, F, Cl, Br, —CH3, —CH2F, —CHF2, —CF3, —CN, —NH2, —NHCH3, —N(CH3)2, —OH, —OCH3, or —OCF3.

[0058] In some embodiments, A is —N(H)— and R a is hydrogen, -CH3, or -CH2CH3, and R b is hydrogen, -CH3, or -CH2CH3, or R a and R b If both exist, R a and R b together with the middle atom to which they are attached to form piperidine, and R 1 is hydrogen, -CH3, or -CH2CH3.

[0059] In some embodiments, the compound of Formula (A) has the structure of Formula (I):

[0060] [ka] During the ceremony, A is —N(H)— or —O—; R a is hydrogen or C1-C6 alkyl, R 2 is hydrogen or C1-C6 alkyl, R 6 is chloro or -C(=O)NH2, L is -L 1 -L 2 - and L 1 is an optional spacer, L 2 is an optional linker, L 1 or L 2 At least one of R d is the payload moiety containing the chemotherapeutic agent.

[0061] In some embodiments, A is —N(H)—.

[0062] In some embodiments, R 2 is hydrogen.

[0063] In some embodiments, R a is hydrogen or methyl. In some embodiments, R a is hydrogen.

[0064] In some embodiments, R 6 is chloro.

[0065] In some embodiments, the compound of Formula (A) has the structure of Formula (Ia): or a pharmaceutically acceptable salt thereof.

[0066] [ka]

[0067] In some embodiments, the compound of Formula (A) has the structure of Formula (Ib): or a pharmaceutically acceptable salt thereof:

[0068] [ka]

[0069] The compounds of the present disclosure also include tautomeric forms. Tautomeric forms result from the swapping of a single bond with an adjacent double bond and the accompanying migration of a proton. Tautomeric forms include prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and cyclic forms in which protons can occupy more than one position in a heterocyclic ring system, such as 1H- and 3H-imidazole, 1H-, 2H-, and 4H-1,2,4-triazole, 2H- and 2H-isoindole, and 2H- and 2H-pyrazole. Tautomeric forms can be in equilibrium or sterically fixed into one form by appropriate substitution.

[0070] Cytotoxic payload / drug-containing payload moiety (R d ) In some embodiments, R d includes chemotherapy drugs.

[0071] In some embodiments, R d teeth,

[0072] [ka]

[0073] [ka] is.

[0074] In some embodiments, R d teeth,

[0075] [ka] is.

[0076] Spacers and Linkers In one embodiment, the SST2R targeting ligand comprises a spacer (L 1 ) and / or linker (L 2 ) is covalently attached to the payload / drug.

[0077] In some embodiments, the spacer (L 1 The spacer has a predetermined length, thereby linking the ligand and the payload or the linker / payload while allowing an appropriate distance between them. The spacer can also modulate the pharmacological activity of the SMDC.

[0078] In some embodiments, the spacer comprises a dendritic spacer for covalently linking two or more drug moieties and / or ligands via a branched polyfunctional moiety, which can increase the molar ratio of ligand to payload, i.e., loading, which correlates with the potency of the conjugate.

[0079] In some embodiments, L 1 is absent or L 1 is a spacer present, and -X 2 -L 3 -L 4 - and X 2 is -C(=O)(CH2) p -, -(CH2) p -, -C(=O)CH(CH2SO3H)NHC(=O)-, or -(X 2a ) p - and each X 2a are independently selected from natural or unnatural amino acids, and any free amine of the amino acid is optionally and independently replaced with —CH; p is 0, 1, 2, 3, 4, 5, or 6; L 3 is absent, unsubstituted or substituted C1-C 10 Alkylene, unsubstituted or substituted C-C 10 Heteroalkylene, C4-C 20 Polyethylene glycol, or -(X 3 CH2CH2) t - and each X 3 are independently O and NR 10 is selected from each t is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; L 4 is absent or -L 4a -(CH2) u -L 4b -(CH2) u -L 4c - and L 4a -O-, -NR 10 -, -NR 10 C(=O)-, -C(=O)NR 10 - or -C(=O)-, L 4b is absent or is an unsubstituted or substituted N-containing 5-10 membered heterocycloalkylene, and any free amine of the N-containing 5-10 membered heterocycloalkylene is optionally and independently substituted with —CHCOH; L 4c -O-, -NR 10 -, -NR 10 C(=O)-, -C(=O)NR 10 -, -C(=O)NR 10 (CH2) u O(CH2) u C(=O)-, CH(CH2SO3H)C(=O)NR 10 (CH2) u O(CH2) u C(=O)-, -C(=O)-, -CH(=N)-, -CH(=N-NH)-, -CCH3(=N)-, -CCH3(=N-NH)-, -C(=O)-(C1-C6 alkylene)-, -C(=O)NR10 -(C1-C6 alkylene)-, -NR 10 C(=O)-(C1-C6 alkylene)-, -NR 10 -(C1-C6 alkylene)- or C1-C6 alkylene-; each u is independently 0, 1, 2, 3, 4, 5, or 6; Each R 10 are independently selected from hydrogen and C1-C6 alkyl.

[0080] In some embodiments, L 4b is absent or

[0081] [ka] is.

[0082] In some embodiments, X 2 is -C(=O)(CH2) p -, p is 0, 1, 2, 3, or 4, and L 3 is unsubstituted or substituted C1-C 10 Heteroalkylene, C4-C 20 Polyethylene glycol, or -(X 3 CH2CH2) t - and each X 3 are independently O and NR 10 and each t is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.

[0083] In some embodiments, L 4 is absent or -L 4a - and L 4a is -NR 10 -, -NR 10 C(=O)-, -C(=O)NR 10 -, or -C(=O)-.

[0084] In some embodiments, L 1 is absent or L 1 is the spacer present,

[0085] [ka]

[0086] [ka]

[0087] [ka] is.

[0088] In some embodiments, L 1 is the spacer present,

[0089] [ka] is.

[0090] In some embodiments, the linker (L 2 ) is the spacer (L 1 ) and payload (R d ) and contains a peptide bond. The peptide bond may comprise L-amino acids and / or D-amino acids. In some embodiments, D-amino acids are preferred to minimize immunogenicity and nonspecific cleavage by background peptidases or proteases. Cellular uptake of oligo-D-arginine sequences is known to be comparable to or better than that of oligo-L-arginine.

[0091] A linker unit may be "self-immolative" or "non-self-immolative." A "non-self-immolative" linker unit is one in which some or all of the linker unit remains attached to the drug moiety upon enzymatic (e.g., proteolytic) cleavage of the conjugate. A "self-immolative" linker unit allows for release of the drug moiety without a separate hydrolysis step.

[0092] In certain embodiments, the linker comprises a p-aminobenzyl unit. In one such embodiment, p-aminobenzyl alcohol is attached to the amino acid unit via an amide bond, and a carbamate, methylcarbamate, or carbonate is created between the benzyl alcohol and the cytotoxic agent. In one embodiment, the linker comprises p-aminobenzyloxycarbonyl (PAB). In certain embodiments, the phenylene moiety of the p-aminobenzyl unit is Q m wherein Q is -C1-C8 alkyl, -O-(C1-C8 alkyl), -halogen, -nitro, or -cyano; and m is an integer ranging from 0 to 4.

[0093] In some embodiments, the linker is cleavable. In some embodiments, the linker is designed to cleave under specific conditions or in the presence of a specific environment, such conditions, or the environment near the target cell, tissue, or region. Cleavable linkers rely on the unique properties of the cytoplasmic compartment of a cell for selective release of a cytotoxic drug. Such linkers primarily include chemically cleavable linkers that respond to low pH (acid-labile linkers) or a reducing environment (disulfide linkers), and enzymatically cleavable linkers that are susceptible to the action of specific lysosomal enzymes (peptide linkers or β-glucuronide linkers).

[0094] In some embodiments, the linker is cleavable under physiological conditions. In some embodiments, the linker is cleavable under intracellular conditions. In some embodiments, the linker is chemically cleavable. In some embodiments, the linker is enzymatically cleavable. In some embodiments, the linker is pH-sensitive, i.e., susceptible to hydrolysis at specific pH values. For example, a pH-sensitive linker may be hydrolyzable under acidic conditions. For example, the linker may be an acid-labile linker (e.g., hydrazone, semicarbazone, thiosemicarbazone, cis-aconitic amide, orthoester, acetal, ketal, etc.) that is hydrolyzable within the lysosome. Such linkers may be relatively stable under neutral pH conditions, such as those found in blood, but are unstable below pH 7.0, such as pH 6.5-4.5, the approximate pH of lysosomes and / or endosomes.

[0095] In some embodiments, the linker comprises one or more disulfide bonds.

[0096] In some embodiments, the linker is cleaved in or near tissues suffering from hypoxia, such as cancer cells and cancerous tissues. In some embodiments, the linker comprises a disulfide bond. In some embodiments, linkers comprising disulfide bonds are preferentially cleaved in hypoxic regions. Hypoxia is thought to enhance cancer cell resistance to radiation and chemotherapy and initiate angiogenesis. In a hypoxic environment, for example, in the presence of leaky or necrotic cells, free thiols and other reducing agents become available outside the cell, but O2, which normally keeps the extracellular environment oxidized, is by definition depleted. In some embodiments, this shift in redox balance promotes the reduction and cleavage of the disulfide bond within the linker.

[0097] In some embodiments, the linker is cleaved by intracellular peptidases or protease enzymes, including, but not limited to, lysosomal or endosomal proteases. In some embodiments, the linker is cleaved by glycosidases, such as glucuronidase. Small peptide sequences such as Val-Cit and Phe-Lys have been developed as linkers for ADCs. These bipeptide linkers exhibit good stability in serum, but after internalization, they can be recognized and rapidly hydrolyzed by specific lysosomal proteases, such as cathepsin B. β-glucuronide linkers can be easily cleaved by the abundant lysosomal enzyme β-glucuronidase, facilitating the easy and selective release of the active drug. In other embodiments, the linker is non-cleavable.

[0098] In some embodiments, the linker is cleaved by a protease, a matrix metalloprotease, a serine protease, or a combination thereof. In some embodiments, the linker is cleaved by a reducing agent. In some embodiments, the linker is cleaved by an oxidizing agent or oxidative stress.

[0099] In some embodiments, the linker is cleaved by a matrix metalloprotease (MMP), whose hydrolytic activity is involved in the invasive migration of metastatic tumor cells.

[0100] In some embodiments, the linker is cleaved by proteolytic enzymes or a reducing environment such as may be found near cancerous cells, where such an environment or such enzymes are not typically found near normal cells.

[0101] In some embodiments, the linker is cleaved by a serine protease, including, but not limited to, thrombin and cathepsin. In some embodiments, the linker is cleaved by cathepsin K, cathepsin S, cathepsin D, cathepsin E, cathepsin W, cathepsin F, cathepsin A, cathepsin C, cathepsin H, cathepsin Z, or any combination thereof. In some embodiments, the linker is cleaved by cathepsin K and / or cathepsin S.

[0102] In some embodiments, the linker is cleaved in a necrotic environment. Necrosis often results in the release of enzymes or other cellular contents that can be used to trigger cleavage of the linker. In some embodiments, cleavage of the linker occurs by necrotic enzymes (e.g., by calpain).

[0103] In some embodiments, L 2 is an optional non-cleavable or cleavable linker.

[0104] In some embodiments, L 2 is an optional cleavable linker that is an acid-sensitive linker, a protease-sensitive linker, or a glutathione-sensitive linker.

[0105] In some embodiments, L 2 is absent or L 2 is -(L 2a ) w -L 2b -or-L 2c - and Each L 2a are independently selected from natural or unnatural amino acids, and any free amine of the amino acid is optionally and independently replaced with —CH; L 2b is absent or -N(R 10 )(unsubstituted or substituted benzyl)-OC(=O)-, wherein substituted benzyl is -C(=O)NHR 12or is substituted with a monosaccharide, Each R 10 are independently selected from hydrogen and C1-C6 alkyl; Each R 12 are independently hydrogen, C4-C 20 and polyethylene glycol, and unsubstituted or substituted C1-C6 alkyl, wherein the substituted C1-C6 alkyl is selected from -NHR 13 , -C(=O)NHR 13 , or -NHC(=O)R 13 is replaced by Each R 13 are independently hydrogen, C4-C 20 Polyethylene glycol and C4-C 20 polyethylene glycol-NH2; or R 13 exists and L 2a If there is at least one free carboxylic group of the amino acid of R 13 and L 2a The free carboxylic groups of the amino acids join together to form a ring, w is 1, 2, 3, 4, 5, or 6; Each L 2c is N-maleimidomethyl-cyclohexane-1-carbonyl (MCC) or -S-.

[0106] In some embodiments, L 2b teeth,

[0107] [ka] is selected from.

[0108] In some embodiments, each L 2aare independently selected from natural or unnatural amino acids, and any free amine of the amino acid is optionally and independently replaced with —CH, and the natural or unnatural amino acid is selected from alanine (Ala), Ala(SOH), 3-(1-piperidinyl)alanine, cyclohexylalanine, arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), homophenylalanine, proline (Pro), serine (Ser), 3-homoserine, tyrosine (Tyr), Tyr(SOH), valine (Val), citrulline, β-alanine, β3-homoserine, β3-homolysine, and β3-homoglutamic acid.

[0109] In some embodiments, each L 2a are independently selected from natural or unnatural amino acids, wherein any free amine of the amino acid is optionally and independently replaced with —CH, and the natural or unnatural amino acid is selected from alanine (Ala), Ala (SO3H), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), serine (Ser), valine (Val), and citrulline.

[0110] In some embodiments, L 2 is -(L 2a ) w -L 2b - and -(L 2a ) w - valine-citrulline, valine-alanine, methionine-valine-lysine, glycine-phenylalanine-glycine-glycine, tyrosine-arginine-valine, arginine-valine, and phenylalanine-lysine.

[0111] In some embodiments, L2 -L 2c - and L 2c is N-maleimidomethyl-cyclohexane-1-carbonyl (MCC) or -S-.

[0112] In some embodiments, L 2 is absent or L 2 is the linker present,

[0113] [ka] , -S-,

[0114] [ka]

[0115] [ka]

[0116] [ka]

[0117] [ka]

[0118] [ka]

[0119] [ka]

[0120] [ka]

[0121] [ka]

[0122] [ka]

[0123] [ka] is.

[0124] In some embodiments, L 2 is absent or L 2 is the linker present,

[0125] [ka] is.

[0126] Representative spacer-linker moieties In some embodiments, L is

[0127] [ka]

[0128] [ka]

[0129] [ka]

[0130] [ka]

[0131] [ka]

[0132]

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[0133]

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[0153] [ka]

[0154] [ka]

[0155] [ka]

[0156] [ka]

[0157] [ka]

[0158] [ka]

[0159] [ka]

[0160] [ka] is.

[0161] Representative spacer / linker and payload moieties In some embodiments, -LR d teeth,

[0162] [ka]

[0163]

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[0164]

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[0165]

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[0166]

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[0167]

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[0194]

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[0195] [ka]

[0196] [ka]

[0197] [ka]

[0198] [ka]

[0199] [ka]

[0200] [ka]

[0201] [ka] is.

[0202] Any combination of the groups described above for the various variables is contemplated herein. Throughout the specification, groups and substituents thereof are chosen by one of ordinary skill in the art to provide stable moieties and compounds.

[0203] Representative SMDC conjugates The compound numbers listed below correspond to those repeated in the examples.

[0204] In some embodiments, the compound of Formula (I) is Compound 6.

[0205] In some embodiments, the compound of Formula (I) is Compound 7.

[0206] In some embodiments, the compound of Formula (I) is Compound 8.

[0207] In some embodiments, the compound of Formula (I) is Compound 9.

[0208] In some embodiments, the compound of Formula (I) is Compound 10.

[0209] In some embodiments, the compound of Formula (I) is Compound 11.

[0210] In some embodiments, the compound of Formula (I) is Compound 13.

[0211] In some embodiments, the compound of Formula (I) is compound 14.

[0212] In some embodiments, the compound of Formula (I) is compound 19.

[0213] In some embodiments, the compound of Formula (I) is Compound 20.

[0214] In some embodiments, the compound of Formula (I) is compound 21.

[0215] In some embodiments, the compound of Formula (I) is compound 22.

[0216] In some embodiments, the compound of Formula (I) is compound 23.

[0217] In some embodiments, the compound of Formula (I) is compound 24.

[0218] In some embodiments, the compound of Formula (I) is Compound 25.

[0219] In some embodiments, the compound of Formula (I) is compound 26.

[0220] In some embodiments, the compound of Formula (I) is compound 27.

[0221] In some embodiments, the compound of Formula (I) is compound 28.

[0222] In some embodiments, the compound of Formula (I) is compound 29.

[0223] In some embodiments, the compound of Formula (I) is compound 30.

[0224] In some embodiments, the compound of Formula (I) is compound 31.

[0225] In some embodiments, the compound of Formula (I) is compound 32.

[0226] In some embodiments, the compound of Formula (I) is compound 33.

[0227] In some embodiments, the compound of Formula (I) is compound 34.

[0228] In some embodiments, the compound of Formula (I) is compound 35.

[0229] In some embodiments, the compound of Formula (I) is compound 36.

[0230] In some embodiments, the compound of Formula (I) is compound 37.

[0231] In some embodiments, the compound of Formula (I) is compound 38.

[0232] In some embodiments, the compound of Formula (I) is compound 39.

[0233] In some embodiments, the compound of Formula (I) is compound 40.

[0234] In some embodiments, the compound of Formula (I) is compound 41.

[0235] In some embodiments, the compound of Formula (I) is compound 42.

[0236] In some embodiments, the compound of Formula (I) is compound 43.

[0237] In some embodiments, the compound of Formula (I) is compound 44.

[0238] In some embodiments, the compound of Formula (I) is compound 45.

[0239] In some embodiments, the compound of Formula (I) is compound 46.

[0240] In some embodiments, the compound of Formula (I) is compound 47.

[0241] In some embodiments, the compound of Formula (I) is compound 48.

[0242] In some embodiments, the compound of Formula (I) is compound 49.

[0243] In some embodiments, the compound of Formula (I) is compound 50.

[0244] In some embodiments, the compound of Formula (I) is compound 51.

[0245] In some embodiments, the compound of Formula (I) is compound 52.

[0246] In some embodiments, the compound of Formula (I) is compound 53.

[0247] In some embodiments, the compound of Formula (I) is compound 54.

[0248] In some embodiments, the compound of Formula (I) is compound 55.

[0249] In some embodiments, the compound of Formula (I) is compound 56.

[0250] In some embodiments, the compound of Formula (I) is compound 57.

[0251] In some embodiments, the compound of Formula (I) is compound 58.

[0252] In some embodiments, the compound of Formula (I) is compound 59.

[0253] In some embodiments, the compound of Formula (I) is compound 60.

[0254] In some embodiments, the compound of Formula (I) is compound 61.

[0255] In some embodiments, the compound of Formula (I) is compound 68.

[0256] In some embodiments, the compound of Formula (I) is compound 69.

[0257] In some embodiments, the compound of Formula (I) is compound 70.

[0258] In some embodiments, the compound of Formula (I) is compound 71.

[0259] In some embodiments, the compound of Formula (I) is compound 72.

[0260] In some embodiments, the compound of Formula (I) is compound 73.

[0261] In some embodiments, the compound of Formula (I) is compound 74.

[0262] In some embodiments, the compound of Formula (I) is Compound 75.

[0263] In some embodiments, the compound of Formula (I) is compound 77.

[0264] In some embodiments, the compound of Formula (I) is compound 78.

[0265] In some embodiments, the compound of Formula (I) is Compound 81.

[0266] In some embodiments, the compound of Formula (I) is Compound 82.

[0267] In some embodiments, the compound of Formula (I) is Compound 83.

[0268] In some embodiments, the compound of Formula (I) is compound 84.

[0269] In some embodiments, the compound of Formula (I) is Compound 85.

[0270] In some embodiments, the compound of Formula (I) is compound 86.

[0271] In some embodiments, the compound of Formula (I) is Compound 87.

[0272] In some embodiments, the compound of Formula (I) is compound 88.

[0273] In some embodiments, the compound of Formula (I) is compound 89.

[0274] In some embodiments, the compound of Formula (I) is Compound 90.

[0275] In some embodiments, the compound of Formula (I) is Compound 91.

[0276] In some embodiments, the compound of Formula (I) is Compound 92.

[0277] In some embodiments, the compound of Formula (I) is Compound 93.

[0278] In some embodiments, the compound of Formula (I) is Compound 94.

[0279] In some embodiments, the compound of Formula (I) is Compound 95.

[0280] In some embodiments, the compound of Formula (I) is Compound 96.

[0281] In some embodiments, the compound of Formula (I) is Compound 97.

[0282] In some embodiments, the compound of Formula (I) is Compound 98.

[0283] In some embodiments, the compound of Formula (I) is Compound 99.

[0284] In some embodiments, the compound of Formula (I) is Compound 100.

[0285] In some embodiments, the compound of Formula (I) is Compound 101.

[0286] In some embodiments, the compound of Formula (I) is Compound 102.

[0287] In some embodiments, the compound of Formula (I) is Compound 103.

[0288] In some embodiments, the compound of Formula (I) is Compound 104.

[0289] In some embodiments, the compound of Formula (I) is Compound 105.

[0290] In some embodiments, the compound of Formula (I) is Compound 106.

[0291] In some embodiments, the compound of Formula (I) is Compound 107.

[0292] In some embodiments, the compound of Formula (I) is Compound 108.

[0293] In some embodiments, the compound of Formula (I) is Compound 109.

[0294] In some embodiments, the compound of Formula (I) is Compound 110.

[0295] In some embodiments, the compound of Formula (I) is Compound 111.

[0296] In some embodiments, the compound of Formula (I) is Compound 112.

[0297] In some embodiments, the compound of Formula (I) is compound 113.

[0298] In some embodiments, the compound of Formula (I) is compound 114.

[0299] In some embodiments, the compound of Formula (I) is Compound 115.

[0300] In some embodiments, the compound of Formula (I) is Compound 116.

[0301] In some embodiments, the compound of Formula (I) is Compound 117.

[0302] In some embodiments, the compound of Formula (I) is Compound 118.

[0303] In some embodiments, the compound of Formula (I) is Compound 119.

[0304] In some embodiments, the compound of Formula (I) is Compound 120.

[0305] In some embodiments, the compound of Formula (I) is Compound 121.

[0306] In some embodiments, the compound of Formula (I) is Compound 122.

[0307] In some embodiments, the compound of Formula (I) is compound 123.

[0308] In some embodiments, the compound of Formula (I) is compound 124.

[0309] In some embodiments, the compound of Formula (I) is Compound 125.

[0310] In some embodiments, the compound of Formula (I) is Compound 126.

[0311] In some embodiments, the compound of Formula (I) is Compound 127.

[0312] In some embodiments, the compound of Formula (I) is Compound 128.

[0313] In some embodiments, the compound of Formula (I) is Compound 129.

[0314] In some embodiments, the compound of Formula (I) is Compound 130.

[0315] In some embodiments, the compound of Formula (I) is Compound 131.

[0316] In some embodiments, the compound of Formula (I) is Compound 132.

[0317] In some embodiments, the compound of Formula (I) is compound 133.

[0318] In some embodiments, the compound of Formula (I) is compound 134.

[0319] In some embodiments, the compound of Formula (I) is Compound 135.

[0320] In some embodiments, the compound of Formula (I) is compound 136.

[0321] In some embodiments, the compound of Formula (I) is Compound 137.

[0322] In some embodiments, the compound of Formula (I) is Compound 138.

[0323] In some embodiments, the compound of Formula (I) is Compound 139.

[0324] In some embodiments, the compound of Formula (I) is Compound 140.

[0325] In some embodiments, the compound of Formula (I) is compound 141.

[0326] In some embodiments, the compound of Formula (I) is compound 142.

[0327] In some embodiments, the compound of Formula (I) is compound 143.

[0328] In some embodiments, the compound of Formula (I) is compound 144.

[0329] In some embodiments, the compound of Formula (I) is Compound 145.

[0330] In some embodiments, the compound of Formula (I) is compound 146.

[0331] In some embodiments, the compound of Formula (I) is compound 147.

[0332] In some embodiments, the compound of Formula (I) is Compound 148.

[0333] In some embodiments, the compound of Formula (I) is Compound 149.

[0334] In some embodiments, the compound of Formula (I) is Compound 150.

[0335] In some embodiments, the compound of Formula (I) is Compound 151.

[0336] In some embodiments, the compound of Formula (I) is compound 152.

[0337] In some embodiments, the compound of Formula (I) is compound 153.

[0338] In some embodiments, the compound of Formula (I) is compound 154.

[0339] In some embodiments, the compound of Formula (I) is compound 155.

[0340] In some embodiments, the compound of Formula (I) is compound 156.

[0341] In some embodiments, the compound of Formula (I) is compound 157.

[0342] In some embodiments, the compound of Formula (I) is compound 158.

[0343] In some embodiments, the compound of Formula (I) is compound 159.

[0344] The payload moiety (R d ) Radiopharmaceuticals are becoming very useful tools for physicians to diagnose, stage, treat, and monitor the progression of several diseases, especially cancer. The main difference between radiopharmaceuticals and other pharmaceuticals is that radiopharmaceuticals contain a radionuclide. The nuclear decay properties of the radionuclide determine whether the radiopharmaceutical is used clinically as a diagnostic or therapeutic agent. Diagnostic radiopharmaceuticals require a radionuclide that emits either a gamma (γ) ray or a positron (β+), which then annihilates with a nearby electron to produce two 511 keV annihilation photons emitted approximately 180° apart from each other. Gamma-emitting radionuclides (e.g., 99m Tc, 111 In, 201 Tl) are useful for single photon emission computed tomography (SPECT), while positron-emitting radionuclides, e.g. 18 F, 89 Zr, 68 Ga) are useful for positron emission tomography (PET).

[0345] In contrast, therapeutic radiopharmaceuticals require radionuclides that emit particulate radiation, such as alpha (α) particles, beta (β) particles, or Auger electrons. These particles interact strongly with target tissue (e.g., cancerous tumors), resulting in widespread localized ionization. These particles can damage chemical bonds within DNA molecules, potentially inducing cytotoxicity.

[0346] In most nuclear medicine applications, a diagnostic radiopharmaceutical is desirably paired with a therapeutic radiopharmaceutical. This concept is commonly known as "theranostics." As a first step in the diagnostic concept, a targeted molecule labeled with a diagnostic radionuclide is used for quantitative imaging of tumor imaging biomarkers using positron emission tomography (PET) or single-photon emission computed tomography (SPECT). Once it has been demonstrated that this targeted molecule can be used to deliver a tumor-killing radiation absorbed dose to tumors and metastases, the second step involves the administration of the same or similar targeted molecule labeled with a therapeutic radionuclide.

[0347] In some embodiments, the chemical and pharmacokinetic behavior of both the diagnostic radiopharmaceutical and the therapeutic radiopharmaceutical are matched. In some embodiments, the diagnostic and therapeutic radionuclides are chemically identical radioisotope pairs (also known as "matched pairs"). One example of a matched pair for therapeutic radiopharmaceutical applications is: 123 I / 131 It is the counterpart of I, 123 I-labeled compounds are used for diagnostic purposes, while 131 I-labeled compounds are used in therapy. Other theranostic matched pairs include, among others: 44 Sc / 47 Sc, 64 Cu / 67 Cu, 72 As / 77 As, 86 Y / 90 Y, and 203 Pb / 212 Alternatively, pairs of radionuclides from different elements may be present because their chemistry is very similar (e.g., 99m Tc / 186 / 188 Re), and there is no significant difference in pharmacokinetic behavior between the diagnostic and therapeutic analogs, they can be used in the development of therapeutic radiopharmaceuticals. 68 Ga / 177 Lu pair, 68 Ga is used for diagnosis, 177Lu is used in therapy. For example, pancreatic endocrine tumors are 68 Ga sst2 ligand conjugate ([ 68 Ga]Ga-DOTA-TATE (NETSPOT trademark) or [ 68 expresses abundant sst receptors that can be targeted with somatostatin receptor scintigraphy for diagnostic purposes using [Ga]Ga-DOTA-TOC (DOTA-(D-Phe1,Tyr3)-octreotide, SomaKit TOC®)) and subsequently for internal radiotherapy. 177 Lu sst2 ligand conjugate ([ 177 Lu]Lu-DOTA-TATE).

[0348] Chelating agents for radionuclides As used herein, "chelating agent" and "chelating moiety" are used interchangeably.

[0349] In some embodiments, the chelator is capable of binding to the radioactive atom. In some embodiments, the binding is direct, e.g., the chelator undergoes hydrogen bonding or electrostatic interactions with the radioactive atom. In some embodiments, the binding is indirect, e.g., the chelator binds to a molecule that includes the radioactive atom. In some embodiments, the chelator is or includes a macrocycle.

[0350] In some embodiments, the chelator contains one or more amine groups. In some embodiments, the metal chelator contains two or more amine groups. In some embodiments, the chelator contains three or more amine groups. In some embodiments, the chelator contains four or more amine groups. In some embodiments, the chelator contains four or more N atoms, four or more carboxylic acid groups, or a combination thereof. In some embodiments, the chelator does not contain S. In some embodiments, the chelator contains a ring. In some embodiments, the ring contains O atoms and / or N atoms. In some embodiments, the chelator is a ring containing three or more N atoms, three or more carboxylic acid groups, or a combination thereof. In some embodiments, the chelator is a multidentate, bidentate, or monodentate ligand. Multidentate ligands range in the number of atoms used to bind to the metal atom or ion. EDTA, a hexadentate ligand, is an example of a multidentate ligand with six donor atoms that have electron pairs that can be used to bind to the central metal atom or ion. Bidentate ligands have two donor atoms, allowing them to bind to the central metal atom or ion at two points. Ethylenediamine (en) and oxalate (ox) are examples of bidentate ligands.

[0351] In some embodiments, the chelating agents described herein include cyclic chelating agents or acyclic chelating agents. In some embodiments, the chelating agents described herein include cyclic chelating agents. In some embodiments, the chelating agents described herein include acyclic chelating agents.

[0352] In some embodiments, the chelator is or includes DOTA, HBED-CC, DOTAGA, DOTA(GA)2, NOTA, and DOTAM, hi some embodiments, the chelator is or includes NODAGA, NOTA, DOTAGA, DOTA(GA)2, TRAP, NOPO, NCTA, DFO, DTPA, and HYNIC.

[0353] In some embodiments, the chelator comprises a macrocycle, e.g., a macrocycle containing O and / or N atoms, DOTA, HBED-CC, DOTAGA, DOTA(GA)2, NOTA, DOTAM, one or more amines, one or more ethers, one or more carboxylic acids, EDTA, DTPA, TETA, DO3A, PCTA, or deferrioxamine.

[0354] In some embodiments, R d teeth, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid (DO3A), 1,4,7,10-tetraazacyclododecane-1,7-diacetic acid (DO2A), α,α',α'',α'''-tetramethyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTMA), 1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetraazacyclododecane (DOTAM), 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrapropionic acid (DOTPA), 2,2',2''-(10-(2-amino-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-tetrapropionic acid) and a chelating moiety selected from the group consisting of 6,6'-(((pyridine-2,6-diylbis(methylene))bis(azanetriyl))-tetrakis(methylene))-tetrapicolinic acid (H4pypa), H4pypa-benzyl, 6,6',6'',6'''-(((pyridine-2,6-diylbis(methylene))bis(azanetriyl))-tetrakis(methylene))-tetrapicolinic acid (H4py4pa), H4py4pa-benzyl, H4octapa-benzyl, 3,6,9,12-tetrakis(carboxymethyl)-3,6,9,12-tetraazatetradecanedioic acid (TTHA), or radionuclide complexes thereof.

[0355] In some embodiments, R dis 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetyl acid (DOTA), or 1,4,7,10-tetraazacyclododecane-1,4,7-triacetyl acid (DO3A), or their radionuclide conjugates.

[0356] In some embodiments, R d teeth,

[0357] [ka] or a radionuclide complex thereof.

[0358] In some embodiments, R d teeth,

[0359] [ka] , or a radionuclide complex thereof.

[0360] radionuclides In some embodiments, the conjugate comprises an Auger electron-emitting radionuclide, an alpha-emitting radionuclide, a beta-emitting radionuclide, or a gamma-emitting radionuclide. In some embodiments, the conjugate comprises 111-indium ( 111 In), 67-gallium ( 67 Ga), 68 gallium ( 68 Ga), 99m-technetium ( 99m Tc), or 195m-Platinum ( 195m In some embodiments, the conjugate comprises an Auger electron emitting radionuclide that is 225-actinium (Pt). 225 Ac), 213-Bismuth ( 213 Bi), 223-radium ( 223 Ra), or 212-lead ( 212 In some embodiments, the conjugate comprises an alpha-emitting radionuclide that is 90-yttrium ( 90 Y), 177-lutetium ( 177Lu), iodine-131( 131 I), 186-rhenium ( 186 Re), 188-rhenium ( 188 Re), 64-Copper( 64 Cu), 67-copper ( 67 Cu), 153-Samarium ( 153 Sm), 89-strontium ( 89 Sr), 198-Kin ( 198 Au), 169-Erbium ( 169 Er), 165-dysprosium ( 165 Dy), 99m-Technetium ( 99m Tc), 89-zirconium ( 89 Zr), or 52-manganese ( 52 In some embodiments, the conjugate comprises a beta-emitting radionuclide that is 60-cobalt ( 60 Co), 103-paldium ( 103 Pd), 137-Cesium ( 137 Cs), 169-ytterbium ( 169 Yb), 192-iridium ( 192 Ir), or 226-radium ( 226 It includes gamma-emitting radionuclides (Ra).

[0361] In some embodiments, the conjugate comprises a radionuclide and a chelator configured to bind to the radionuclide, wherein the radionuclide is suitable for positron emission tomography (PET) analysis, single photon emission computed tomography (SPECT), or magnetic resonance imaging (MRI). In some embodiments, the radionuclide is copper-64 ( 64 Cu), Gallium-68( 68 Ga), 111-Indium ( 111 In), or technetium-99m ( 99m Tc).

[0362] Auger electrons (AE) are very low energy electrons emitted by radionuclides that decay by electron capture (EC) (e.g., 111 In, 67 Ga, 99m Tc,195m Pt, 125 I, and 123 I). This energy is deposited over nanometer-to-micrometer distances, resulting in a powerful high-linear energy transfer that causes lethal damage to cancer cells. Therefore, AE-emitting radiotherapeutic agents have great potential for the treatment of cancer.

[0363] Beta particles are electrons emitted from the nucleus. They typically have a longer range (about 1-5 mm) and are the most frequently used term.

[0364] Alpha particles are helium nuclei (two protons and two neutrons) emitted from the nuclei of radioactive atoms. Depending on the energy emitted, they can travel 50–100 μm in tissue. They are positively charged and several orders of magnitude greater than electrons. The amount of energy deposited per path length of an alpha particle (called "linear energy transfer") is approximately 400 times greater than that of an electron. This results in substantially more damage along their path than that caused by electrons. Alpha particle tracks result in the predominance of complex and largely irreparable DNA double-strand breaks. The absorbed dose required to achieve cytotoxicity is related to the number of alpha particles traversing the cell nucleus. Using this as a measure, cytotoxicity can be achieved within a range of 1–20 alpha particle traversals of the cell nucleus. The resulting high potency, combined with the short range of alpha particles (which reduces normal organ toxicity), has led to substantial interest in the development of alpha particle-emitting agents. Typically used alpha particle emitters include bismuth-212, lead-212, bismuth-213, actinium-225, radium-223, and thorium-227.

[0365] In some embodiments, the conjugate comprises a diagnostic or therapeutic radionuclide.

[0366] [Table 1]

[0367] Radionuclides are used in single photon emission computed tomography (SPECT, e.g. 67 Ga, 99m Tc, 111 In, 177 Lu) and positron emission tomography (PET, e.g., 68 Ga, 64 Cu, 44 Sc, 86 Y, 89 Zr), as well as therapeutic applications (e.g., 47 Sc, 114 mIn, 177 Lu, 90 Y, 212 / 213 Bi, 212 Pb, 225 Ac, 186 / 188 Radiometals have useful emission properties that allow them to be used in diagnostic imaging techniques such as radiopharmaceuticals (e.g., Re). The basic building block of radiometal-based radiopharmaceuticals is a chelator, a ligand system that binds to the radiometal ion in a tight, stable coordination complex so that it can be appropriately targeted to the desired molecular target in vivo. Guidance for selecting the optimal match between a chelator and a radiometal for a particular use is provided in the art (e.g., Price et al., "Matching chelators to radiometals for radiopharmaceuticals", Chem. Soc. Rev., 2014, 43, 260-290).

[0368] In some embodiments, R d comprises a chelated radionuclide that is suitable for positron emission tomography (PET) analysis or single photon emission computed tomography (SPECT). In some embodiments, R d comprises a chelated radionuclide that is suitable for single photon emission computed tomography (SPECT). In some embodiments, R d comprises a chelated radionuclide that is suitable for positron emission tomography (PET) analysis. In some embodiments, R dcomprises a chelated radionuclide that is suitable for positron emission tomography imaging, positron emission tomography with computed tomography imaging, or positron emission tomography with magnetic resonance imaging.

[0369] Response and toxicity prediction is essential for the rational implementation of cancer therapy. The biological effect of radionuclide therapy is mediated by a well-defined physical quantity: the absorbed dose (D), defined as the energy absorbed per unit mass of tissue.

[0370] Radiation dosimetry is the measurement, calculation, and assessment of the amount of ionizing radiation absorbed by an object, usually the human body, and can be thought of as the ability to perform the equivalent of a pharmacodynamic study in real time on treated patients. This applies both internally, via ingested or inhaled radioactive material, or externally, via irradiation by a radioactive source. Dosimetry analyses can be performed as part of patient treatment to calculate tumor versus normal organ absorbed doses and therefore the likelihood of treatment success.

[0371] In some embodiments, R d teeth,

[0372] [ka] , or a radionuclide complex thereof.

[0373] In some embodiments, -LR d is -CH2CH2NH-R d , -C(=O)CH2NH-R d , -C(=O)CH2CH2NH-R d , -CH2CH2(OCH2CH2)2NH-R d , -CH2CH2(OCH2CH2)3NH-R d , -CH2CH2(OCH2CH2)4NH-R d , -CH2CH2(OCH2CH2)5NH-R d , -CH2CH2(OCH2CH2)6NH-R d, -CH2CH2(OCH2CH2)7NH-R d , -CH2CH2(OCH2CH2)8NH-R d , -C(=O)CH2CH2(OCH2CH2)2NH-R d , -C(=O)CH2CH2(OCH2CH2)3NH-R d , -C(=O)CH2CH2(OCH2CH2)4NH-R d , -C(=O)CH2CH2(OCH2CH2)5NH-R d , -C(=O)CH2CH2(OCH2CH2)6NH-R d , -C(=O)CH2CH2(OCH2CH2)7NH-R d , or -C(=O)CH2CH2(OCH2CH2)8NH-R d and R d teeth,

[0374] [ka] , or a radionuclide complex thereof.

[0375] In some embodiments, -L 1 -R d is -(PEG2)NH-R d , -(PEG3)NH-R d , -(PEG4)NH-R d , -(PEG5)NH-R d , -(PEG6)NH-R d , -(PEG7)NH-R d , -(PEG8)NH-R d , -C(=O)(PEG2)NH-R d , -C(=O)(PEG3)NH-R d , -C(=O)(PEG4)NH-R d , -C(=O)(PEG5)NH-R d , -C(=O)(PEG6)NH-R d , -C(=O)(PEG7)NH-R d , or -C(=O)(PEG8)NH-R d and Rd teeth,

[0376] [ka] , or a radionuclide complex thereof.

[0377] In some embodiments, -L 1 -R d teeth,

[0378] [ka] or a radionuclide complex thereof.

[0379] In some embodiments, the radionuclide of the radionuclide conjugate is a lanthanide or an actinide.

[0380] In some embodiments, the radionuclide of the radionuclide conjugate is actinium, bismuth, cesium, cobalt, copper, dysprosium, erbium, gold, indium, iridium, gallium, lead, lutetium, manganese, palladium, platinum, radium, rhenium, samarium, strontium, technetium, ytterbium, yttrium, or zirconium.

[0381] In some embodiments, the radionuclide of the radionuclide conjugate is a diagnostic or therapeutic radionuclide.

[0382] In some embodiments, the radionuclide of the radionuclide conjugate is an Auger electron-emitting radionuclide, an alpha-emitting radionuclide, a beta-emitting radionuclide, or a gamma-emitting radionuclide.

[0383] In some embodiments, the radionuclide of the radionuclide conjugate is copper-64 ( 64 Cu), 67-copper ( 67 Cu), 111-Indium ( 111 In), 115-indium ( 115 In), 67-gallium (67 Ga), 68-gallium ( 68 Ga), 70-gallium ( 70 Ga), 225-actinium ( 225 Ac), 175-lutetium ( 175 Lu), 177-lutetium ( 177 Lu), or 212-lead ( 212 Pb).

[0384] In some embodiments, the radionuclide of the radionuclide conjugate is 111-indium ( 111 In), 115-indium ( 115 In), 67-gallium ( 67 Ga), 68-gallium ( 68 Ga), 70-gallium ( 70 Ga), 225-actinium ( 225 Ac), 175-lutetium ( 175 Lu), or 177-lutetium ( 177 Lu).

[0385] Compound synthesis The compounds described herein are synthesized using standard synthetic techniques, or using methods known in the art in combination with the methods described herein.

[0386] Unless otherwise indicated, conventional methods of mass spectrometry, NMR, and HPLC are used.

[0387] The compounds are described, for example, in March's Advanced Organic Chemistry, 6 th Edition, John Wiley and Sons, Inc. Alternative reaction conditions for the synthetic transformations described herein may be used, such as variations in solvents, reaction temperatures, reaction times, as well as different chemical reagents and other reaction conditions.

[0388] In one aspect, the compounds described herein are in the form of pharmaceutically acceptable salts.

[0389] The term "pharmaceutically acceptable salt" refers to a form of a therapeutically active agent consisting of the cationic form of the therapeutically active agent in combination with a suitable anion, or in an alternative embodiment, the anionic form of the therapeutically active agent in combination with a suitable cation. Handbook of Pharmaceutical Salts: Properties, Selection and Use. International Union of Pure and Applied Chemistry, Wiley-VCH 2002. S. M. Berge, L. D. Bighley, D. C. Monkhouse, J. Pharm. Sci. 1977, 66, 1-19.

[0390] In some embodiments, a pharmaceutically acceptable salt is obtained by reacting a compound of Formula (I) with an acid. In some embodiments, a compound of Formula (I) (i.e., the free base form) is basic and is reacted with an organic or inorganic acid.

[0391] In some embodiments, a pharmaceutically acceptable salt is obtained by reacting a compound of Formula (I) with a base. In some embodiments, the compound of Formula (I) is acidic and reacts with a base. In such a situation, the acidic proton of the compound of Formula (I) is replaced by a metal ion.

[0392] In some embodiments, compounds of Formula (I) have one or more stereocenters, and each stereocenter exists independently in either the R or S configuration. In some embodiments, compounds of Formula (I) exist in the R configuration. In some embodiments, compounds of Formula (I) exist in the S configuration. The compounds provided herein include all diastereomeric, individual enantiomeric, atropisomeric, and epimeric forms, and the appropriate mixtures thereof. The compounds and methods provided herein include all cis, trans, syn, anti, entogen (E), and tusamen (Z) isomers, and the appropriate mixtures thereof.

[0393] If desired, individual stereoisomers can be obtained by methods such as stereoselective synthesis and / or separation of stereoisomers by chiral chromatographic columns, or separation of diastereomers by either non-chiral or chiral chromatographic columns, or crystallization and recrystallization in an appropriate solvent or mixture of solvents. In certain embodiments, compounds of Formula (I) are prepared as their individual stereoisomers by reacting a racemic mixture of the compound with an optically active resolving agent to form diastereomeric compound / salt pairs, separating the diastereomers, and recovering the optically pure individual enantiomers. In some embodiments, resolution of the individual enantiomers is achieved using covalent diastereomeric derivatives of the compounds described herein. In other embodiments, diastereomers are separated by separation / resolution techniques based on differences in solubility. In other embodiments, separation of stereoisomers is achieved by chromatography, or by forming diastereomeric salts and separating them by recrystallization, chromatography, or any combination thereof. Jean Jacques, Andre Collet, Samuel H. Wilen, "Enantiomers, Racemates and Resolutions", John Wiley and Sons, Inc., 1981. In some embodiments, stereoisomers are obtained by stereoselective synthesis.

[0394] In some embodiments, the cytotoxic SMDCs of the invention described herein are prepared as depicted in Schemes A through F. In the following schemes, "X" and the carbonyl moieties on either side of "X" are linked to a spacer moiety L 1 Represents.

[0395] Scheme A:

[0396] [ka]

[0397] Scheme B:

[0398] [ka]

[0399] Scheme C:

[0400] [ka]

[0401] Scheme D:

[0402] [ka]

[0403] Scheme E

[0404] [ka]

[0405] Scheme F:

[0406] [ka]

[0407] In some embodiments, the radionuclide SMDCs of the invention described herein are prepared as depicted in Schemes GH.

[0408] Scheme G:

[0409] [ka]

[0410] Scheme H:

[0411] [ka]

[0412] Pharmaceutical Compositions In some embodiments, the compounds described herein are formulated into pharmaceutical compositions.

[0413] In some embodiments, the compounds described herein are administered in a pharmaceutical composition, either alone or in combination with a pharmaceutically acceptable carrier, excipient, or diluent.

[0414] Treatment methods In some embodiments, the method includes administering a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to the subject. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered in a pharmaceutical composition. In some embodiments, the subject has cancer. In some embodiments, the cancer is a solid tumor or a blood cancer. In some embodiments, the subject has a non-cancerous tumor. In some embodiments, the subject has an adenoma.

[0415] In embodiments, treatment is sufficient to reduce or inhibit tumor growth, reduce the number or size of metastatic lesions, reduce tumor burden, reduce primary tumor burden, reduce invasiveness, prolong survival, or maintain or improve quality of life in a subject, or a combination thereof.

[0416] In some embodiments, provided herein are methods for killing tumor cells, comprising contacting tumor cells with a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0417] In one aspect, methods and compositions for treating cancer are provided herein. Cancers include tissue and organ cancer development, including metastases, such as gastrointestinal cancer (e.g., gastric cancer, esophageal cancer, pancreatic cancer, colorectal cancer, intestinal cancer, anal cancer, liver cancer, gallbladder cancer, or colon cancer), lung cancer, thyroid cancer, skin cancer (e.g., melanoma), oral cancer, urinary tract cancer (e.g., bladder cancer or kidney cancer), blood cancer (e.g., myeloma or leukemia), or prostate cancer. In some embodiments, the present disclosure provides methods and compositions for treating gastrointestinal cancer in a subject in need thereof by administering to the subject an effective amount of a non-peptide targeted therapeutic compound disclosed herein. Non-limiting examples of gastrointestinal cancers that can be treated according to the methods of the present disclosure include gastric cancer, esophageal cancer, pancreatic cancer, lung cancer (small cell lung cancer and / or non-small cell lung cancer), colorectal cancer, intestinal cancer, anal cancer, liver cancer, gallbladder cancer, or colon cancer. In some embodiments, the cancer is Hodgkin's lymphoma or B-cell lymphoma.

[0418] In one aspect, provided herein are methods and compositions for treating adenoma.

[0419] In one aspect, methods and compositions for treating peptide hormone G protein-coupled receptor-expressing cancers are provided herein. In some embodiments, the peptide hormone G protein-coupled receptor-expressing cancer being treated is a primary or metastatic cancer of gastrointestinal origin, such as colorectal cancer, gastric cancer, small intestine cancer, or esophageal cancer. In some embodiments, the peptide hormone G protein-coupled receptor-expressing cancer being treated is a primary or metastatic pancreatic cancer. In some embodiments, the peptide hormone G protein-coupled receptor-expressing cancer being treated is a primary or metastatic lung cancer, such as squamous cell carcinoma, adenosquamous carcinoma, or adenocarcinoma. In some embodiments, the peptide hormone G protein-coupled receptor-expressing cancer being treated is a sarcoma, such as a leiomyosarcoma or a rhabdomyosarcoma. In some embodiments, the peptide hormone G protein-coupled receptor-expressing cancer being treated is a primary or metastatic neuroectodermal tumor, such as a pheochromocytoma or a paraganglioma. In some embodiments, the peptide hormone G protein-coupled receptor-expressing cancer being treated is a primary or metastatic bronchopulmonary tumor or a gastrointestinal neuroendocrine tumor. In some embodiments, the cancer is colorectal cancer.

[0420] In another aspect, described herein are methods for treating cancer in a mammal, comprising administering a non-peptide targeted therapeutic compound disclosed herein to a mammal in need thereof. In some embodiments, the cancer expresses one or more peptide hormone G protein-coupled receptors. In some embodiments, the cancer comprises a peptide hormone G protein-coupled receptor-positive cancer. In some embodiments, the cancer comprises a solid tumor. In some embodiments, the cancer comprises a sarcoma, carcinoma, or lymphoma. In some embodiments, the cancer comprises a neuroendocrine tumor. In some embodiments, the cancer comprises an insulinoma. In some embodiments, the cancer comprises a peptide hormone G protein-coupled receptor-positive (e.g., somatostatin receptor-positive) gastrointestinal pancreatic neuroendocrine tumor (GEP-NET).

[0421] Methods of Administration and Treatment Regimen In one embodiment, a compound of formula (I), or a pharmaceutically acceptable salt thereof, is used in the preparation of a medicament for the treatment of a tumor in a mammal. A method for treating any of the diseases or conditions described herein in a mammal in need of such treatment comprises administering to the mammal a therapeutically effective amount of a pharmaceutical composition comprising at least one compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0422] In some embodiments, the conjugate or a pharmaceutically acceptable salt thereof is administered in combination with another therapeutic agent. In certain embodiments, the therapeutic agent is a chemotherapeutic agent, including, but not limited to, lutetium Lu 177 dotatate (LUTATHERA®).

[0423] Specific Terms Unless otherwise stated, the following terms used in this application have the definitions set forth below. The term "including," as well as other forms of "include," "includes," and "included," are non-limiting. The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described.

[0424] As used herein, C1-C x is C1-C2, C1-C3...C1-C x By way of example only, a group designated "C1-C6" indicates that there are from 1 to 6 carbon atoms in the moiety, i.e., 1 carbon atom, 2 carbon atoms, 3 carbon atoms, or 4 carbon atoms in the group. Thus, by way of example only, "C1-C4 alkyl" indicates that there are from 1 to 4 carbon atoms in the alkyl group, i.e., the alkyl group is selected from among methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and t-butyl.

[0425] An "alkyl" group refers to an aliphatic hydrocarbon group. An alkyl group is branched or straight-chain. In some embodiments, an "alkyl" group contains 1 to 10 carbon atoms, i.e., C1-C 10 It has alkyl. Whenever it appears herein, a numerical range such as "1 to 10" refers to each integer within the given range; for example, "1 to 10 carbon atoms" means that the alkyl group consists of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms, although this definition also encompasses occurrences of the term "alkyl" without a specified numerical range. In some embodiments, alkyl is C1-C6 alkyl. In one aspect, alkyl is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or t-butyl. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tertiary butyl, pentyl, neopentyl, or hexyl.

[0426] An "alkylene" group refers to a divalent alkyl radical. Any of the above monovalent alkyl groups can be alkylene by abstracting a second hydrogen atom from the alkyl. In some embodiments, the alkylene is a C1-C6 alkylene. In other embodiments, the alkylene is a C1-C4 alkylene. Typical alkylene groups include, but are not limited to, -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2-, and the like. In some embodiments, the alkylene is -CH2-.

[0427] An "alkoxy" group refers to a (alkyl)O- group, where alkyl is as defined herein.

[0428] The term "alkenyl" refers to a type of alkyl group in which at least one carbon-carbon double bond is present. In one embodiment, an alkenyl group has the formula -C(R)=CR2, where R refers to the remainder of the alkenyl group, which can be the same or different. In some embodiments, R is H or alkyl. In some embodiments, alkenyl is selected from ethenyl (i.e., vinyl), propenyl (i.e., allyl), butenyl, pentenyl, pentadienyl, and the like. Non-limiting examples of alkenyl groups include -CH=CH2, -C(CH3)=CH2, -CH=CHCH3, -C(CH3)=CHCH3, and -CH2CH=CH2.

[0429] The term "alkynyl" refers to a type of alkyl group in which at least one carbon-carbon triple bond is present. In one embodiment, an alkenyl group has the formula -C≡CR, where R refers to the remainder of the alkynyl group. In some embodiments, R is H or alkyl. In some embodiments, alkynyl is selected from ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Non-limiting examples of alkynyl groups include -C≡CH, -C≡CCH3-C≡CCH2CH3, and -CH2C≡CH.

[0430] The term "heteroalkyl" refers to an alkyl group in which one or more skeletal atoms of the alkyl is selected from an atom other than carbon, e.g., oxygen, nitrogen (e.g., -NH-, -N(alkyl)-, sulfur, or combinations thereof. The heteroalkyl is attached to the remainder of the molecule at a carbon atom of the heteroalkyl. In one aspect, the heteroalkyl is a C1-C6 heteroalkyl.

[0431] The term "carbocyclic" or "carbocyclic" refers to a ring or ring system in which the atoms forming the backbone of the ring are all carbon atoms. Thus, this term distinguishes carbocycles from "heterocyclic" rings or "heterocycles" in which the ring backbone contains at least one atom other than carbon. In some embodiments, at least one of the two rings in a bicyclic carbocycle is aromatic. In some embodiments, both rings in a bicyclic carbocycle are aromatic. Carbocycles include aryl and cycloalkyl.

[0432] As used herein, the term "aryl" refers to an aromatic ring in which each of the atoms forming the ring is a carbon atom. In one aspect, an aryl is phenyl or naphthyl. In some embodiments, an aryl is phenyl. In some embodiments, an aryl is phenyl, naphthyl, indanyl, indenyl, or tetrahydronaphthyl. In some embodiments, an aryl is C6-C 10 Aryl. Depending on the structure, an aryl group can be a monoradical or a diradical (i.e., an arylene group).

[0433] The term "cycloalkyl" refers to a monocyclic or polycyclic aliphatic, non-aromatic radical in which each of the atoms forming the ring (i.e., skeletal atoms) is a carbon atom. In some embodiments, the cycloalkyl is a spirocyclic or bridged compound. In some embodiments, the cycloalkyl is optionally fused to an aromatic ring, and the point of attachment is to a carbon that is not an aromatic ring carbon atom. Cycloalkyl groups include groups having 3 to 10 ring atoms. In some embodiments, the cycloalkyl group is selected from among cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, spiro[2.2]pentyl, norbornyl, and bicyclo[1.1.1]pentyl. In some embodiments, the cycloalkyl is a C3-C6 cycloalkyl. In some embodiments, the cycloalkyl is a C3-C4 cycloalkyl.

[0434] The term "halo" or, alternatively, "halogen" or "halide" means fluoro, chloro, bromo, or iodo. In some embodiments, halo is fluoro, chloro, or bromo.

[0435] The term "fluoroalkyl" refers to an alkyl in which one or more hydrogen atoms have been replaced by a fluorine atom. In one embodiment, the fluoroalkyl is a C1-C6 fluoroalkyl.

[0436] The term "heterocycle" or "heterocyclic" refers to heteroaromatic (also known as heteroaryl) and heterocycloalkyl rings containing 1 to 4 heteroatoms in the ring, where each heteroatom in the ring is selected from O, S, and N, with the proviso that each heterocyclic group has 3 to 10 atoms in its ring system and no ring contains two adjacent O or S atoms. Non-aromatic heterocyclic groups (also known as heterocycloalkyl) include rings having 3 to 10 atoms in their ring system, and aromatic heterocyclic groups include rings having 5 to 10 atoms in their ring system. Heterocyclic groups include benzo-fused ring systems. Examples of non-aromatic heterocyclic groups are pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, oxazolidinonyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, thioxanyl, piperazinyl, aziridinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, pyrrolin-2-yl, pyrrolin-3-yl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiazole ... oranyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, 3H-indolyl, indolin-2-onyl, isoindolin-1-onyl, isoindolin-1,3-dionyl, 3,4-dihydroisoquinolin-1(2H)-onyl, 3,4-dihydroquinolin-2(1H)-onyl, isoindolin-1,3-dithionyl, benzo[d]oxazol-2(3H)-onyl, 1H-benzo[d]imidazol-2(3H)-onyl, benzo[d]thiazol-2(3H)-onyl, and quinolidinyl.Examples of aromatic heterocyclic groups are pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. The foregoing groups are either C-linked or N-linked where possible. For example, groups derived from pyrrole include both pyrrol-1-yl (N-linked) and pyrrol-3-yl (C-linked). Further, groups derived from imidazole include imidazol-1-yl or imidazol-3-yl (both N-linked) or imidazol-2-yl, imidazol-4-yl, or imidazol-5-yl (all C-linked). Heterocyclic groups include benzo-fused ring systems. Non-aromatic heterocycles are optionally substituted with one or two oxo (=O) moieties, such as pyrrolidin-2-one. In some embodiments, at least one of the two rings of a bicyclic heterocycle is aromatic. In some embodiments, both rings of a bicyclic heterocycle are aromatic.

[0437] The term "heteroaryl" or, alternatively, "heteroaromatic" refers to an aryl group containing one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur. Illustrative examples of heteroaryl groups include monocyclic heteroaryls and bicyclic heteroaryls. Monocyclic heteroaryls include pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, pyridazinyl, triazinyl, oxadiazolyl, thiadiazolyl, and furazanyl. Monocyclic heteroaryls include indolizine, indole, benzofuran, benzothiophene, indazole, benzimidazole, purine, quinolizine, quinoline, isoquinoline, cinnoline, phthalazine, quinazoline, quinoxaline, 1,8-naphthyridine, and pteridine. In some embodiments, heteroaryls contain 0 to 4 N atoms in the ring. In some embodiments, the heteroaryl contains 1-4 N atoms in the ring. In some embodiments, the heteroaryl contains 0-4 N atoms, 0-1 O atoms, and 0-1 S atoms in the ring. In some embodiments, the heteroaryl contains 1-4 N atoms, 0-1 O atoms, and 0-1 S atoms in the ring. In some embodiments, the heteroaryl is a C1-C9 heteroaryl. In some embodiments, the monocyclic heteroaryl is a C1-C5 heteroaryl. In some embodiments, the monocyclic heteroaryl is a 5- or 6-membered heteroaryl. In some embodiments, the bicyclic heteroaryl is a C6-C9 heteroaryl.

[0438] A "heterocycloalkyl" group refers to a cycloalkyl group containing at least one heteroatom selected from nitrogen, oxygen, and sulfur. In some embodiments, a heterocycloalkyl is fused with an aryl or heteroaryl. In some embodiments, a heterocycloalkyl is oxazolidinonyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, piperidin-2-onyl, pyrrolidine-2,5-dithionyl, pyrrolidine-2,5-dionyl, pyrrolidinonyl, imidazolidinyl, imidazolidin-2-onyl, or thiazolidin-2-onyl. In one aspect, a heterocycloalkyl is a C2-C 10 In another embodiment, heterocycloalkyl is C-C 10 Heterocycloalkyl. In some embodiments, the heterocycloalkyl is monocyclic or bicyclic. In some embodiments, the heterocycloalkyl is monocyclic and has a 3-, 4-, 5-, 6-, 7-, or 8-membered ring. In some embodiments, the heterocycloalkyl is monocyclic and has a 3-, 4-, 5-, or 6-membered ring. In some embodiments, the heterocycloalkyl is monocyclic and has a 3- or 4-membered ring. In some embodiments, the heterocycloalkyl contains 0-2 N atoms in the ring. In some embodiments, the heterocycloalkyl contains 0-2 N atoms, 0-2 O atoms, and 0-1 S atoms in the ring.

[0439] The term "bond" or "single bond" refers to a chemical bond between two atoms, or two moieties when the atoms connected by the bond are considered to be part of a larger substructure. In one aspect, when a group described herein is a bond, the reference group is absent, thereby allowing a bond to be formed between the remaining identified groups.

[0440] The term "moiety" refers to a specific segment or functional group of a molecule. A chemical moiety is often recognized as a chemical entity embedded in or appended to a molecule.

[0441] The terms "optionally substituted" or "substituted" mean that the referenced group is optionally substituted with one or more additional groups individually and independently selected from halogen, -CN, -NH, -NH(alkyl), -N(alkyl), -OH, -COH, -COalkyl, -C(=O)NH, -C(=O)NH(alkyl), -C(=O)N(alkyl), -S(=O)NH, -S(=O)NH(alkyl), -S(=O)N(alkyl), alkyl, cycloalkyl, fluoroalkyl, heteroalkyl, alkoxy, fluoroalkoxy, heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkylsulfoxide, arylsulfoxide, alkylsulfone, and arylsulfone. In some other embodiments, the optional substituents are independently selected from halogen, —CN, —NH, —NH(CH), —N(CH), —OH, —COH, —CO(C-C alkyl), —C(═O)NH, —C(═O)NH(C-C alkyl), —C(═O)N(C-C alkyl), —S(═O)NH, —S(═O)NH(C-C alkyl), —S(═O)N(C-C alkyl), C-C alkyl, C-C cycloalkyl, C-C fluoroalkyl, C-C heteroalkyl, C-C alkoxy, C-C fluoroalkoxy, —SC-C alkyl, —S(═O)C-C alkyl, and —S(═O)C-C alkyl. In some embodiments, optional substituents are independently selected from halogen, -CN, -NH, -OH, -NH(CH), -N(CH), -CH, -CHCH, -CHF, -CF, -OCH, -OCHF, and -OCF. In some embodiments, a substituent is substituted with one or two of the foregoing groups. In some embodiments, an optional substituent on an aliphatic carbon atom (acyclic or cyclic) comprises oxo (=O).

[0442] The term "modulate," as used herein, means to directly or indirectly interact with the activity of a target so as to alter the activity of the target, including, by way of example only, enhancing the activity of the target, inhibiting the activity of the target, limiting the activity of the target, or prolonging the activity of the target.

[0443] As used herein, the term "modulator" refers to a molecule that interacts directly or indirectly with a target. Interactions include, but are not limited to, those of an agonist, partial agonist, inverse agonist, antagonist, degrader, or combinations thereof. In some embodiments, the modulator is an agonist.

[0444] The terms "administer," "administering," "administration," and the like, as used herein, refer to methods that can be used to enable delivery of a compound or composition to a desired site of biological action. These methods include, but are not limited to, oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular, or infusion). Those of skill in the art are familiar with administration techniques that can be used with the compounds and methods described herein.

[0445] "Concurrent administration" and like terms, as used herein, are meant to encompass administration of selected therapeutic agents to a single patient and are intended to include therapeutic regimens in which agents are administered by the same or different routes of administration or at the same or different times.

[0446] The terms "effective amount" or "therapeutically effective amount," as used herein, refer to a sufficient quantity of an agent or compound administered, which relieves to some extent one or more of the symptoms of the disease or condition being treated. The result includes reduction and / or alleviation of the signs, symptoms, or causes of the disease, or any other desired alteration of a biological system. For example, an "effective amount" for therapeutic use is the quantity of a composition comprising a compound disclosed herein required to result in a clinically significant reduction in a disease symptom. An appropriate "effective" amount in any individual case is optionally determined using techniques such as a dose escalation study.

[0447] The terms "enhance" or "enhancing," as used herein, means to increase or prolong either in potency or duration a desired effect. Thus, in regard to enhancing the effect of therapeutic agents, the term "enhancing" refers to the ability to increase or prolong, either in potency or duration, the effect of other therapeutic agents on a system. An "enhancing-effective amount," as used herein, refers to an amount sufficient to enhance the effect of another therapeutic agent in a desired system.

[0448] As used herein, the term "pharmaceutical combination" refers to a product resulting from the mixing or combining of two or more active ingredients, and includes both fixed and non-fixed combinations of active ingredients. The term "fixed combination" means that both the active ingredients, e.g., a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and the adjuvant are administered to a patient simultaneously in the form of a single entity or dosage. The term "non-fixed combination" means that the active ingredients, e.g., a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and the adjuvant are administered to a patient simultaneously, concurrently, or sequentially as separate entities without any specific intervening time restriction, such that such administration provides effective levels of the two compounds in the patient's body. The latter also applies to cocktail therapy, e.g., the administration of three or more active ingredients.

[0449] The terms "article of manufacture" and "kit" are used synonymously.

[0450] The term "subject" or "patient" includes mammals. Examples of mammals include, but are not limited to, any member of the mammalian class, humans, non-human primates such as chimpanzees, and other ape and monkey species, farm animals such as cows, horses, sheep, goats, pigs, domestic animals such as rabbits, dogs, and cats, and laboratory animals including rodents such as rats, mice, and guinea pigs. In one aspect, the mammal is a human.

[0451] The terms "treat," "treating," or "treatment," as used herein, include improving, alleviating, or ameliorating at least one symptom of a disease or condition, preventing additional symptoms, inhibiting a disease or condition, e.g., preventing the onset of a disease or condition, relieving a disease or condition, causing regression of a disease or condition, alleviating symptoms caused by a disease or condition, or arresting the symptoms of a disease or condition, either prophylactically and / or therapeutically. [Example]

[0452] The following examples are provided for illustrative purposes only and are not intended to limit the scope of the claims provided herein.

[0453] Compound synthesis Example A. 2-{7-[({17-[(2-{[4-(4-aminopiperidin-1-yl)-3-(5-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]-3,6,9,12,15-pentaoxaheptadecan-1-yl}carbamoyl)methyl]-4,10-bis(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl}acetic acid (Compound 1)

[0454] [ka] Step A-1, Preparation of benzyl (1-(3-(5-chloro-1H-benzo[d]imidazol-2-yl)-2-((2,2-dimethyl-4,23-dioxo-3,8,11,14,17,20-hexaoxa-5,24-diazahexacosan-26-yl)amino)-5-(3-fluoro-5-methylphenyl)pyridin-4-yl)piperidin-4-yl)carbamate: 2,2-dimethyl-4-oxo-3,8,11,14,17,20-hexaoxa-5-azatricosane-23-o in dimethylformamide (DMF) (2 mL) To a mixture of benzoic acid (150 mg, 1.29 equiv., 366 μmol), perfluorophenyl diphenylphosphinate (150 mg, 1.38 equiv., 390 μmol), and N-methylmorpholine (NMM) (70 mg, 2.4 equiv., 0.69 mmol) was added benzyl (1-(2-((2-aminoethyl)amino)-3-(5-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-4-yl)piperidin-4-yl)carbamate (178 mg, 1 equiv., 283 μmol). The resulting reaction mixture was stirred at 25° C. for 2 hours. The crude product was purified by preparative HPLC (Prep HPLC-013) under the following conditions: column, SunFire Prep C18 OBD column, 120 g; mobile phase, water (0.1% trifluoroacetic acid (TFA)) and acetonitrile (ACN) (30.0% ACN in 7 min - max 98%); total flow rate, 70 mL / min; detector, UV 220 nm. The collected fractions were combined and concentrated in vacuo to give benzyl (1-(3-(5-chloro-1H-benzo[d]imidazol-2-yl)-2-((2,2-dimethyl-4,23-dioxo-3,8,11,14,17,20-hexaoxa-5,24-diazahexacosan-26-yl)amino)-5-(3-fluoro-5-methylphenyl)pyridin-4-yl)piperidin-4-yl)carbamate (120 mg, 41.5%). MS (M+H) + =1019.5.

[0455] Step A-2, Preparation of benzyl (1-(2-((1-amino-18-oxo-3,6,9,12,15-pentaoxa-19-azaheneicosan-21-yl)amino)-3-(5-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-4-yl)piperidin-4-yl)carbamate: Dissolve benzyl (1-(3-(5-chloro)-2-((1-amino-18-oxo-3,6,9,12,15-pentaoxa-19-azaheneicosan-21-yl)amino)-3-(5-chloro-1H-benzo[d]imidazol-2-yl)- A mixture of 1H-benzo[d]imidazol-2-yl)-2-((2,2-dimethyl-4,23-dioxo-3,8,11,14,17,20-hexaoxa-5,24-diazahexacosan-26-yl)amino)-5-(3-fluoro-5-methylphenyl)pyridin-4-yl)piperidin-4-yl)carbamate (118 mg, 1 equivalent, 116 μmol) and TFA (1 mL) was stirred at 25° C. for 1 hour. The reaction mixture was concentrated, and the crude product was adjusted to pH 8 with ammonium bicarbonate (NHHCO) and extracted with ethyl acetate (3 × 30 mL). The collected fractions were combined and concentrated in vacuo to give benzyl (1-(2-((1-amino-18-oxo-3,6,9,12,15-pentaoxa-19-azaheneicosan-21-yl)amino)-3-(5-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-4-yl)piperidin-4-yl)carbamate (95 mg, 89%). MS (M+H) + =919.3.

[0456] Step A-3, tri-tert-butyl 2,2',2''-(10-(24-((4-(4-(((benzyloxy)carbonyl)amino)piperidin-1-yl)-3-(5-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-2,21-dioxo-6,9,12,15,1 Preparation of 8-pentaoxa-3,22-diazatetracosyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate: 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)acetic acid (70 mg, 1.2 equiv., 0.12 mL) in DMF (1 mL) A mixture of N,N-diisopropylethylamine (DIEA) (40 mg, 54 μL, 3.2 equivalents, 0.31 mmol), N,N,N′,N′-tetramethyl-O-(N-succinimidyl)uronium hexafluorophosphate (HSTU) (45 mg, 1.3 equivalents, 0.13 mmol) was stirred at 25° C. for 10 minutes, followed by the addition of benzyl (1-(2-((1-amino-18-oxo-3,6,9,12,15-pentaoxa-19-azaheneicosan-21-yl)amino)-3-(5-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-4-yl)piperidin-4-yl)carbamate (90 mg, 1 equivalent, 98 μmol). The resulting reaction mixture was stirred for 1 hour at 25° C. The mixture was purified by preparative HPLC under the following conditions: column, SunFire Prep C18 OBD column, 19×150 mm, 5 μm; mobile phase, water (0.1% TFA) and ACN (30% ACN in 7 min - max 65%); total flow rate, 20 mL / min; detector, UV 220 nm.The collected fractions were combined and concentrated in vacuo to give tri-tert-butyl 2,2′,2″-(10-(24-((4-(4-(((benzyloxy)carbonyl)amino)piperidin-1-yl)-3-(5-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-2,21-dioxo-6,9,12,15,18-pentaoxa-3,22-diazatetracosyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate (62 mg, 43%). MS (M / 2+H). + =737.9.

[0457] Step A-4, Preparation of 2-{7-[({17-[(2-{[4-(4-aminopiperidin-1-yl)-3-(5-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]-3,6,9,12,15-pentaoxaheptadecan-1-yl}carbamoyl)methyl]-4,10-bis(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl}acetic acid: tri-tert-butyl 2,2',2''-(10-(2 A mixture of 4-((4-(4-(((benzyloxy)carbonyl)amino)piperidin-1-yl)-3-(5-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-2,21-dioxo-6,9,12,15,18-pentaoxa-3,22-diazatetracosyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate (60 mg, 1 equivalent, 41 μmol) and TFA (1 mL) was stirred at 60° C. for 2 hours. The reaction mixture was concentrated, and the crude product was purified by preparative HPLC (Prep HPLC-013) using the following conditions: column, SunFire Prep C18 OBD column, 19 × 150 mm, 5 μm; mobile phase, water (0.05% TFA) and ACN (30.0% ACN in 7 min - max. 50.0%); total flow rate, 20 mL / min; detector, UV 220 nm. The collected fractions were combined and concentrated under vacuum. The collected fractions were combined and concentrated in vacuo to give 2-{7-[({17-[(2-{[4-(4-aminopiperidin-1-yl)-3-(5-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]-3,6,9,12,15-pentaoxaheptadecan-1-yl}carbamoyl)methyl]-4,10-bis(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl}acetic acid (30 mg, 53%). MS (M+H) + =1171.7.

[0458] 111 Radiochemical synthesis of In[In]-compound 1 [ 111 [In]InCl (20.0 MBq, 40.0 μL, 0.1 M HCl) and compound 1 (2.9 nmol, 2.9 μL, 1.0 mM in DI water) were added to a NH4OAc solution (4.0 μL, 1.0 M). The resulting mixture was heated at 85 °C in a thermal mixer for 30 min. At the end of labeling, Ca-DTPA (4.0 μL, 4 mM) was added. The radiochemical purity was 97.5% as determined by RP-HPLC. The radiotracer solution for in vivo studies was prepared by diluting with 0.9% saline.

[0459] The following conjugates were prepared similarly to Example A using appropriate substitution reagents and substrates at different steps, and may require additional functional group modifications via well-known chemistry with appropriate reagents.

[0460] [Table 2-1]

[0461] [Table 2-2]

[0462] [Table 2-3]

[0463] [Table 2-4]

[0464] [Table 2-5]

[0465] [Table 2-6]

[0466] Example B N-(2-{[4-(4-aminopiperidin-1-yl)-3-(5-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)-1-(2-{3,16,19-trioxo-2,17,18-trioxa-5,8,11,14-tetraaza-1-indatricyclo[9.6.3.2 5 , 14 ]docosan-8-yl}acetamido)-3,6,9,12,15-pentaoxaoctadecan-18-amide (compound 15)

[0467] [ka] N-(2-{[4-(4-aminopiperidin-1-yl)-3-(5-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)-1-(2-{3,16,19-trioxo-2,17,18-trioxa-5,8,11,14-tetraaza-1-indatricyclo[9.6.3.2 5 , 14]docosane-8-yl}acetamido)-3,6,9,12,15-pentaoxaoctadecan-18-amide: Preparation of 2,2',2''-(10-(24-((4-(4-aminopiperidin-1-yl)-3-(5-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)- in ACN (0.2 mL) and water (0.1 mL) A mixture of 2,21-dioxo-6,9,12,15,18-pentaoxa-3,22-diazatetracosyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (16 mg, 1 equivalent, 14 μmol), indium trichloride (8 mg, 2 μL, 3 equivalents, 0.04 mmol), and sodium bicarbonate (5 mg, 2 μL, 4 equivalents, 0.06 mmol) was stirred at 80° C. for 2 hours. The mixture was diluted with 4 mL of dimethyl sulfoxide (DMSO), filtered, and the filtrate was purified by preparative HPLC (Prep HPLC-007) under the following conditions: column, SunFire Prep C18 OBD column, 19 × 150 mm 5 um 10 nm; mobile phase, water (0.05% TFA) and ACN (30% ACN in 15 min - max 75%); total flow rate, 20 mL / min; detector, UV 220 nm. The collected fractions were combined and concentrated under vacuum to give N-(2-{[4-(4-aminopiperidin-1-yl)-3-(5-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)-1-(2-{3,16,19-trioxo-2,17,18-trioxa-5,8,11,14-tetraaza-1-indatricyclo[9.6.3.2] 5 , 14 ]docosan-8-yl}acetamido)-3,6,9,12,15-pentaoxaoctadecan-18-amide (8.9 mg, 43%). MS (M+H) + =1283.6.

[0468] The following conjugates were prepared similarly to Example B using appropriate substitution reagents and substrates at different steps, and may require additional functional group modifications via well-known chemistry with appropriate reagents.

[0469] [Table 3]

[0470] Example C. (2S,5S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.1 10 , 14 .0 3 , 5 ]Hexacosa-10,12,14(26),16,18-pentaen-6-yl(2R)-2-{1-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]-N-methyl-3,6,9-trioxa-12,13-dithiahexadecan-16-amido}propanoate (Compound 19)

[0471] [ka] Step C-1, (14S,33S,2S,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenacyclotetradecaphane-10,12-dien-4-yl Preparation of N-methyl-N-(3-(pyridin-2-yldisulfanyl)propanoyl)-D-alaninate: (14S,33S,2S,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenacyclotetradecaphane-10,12-dien-4-yl A mixture of N-(3-mercaptopropanoyl)-N-methyl-D-alaninate (50 mg, 1 equivalent, 68 μmol) and 1,2-di(pyridin-2-yl)disulfane (45 mg, 3.0 equivalents, 0.20 mmol) was stirred at 25° C. for 20 minutes. Sodium acetate (0.5 mL, 0.2 M) was then added to the reaction mixture. The crude product was purified by preparative HPLC (Prep HPLC-013) under the following conditions: column, Atlantis Prep T3 OBD column, 19 × 150 mm, 5 μm; mobile phase, water (0.05% TFA) and ACN (28% phase B in 6 min - max. 53%); 20 mL / min, detector, UV 220, 254 nm. The collected fractions were combined and concentrated in vacuo to give (14S,33S,2S,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenacyclotetradecaphane-10,12-dien-4-yl N-methyl-N-(3-(pyridin-2-yldisulfanyl)propanoyl)-D-alaninate (52 mg, 91%). MS (M+H) + =847.

[0472] Step C-2, Preparation of (2R)-1-(((14S,33S,2S,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenacyclotetradecaphane-10,12-dien-4-yl)oxy)-2,3-dimethyl-1,4-dioxo-11,14,17-trioxa-7,8-dithia-3-azaicosan-20-oic acid: Dissolve (14S,33S,2S,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenacyclotetradecaphane-10,12-dien-4-yl)oxy) in methanol (MeOH) (2 mL) A mixture of 2E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenacyclotetradecaphane-10,12-dien-4-yl N-methyl-N-(3-(pyridin-2-yldisulfanyl)propanoyl)-D-alaninate (25 mg, 1 equivalent, 30 μmol) and 3-(2-(2-(2-mercaptoethoxy)ethoxy)ethoxy)propanoic acid (14.0 mg, 2.0 equivalents, 58.7 μmol) was stirred at 25° C. for 1 hour. The crude product was purified by preparative HPLC (Prep HPLC-013) under the following conditions: Column, Atlantis Prep T3 OBD column, 19 x 150 mm 5 um; Mobile phase, water (0.05% TFA) and ACN (28% Phase B in 6 min - max 53%); 20 mL / min, Detector, UV220, 254 nm. The collected fractions were combined and concentrated in vacuo to give (2R)-1-(((14S,33S,2S,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenacyclotetradecaphane-10,12-dien-4-yl)oxy)-2,3-dimethyl-1,4-dioxo-11,14,17-trioxa-7,8-dithia-3-azaicosan-20-oic acid (31 mg, 97.8%). MS (M+H) + =975.

[0473] Step C-3, (14S,33S,2S,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenacyclotetradecaphane-10,12-dien-4-yl(2R)-23-((4-(4-((tertbutoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-2,3-dimethyl-4,20-dioxo-11,14,17-tetramethyl- Preparation of trioxa-7,8-dithia-3,21-diazatricosanoate: (2R)-1-(((14S,33S,2S,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenacyclotetradecaphane-10,12-dien-4-yl)oxy)-2,3-dimethyl-1,4-dioxo-11,14,17-trioxa-7,8-dithia-3-azaicosan-20-oic acid (31 mg, 1 equivalent, 32 μmol) and hexafluorophosphate in DMF (2 mL). A mixture of azabenzotriazole tetramethyluronium (HATU) (24 mg, 2.0 equivalents, 63 μmol) was stirred at 25° C. for 10 minutes. Then, tert-butyl (1-(2-((2-aminoethyl)amino)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-4-yl)piperidin-4-yl)carbamate (21 mg, 1.1 equivalents, 35 μmol) was added to the reaction mixture. The crude product was purified by preparative HPLC (Preparative HPLC-013) using the following conditions: column, Atlantis Prep T3 OBD column, 19×150 mm, 5 μm; mobile phase, water (0.05% TFA) and ACN (28% phase B in 6 minutes—maximum 53%); 20 mL / min, detector, UV220, 254 nm.The collected fractions were combined and concentrated under vacuum to give (14S,33S,2S,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenacyclotetradecaphane-10,12-dien-4-yl(2R)-2 3-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-2,3-dimethyl-4,20-dioxo-11,14,17-trioxa-7,8-dithia-3,21-diazatricosanoate (25 mg, 51%) was obtained. MS (M+H). + =1550.7.

[0474] Step C-4, (2S,5S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.1 10 , 14 .0 3 , 5] Preparation of hexacosa-10,12,14(26),16,18-pentaen-6-yl(2R)-2-{1-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]-N-methyl-3,6,9-trioxa-12,13-dithiahexadecan-16-amido}propanoate: (14S,33S,2S,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza- in DCM (2 mL) A mixture of 1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenacyclotetradecaphane-10,12-dien-4-yl(2R)-23-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-2,3-dimethyl-4,20-dioxo-11,14,17-trioxa-7,8-dithia-3,21-diazatricosanoate (17 mg, 1 equivalent, 11 μmol) and zinc(II) chloride (12 mg, 8.0 equivalents, 88 μmol) was stirred at 25° C. for 3 hours. The crude product was purified by preparative HPLC (Prep HPLC-013) using the following conditions: Column, Atlantis Prep T3 OBD column, 19 x 150 mm 5 μm; Mobile phase, water (0.05% TFA) and ACN (28% Phase B in 6 min - max 53%); 20 mL / min; Detector, UV 220, 254 nm. The collected fractions were combined and concentrated under vacuum to give (2S,5S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.1]. 10 , 14 .0 3 , 5]Hexacosa-10,12,14(26),16,18-pentaen-6-yl(2R)-2-{1-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]-N-methyl-3,6,9-trioxa-12,13-dithiahexadecan-16-amido}propanoate (2.1 mg, 11%) was obtained. MS (M+H) + =1451.5.

[0475] The following conjugates were prepared similarly to Example C using appropriate substitution reagents and substrates at different steps, and may require additional functional group modifications via well-known chemistry with appropriate reagents.

[0476] [Table 4-1]

[0477] [Table 4-2]

[0478] [Table 4-3]

[0479] Example D. {4-[(2S)-2-[(2S)-2-[3-(2-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)-139-yl chloro-2-yl]amino}ethyl)carbamoyl]ethoxy}ethoxy)propanamido]-3-methylbutanamido]-5-(carbamoylamino)pentanamido]phenyl}methyl N-[(1S) -1-{[(1S)-1-{[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]-139-pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl]-N-methylcarbamate (Compound 24)

[0480] [ka] Step D-1, 4-((S)-2-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)benzyl((2R)-1-((1-(((3S,4R,5R)-1-((R)-2-((1S,2S)-3-(((1R,2S)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopeptidase Preparation of N-((3S,4R,5R)-1-((R)-2-((1S,2S)-3-(((1R,2S)-1-hydroxy-1-phenylpropane-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate: N-((3S,4R,5R)-1-((R)-2-((1S,2S)-3-(((1R,2S)-1-hydroxy-1-phenylpropane-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate -yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)-N,3-dimethyl-2-((R)-3-methyl-2-(methylamino)butanamide)butanamide (100 mg, 1 equivalent, 139 μmol), tert-butyl((S)-3-methyl-1-(((S)-1-((4-((((4-ni To a solution of 1-(2 ...The reaction mixture was then diluted with ethyl acetate, washed with water and brine, and concentrated to give crude product 4-((S)-2-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)benzyl ((2R)-1-((1-(((3S,4R,5R)-1-((R)-2-((1S,2S)-3-(((1R,2S)-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. MS (M+H). + = 1223.9. This material was used in the next step without further purification.

[0481] Step D-2, Preparation of 4-((S)-2-((S)-2-amino-3-methylbutanamido)-5-ureidopentanamido)benzyl ((2R)-1-((1-(((3S,4R,5R)-1-((R)-2-((1S,2S)-3-(((1R,2S)-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: 4-((S)-2-((S)-2-((tert-butoxycarbonyl)amino)-3-methyl To a solution of (2R)-1-((1-(((3S,4R,5R)-1-((R)-2-((1S,2S)-3-(((1R,2S)-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 (170 mg, 1 equivalent, 139 μmol) in DCM (0.6 mL) was added 2,2,2-trifluoroacetic acid (15.8 mg, 1 equivalent, 139 μmol). The resulting mixture was stirred for 30 minutes at 0° C. The reaction crude was concentrated and then purified by C18 reverse phase chromatography eluting with MeCN (0.1% TFA) / water (0.1% TFA).The collected fractions were combined and concentrated in vacuo to give 4-((S)-2-((S)-2-amino-3-methylbutanamido)-5-ureidopentanamido)benzyl ((2R)-1-((1-(((3S,4R,5R)-1-((R)-2-((1S,2S)-3-(((1R,2S)-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 (71.8 mg, 46.0%). MS (M+H). + =1124.1.

[0482] Step D-3, tert-butyl(6S,9S)-1-amino-6-((4-((5R,11R,12S)-11-((R)-sec-butyl)-12-(2-((R)-2-((1S,2S)-3-(((1R,2S)-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, Preparation of 13-dioxa-4,7,10-triazatetradecyl)phenyl)carbamoyl)-9-isopropyl-1,8,11-trioxo-14,17-dioxa-2,7,10-triazaicosan-20-oate: 3-(2-(3-(tert-butoxy)-3-oxopropoxy)ethoxy)propanoic acid (6.0 mg, 1 equivalent, 23 μmol) and 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethyl To a solution of isouronium hexafluorophosphate (V) (13 mg, 1.5 equiv., 34 μmol) in DMF (0.5 mL) was added N-ethyl-N-isopropylpropan-2-amine (12 mg, 16 μL, 4 equiv., 91 μmol) and 4-((S)-2-((S)-2-amino-3-methylbutanamido)-5-ureidopentanamido)benzyl ((2R)-1-((1-(((3S,4R,5R)-1-((R)-2-((1S,2S)-3-(((1R,2S )-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-2,2,2-trifluoroacetaldehyde (1 / 1) (28 mg, 1 equivalent, 23 μmol) was added. The reaction mixture was stirred at 20° C. for 2 hours.The reaction mixture was diluted with ethyl acetate, washed with water and brine, and concentrated to give the crude product tert-butyl(6S,9S)-1-amino-6-((4-((5R,11R,12S)-11-((R)-sec-butyl)-12-(2-((R)-2-((1S,2S)-3-(((1R,2S)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methylpropan-2-yl) MS (M+H) was obtained. + =1369.4. This material was used in the next step without further purification.

[0483] Step D-4, (6S,9S)-1-amino-6-((4-((5R,11R,12S)-11-((R)-sec-butyl)-12-(2-((R)-2-((1S,2S)-3-(((1R,2S)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2-oxoethyl)-5, Preparation of 8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)carbamoyl)-9-isopropyl-1,8,11-trioxo-14,17-dioxa-2,7,10-triazaicosan-20-oic acid: tert-butyl(6S,9S)-1-amino-6-((4-((5R, 11R,12S)-11-((R)-sec-butyl)-12-(2-((R)-2-((1S,2S)-3-(((1R,2S)-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 This reaction mixture was prepared by adding TFA (0.7 g, 0.5 mL, 3e+2 equiv, 6 mmol) to a solution of -2,13-dioxa-4,7,10-triazatetradecyl)phenyl)carbamoyl)-9-isopropyl-1,8,11-trioxo-14,17-dioxa-2,7,10-triazaicosan-20-oate (31 mg, 1 equiv, 23 μmol) in DCM (0.6 mL). The resulting mixture was stirred at 20° C. for 30 minutes. The reaction crude was concentrated and purified by C18 reverse-phase chromatography eluting with ACN (0.1% TFA) / water (0.1% TFA).The collected fractions were combined and concentrated under vacuum to give (6S,9S)-1-amino-6-((4-((5R,11R,12S)-11-((R)-sec-butyl)-12-(2-((R)-2-((1S,2S)-3-(((1R,2S)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidine- (1-yl)-2-oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)carbamoyl)-9-isopropyl-1,8,11-trioxo-14,17-dioxa-2,7,10-triazaicosan-20-oic acid (13.6 mg, 46%) was obtained. MS (M+H). + =1312.3

[0484] Step D-5, 4-((2S,5S)-19-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-4,7,16-trioxo-2-(3-ureidopropyl)-10,13-dioxa-3,6,17-triazanonadecanamide)benzyl((2R)-1-((1-(((3S,4R,5R Preparation of (6S,9S)-1-amino-6-((4-((5R,11R ,12S)-11-((R)-sec-butyl)-12-(2-((R)-2-((1S,2S)-3-(((1R,2S)-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)carbamoyl)-9-isopropyl-1,8,11 To a solution of -trioxo-14,17-dioxa-2,7,10-triazaicosan-20-oic acid (13.6 mg, 1 equiv., 9.94 μmol) and 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisouronium hexafluorophosphate (V) (5.67 mg, 1.5 equiv., 14.9 μmol) in DMF (0.2 mL) was added N-ethyl-N-isopropylpropan-2-amine (5.14 mg, 6.93 μL, 4 equiv., 39.8 μmol), followed by tert-butyl (1-(2-((2-aminoethyl)amino)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-4-yl)piperidin-4-yl)carbamate (6.50 mg, 1.1 equivalents, 10.9 μmol). The reaction mixture was stirred at 20° C. for 2 hours. The reaction mixture was diluted with ethyl acetate, washed with water and brine, and concentrated to give 4-((2S,5S)-19-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-4,7,16-trioxo-2-(3-ureidopropyl)-10,13-dioxa-3,6,17-triazanonadecanamide ) benzyl ((2R)-1-((1-(((3S,4R,5R)-1-((R)-2-((1S,2S)-3-(((1R,2S)-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. MS (M+H). + =1888.0. This material was used in the next step without further purification.

[0485] Step D-6, {4-[(2S)-2-[(2S)-2-[3-(2-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}ethoxy)propanamido]-3-methylbutanamido]-5-(carbamoylamino)pentanamido]phenyl}methyl N-[(1S)-1-{[(1S)-1-{[(3 Preparation of R,4S,5S)-1-[(2S)-2-[(1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl]-N-methylcarbamate: 4-((2S,5S)-19-((4-(4-((tert-butoxycarbamoyl) Nyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-4,7,16-trioxo-2-(3-ureidopropyl)-10,13-dioxa-3,6,17-triazanonadecanamido)benzyl((2R)-1-((1-(((3S,4R,5R)-1-((R)-2-((1S,2S)-3-(((1R,2S)-1-hydroxy- To a solution of 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 (18.8 mg, 1 equivalent, 9.96 μmol) in DCM (0.6 mL) was added TFA (0.7 g, 0.5 mL, 7e+2 equivalents, 6 mmol). The resulting mixture was stirred at 0° C. for 30 minutes. The reaction crude was concentrated and then purified by C18 reverse-phase chromatography eluting with ACN (0.1% TFA) / water (0.1% TFA).The collected fractions were combined and concentrated under vacuum to give {4-[(2S)-2-[(2S)-2-[3-(2-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}ethoxy)propanamido]-3-methylbutanamido]-5-(carbamoylamino)pentanamido]phenyl}methyl N-[(1 S)-1-{[(1S)-1-{[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl]-N-methylcarbamate (10.5 mg, 59.0%) was obtained. MS (M+H). + =1788.3.

[0486] The following conjugates were prepared similarly to Example D using appropriate substitution reagents and substrates at different steps, and may require additional functional group modifications via well-known chemistry with appropriate reagents.

[0487] [Table 5-1]

[0488] [Table 5-2]

[0489] [Table 5-3]

[0490] [Table 5-4]

[0491] [Table 5-5]

[0492] Example E. (4S)-4-(3-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}propanamide)-4-{[(1S)-1-{[(1S)-1-({4-[({[(1S)-1-{[(1S)-1-{[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-2 -{[(1S,2R)-1-Hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl](methyl)carbamoyl}oxy)methyl]phenyl}carbamoyl)ethyl]carbamoyl}-2-methylpropyl]carbamoyl}butanoic acid (Compound 32)

[0493] [ka] Step E-1, 4-((S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)propanamido)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)pyrrolyl Preparation of (S)-N-((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate: (S)-N-((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate in DMF (1.5 mL) To a mixture of (9H-fluoren-9-yl)methyl((S)-3-methyl-1-(((S)-1-((4-(((4-nitrophenoxy)methyl)-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)-N,3-dimethyl-2-((S)-3-methyl-2-(methylamino)butanamide)butanamide (150 mg, 1 equivalent, 209 μmol), (I)carbonyl)oxy)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-1-oxobutan-2-yl)carbamate (160 mg, 1.13 equivalents, 235 μmol), 1H-benzo[d][1,2,3]triazol-1-ol hydrate (50 mg, 1.6 equivalents, 0.33 mmol), and N-ethyl-N-isopropylpropan-2-amine (100 mg, 3.70 equivalents, 774 μmol) were added to the reaction mixture, which was stirred at 20° C. for 5 hours. The mixture was directly purified by MPLC under the following conditions: column, C18 120 g, spherical 20–40 μm; mobile phase, water (0.05% FA) and ACN (5% ACN-5% ACN in 1 min, 30% ACN-up to 90% in 10 min, 95% ACN-95% in 2 min); total flow rate, 70 mL / min; detector, UV 220 nm.The collected fractions were combined and concentrated under vacuum to give 4-((S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)propanamido)benzyl((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-furan) (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 (130 mg, 49.4%) was obtained. MS (M+H). + =1259.8.

[0494] Step E-2, 4-((S)-2-((S)-2-amino-3-methylbutanamido)propanamido)-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)pyrrolidine Preparation of 4-((S)-2-(((S)-2-(((9H-fluoren-9-yl)methoxy)carbonyl)-amino)-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 in ACN (1.5 mL) )-3-Methylbutanamido)propanamido)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-oxo To a mixture of (heptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate (125 mg, 1 equivalent, 99.2 μmol), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (50 mg, 50 μL, 3.3 equivalents, 0.33 mmol) was added. The reaction mixture was stirred at 20° C. for 1 hour. The mixture was directly purified by MPLC under the following conditions: column, WelFlash™, C18 120 g, Spherical 20-40 μm; mobile phase, water (0.05% FA) and ACN (5% ACN-5% ACN in 1 min, 30% ACN-up to 90% in 10 min, 95% ACN-95% in 2 min); total flow rate, 70 mL / min; detector, UV 220 nm.The collected fractions were combined and concentrated in vacuo to give 4-((S)-2-((S)-2-amino-3-methylbutanamido)-propanamido)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 (110 mg, 91%, 85% pure). MS (M+H). + =1037.8.

[0495] Step E-3, tert-butyl(S)-4-((((9H-fluoren-9-yl)methoxy)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-2,1 Preparation of 3-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate: To a mixture of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanoic acid (35 mg, 1.1 equiv., 82 μmol) in DMF (1 mL) was added HATU (33 mg, 1.1 equiv., 87 μmol) and DIEA (30 mg, 40 μL, 3.0 equiv., 0.23 mmol). The reaction mixture was stirred at 20° C. for 10 minutes, then 4-((S)-2-((S)-2-amino-3-methylbutanamido)propanamido)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 (80 mg, 1 equivalent, 77 μmol) was added and the reaction mixture was stirred at 20° C. for an additional 1.5 hours. The mixture was directly purified by MPLC under the following conditions: column, C18 120 g, spherical 20–40 μm; mobile phase, water (0.05% TFA) and ACN (5% ACN in 1 min, 30% ACN up to 78% in 9 min, 90% ACN up to 90% in 3 min); total flow rate, 70 mL / min; detector, UV 220 nm.The collected fractions were combined and concentrated under vacuum to give tert-butyl(S)-4-((((9H-fluoren-9-yl)methoxy)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-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (60 mg, 54%) was obtained. MS (M+H). + =1445.0.

[0496] Step E-4, tert-butyl (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-oxopropyl Preparation of tert-butyl (S)-4-(((9H-fluoren-9-yl)methyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate: Preparation of tert-butyl (S)-4-(((9H-fluoren-9-yl)methyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate in ACN (1 mL) 5,8-trimethoxy)carbonyl)amino)-5-(((S)-1-(((S)-1-((4-((5S,8S,11S,12R)-11-((S)-secbutyl)-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- To a mixture of diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (55 mg, 1 equivalent, 38 μmol) was added DBU (20 mg, 20 μL, 3.5 equivalents, 0.13 mmol). The reaction mixture was stirred at 20° C. for 1 hour. The mixture was directly purified by MPLC under the following conditions: column, WelFlash™, C18 120 g, spherical 20-40 μm; mobile phase, water (0.05% TFA) and ACN (5% ACN-5% ACN in 1 min, 30% ACN-up to 80% in 9 min, 90% ACN-90% in 3 min); total flow rate, 70 mL / min; detector, UV 220 nm. The collected fractions were dried by lyophilization.The collected fractions were combined and concentrated under vacuum to give tert-butyl(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-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (40 mg, 86%) was obtained. MS (M+H). + =1223.2.

[0497] Step E-5, Preparation of 3-(3-((2-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)ethyl)amino)-3-oxopropoxy)propanoic acid: 3,3′-oxydipropionic acid (150 mg, 1.10 equiv., 925 μmol), HATU (384 mg, 1.20 equiv.), HCl (3.25 ... A mixture of 1-(2-((2-aminoethyl)amino)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-4-yl)piperidin-4-yl)carbamate (500 mg, 1 equiv., 842 μmol) was added to the reaction mixture. The resulting reaction mixture was stirred at 25° C. for 2 hours. The reaction was then quenched with water (200 mL). The resulting solution was extracted with ethyl acetate (3×200 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied to a silica gel column with ethyl acetate / petroleum ether (1:1). The collected fractions were combined and concentrated in vacuo to give 3-(3-((2-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)ethyl)amino)-3-oxopropoxy)propanoic acid (290 mg, 46.7%). MS (M+H) + =738.5.

[0498] Step E-6, tert-butyl (S)-4-(3-(3-((2-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)ethyl)amino)-3-oxopropoxy)propanamide)-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-tetramethyl- Preparation of rioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate: 3-(3-((2-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[ d] To a mixture of (imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)ethyl)amino)-3-oxopropoxy)propanoic acid (25 mg, 1.0 equiv., 34 μmol), 4-methylmorpholine (12 mg, 3.6 equiv., 0.12 mmol) and perfluorophenyl diphenylphosphinate (16 mg, 1.3 equiv., 42 μmol) was added.The reaction mixture was stirred at 20° C. for 10 minutes, and then tert-butyl (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)pyrrolidine (1-yl)-2-oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (40 mg, 1 equiv., 33 μmol) was added and the reaction mixture was stirred at 20° C. for another 2 h. The mixture was diluted with 20 mL of water and extracted with ethyl acetate (20 mL × 3), and the combined organic layers were washed with water (10 mL × 2) and brine (20 mL), dried over anhydrous NaSO and concentrated under reduced pressure to give tert-butyl (S)-4-(3-(3-((2-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)ethyl)amino)-3-oxopropoxy)propanamido)-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-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (50 mg, 79%). MS (M+H). + =1942.2. This material was used in the next step without further purification.

[0499] Step E-7, (4S)-4-(3-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}propanamide)-4-{[(1S)-1-{[(1S)-1-({4-[({[(1S)-1-{[(1S)-1-{[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenyl Preparation of tert-butyl (S)-4-(3-(3-((2-((4-(4-((tert -butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)ethyl)amino)-3-oxopropoxy)propanamido)-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) To a mixture of (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-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (45 mg, 1 equivalent, 23 μmol), TFA (0.2 mL) was added at 0° C., and the reaction mixture was stirred for 30 minutes. The pH of the reaction mixture was adjusted to 7.0 with DIEA at 0° C., and the mixture was dried with a nitrogen drier.The crude product was purified by preparative HPLC under the following conditions: Column: SunFire prep OBD 19 x 150 mm 5 um; Mobile phase A: Water (0.05% TFA); Mobile phase B: ACN; Gradient: 20% B - 53% B in 15 min; Flow rate: 20 mL / min; Wavelength: 220 nm. The collected fractions were dried by lyophilization to give (4S)-4-(3-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}propanamide)-4-{[(1S)-1-{[(1S)-1-({4-[({[(1S)-1-{[(1S)-1-{[(3R,4S,5S)-1-[(2S)-2-[(1R,2 R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl](methyl)carbamoyl}oxy)methyl]phenyl}carbamoyl)ethyl]carbamoyl}-2-methylpropyl]carbamoyl}butanoic acid (8.4 mg, 17%). MS (M+H). + =1790.

[0500] The following conjugates were prepared similarly to Example E using appropriate substitution reagents and substrates at different steps, and may require additional functional group modifications via well-known chemistry with appropriate reagents.

[0501] [Table 6-1]

[0502] [Table 6-2]

[0503] [Table 6-3]

[0504] Table 6-4

[0505] Table 6-5

[0506] Table 6-6

[0507] Table 6-7

[0508] Table 6-8

[0509] Table 6-9

[0510] Table 6-10

[0511] Table 6-11

[0512] Table 6-12

[0513] Table 6-13

[0514] Table 6-14

[0515] Table 6-15

[0516] Table 6-16

[0517] Table 6-17

[0518] Table 6-18

[0519] Table 6-19

[0520] Table 6-20

[0521] Table 6-21

[0522] Table 6-22

[0523] Table 6-23

[0524] Table 6-24

[0525] Table 6-25

[0526] Table 6-26

[0527] Table 6-27

[0528] Table 6-28

[0529] Table 6-29

[0530] Table 6-30

[0531] Table 6-31

[0532] Table 6-32

[0533] Table 6-33

[0534] Table 6-34

[0535] Table 6-35

[0536] Table 6-36

[0537] Table 6-37

[0538] Table 6-38

[0539] Table 6-39

[0540] Table 6-40

[0541] Table 6-41

[0542] Example F. {4-[(2S)-2-[(2S)-2-[(2S)-2-(3-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}propanamido)-4-carbamoylbutanamido]-3-methylbutanamido]-5-(carbamoylamino)pentanamido]phenyl}methyl N-[(1S)-1-{[(1S)-1-{[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl]-N-methylcarbamate (Compound 56)

[0543] [ka] Step F-1, tert-butyl(4S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(((2S)-1-(((2S)-1-((4-((5S,11S,14S,17S,18R)-17-((S)-sec-butyl)-18-(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 (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(tert-butyl)-5,8,11,14-tetraisopropyl-4,10,16-trimethyl-3,6,9,12,15-pentaoxo-2,19-dioxa-4,7,10,13,16-pentaazaicosyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate in DMF (1 mL). A mixture of 4-((S)-2-((S)-2-amino-3-methylbutanamido)-5-ureidopentanamido)benzyl((3R,4S,7S,10S,16S)-4-((S)-sec-butyl)-3-(2-((S)-2-((1R,2 R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2-oxoethyl)-7,10,13-triisopropyl-5,11,17-trimethyl-6,9,12,15-tetraoxo-2-oxa-5,8,11,14-tetraazaoctadecan-16-yl)(methyl)carbamate (80 mg, 1 equivalent, 60 μmol) was added and the reaction mixture was stirred at 20° C. for 1 hour.The mixture was directly purified by MPLC under the following conditions: column, WelFlash™, C18 120 g, Spherical 20-40 μm; mobile phase, water (0.05% FA) and ACN (5% ACN-5% ACN in 1 min, 30% ACN-up to 80% in 7 min, 95% ACN-95% in 3 min); total flow rate, 70 mL / min; detector, UV 220 nm. The collected fractions were combined and concentrated under vacuum to give tert-butyl(4S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(((2S)-1-(((2S)-1-((4-((5S,11S,14S,17S,18R)-17-((S)-sec-butyl)-18-(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)-5,8,11,14-tetraisopropyl-4,10,16-trimethyl-3,6,9,12,15-pentaoxo-2,19-dioxa-4,7,10,13,16-pentaazaicosyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (90 mg, 86%). MS (M+H). + =1531.6.

[0544] Step F-2, tert-butyl(4S)-4-amino-5-(((2S)-1-(((2S)-1-((4-((5S,11S,14S,17S,18R)-17-((S)-secbutyl)-18-(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)-5,8,11,14-tetraisopropyl Preparation of propyl-4,10,16-trimethyl-3,6,9,12,15-pentaoxo-2,19-dioxa-4,7,10,13,16-pentaazaicosyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate: tert-butyl (4S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(( (2S)-1-(((2S)-1-((4-((5S,11S,14S,17S,18R)-17-((S)-sec-butyl)-18-(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)-5,8,11,14-tetraisopropyl-4,10,16-trimethyl-3,6,9,12,1 A mixture of 5-pentaoxo-2,19-dioxa-4,7,10,13,16-pentaazaicosyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (85 mg, 1 equivalent, 49 μmol) and 2,3,4,6,7,8,9,10-octahydropyrimido[1,2-a]azepine (25 mg, 3.4 equivalents, 0.16 mmol) was stirred at 20° C. for 1 hour.The mixture was directly purified by MPLC under the following conditions: column, WelFlash™, C18 120 g, Spherical 20-40 μm; mobile phase, water (0.05% FA) and ACN (5% ACN-5% ACN in 1 min, 20% ACN-up to 70% in 7 min, 95% ACN-95% in 3 min); total flow rate, 70 mL / min; detector, UV 220 nm. The collected fractions were combined and concentrated under vacuum to give tert-butyl(4S)-4-amino-5-(((2S)-1-(((2S)-1-((4-((5S,11S,14S,17S,18R)-17-((S)-sec-butyl)-18-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidine. (1-oxo-2-yl)-5,8,11,14-tetraisopropyl-4,10,16-trimethyl-3,6,9,12,15-pentaoxo-2,19-dioxa-4,7,10,13,16-pentaazaicosyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (70 mg, 94%). MS (M+H). + =1531.6.

[0545] Step F-3, tert-butyl (S)-4-(3-(3-((2-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)ethyl)amino)-3-oxopropoxy)propanamide)-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 Preparation of 3-(3-((2-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate in DMF (1 mL) -3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)ethyl)amino)-3-oxopropoxy)propanoic acid (40 mg, 1.3 equivalents, 54 μmol), 4-methylmorpholine (20 mg, 4.6 equivalents, 0.20 mmol), and pentafluorophenyl diphenylphosphinate (25 mg, 1.5 equivalents, 65 μmol) was stirred at 20° C. for 10 minutes, and then tert-butyl(4S)-4-amino-5-(((2S)-1-(((2S)-1-((4-((5S,11S,14S,17S,18R)-17-((S)-sec-butyl)-18-(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)-5,8,11,14-tetraisopropyl-4,10,16-trimethyl-3,6,9,12,15-pentaoxo-2,19-dioxa-4,7,10,13,16-pentaazaicosyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentanoate (65 mg, 1 equivalent, 43 μmol) was added and the reaction mixture was stirred at 20° C. for 1 hour. The mixture was diluted with 20 mL of water and extracted with EtOAc (20 mL × 3), and the combined organic layers were washed with water (10 mL × 2) and brine (20 mL), dried over anhydrous NaSO and concentrated under reduced pressure to give tert-butyl (S)-4-(3-(3-((2-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)ethyl)amino)-3-oxopropoxy)propanamido)-5-(((S)-1-(((S)-1-((4-((5S,8S,11 S,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-oxopentanoate (70 mg, 81%). MS (M / 2+H).+ = 1015.2. This material was used in the next step without further purification.

[0546] Step F-4, 4-((S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)propanamido)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)amido Preparation of (4S)-4-(3-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}propanamido)-4-{[(1S)-1-{[(1S)-4-(carbamoylamino)-1-( {4-[({[(1S)-1-{[(1S)-1-{[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl](methyl)carbamoyl}oxy)methyl]phenyl}carbamoyl)butyl]carbamoyl To a mixture of {N,N-dimethyl}-2-methylpropyl]carbamoyl}butanoic acid (35 mg, 80 wt%, 1 equiv., 15 μmol), ammonium chloride (20 mg, 25 equiv., 0.37 mmol), 1H-benzo[d][1,2,3]triazol-1-ol hydrate (3 mg, 1 equiv., 0.02 mmol), 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (5 mg, 2 equiv., 0.03 mmol), and N,N-dimethylpyridin-4-amine (3 mg, 2 equiv., 0.02 mmol) was added. The reaction mixture was stirred at 20° C. for 6 h.The crude product was purified by preparative HPLC under the following conditions: Column: SunFire prep OBD 19 x 150 mm 5 um; Mobile phase A: Water (0.05% TFA); Mobile phase B: ACN; Gradient: 25% B - 55% B in 16 min; Flow rate: 20 mL / min; Wavelength: 220 nm. The collected fractions were dried by lyophilization to give {4-[(2S)-2-[(2S)-2-[(2S)-2-(3-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}propanamido)-4-carbamoylbutanamido]-3-methylbutanamido]-5-(carbamoylamino)pentanamido]phenyl To give {(1S)-1-{[(1S)-1-{[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl]-N-methylcarbamate (10.3 mg, 31%). MS (M+H). + =1872.1.

[0547] The following conjugates were prepared similarly to Example F using appropriate substitution reagents and substrates at different steps, and may require additional functional group modifications via well-known chemistry with appropriate reagents.

[0548] [Table 7-1]

[0549] [Table 7-2]

[0550] Example G. (2S,3S,4S,5R,6S)-6-{2-[(2S)-2-[(2S)-2-(3-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}propanamido)-3-methylbutanamido]-5-(carbamoylamino)pentanamido]-5-[({[(1S)-1-{[(1S)-1-{[(3R ,4S,5S)-1-[(2S)-2-[(1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl](methyl)carbamoyl}oxy)methyl]phenoxy}-3,4,5-trihydroxyoxane-2-carboxylic acid (Compound 61)

[0551] [ka] Step G-1, Preparation of (2S,3R,4S,5S,6S)-2-(5-formyl-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate: A mixture of 3-hydroxy-4-nitrobenzaldehyde (2.334 g, 1 equivalent, 13.97 mmol), (2R,3R,4S,5S,6S)-2-bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (16.64 g, 3.000 equivalents, 41.90 mmol), and silver(I) oxide (25.9 g, 8.00 equivalents, 112 mmol) in ACN (140 mL) was stirred in the dark at 25° C. for 24 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by MPLC using the following conditions: 120 g silica gel column, petroleum ether / ethyl acetate (PE / EtOAc) system, 0%-85% EtOAc in 15 min, flow rate: 70 mL / min; wavelength: 254 nm. The collected fractions were combined and concentrated in vacuo to give (2S,3R,4S,5S,6S)-2-(5-formyl-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (3.8 g, 56%). MS (M+H) + =484.4.

[0552] Step G-2, Preparation of (2S,3R,4S,5S,6S)-2-(2-amino-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate: To a mixture of (2S,3R,4S,5S,6S)-2-(5-formyl-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (3.8 g, 1 equivalent, 7.9 mmol) and triethylamine (0.16 g, 0.22 mL, 0.20 equivalents, 1.6 mmol) in ethyl acetate (210 mL) under an inert atmosphere of nitrogen, Pearlman's catalyst (85 mg, 0.10 equivalents, 0.80 mmol) was added. The reaction mixture was flushed with hydrogen three times, followed by flushing with hydrogen and stirring under a pressure of H gas at 25° C. for 24 hours. The reaction mixture was filtered through a diatomaceous earth cushion, and the filtrate was concentrated under reduced pressure to give (2S,3R,4S,5S,6S)-2-(2-amino-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (2.7 g, 75%). MS (M+H) + =456.4.

[0553] Step G-3, Preparation of (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate: (2S,3R,4S,5S,6S)-2-(2-amino-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate in DCM (10 mL) To a mixture of hydro-2H-pyran-3,4,5-triyl triacetate (1.05 g, 1 equivalent, 2.31 mmol) and 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) (685 mg, 1.20 equivalents, 2.77 mmol), (S)-2-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamido)-5-ureidooxopentanoic acid (865 mg, 1.00 equivalents, 2.31 mmol) was added at room temperature, and the reaction mixture was stirred in the dark at 25 °C for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by MPLC under the following conditions: 120 g silica gel column, PE / EtOAc system, 0% to 85% EtOAc in 15 min, flow rate: 70 mL / min; wavelength: 254 nm. The collected fractions were combined and concentrated in vacuo to give (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (1.36 g, 72.7%). MS (M+H) + =812.2.

[0554] Step G-4, Preparation of (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)-5-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate: (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((t A mixture of (ert-butoxycarbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (460 mg, 1 equiv., 567 μmol), carbonic acid, bis(4-nitrophenyl) ester (260 mg, 1.51 equiv., 855 μmol), and N-ethyl-N-isopropylpropan-2-amine (150 mg, 2.05 equiv., 1.16 mmol) was stirred at room temperature for 24 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by MPLC using the following conditions: 120 g silica gel column, PE / EtOAc system, 0% to 85% EtOAc in 15 min, flow rate: 70 mL / min, wavelength: 254 nm. The collected fractions were combined and concentrated in vacuo to give (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)-5-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (400 mg, 72.3%). MS (M+H) + =977.1.

[0555] Step G-5, (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)-5-((5S,8S,11S,12R)-11-((S)-secbutyl)-12-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidone Preparation of (lysin-1-yl)-2-oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate: (2S,3R,4S,5S,6S)-2-(2-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanal) in DMF (1.7 mL) (S)-N-((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxo- A mixture of (isopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)-N,3-dimethyl-2-((S)-3-methyl-2-(methylamino)butanamido)butanamide (100 mg, 0.800 equiv., 139 μmol), 1H-benzo[d][1,2,3]triazol-1-ol (30 mg, 1.3 equiv., 0.22 mmol), and DIEA (70 mg, 94 μL, 3.1 equiv., 0.54 mmol) was stirred at 25° C. for 2 hours.The mixture was directly purified by MPLC under the following conditions: column, WelFlash™, C18 120 g, Spherical 20-40 μm; mobile phase, water (0.05% NH.HO) and ACN (5% ACN-5% ACN in 1 min, 30% ACN-up to 98% in 6 min, 98% ACN-98% in 3 min); total flow rate, 70 mL / min; detector, UV 254 nm. The collected fractions were combined and concentrated under vacuum to give (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((tertbutoxycarbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)-5-((5S,8S,11S,12R)-11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenyl)-2-propan-1-yl)-2-methylbutan ...methylbutanamido)-5-((5S,8S,11S,12R)-11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenyl)-2-methylbutanamido)-5-((5S,8S,11S,12R)-11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenyl)-2 (Nylpropan-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)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (80 mg, 30%) was obtained. MS (M+H). + =1556.5.

[0556] Step G-6, (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-amino-3-methylbutanamido)-5-ureidopentanamido)-5-((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 Preparation of (2S,3R,4S,5S,6S)-2-(2-oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate: (2S,3R,4S,5S,6S)-2-(2-oxoethyl)-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy) in DCM (0.5 mL) -((S)-2-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamido)-5-ureidopentanamido)-5-((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-oxopropan-2-yl)amino) A mixture of (pyr)pyrrolidin-1-yl)-2-oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (76 mg, 1 equivalent, 49 μmol) and TFA (0.5 mL) was stirred at 0° C. for 2 minutes.The mixture was adjusted to pH 9.0 with DIEA and dried with a nitrogen drier to give (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-amino-3-methylbutanamido)-5-ureidopentanamido)-5-((5S,8S,11S,12R)-11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropionamido)). (2-oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (60 mg, 84%) was obtained. MS (M+H). + =1456.5. This material was used in the next step without further purification.

[0557] Step G-7, (2S,3R,4S,5S,6S)-2-(2-((2S,5S)-16-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-4,7,13-trioxo-2-(3-ureidopropyl)-10-oxa-3,6,14-triazahexadecanamide)-5-((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)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyra Preparation of benzophenone-3,4,5-triyl triacetate: (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-amino-3-methylbutanamido)-5-ureidopentanamido)-5-((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) in DMF (0.6 mL) (pyrrolidin-1-yl)-2-oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (60 mg, 1 equivalent, 41 μmol), pentafluorophenyl diphenylphosphinate (20 mg, 1.3 equivalents, 52 μmol), 4-methylmorpholine (13 mg, 14 μL, 3.1 equivalents, 0.A mixture of 13 mmol) and 3-(3-((2-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)ethyl)amino)-3-oxopropoxy)propanoic acid (30 mg, 0.99 equivalents, 41 μmol) was stirred at 25° C. for 2 hours. The mixture was directly purified by MPLC under the following conditions: column, WelFlash™, C18 120 g, Spherical 20-40 μm; mobile phase, water (0.05% NH.HO) and ACN (5% ACN-5% ACN in 2 min, 30% ACN-up to 98% in 12 min, 98% ACN-98% in 1 min); total flow rate, 70 mL / min; detector, UV 220 nm. The collected fractions were combined and concentrated under vacuum to give (2S,3R,4S,5S,6S)-2-(2-((2S,5S)-16-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-4,7,13-trioxo-2-(3-ureidopropyl)-10-oxa-3,6,14-triazahexadecanamido)-5-((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)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (64 mg, 71%). MS (M / 2+H). + =1088.6.

[0558] Step G-8, (2S,3S,4S,5R,6S)-6-(2-((2S,5S)-16-((4-(4-((tertbutoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-4,7,13-trioxo-2-(3-ureidopropyl)-10-oxa-3,6,14-triazahexadecanamide)-5-((5S,8S,11S,12R)-11-((S)-sec-butyl)- Preparation of 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)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid: (2S,3R, 4S,5S,6S)-2-(2-((2S,5S)-16-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-4,7,13-trioxo-2-(3-ureidopropyl)-10-oxa-3,6,14-triazahexadecanamido)-5-((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)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (64 mg, 1 equivalent, 29 μmol), and lithium hydroxide (4 mg, 6 equivalents, 0.A mixture of 2 mmol) of 1,2-dichloro-2 ... The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum to give (2S,3S,4S,5R,6S)-6-(2-((2S,5S)-16-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-4,7,13-trioxo-2-(3-ureidopropyl)-10-oxa-3,6,14-triazahexadecanamido)-5-((5S,8S,11S,12R)-1 1-((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)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (50 mg, 15 μmol, 52%, purity 62%) was obtained. MS (M / 2+H). + = 1019.2. This material was used in the next step without further purification.

[0559] Step G-9, (2S,3S,4S,5R,6S)-6-{2-[(2S)-2-[(2S)-2-(3-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}propanamide)-3-methylbutanamide]-5-(carbamoylamino)pentanamide]-5-[({[(1S)-1-{[(1S)-1-{[(3R,4S,5S)-1-[(2S Preparation of (2S,3S,4S,5R,6S)-6-((2S,3S,4S,5R,6S)-2-[(1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl](methyl)carbamoyl}oxy)methyl]phenoxy}-3,4,5-trihydroxyoxane-2-carboxylic acid: (2S,3S,4S,5R,6S)-6-( 2-((2S,5S)-16-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-4,7,13-trioxo-2-(3-ureidopropyl)-10-oxa-3,6,14-triazahexadecanamido)-5-((5S,8S,11S,12R)-11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2R) A mixture of -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)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (50 mg, 52% by weight, 1 equivalent, 13 μmol) and TFA (0.2 mL) was stirred at 0° C. for 30 minutes.The crude product was purified by preparative HPLC under the following conditions: Column: SunFire prep OBD 19 x 150 mm 5 um; Mobile phase A: Water (0.05% NH.HO); Mobile phase B: ACN; Gradient: 25% B - 65% B in 8 min; Flow rate: 20 mL / min; Wavelength: 220 nm. The collected fractions were dried by lyophilization to give (2S,3S,4S,5R,6S)-6-{2-[(2S)-2-[(2S)-2-(3-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}propanamido)-3-methylbutanamido]-5-(carbamoylamino)pentanamido]-5-[({[(1S)-1-{[(1S)-1- Obtained {[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl](methyl)carbamoyl}oxy)methyl]phenoxy}-3,4,5-trihydroxyoxane-2-carboxylic acid (8.1 mg, 33%). MS (M+H). + =1936.0.

[0560] The following conjugates were prepared similarly to Example G using appropriate substitution reagents and substrates at different steps, and may require additional functional group modifications via well-known chemistry with appropriate reagents.

[0561] [Table 8-1]

[0562] [Table 8-2]

[0563] Table 8-3

[0564] Table 8-4

[0565] Table 8-5

[0566] Table 8-6

[0567] Table 8-7

[0568] Table 8-8

[0569] Table 8-9

[0570] Table 8-10

[0571] Table 8-11

[0572] Table 8-12

[0573] Table 8-13

[0574] [Table 8-14]

[0575] [Table 8-15]

[0576] [Table 8-16]

[0577] [Table 8-17]

[0578] [Table 8-18]

[0579] Example H. (2S,3S,4S,5R,6S)-6-{2-[(2S)-2-[(2S)-2-(3-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}propanamido)-3-methylbutanamido]propanamido]-5-[({[(1S)-1-{[(1S)-1-{[(3R,4S,5S) -1-[(2S)-2-[(1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl](methyl)carbamoyl}oxy)methyl]phenoxy}-3,4,5-trihydroxyoxane-2-carboxylic acid (Compound 68)

[0580] [ka] Step H-1, Preparation of (2S,3R,4S,5S,6S)-2-(2-((S)-2-((tert-butoxycarbonyl)amino)propanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate: (2S,3R,4S,5S,6S)-2-(2-amino-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate in DMF (10 mL) A mixture of (tert-butoxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (800 mg, 1 equivalent, 1.76 mmol), (tert-butoxycarbonyl)-L-alanine (400 mg, 1.20 equivalents, 2.11 mmol), and EEDQ (520 mg, 1.20 equivalents, 2.10 mmol) was stirred at 25 °C for 24 h. The reaction was then quenched with water (100 mL), and the resulting solution was extracted with ethyl acetate (3 × 100 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate / petroleum ether (3:1). The collected fractions were combined and concentrated in vacuo to give (2S,3R,4S,5S,6S)-2-(2-((S)-2-((tert-butoxycarbonyl)amino)propanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (770 mg, 70.0%). MS (M+H) + =627.5.

[0581] Step H-2, Preparation of (2S,3R,4S,5S,6S)-2-(2-((S)-2-aminopropanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate: A mixture of (2S,3R,4S,5S,6S)-2-(2-((S)-2-((tert-butoxycarbonyl)amino)propanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (770 mg, 1 equiv., 1.23 mmol) and TFA (2 mL) in DCM (6 mL) was stirred at 25° C. for 30 minutes. The resulting solution was concentrated in vacuo. The resulting mixture was dissolved in 20 mL of DCM and washed with 2 mL of saturated NaHCO. The organic layer was dried over anhydrous sodium sulfate and concentrated to give (2S,3R,4S,5S,6S)-2-(2-((S)-2-aminopropanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (650 mg, 100%). MS (M+H) + =527.2. This material was used in the next step without further purification.

[0582] Step H-3, Preparation of (2S,3R,4S,5S,6S)-2-(2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)propanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate: (2S,3R,4S,5S,6S)-2-( A mixture of 2-((S)-2-aminopropanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (650 mg, 1 equiv., 1.23 mmol), HATU (580 mg, 1.24 equiv., 1.53 mmol), and diisopropylethylamine (480 mg, 642 μL, 3.01 equiv., 3.71 mmol) was stirred at 25° C. for 10 minutes. (((9H-Fluoren-9-yl)methoxy)carbonyl)-L-valine (420 mg, 1.00 equiv., 1.24 mmol) was then added, and the resulting reaction mixture was stirred at 25° C. for 1 hour. The reaction was then quenched with water (100 mL), and the resulting solution was extracted with ethyl acetate (3×100 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied to a silica gel column with ethyl acetate / petroleum ether (2:1). The collected fractions were combined and concentrated under vacuum to give (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)propanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (620 mg, 59.2%). MS (M+H) + =848.3.

[0583] Step H-4, Preparation of (2S,3R,4S,5S,6S)-2-(2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)propanamido)-5-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate: (2S,3R,4S,5S,6S)-2-(2-((S A mixture of )-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)propanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (520 mg, 1 eq, 613 μmol) and carbonic acid, bis(4-nitrophenyl) ester (280 mg, 1.50 eq, 920 μmol) was stirred at 25° C. under a nitrogen atmosphere for 24 hours. 70% product was detected by LCMS. The reaction mixture was worked up in the next batch. MS (M+H) + =1013.

[0584] Step H-5, (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-(((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)propanamido)-5-((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)pyrrolidone Preparation of (lysin-1-yl)-2-oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate: (2S,3R,4S,5S,6S)-2-(2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methyl)-2H-pyran-3,4,5-triyl triacetate in DMF (3.5 mL) (S)-N-((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidine A mixture of (lysin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)-N,3-dimethyl-2-((S)-3-methyl-2-(methylamino)butanamido)butanamide (205 mg, 0.904 equiv., 286 μmol), 1-hydroxy-1H-benzotriazole (52 mg, 53 μL, 1.2 equiv., 0.38 mmol), and N-ethyl-N-isopropylpropan-2-amine (123 mg, 3.01 equiv., 952 μmol) was stirred at 25° C. for 16 hours. 68% product was detected by LCMS. The reaction mixture was worked up in the next batch. MS (M+H) + =1591.6.

[0585] Step H-6, (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-amino-3-methylbutanamido)propanamido)-5-((5R,11R,12S)-11-((R)-sec-butyl)-12-(2-((R)-2-((1S,2S)-3-(((1R,2S)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidine-1 Preparation of (2S,3R,4S,5S,6S)-2-(2-((S)-2-(((9H-fluorene-9 -yl)methoxy)carbonyl)amino)-3-methylbutanamido)propanamido)-5-((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 To a mixture of pyr-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (120 mg, 1 equivalent, 75.4 μmol), piperidine (45.0 mg, 52.0 μL, 7.01 equivalents, 528 μmol) was added at 0° C., and the resulting reaction mixture was stirred at 0° C. for 5 hours. The mixture was directly purified by MPLC under the following conditions: column, WelFlash™, C18 120 g, Spherical 20-40 μm; mobile phase, water (0.05% NH.HO) and ACN (5% ACN-5% ACN in 1 min, 20% ACN-up to 98% in 8 min, 98% ACN-98% in 1 min); total flow rate, 70 mL / min; detector, UV 220 nm.The collected fractions were combined and concentrated under vacuum to give (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-amino-3-methylbutanamido)propanamido)-5-((5R,11R,12S)-11-((R)-sec-butyl)-12-(2-((R)-2-((1S,2S)-3-(((1R,2S)-1-hydroxy-1-phenylpropan-2-yl)amido) (120 mg, 58%) of (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)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate was obtained. MS (M+H). + =1369.6.

[0586] Step H-7, (2S,3R,4S,5S,6S)-2-(2-((2S,5S)-16-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-2-methyl-4,7,13-trioxo-10-oxa-3,6,14-triazahexadecanamide)-5-((5R,11R,12S)-11-((R)-sec-butyl)-12-(2-((R)- Preparation of 2-((1S,2S)-3-(((1R,2S)-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)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate: (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S) -2-amino-3-methylbutanamido)propanamido)-5-((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)phenoxy)-6-(methox- (tert-Butoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (100 mg, 1 equivalent, 73.0 μmol), 3-(3-((2-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)ethyl)amino)-3-oxopropoxy)propanoic acid (55 mg, 1.0 equivalent, 74 μmol), perfluorophenyl diphenylphosphinate (34 mg, 1.A mixture of (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-amino-3-methylbutanamido)propanamido)-5-((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-methyl-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)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (100 mg, 1 equivalent, 73.0 μmol) was added. The resulting reaction mixture was stirred at 25 °C for 1 h. The mixture was directly purified by MPLC with the following conditions: Column, Flash™, C18 120 g, Spherical 20-40 μm; Mobile phase, water (0.05% NH ).HO) and ACN (5% ACN-5% ACN in 2 min, 30% ACN-up to 98% in 12 min, 98% ACN-98% in 1 min); total flow rate, 70 mL / min; detector, UV 220 nm. The collected fractions were combined and concentrated under vacuum to give (2S,3R,4S,5S,6S)-2-(2-((2S,5S)-16-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-2-methyl-4,7,13-trioxo-10-oxa-3,6,14-triazahexadecanamido)-5-((5R,11R,12S)-11-((R)- secbutyl)-12-(2-((R)-2-((1S,2S)-3-(((1R,2S)-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)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (20 mg, 13%). MS (M / 2+H). + =1050.5; MS(M+H) + =2099.5.

[0587] Step H-8, (2S,3S,4S,5R,6S)-6-(2-((2S,5S)-16-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-2-methyl-4,7,13-trioxo-10-oxa-3,6,14-triazahexadecanamide)-5-((5S,8S,11S,12R)-11-((S)-sec-butyl)-12-(2-( Preparation of (S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)allyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid: (2S,3R,4S,5S,6S)-2-( 2-((2S,5S)-16-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-2-methyl-4,7,13-trioxo-10-oxa-3,6,14-triazahexadecanamide)-5-((5S,8S,11S,12R)-11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1 A mixture of (hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)allyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (18 mg, 1 equivalent, 8.6 μmol) and lithium hydroxide (2 mg, 1e+1 equivalent, 0.08 mmol) was stirred at 25° C. for 30 minutes.The residue was diluted with water (10 ml) and then adjusted to pH 6-7 with AcOH (1 M). The resulting solution was extracted with ethyl acetate (3 × 2 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo to give (2S,3S,4S,5R,6S)-6-(2-((2S,5S)-16-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-2-methyl-4,7,13-trioxo-10-oxa-3,6,14-triazahexadecanamido)-5-((5S,8S,11S,1 2R)-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)allyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (20 mg, 84%) was obtained. MS (M / 2+H). + =975;MS(M+H) + = 1949. This material was used in the next step without further purification.

[0588] Step H-9, (2S,3S,4S,5R,6S)-6-{2-[(2S)-2-[(2S)-2-(3-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}propanamide)-3-methylbutanamide]propanamide]-5-[({[(1S)-1-{[(1S)-1-{[(3R,4S,5S)-1-[(2S)-2-[ Preparation of (1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl](methyl)carbamoyl}oxy)methyl]phenoxy}-3,4,5-trihydroxyoxane-2-carboxylic acid: (2S,3S,4S,5R,6S)- in DCM (0.5 mL) 6-(2-((2S,5S)-16-((4-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-(6-chloro-1H-benzo[d]imidazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl)amino)-5-isopropyl-2-methyl-4,7,13-trioxo-10-oxa-3,6,14-triazahexadecanamide)-5-((5S,8S,11S,12R)-11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2 A mixture of R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)allyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (20 mg, 1 equivalent, 10 μmol) and TFA (0.1 mL) was stirred at 25° C. for 30 minutes.The crude product was purified by preparative HPLC (Prep HPLC-013) under the following conditions: Column, Atlantis Prep T3 OBD column, 19 x 150 mm 5 um; Mobile phase, water (0.1% TFA) and ACN (28% Phase B in 10 min - max 53%); 20 mL / min, Detector, UV220, 254 nm. The collected fractions were combined and concentrated under vacuum to give (2S,3S,4S,5R,6S)-6-{2-[(2S)-2-[(2S)-2-(3-{2-[(2-{[4-(4-aminopiperidin-1-yl)-3-(6-chloro-1H-1,3-benzodiazol-2-yl)-5-(3-fluoro-5-methylphenyl)pyridin-2-yl]amino}ethyl)carbamoyl]ethoxy}propanamido)-3-methylbutanamido]propanamido]-5-[({[(1S)-1-{[(1S)-1-{[(3R,4S ,5S)-1-[(2S)-2-[(1R,2R)-2-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]carbamoyl}-1-methoxy-2-methylethyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl](methyl)carbamoyl}-2-methylpropyl]carbamoyl}-2-methylpropyl](methyl)carbamoyl}oxy)methyl]phenoxy}-3,4,5-trihydroxyoxane-2-carboxylic acid (4.7 mg, 21%). MS (M+H). + =1850.0.

[0589] Biological assays Example I: SSTR Assay General Overview: All five SSTR subtypes are G-coupled G protein-coupled receptors (GPCRs) that, when activated by agonists, result in a decrease in intracellular cyclic AMP (cAMP). Therefore, measuring intracellular cAMP levels can be used to evaluate whether the compounds of the present invention are agonists of SSTR subtypes (John Kelly, Troy Stevens, W. Joseph Thompson, and Roland Seifert, Current Protocols in Pharmacology, 2005, 2.2.1-2.2). An example of an intracellular cAMP assay is described below.

[0590] Functional assay for SSTR2 agonists Four days prior to the assay, 2,000 Chinese hamster ovary cells (CHO-K1, ATCC #CCL-61) stably expressing human somatostatin receptor subtype 2 are plated into each well of a 96-well tissue culture-treated plate in Ham's F12 growth medium (ThermoFisher #10-080-CM) supplemented with 10% donor bovine serum (Gemini Bio-Products #100-506), 100 U / mL penicillin, 100 μg / mL streptomycin, 2 mM L-glutamine (Gemini Bio-Products #400-110), and 0.2 mg / mL hygromycin B (GoldBio #31282-04-9). Cells are cultured at 37°C, 5% CO2, and 95% humidity. cAMP is measured using an HTRF kinetic cAMP assay (Cisbio, #62AM5PEJ) according to the manufacturer's instructions. On the day of the assay, the medium is aspirated and the cells are treated with 50 μL of stimulation buffer supplemented with 10.2 mM 3-isobutyl-1-methylxanthine (IBMX, Millipore Sigma #I5879) and 1.6 μM NKH477 (Tocris #1603) along with various dilutions of the SMDCs of the present invention. The cells are incubated for 20 minutes at 37°C (final concentrations of the compounds of the present invention are typically 0-10,000 nM). The cells are treated with 50 μL of lysis buffer (HRTF cAMP kit, Cisbio) and incubated for 30 minutes at room temperature with rotary shaking at 600 rpm, then diluted with 150 μL of stimulation buffer and shaken at 300 rpm for an additional 5 minutes. Lysates are transferred to 384-well plates and incubated at room temperature for 1-24 hours, and cAMP accumulation is detected with d2-labeled cAMP and anti-cAMP-cryptate. Time-resolved fluorescence signals are read using an m1000 Pro (Tecan) or CLARIOStar (BMG Labtech) microplate reader; samples are excited at 340 nm and emission is measured at 620 nm and 665 nm. Data are expressed as calculated fluorescence ratios (665 nm / 620 nm). Intracellular cAMP concentrations are calculated by regression to a standard curve and plotted against the concentration of compounds of the invention. EC of compounds is calculated. 50is calculated using standard methods. All data manipulations are in GraphPad Prism v9 (GraphPad, San Diego, CA).

[0591] Example J: Internalization Assay Protocol The SMDCs of the present invention were evaluated in vitro using DiscoverX's Human PathHunter CHO-K1 SST2 β-Arrestin2 and PathHunter CHO-K1 SST2 Internalization Assays. CHO-K cells were seeded at a density of 2500 cells / well in 20 μL of cell culture medium in white-walled 384-well tissue culture-compatible plates. After 48 hours of incubation, cells were treated with 5 μL of 1:3 serially diluted SST2 SMDCs prepared at 5x concentrations, for 90 minutes at a final top compound concentration of 10 μM, at 37°C and 5% CO2. Eleven dilutions and one treatment without compound were tested. After incubation, the medium containing the treatments was discarded, and 20 μL of Beta-Glo working solution from the Promega Beta-Glo Assay System was added to each well. The plate was then incubated in the dark at room temperature for 60 minutes. Luminescence was measured using a Tecan plate reader. I C 50 Curves were generated using nonlinear regression analysis (4 parameters) with GraphPad Prism 9. Results are the average of duplicates and corrected for background.

[0592] Example K: H524 Assay Protocol The SMDCs of the present invention were evaluated in an in vitro assay to assess cell proliferation inhibition. NCI-H524 (ATCC) human small cell lung cancer cells were plated at a concentration of 5,000 cells / well in a 96-well V-bottom plate (Costar). 24 hours later, the cells were treated with SMDCs for 2 hours. The starting dose of SMDC was 1 μM, and two-fold serial dilutions were performed for a total of 11 points. After 2 hours of treatment, the cells were spun down, the drug-containing medium was removed, and fresh complete medium was added and used to resuspend the cells, which were then spun again. After removing the medium, the cells were resuspended in complete medium and incubated for an additional 70 hours. Cell proliferation was assessed using the CellTiter Glo assay (Promega) according to standard protocols. Luminescence was measured using a TECAN plate reader. Percent growth inhibition was calculated using the following formula: % Inhibition = (Luminescence Control - Luminescence Treatment) / Luminescence Control x 100 IC 50 Curves were generated using nonlinear regression analysis (4 parameters) with GraphPad Prism 9. Exemplary biological activities of the compounds are shown in the table below.

[0593] [Table 9-1]

[0594] [Table 9-2]

[0595] [Table 9-3]

[0596] [Table 9-4]

[0597] [Table 9-5]

[0598] comparative activity The compounds of the present invention (25 and 68) use an ethylenediamine moiety to connect the targeting ligand to the linker or spacer of the SMDC. The inventors unexpectedly discovered that the use of an ethylenediamine moiety significantly improves the internalization and H524 activity of the conjugate compared to conjugates that utilize a piperazine moiety to connect the targeting ligand to the linker or spacer. This is demonstrated in the comparative examples below, where, as shown in Table 2, the SMDC with an ethylenediamine moiety exhibits a 5- to 10-fold increase in internalization compared to the corresponding conjugate with a piperazine moiety.

[0599] The structures of comparative compounds 1 (Compound 1) and 2 (Compound 2) having a piperazine moiety are shown below.

[0600] [ka]

[0601] The structures of their corresponding SMDCs of the present invention bearing an ethylenediamine moiety are shown below.

[0602] [ka]

[0603] [Table 10]

[0604] Example L: H524 Tumor Biodistribution Model In this example, 5 million H524 (small cell lung cancer) cells in 1:1 Matrigel:cold medium (RPMI1640) were injected subcutaneously (SQ) into the flanks of female athymic nude mice. Tumor volumes and body weights were measured twice weekly. Tumors were grown to 250-500 mm before randomizing the animals into groups. 3The tumors were allowed to grow until they reached a size of 100 nmol / kg (n=3 / time point). Animals were intravenously injected with a single dose of 500 nmol / kg of the SMDCs of the present invention. Plasma and tissues were collected 1, 4, 24, and 72 hours after compound administration after whole-body perfusion with saline to remove any vascular signals, and the concentrations of the conjugate and MMAE were measured by LC / MS-MS. The tumor and plasma levels of SMDC and MMAE are shown in Table 4. BLQ: below limit of quantification.

[0605] [Table 11]

[0606] [Table 12]

[0607] [Table 13]

[0608] [Table 14]

[0609] Example M: ​​In female Swiss nude mice bearing tumors derived from the AR42J rat pancreatic cancer cell line 111 Biodistribution of In-compound 1. Study Summary: 24 hours prior to the start of the biodistribution study: 111 In-Compound 1 was radiolabeled as described herein. On the day of the study, animals were radiolabeled as described in Table 2. 111 In-labeled compound 1 (1 nmol) was administered via a catheter into the tail vein with a single IV injection of 200 μL. Block studies (group 6) were performed with 1 nmol of 111 100 nmol of In-compound 1 115 In-combined with Compound 1.

[0610] [Table 15]

[0611] After drug administration, animals were euthanized at specific time points (0.5, 1, 2, 6, and 22 hours), and organs (blood, tumor, heart, kidney, pituitary gland, brain, liver, spleen, lung, intestine, adrenal gland, pancreas, stomach, bone (femur), and tail) were collected and weighed, and radioactivity was assessed in each organ / tissue. Activity was quantified and expressed as %ID / g (percentage of initial dose / gram of tissue).

[0612] Biodistribution Results: In the AR42J xenograft tumor model, 111 In-Compound 1 showed high and sustained uptake in SST2R-positive tumors. 111 Incorporation of In-compound 1 was measured using a 100-fold molar excess of non-radioactive 115 Co-administration of In-Compound 1 111 The tumor uptake of In-labeled compound-15 was significantly reduced by competition studies, demonstrating its specificity for SST2R (Figure 1). Only the kidneys showed excess non-radioactive 115 was not blocked by co-administration of In-compound 1 111 In-Compound 1 showed nonspecific uptake and urinary excretion 111 This suggests that In- is the major pathway for compound 1 elimination.

[0613] The examples and embodiments described herein are for illustrative purposes only, and various modifications or changes suggested to those skilled in the art are to be included within the spirit and scope of this application and the appended claims.

Claims

1. A compound having the structure of formula (I): 【Chemistry 1】 During the ceremony, A is —N(H)— or —O—; R ａ is hydrogen or C １ -C ６ is alkyl, R ２ is hydrogen or C １ -C ６ is alkyl, R ６ is chloro or -C(=O)NH ２ and L is -L １ -L ２ - and L １ is an optional spacer, L ２ is an optional linker, L １ or L ２ and at least one of R ｄ is a payload moiety comprising a chemotherapeutic agent, or a pharmaceutically acceptable salt thereof.

2. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein A is -N(H)-.

3. R ２ 3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein is hydrogen.

4. R ａ The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein is hydrogen or methyl.

5. R ａ The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, wherein is hydrogen.

6. R ６ The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein is chloro.

7. L ２ exists, and -(L ２ａ ) ｗ -L ２ｂ -or-L ２ｃ - and Each L ２ａ are independently selected from natural or unnatural amino acids, and any free amine of the amino acid is optionally independently selected from -CH ３ is replaced by L ２ｂ However, absent or -N(R １０ ) (unsubstituted or substituted benzyl)-OC(=O)-, wherein the substituted benzyl is -C(=O)NHR １２ or is substituted with a monosaccharide, Each R １０ are independently hydrogen and C １ -C ６ alkyl, Each R １２ are independently hydrogen, C ４ -C ２０ Polyethylene glycol, and unsubstituted or substituted C １ -C ６ alkyl, wherein the substituted C １ -C ６ The alkyl is —NHR １３ , -C(=O)NHR １３ , or —NHC(═O)R １３ is replaced by Each R １３ are independently hydrogen, C ４ -C ２０ Polyethylene glycol, and C ４ -C ２０ Polyethylene glycol-NH ２ Selected from or or R １３ exists and L ２ａ If there is at least one free carboxylic group of the amino acid of １３ and L ２ａ the free carboxylic groups of the amino acids are joined together to form a ring, w is 1, 2, 3, 4, 5, or 6; Each L ２ｃ The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, wherein is N-maleimidomethyl-cyclohexane-1-carbonyl (MCC) or -S-.

8. L ２ｂ but, 【Chemistry 2】 8. The compound of claim 7, selected from:

9. 9. The compound of claim 7 or 8, wherein the compound has the structure of Formula (Ia), or a pharmaceutically acceptable salt thereof. 【Transformation 3】

10. Each L ２ａ are independently selected from natural or unnatural amino acids, and any free amine of the amino acid is optionally independently selected from -CH ３ and the natural or unnatural amino acid is substituted with alanine (Ala), Ala(SO ３ H), 3-(1-piperidinyl)alanine, cyclohexylalanine, arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), homophenylalanine, proline (Pro), serine (Ser), 3-homoserine, tyrosine (Tyr), Tyr(SO ３ 10. The compound according to any one of claims 7 to 9, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of β-H, valine (Val), citrulline, β-alanine, β3-homoserine, β3-homolysine, and β3-homoglutamic acid.

11. Each L ２ａ are independently selected from natural or unnatural amino acids, and any free amine of the amino acid is optionally independently selected from -CH ３ and the natural or unnatural amino acid is substituted with alanine (Ala), Ala(SO ３ 11. The compound according to any one of claims 7 to 10, or a pharmaceutically acceptable salt thereof, wherein the amino acid is selected from the group consisting of arginine (Arg), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), serine (Ser), valine (Val), and citrulline.

12. L ２ But, -(L ２ａ ) ｗ -L ２ｂ - and -(L ２ａ ) ｗ 12. The compound according to any one of claims 7 to 11, or a pharmaceutically acceptable salt thereof, wherein - is valine-citrulline, valine-alanine, methionine-valine-lysine, glycine-phenylalanine-glycine-glycine, tyrosine-arginine-valine, arginine-valine, and phenylalanine-lysine.

13. 8. The compound of claim 7, wherein the compound has the structure of formula (Ib): 【Chemistry 4】

14. L ２ is absent, or 【Transformation 5】 、 -S-, 【Chemistry 6-1】 【Chemistry 6-2】 【Transformation 6-3】 【Chemistry 6-4】 【Transformation 6-5】 【Transformation 6-6】 [Transformation 6-7] [Transformation 6-8] 2. The compound of claim 1, wherein:

15. L ２ is absent, or 【Transformation 7】 2. The compound of claim 1, wherein:

16. L １ exists and -X ２ -L ３ -L ４ - and X ２ is -C(=O)(CH ２ ) ｐ -, -(CH ２ ) ｐ -, -C(=O)CH(CH ２ SO ３ H) NHC(=O)-, or -(X ２ａ ) ｐ - and each X ２ａ are independently selected from natural or unnatural amino acids, and any free amine of the amino acid is optionally independently selected from -CH ３ is replaced by p is 0, 1, 2, 3, 4, 5, or 6; L ３ is absent, unsubstituted or substituted C １ -C １０ Alkylene, unsubstituted or substituted C １ -C １０ Heteroalkylene, C ４ -C ２０ Polyethylene glycol, or -(X ３ CH ２ CH ２ ) ｔ - and each X ３ are independently O and NR １０ is selected from each t is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; L ４ But absent or -L ４ａ - (CH ２ ) ｕ -L ４ｂ - (CH ２ ) ｕ -L ４ｃ - and L ４ａ But absent, -O-, -NR １０ -, -NR １０ C(=O)-, -C(=O)NR １０ - or -C(=O)-, L ４ｂ is absent or is an unsubstituted or substituted N-containing 5-10 membered heterocycloalkylene, and any free amine of said N-containing 5-10 membered heterocycloalkylene is optionally independently selected from -CH ２ CO ２ is substituted with H, L ４ｃ But absent, -O-, -NR １０ -, -NR １０ C(=O)-, -C(=O)NR １０ -, -C(=O)NR １０ (CH ２ ) ｕ O (CH ２ ) ｕ C(=O)-, CH(CH ２ SO ３ H)C(=O)NR １０ (CH ２ ) ｕ O (CH ２ ) ｕ C(=O)-, -C(=O)-, -CH(=N)-, -CH(=N-NH)-, -CCH ３ (=N)-,-CCH ３ (=N-NH)-, -C(=O)-(C １ -C ６ alkylene)-, -C(=O)NR １０ -(C １ -C ６ alkylene)-, -NR １０ C(=O)-(C １ -C ６ alkylene)-, -NR １０ -(C １ -C ６ alkylene)-, or C １ -C ６ alkylene-, each u is independently 0, 1, 2, 3, 4, 5, or 6; Each R １０ are independently hydrogen and C １ -C ６ 11. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein:

17. X ２ is -C(=O)(CH ２ ) ｐ - and p is 0, 1, 2, 3, or 4; L ３ is unsubstituted or substituted C １ -C １０ Heteroalkylene, C ４ -C ２０ Polyethylene glycol, or -(X ３ CH ２ CH ２ ) ｔ - and each X ３ are independently O and NR １０ is selected from 12. The compound of claim 11, or a pharmaceutically acceptable salt thereof, wherein each t is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.

18. L ４ But absent or -L ４ａ - and L ４ａ But, -NR １０ -, -NR １０ C(=O)-, -C(=O)NR １０ 12. The compound according to claim 10 or 11, wherein the aryl group is -, or -C(=O)-, or a pharmaceutically acceptable salt thereof.

19. L １ is absent, or 【Chemistry 8-1】 【Chemistry 8-2】 【Chemistry 8-3】 11. The compound of claim 10, wherein:

20. L １ is absent, or 【Chemistry 9】 11. The compound of claim 10, wherein:

21. L, 【Chemistry 10-1】 【Chemistry 10-2】 【Chemistry 10-3】 [Chemistry 10-4] [Transformation 10-5] 【Chemistry 10-6】 【Chemistry 10-7】 [Transformation 10-8] 【Chemistry 10-9】 【Chemistry 10-10】 【Chemistry 10-11】 [Chemistry 10-12] [Chemistry 10-13] [Chemistry 10-14] [Chemistry 10-15] [Chemistry 10-16] 【Chemistry 10-17】 [Chemistry 10-18] [Chemistry 10-19] [Chemistry 10-20] [Chemistry 10-21] [Chemistry 10-22] [Chemistry 10-23] [Chemistry 10-24] [Chemistry 10-25] [Chemistry 10-26] [Chemistry 10-27] [Chemistry 10-28] 2. The compound of claim 1, wherein:

22. R ｄ but, 【Chemistry 11】 17. The compound according to any one of claims 1 to 16, wherein:

23. R ｄ but, 【Chemistry 12】 18. The compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, wherein:

24. -L-R ｄ but, 【Chemistry 13-1】 【Chemistry 13-2】 【Chemistry 13-3】 【Chemistry 13-4】 【Chemistry 13-5】 【Chemistry 13-6】 【Chemistry 13-7】 【Chemistry 13-8】 【Chemistry 13-9】 【Chemistry 13-10】 【Chemistry 13-11】 [Chemistry 13-12] 【Chemistry 13-13】 [Chemistry 13-14] [Chemistry 13-15] [Chemistry 13-16] [Chemistry 13-17] [Chemistry 13-18] [Chemistry 13-19] [Chemistry 13-20] [Chemistry 13-21] [Chemistry 13-22] [Chemistry 13-23] [Chemistry 13-24] [Chemistry 13-25] [Chemistry 13-26] [Chemistry 13-27] [Chemistry 13-28] [Chemistry 13-29] 【Chemistry 13-30】 【Chemistry 13-31】 [Chemistry 13-32] 【Chemistry 13-33】 [Chemistry 13-34] [Chemistry 13-35] 16. The compound according to any one of claims 1 to 15, wherein: 【Request Item 25】 【Chemistry 14-1】 【Chemistry 14-2】 【Chemistry 14-3】 【Chemistry 14-4】 【Chemistry 14-5】 【Chemistry 14-6】 【Chemistry 14-7】 【Chemistry 14-8】 【Chemistry 14-9】 【Chemistry 14-10】 【Chemistry 14-11】 【Chemistry 14-12】 [Chemistry 14-13] 【Chemistry 14-14】 [Chemistry 14-15] [Chemistry 14-16] [Chemistry 14-17] [Chemistry 14-18] [Chemistry 14-19] 【Chemistry 14-20】 [Chemistry 14-21] [Chemistry 14-22] [Chemistry 14-23] [Chemistry 14-24] [Chemistry 14-25] [Chemistry 14-26] [Chemistry 14-27] [Chemistry 14-28] [Chemistry 14-29] 【Chemistry 14-30】 【Chemistry 14-31】 [Chemistry 14-32] [Chemistry 14-33] [Chemistry 14-34] [Chemistry 14-35] or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

26. A compound having the structure of formula (A): 【Chemistry 15】 During the ceremony, A is —N(H)— or —O—; X １ But -N(R ａ )-, -O-, -C(=O)-, -C(=O)N(R ａ )-, -S(=O)-,-(CH ２ ) C (R ａ )=N-O-(CH ２ ) - and R ａ is hydrogen or C １ -C ６ is alkyl, R ｂ is hydrogen or C １ -C ６ Is it alkyl? or R ａ and R ｂ If both R ａ and R ｂ together with the middle atom to which they are attached form piperidine or pyrrolidine, q is 1, 2, or 3; R １ is hydrogen, R ２ is hydrogen or C １ -C ６ is alkyl, R ３ is hydrogen, -OR ８ , -N(R ８ ) ２ , -CN, halogen, C １ -C ６ Alkyl or C １ -C ６ fluoroalkyl or or R ２ and R ３ together with the middle atom to which they are attached to form morpholine, Each R ４ and R ５ are independently hydrogen, halogen, C １ -C ６ Alkyl, C １ -C ６ Fluoroalkyl, substituted or unsubstituted C １ -C ６ Heteroalkyl, —CN, —N(R ８ ) ２ , or -OR ８ and m is 1, 2, or 3; R ｃ but, 【Chemistry 16】 and Each R ６ and R ７ are independently hydrogen, halogen, C １ -C ４ Alkyl, C １ -C ４ Fluoroalkyl, C ２ -C ４ Alkenyl, C ２ -C ４ Alkynyl, substituted or unsubstituted C １ -C ６ heteroalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted C ３ -C ６ Cycloalkyl, —CN, —OR ８ , -CO ２ R ８ , -C(=O)N(R ８ ) ２ , -N(R ８ ) ２ , -NR ８ C(=O)R ９ , -NR ８ C(=O)OR ９ , -SR ８ , -S(=O)R ９ , -SO ２ R ９ , or -SO ２ N (R ８ ) ２ and n is 1, 2, or 3; Each R ８ are independently hydrogen, C １ -C ４ Alkyl, C １ -C ４ Fluoroalkyl, substituted or unsubstituted C １ -C ４ is heteroalkyl, Each R ９ became independent and C １ -C ４ Alkyl, C １ -C ４ Fluoroalkyl, substituted or unsubstituted C １ -C ４ is heteroalkyl, L is -L １ -L ２ - and L １ is an optional spacer, L ２ is an optional linker, L １ Or L ２ and at least one of R ｄ is a payload moiety comprising a chelating moiety or a radionuclide conjugate thereof, or a pharmaceutically acceptable salt thereof.

27. A compound having the structure of formula (A): 【Chemistry 17】 During the ceremony, A is —N(H)— or —O—; X １ But -N(R ａ )-, -O-, -C(=O)-, -C(=O)N(R ａ )-, -S(=O)-,-(CH ２ ) C (R ａ )=N-O-(CH ２ ) - and R ａ is hydrogen or C １ -C ６ is alkyl, R ｂ is hydrogen or C １ -C ６ Is it alkyl? or R ａ and R ｂ If both R ａ and R ｂ together with the middle atom to which they are attached form piperidine or pyrrolidine, q is 1, 2, or 3; R １ is hydrogen, R ２ is hydrogen or C １ -C ６ is alkyl, R ３ is hydrogen, -OR ８ , -N(R ８ ) ２ , -CN, halogen, C １ -C ６ Alkyl or C １ -C ６ fluoroalkyl or or R ２ and R ３ together with the middle atom to which they are attached to form morpholine, Each R ４ and R ５ are independently hydrogen, halogen, C １ -C ６ Alkyl, C １ -C ６ Fluoroalkyl, substituted or unsubstituted C １ -C ６ Heteroalkyl, —CN, —N(R ８ ) ２ , or -OR ８ and m is 1, 2, or 3; R ｃ but, [Chemistry 18] and Each R ６ and R ７ are independently hydrogen, halogen, C １ -C ４ Alkyl, C １ -C ４ Fluoroalkyl, C ２ -C ４ Alkenyl, C ２ -C ４ Alkynyl, substituted or unsubstituted C １ -C ６ heteroalkyl, substituted or unsubstituted phenyl, substituted or unsubstituted C ３ -C ６ Cycloalkyl, —CN, —OR ８ , -CO ２ R ８ , -C(=O)N(R ８ ) ２ , -N(R ８ ) ２ , -NR ８ C(=O)R ９ , -NR ８ C(=O)OR ９ , -SR ８ , -S(=O)R ９ , -SO ２ R ９ , or -SO ２ N (R ８ ) ２ and n is 1, 2, or 3; Each R ８ are independently hydrogen, C １ -C ４ Alkyl, C １ -C ４ Fluoroalkyl, substituted or unsubstituted C １ -C ４ is heteroalkyl, Each R ９ became independent and C １ -C ４ Alkyl, C １ -C ４ Fluoroalkyl, substituted or unsubstituted C １ -C ４ is heteroalkyl, L is -L １ -L ２ - and L １ is an optional spacer, L ２ is an optional linker, L １ Or L ２ and at least one of R ｄ is a payload moiety comprising a chelating moiety or a radionuclide conjugate thereof, or a pharmaceutically acceptable salt thereof.

28. R ２ is hydrogen, and R ３ is hydrogen, —OH, or —OCH ３ or R ２ and R ３ 28. The compound of claim 26 or 27, or a pharmaceutically acceptable salt thereof, wherein:

29. Each R ４ and R ５ are independently hydrogen, F, Cl, Br, C １ -C ４ Alkyl, C １ -C ４ Fluoroalkyl, —CN, —N(R ７ ) ２ , or -OR ７ 29. The compound according to any one of claims 26 to 28, wherein:

30. Each R ４ and R ５ are independently hydrogen, F, Cl, Br, -CH ３ , -CH ２ F, -CHF ２ , -CF ３ , -CN, -NH ２ , -NHCH ３ , -N(CH ３ ) ２ , —OH, —OCH ３ , or -OCF ３ 30. The compound according to any one of claims 26 to 29, or a pharmaceutically acceptable salt thereof, wherein:

31. A is —N(H)—, and R ａ is hydrogen, -CH ３ , or -CH ２ CH ３ and R ｂ is hydrogen, -CH ３ , or -CH ２ CH ３ or R ａ and R ｂ If both R ａ and R ｂ together with the middle atom to which they are attached to form a piperidine, and R １ is hydrogen, -CH ３ , or -CH ２ CH ３ 31. The compound according to any one of claims 26 to 30, wherein:

32. 32. The compound of any one of claims 26-31, wherein the compound has the structure of formula (II), or a pharmaceutically acceptable salt thereof: 【Chemistry 19】 【Request Item 33】 【Chemistry 20】 but, 【Chemistry 21】 33. The compound according to any one of claims 26 to 32, or a pharmaceutically acceptable salt thereof, wherein:

34. Each R ６ and R ７ are independently hydrogen, F, Cl, Br, -CH ３ , -CH ２ CH ３ , -CO ２ H, -CO ２ CH ３ , -CO ２ CH ２ CH ３ , -CH ２ F, -CHF ２ , -CF ３ , -CH=CH ２ , -C≡CH, -CN, -OH, -OCH ３ , -OCH ２ CH ３ , -OCF ３ , —C(═O)NH ２ , -C(=O)NHCH ３ , -C(=O)N(CH ３ ) ２ , -NH ２ , -NHCH ３ , -N(CH ３ ) ２ , -NHC(=O)CH ３ , -NCH ３ C(=O)CH ３ , -SO ２ CH ３ , -SO ２ NH ２ , -SO ２ NHCH ３ , or -SO ２ N (CH ３ ) ２ 34. The compound according to any one of claims 26 to 33, wherein: 【Request Item 35】 【Chemistry 22】 but, 【Chemistry 23】 35. The compound according to any one of claims 26 to 34, wherein:

36. R ｄ but, 【Chemistry 24】 36. The compound of any one of claims 26 to 35, or a radionuclide complex thereof, or a pharmaceutically acceptable salt thereof.

37. L-R ｄ が、-CH ２ CH ２ NH-R ｄ 、-C(=O)CH ２ NH-R ｄ 、-C(=O)CH ２ CH ２ NH-R ｄ 、-CH ２ CH ２ (OCH) ２ CH ２ ) ２ NH-R ｄ 、-CH ２ CH ２ (OCH) ２ CH ２ ) ３ NH-R ｄ 、-CH ２ CH ２ (OCH) ２ CH ２ ) ４ NH-R ｄ 、-CH ２ CH ２ (OCH) ２ CH ２ ) ５ NH-R ｄ 、-CH ２ CH ２ (OCH) ２ CH ２ ) ６ NH-R ｄ 、-CH ２ CH ２ (OCH) ２ CH ２ ) ７ NH-R ｄ 、-CH ２ CH ２ (OCH) ２ CH ２ ) ８ NH-R ｄ 、-C(=O)CH ２ CH ２ (OCH) ２ CH ２ ) ２ NH-R ｄ 、-C(=O)CH ２ CH ２ (OCH) ２ CH ２ ) ３ NH-R ｄ 、 -C(=O)CH ２ CH ２ (OCH ２ CH ２ ) ４ NH-R ｄ 、 -C(=O)CH ２ CH ２ (OCH ２ CH ２ ) ５ NH-R ｄ 、 -C(=O)CH ２ CH ２ (OCH ２ CH ２ ) ６ NH-R ｄ 、 -C(=O)CH ２ CH ２ (OCH ２ CH ２ ) ７ NH-R ｄ 、 or -C(=O)CH ２ CH ２ (OCH ２ CH ２ )<> ８ NH-R ｄ and R ｄ but, 【Chemistry 25】 、 36. The compound of any one of claims 26 to 35, or a radionuclide complex thereof, or a pharmaceutically acceptable salt thereof.

38. -L １ -R ｄ is -(PEG2)NH-R ｄ -(PEG3)NH-R ｄ -(PEG4)NH-R ｄ -(PEG5)NH-R ｄ -(PEG6)NH-R ｄ -(PEG7)NH-R ｄ -(PEG8)NH-R ｄ -C(=O)(PEG2)NH-R ｄ -C(=O)(PEG3)NH-R ｄ -C(=O)(PEG4)NH-R ｄ -C(=O)(PEG5)NH-R ｄ -C(=O)(PEG6)NH-R ｄ -C(=O)(PEG7)NH-R ｄ or -C(=O)(PEG8)NH-R ｄ and R ｄ but, 【Chemistry 26】 、 or a radionuclide complex thereof.

36. The compound according to any one of claims 26 to 35, or a pharmaceutically acceptable salt thereof.

39. -L １ -R ｄ but, 【Chemistry 27】 36. The compound of any one of claims 26 to 35, or a radionuclide complex thereof, or a pharmaceutically acceptable salt thereof.

40. The radionuclide of the radionuclide conjugate is copper-64 ( ６４ Cu), 67-copper ( ６７ Cu), 111-indium ( １１１ In), 115-indium ( １１５ In), 67-gallium ( ６７ Ga), 68-gallium ( ６８ Ga), 225-actinium ( ２２５ Ac), 175-lutetium ( １７５ Lu), 177-lutetium ( １７７ Lu), or 212-lead ( ２１２ Pb), or a pharmaceutically acceptable salt thereof.

41. The radionuclide of the radionuclide conjugate is 111-indium ( １１１ In), 115-indium ( １１５ In), 67-gallium ( ６７ Ga), 68-gallium ( ６８ Ga), 225-actinium ( ２２５ Ac), 175-lutetium ( １７５ Lu), or 177-lutetium ( １７７ Lu), or a pharmaceutically acceptable salt thereof. 【Request Item 42】 【Chemistry 28】 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

43. A pharmaceutical composition comprising a compound according to any one of claims 1 to 42, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

44. 43. A method for treating cancer, comprising administering an effective amount of a compound according to any one of claims 1 to 42, or a pharmaceutically acceptable salt thereof, to a mammal having cancer.

45. 43. A method for treating a tumor, comprising administering an effective amount of a compound according to any one of claims 1 to 42, or a pharmaceutically acceptable salt thereof, to a mammal having a tumor.

46. 46. ​​The method of claim 44 or 45, wherein the mammal has anal cancer, bladder cancer, intestinal cancer, brain cancer, breast cancer, colon cancer, colorectal cancer, endometrial cancer, esophageal cancer, gallbladder cancer, stomach cancer, heart cancer, kidney cancer, lung cancer, liver cancer, melanoma, uterine cancer, lymphoma, ovarian cancer, pancreatic cancer, prostate cancer, thymic cancer, pheochromocytoma, medullary thyroid cancer, or head and neck cancer.

47. 46. ​​The method of claim 44 or 45, wherein the mammal has an endocrine cancer.

48. 48. The method of claim 47, wherein the endocrine cancer comprises an adrenal tumor, a neuroendocrine tumor, a parathyroid tumor, a pituitary tumor, or a thyroid tumor.

49. 46. ​​The method of claim 44 or 45, wherein the mammal has a neuroendocrine tumor.

50. 46. ​​The method of claim 44 or 45, wherein the mammal has a somatostatin receptor-positive entero-pancreatic neuroendocrine tumor (GEP-NET).

51. 43. A method for targeting the delivery of a chemotherapeutic agent to a tumor in a mammal, comprising administering to a mammal having a tumor a compound of any one of claims 1 to 42, or a pharmaceutically acceptable salt thereof.

52. 43. A method for killing a tumor that overexpresses the somatostatin subtype-2 receptor (SST2R), comprising administering to a mammal bearing said tumor a compound according to any one of claims 1 to 42, or a pharmaceutically acceptable salt thereof.

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