Antibody-drug conjugates

Abstract

The present disclosure provides antibody-drug conjugates (ADCs), linkers, and compounds thereof that are useful in the treatment of cancer.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 389,743, filed on July 15, 2022, U.S. Provisional Application No. 63 / 400,703, filed on August 24, 2022, U.S. Provisional Application No. 63 / 489,473, filed on March 10, 2023, and U.S. Provisional Application No. 63 / 489,474, filed on March 10, 2023, all of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present disclosure generally relates to antibody-drug conjugates and methods for using the same to treat cancer and other diseases. Background Art

[0004] Although various chemotherapeutic agents have been developed, many chemotherapeutic agents often exhibit unacceptable toxicity and or lack specificity for cancer cells compared to their specificity for non-cancerous tissues. In order to avoid the nonspecific cytotoxic effects of chemotherapeutic agents, targeted antibody therapy has revolutionized cancer treatment through several monoclonal antibodies that have shown clinical potential. Because antibodies against tumor-specific antigens generally lack therapeutic activity, the antibodies have been conjugated to cytotoxic agents to combine the effectiveness of chemotherapy with the targeting properties of antibodies. In principle, the selective delivery of cytotoxic agents to specific tumor tissues through antibody binding should reduce the systemic toxicity of traditional small molecule chemotherapeutic agents.

[0005] Since a successful antibody drug conjugate (ADC) approach must successfully bind to the target antigen to deliver the toxic payload to the target cell without significant binding to non-target cells, it is crucial that the ADC is able to deliver the toxic payload to the target cell, be internalized thereby, and then release the payload once inside the appropriate compartment within the cell.

[0006] Despite an increased understanding of tumor-specific proteins for targeting using ADC therapy, there remains an unmet need in the art for ADCs that can be used for therapeutic purposes in the treatment of cancer. Summary of the Invention

[0007] In various aspects, the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0008] LD

[0009] Formula (III)

[0010] Wherein in formula (III):

[0011] L is of the formula R*-L1-L A - connector;

[0012] R* is maleimide;

[0013] L1 is -[CH2] 1-3 -C(O)NH-;

[0014] L A is -[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA -, wherein p is an integer from 5 to 10, and X AA is an amino acid sequence having two amino acid portions; and

[0015] D is selected from

[0016]

[0017]

[0018] In some embodiments, L1 is -[CH2]2-C(O)NH-. In some embodiments, p is 7 or 8. In some embodiments, p is 8. In some embodiments, X AA is selected from the group consisting of Val-Ala, Tyr-Arg, Phe-Arg, Val-Gln, Val-Cit, Tyr-Met, Leu-Gln, Val-Arg, Met-Thr, Phe-Gln, Thr-Thr, Val-Thr, Ala-Ala, Val-Met, Leu-Met, Ala-Asn, D-Val-D-Gln, D-Ala-D-Ala, and Phe-Met. In some embodiments, X AA In some embodiments, L A is -[CH2CH2O] p -(CH2) 1-3 -C(O)-X AA -. In some embodiments, L A is -[CH2CH2O] p -(CH2)2-C(O)-X AA -. In some embodiments, D is In some embodiments, the linker L has the formula:

[0019]

[0020] In some embodiments, D is In some embodiments, the compound has the formula:

[0021]

[0022] In various aspects, the present disclosure provides an antibody-drug conjugate (ADC) having formula (I):

[0023] Ab-[LD] n

[0024] Formula (I)

[0025] Wherein in formula (I):

[0026] Ab comprises an antibody or a binding fragment thereof;

[0027] L is the formula -R * -L1-L A - connector;

[0028] R * It is succinimide;

[0029] L1 is -[CH2] 1-3 -C(O)NH-;

[0030] L A is -[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA -, wherein p is an integer from 5 to 10, and X AA is an amino acid sequence having two amino acid portions;

[0031] D is selected from

[0032]

[0033] and

[0034] n is an integer from 1 to 20. In some embodiments, L1 is In some embodiments, p is 7 or 8. In some embodiments, p is 8. In some embodiments, X AAis selected from the group consisting of Val-Ala, Tyr-Arg, Phe-Arg, Val-Gln, Val-Cit, Tyr-Met, Leu-Gln, Val-Arg, Met-Thr, Phe-Gln, Thr-Thr, Val-Thr, Ala-Ala, Val-Met, Leu-Met, Ala-Asn, D-Val-D-Gln, D-Ala-D-Ala, and Phe-Met. In some embodiments, X AA In some embodiments, L A is -[CH2CH2O] p -(CH2) 1-3 -C(O)-X AA -. In some embodiments, L A is -[CH2CH2O] p -(CH2)2-C(O)-X AA In some embodiments, the linker L has the formula:

[0035]

[0036] In some embodiments, D is In some embodiments, LD has the formula:

[0037]

[0038] In some embodiments, n is an integer from 4 to 8. In some embodiments, n is 4. In some embodiments, n is 8. In some embodiments, the antibody-drug conjugate has a drug-antibody ratio (DAR) in the range of about 1 to about 10, optionally wherein the DAR is about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10, optionally the DAR is about 4, optionally the DAR is about 8.

[0039] In various aspects, the present disclosure provides a pharmaceutical composition comprising an antibody drug conjugate of formula (I) and a pharmaceutically acceptable carrier.

[0040] In various aspects, the present disclosure provides a method of treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of an antibody drug conjugate of Formula (I).

[0041] In various aspects, the present disclosure provides a method for treating cancer, comprising administering a therapeutically effective amount of a pharmaceutical composition of the present disclosure to a subject in need thereof. In some embodiments, about 24 hours after administering the therapeutically effective amount of the antibody-drug conjugate to the subject, less than about 50% of the antibody-drug conjugate is converted into a metabolite. In some embodiments, about 96 hours after administering the therapeutically effective amount of the antibody-drug conjugate to the subject, about 50% of the antibody-drug conjugate is converted into a metabolite. In some embodiments, the antibody-drug conjugate is converted into a metabolite of Formula 300:

[0042]

[0043] Formula 300. In some embodiments, the antibody-drug conjugate is converted to a metabolite of Formula 301:

[0044]

[0045] In some embodiments, the antibody-drug conjugate is converted to a metabolite of Formula 302:

[0046]

[0047] In some embodiments, the method comprises:

[0048] (a) converting the metabolite of formula 300 into the metabolite of formula 301;

[0049] (b) converting the metabolite of formula 300 into the metabolite of formula 302;

[0050] (c) converting the metabolite of formula 301 into the metabolite of formula 302; or

[0051] (d) converting the metabolite of formula 300 into the metabolite of formula 301, and converting the metabolite of formula 301 into the metabolite of formula 302.

[0052] In some embodiments, the antibody-drug conjugate is converted into a metabolite in vivo. In some embodiments, the antibody-drug conjugate is converted into a metabolite in vitro. In some embodiments, the cancer is selected from the group consisting of pancreatic cancer, breast cancer, prostate cancer, lymphoma, skin cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain cancer, head-neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck carcinoma, breast carcinoma, ovarian carcinoma, lung cancer, small-cell lung cancer, Wilms' tumor, cervical cancer, testicular cancer, bladder cancer, pancreatic cancer, gastric cancer, colon cancer, prostatic cancer, carcinoma), genitourinary cancer, thyroid cancer, esophageal cancer, myeloma, multiple myeloma, adrenal cancer, renal cell carcinoma, endometrial cancer, adrenocortical carcinoma, malignant pancreatic insulinoma, malignant carcinoid, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic granulocytic leukemia, acute myeloid leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, polycythemia vera, essential thrombocythemia, Hodgkin's disease, non-Hodgkin's lymphoma lymphoma), soft tissue sarcoma, osteosarcoma, primary macroglobulinemia or retinoblastoma, etc.In other embodiments, the cancer is an acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, triple negative breast cancer (TNBC), bronchogenic carcinoma, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, embryonal carcinoma, endothelial carcinoma, ependymoma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer, cancer), glioblastoma multiforme, glioma, head and neck cancer, hemangioblastoma, hepatoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelioma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, stomach cancer cancer), sweat gland cancer, synovioma, testicular cancer, small cell lung cancer, laryngeal cancer, uterine cancer, Wilms' tumor, blood cancer, acute erythroleukemic leukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute lymphoblastic leukemia, acute megakaryocytic leukemia, acute monocytic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute nonlymphocytic leukemia, acute promyelocytic leukemia, acute undifferentiated leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma, polycythemia vera, or Waldenstrom's macroglobulinemia.

[0053] In various aspects, the present disclosure provides an antibody-drug conjugate having formula (I):

[0054] Ab-[LD] n

[0055] Formula (I)

[0056] Wherein in formula (I):

[0057] Ab comprises an antibody or a binding fragment thereof;

[0058] n is an integer from 1 to 20; and

[0059] LD has the formula:

[0060]

[0061] In some embodiments, n is an integer from 1 to 10, from 2 to 8, or from 4 to 8, optionally, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n is 4 or 8. In some embodiments, n is 4. In some embodiments, n is 8. In some embodiments, the antibody-drug conjugate has a drug-antibody ratio (DAR) in the range of about 1 to about 10, optionally wherein the DAR is about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10, optionally the DAR is about 4, optionally the DAR is about 8.

[0062] In various aspects, the present disclosure provides a compound having Formula 30, any one of Formulas 3031-3064, or 3101-3118, or a salt, solvate, tautomer, isomer, or mixture thereof.

[0063] In various aspects, the present disclosure provides a compound having the following formula, or a salt, solvate, tautomer, isomer, or mixture thereof:

[0064]

[0065] In various aspects, the present disclosure provides an antibody-drug conjugate having any one of Formulas 1030-1064 or 1100-1118.

[0066] In various aspects, the present disclosure provides a compound of any one of Embodiments (CI)-(CXVI).

[0067] In various aspects, the present disclosure provides an antibody-drug conjugate of any one of embodiments (I)-(XVII).

[0068] In various aspects, the present disclosure provides an antibody-drug conjugate having formula (I):

[0069] Ab-[LD] n

[0070] Formula (I)

[0071] Wherein in formula (I):

[0072] Ab comprises an antibody or a binding fragment thereof;

[0073] n is 1; and

[0074] LD has the formula:

[0075]

[0076] In various aspects, the present disclosure provides an antibody-drug conjugate having formula (I):

[0077] Ab-[LD] n

[0078] Formula (I)

[0079] Wherein in formula (I):

[0080] Ab comprises an antibody or a binding fragment thereof;

[0081] n is 4; and

[0082] LD has the formula:

[0083]

[0084] In various aspects, the present disclosure provides an antibody-drug conjugate having formula (I):

[0085] Ab-[LD] n

[0086] Formula (I)

[0087] Wherein in formula (I):

[0088] Ab comprises an antibody or a binding fragment thereof;

[0089] n is 8; and

[0090] LD has the formula:

[0091]

[0092] In various aspects, the present disclosure provides a method of treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of an antibody drug conjugate of the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

[0093] The foregoing summary, as well as the following detailed description of embodiments of the present disclosure, will be better understood when read in conjunction with the appended drawings.

[0094] Figure 1 Shown are spectra showing the SEC profile of the mAbs.

[0095] Figure 2 Shown are spectra illustrating the HIC profiles of the mAbs.

[0096] Figure 3 Shown is a spectrum showing the PLRP trace of mAb. The heavy (H0) chain peak and the light (L0) chain peak are indicated as peaks.

[0097] Figure 4Spectra are shown, which show the spectrum of the PLRP analysis used to assign DAR to ADC3 based on the target antigen. The light and heavy chains of the mAb are shown (L label and H label). The average DAR was calculated to be 7.3.

[0098] Figure 5 Shown is a spectrum showing SEC analysis for purification of ADC3 based on target antigen. SEC analysis showed that the monomer purity was 97.5%.

[0099] Figure 6 Shown is a spectrum showing the HIC profile of ADC 4. With the DAR class assignment as indicated, the average DAR was calculated to be 4.

[0100] Figure 7 Shown is a spectrum showing SEC analysis for purification of ADC4 based on the target antigen. SEC analysis showed that the monomer purity was 97.6%.

[0101] Figure 8 Spectra are shown, which show the PLRP analysis used to assign DARs to ADC5 based on the target antigen. Different colors represent different conjugation times (30 minutes, 60 minutes, and 90 minutes), and little difference in the profiles was observed. The average DAR was calculated to be 6.7.

[0102] Figure 9 Shown is a spectrum showing SEC analysis for purification of target antigen-based ADC5. Different colors represent different conjugation times (30 minutes, 60 minutes, and 90 minutes), where little difference in profile is observed and unconjugated payload is minimal.

[0103] Figure 10 is a graph showing experimental data indicating mean tumor volume versus time following a single dose of ADC3 at 10 mg / kg and 6 mg / kg for MDA-MB-231.

[0104] Figure 11 is a graph showing experimental data indicating mean tumor volume versus time following three doses of ADC3 at 10 mg / kg, 6 mg / kg, and 3 mg / kg for MDA-MB-231.

[0105] Figure 12A is a graph showing experimental data demonstrating the PK profiles of mAb and ADC3 in male CD1 mouse plasma. Figure 12B A comparison between multiple doses of ADC3 and a single dose of ADC3 is shown.

[0106] Figure 13is a graph showing experimental data indicating mean tumor volume versus time following a single dose of ADC4 at 10 mg / kg for MDA-MB-231.

[0107] Figure 14 is a graph showing experimental data indicating mean tumor volume versus time following three doses of ADC4 at 10 mg / kg and 6 mg / kg for MDA-MB-231.

[0108] Figure 15A : is a graph showing experimental data demonstrating the PK profile of ADC4 at 10 mg / kg relative to the PK profile of an unconjugated mAb at the same dose in the plasma of male CD-1 mice. The PK profile of the ADC was favorable, with little difference in clearance observed between the mAb and the ADC. Figure 15B Experimental data demonstrating multiple dose administration of ADC4 in MDA-MB-231 are shown.

[0109] Figure 16 Experimental data are shown, which show that 10A-MMAE (DAR=4) produces complete regression on a multiple dose basis in MDA-MB-231. The dose was increased to 5 mg / kg (multiple doses), and complete regression was observed. The number of antigen copies of target 10 = approximately 89,438.

[0110] Figure 17 is an image showing the sequence of the labeled strands of the TyrT DNA fragment used for cross-linking studies.

[0111] Figure 18 is an autoradiograph of a denaturing polyacrylamide gel investigating the mechanism of DNA interaction of 26 with linear 32P end-labeled TyrT DNA after overnight incubation at 37°C at various concentrations.

[0112] Figure 19 is an autoradiograph of a denaturing polyacrylamide gel showing DNA interstrand crosslinks of the PBD dimer Talirine with linear 32P end-labeled TyrT DNA after overnight incubation at 37°C at various concentrations.

[0113] Figure 20 Images of DNA footprints showing the interaction of various G alkylating agents, including 19 and 26, with MS1 DNA fragments (left and left center) and HexA (right center), as well as DNA footprints showing the interaction of the PBD dimer talin with MS1 (right). Ligand concentrations are indicated at the top of the gel. Tracks labeled "GA" are markers for specific purines.

[0114] Figure 21 The sequence of the HexA DNA fragment is shown, which shows the possible monoalkylated adducts produced by the analyzed compounds. Strong DNA footprints are represented by solid lines, and weaker footprints are represented by cross-hatching.

[0115] Figure 22 Shown is a spectrum showing the HIC profile of ADC 1. With the DAR (drug-to-antibody ratio) class assignments as indicated, the average DAR was calculated to be 1.8.

[0116] Figure 23 Shown is a spectrum showing the SEC profile of ADC1; 94.3% monomer.

[0117] Figure 24 Shown is a spectrum showing the free toxin linker trace of the ADC1 sample. <2% free toxin linker could be detected in the ADC trace. Red: 100 pmol NAC-FGX11. Blue: target antigen-27 after protein precipitation; the identified peaks indicate residual protein material.

[0118] Figure 25 Shown is a spectrum showing the HIC profile of ADC2. With the DAR class assignments as indicated, the average DAR was calculated to be 4.2.

[0119] Figure 26 Shown is a spectrum showing the SEC profile of ADC2; 94.4% monomer.

[0120] Figure 27 Shown is a spectrum showing the free toxin linker trace of the ADC2 sample. 0.4% of free toxin linker could be detected in the ADC trace. Red: 100 pmol NAC product, blue: ADC2 after protein precipitation; identified peaks indicate residual protein material.

[0121] Figure 28 Shown is a spectrum showing the HIC profile of ADC 7. With the DAR class assignments as indicated, the average DAR was calculated to be 2.1.

[0122] Figure 29 Shown is a spectrum showing an example of SEC analysis used for the purification of ADC 7. The ADC contained 97.5% monomer.

[0123] Figure 30Figure 1 is a graph of experimental data demonstrating binding of ADCs to an antigen-positive cell line (A427). All ADCs had similar binding affinities to unconjugated mAbs. The data also demonstrate that unconjugated non-targeted isotype control mAbs did not bind to the antigen.

[0124] Figure 31 is a graph of experimental data showing mean tumor volume versus time after a single dose of ADC1 (Day 1) for K562. Dose-dependent regression was observed.

[0125] Figure 32 is a graph of experimental data showing the PK profiles of mAb and ADC1 in male CD1 mouse plasma. The PK profile of ADC was favorable, with little difference in clearance observed between mAb and ADC.

[0126] Figure 33 is a graph of experimental data showing mean tumor volume versus time following a single dose of ADC2 to K562 (Day 1). Regression was observed with no weight loss.

[0127] Figure 34 Figure 2 is a graph of experimental data showing the mean tumor volume versus time after a single dose of ADC2 (day 1) at both 5 mg / kg and 10 mg / kg for MDA-MB-231. Complete regression with no weight loss was observed at the higher dose. The effect of the unconjugated mAb was negligible, demonstrating the targeted cell killing ability of the ADC.

[0128] Figure 35 Figure 2 is a graph of experimental data showing the mean tumor volume versus time after multiple doses of ADC2 (days 1, 8, and 15 or day 1, 22, and 43) at both 5 mg / kg and 10 mg / kg for MDA-MB-231. Complete regression with no weight loss was observed at the higher doses. The effect of the unconjugated mAb was negligible, demonstrating the targeted cell killing ability of the ADC.

[0129] Figure 36 is a graph of experimental data showing mean tumor volume versus time following a single dose of ADC2 at doses ranging from 1 mg / kg to 10 mg / kg against PC3 (Day 1). Concentration-dependent regression was observed with no weight loss.

[0130] Figure 37 is a graph of experimental data showing mean tumor volume versus time following three doses of ADC2 at 10 mg / kg (Day 1, Day 7, and Day 14) for A427.

[0131] Figure 38 is a graph of experimental data showing the PK profiles of mAb and ADC2 in male CD1 mouse plasma. The PK profile of ADC was favorable, with little difference in clearance observed between mAb and ADC.

[0132] Figure 39A is a graph of experimental data showing mean tumor volume versus days after first dose (Q7dx3) of 3 mg / kg, 6 mg / kg and 10 mg / kg of ADC3 (DAR of 8) and 1 mg / kg, 3 mg / kg and 6 mg / kg of mAb-MMAE (DAR of 4) against NSCLC CALU-6. Figure 39B are immunohistochemistry (IHC) images showing the expression of target antigens in NSCLC cell lines.

[0133] Figure 40A is a graph of experimental data showing mean tumor volume versus days after first dose (Q7dx3) of 3 mg / kg, 6 mg / kg and 10 mg / kg of ADC3 (DAR of 8) and 1 mg / kg, 3 mg / kg and 6 mg / kg of mAb-MMAE (DAR of 4) against TNBC MDA-MB-231. Figure 40B are IHC images showing the expression of target antigens in TNBC cell lines pre-treated.

[0134] Figure 41 Summary of data from dose-range finding (DRF) and PK studies in cynomolgus monkeys using 4 mg / kg, 6 mg / kg, and 8 mg / kg of mAb-vcMMAE (DAR of 4) and 15 mg / kg, 30 mg / kg, and 45 mg / kg of ADC3 (DAR of 8).

[0135] Figure 42 is a framework for the design of a non-restricted Good Laboratory Practice (GLP) toxicology study examining ADC3 (DAR of 8) in cynomolgus monkeys.

[0136] Figure 43A is a graph of experimental data showing the PK profile of mAb-vcMMAE following three doses at 6 mg / kg in cynomolgus monkeys compared to the PK profiles of unconjugated mAb and unconjugated payload at the same doses. Figure 43B is a table summarizing additional PK data in cynomolgus monkeys (in cynomolgus plasma) following three doses of mAb-vcMMAE at 8 mg / kg, 6 mg / kg, and 4 mg / kg.

[0137] Figure 44A is a graph of experimental data showing binding of unconjugated mAb to huTR(F30-T667Q525)-8×His_T3. Figure 44B is a graph of experimental data showing the binding of mAb-vcMMAE (DAR of 4) to huTR(F30-T667 Q525)-8×His_T3. Figure 44C is a graph of experimental data showing the binding of ADC2 (DAR of 4) to huTR(F30-T667 Q525)-8×His_T3. Figure 44D is a graph of experimental data showing the binding of ADC4 (DAR of 4) to huTR(F30-T667 Q525)-8×His_T3. Figure 44E is a graph of experimental data showing the binding of ADC3 (DAR of 8) to huTR(F30-T667 Q525)-8×His_T3. Figure 44F TABLE 1 is a table summarizing additional binding data for unconjugated mAbs and ADCs to the recombinant target antigen ECD huTR(F30-T667 Q525)-8×His_T3. mAb = sequence 1. TR = target antigen.

[0138] Figure 45A is a graph of experimental data showing binding of unconjugated mAb and mAb-vcMMAE to MDA-MB-468 cells after target antigen+cleavage. Figure 45B is a graph of experimental data showing binding of unconjugated mAb and mAb-vcMMAE to PC3 cells after target antigen++ cleavage. Figure 45C is a graph of experimental data showing binding of unconjugated mAb and mAb-vcMMAE to DU145 cells after +++ cleavage of the target antigen. Figure 45D is a graph of experimental data showing binding of unconjugated mAb and mAb-vcMMAE to OVMZ-6 cells after target antigen cleavage. Figure 45E is an image of a polyacrylamide gel binding assay.

[0139] Figure 46A is a graph of experimental data demonstrating the binding affinity of unconjugated mAb to target antigen+cells. Figure 46B is a graph of experimental data demonstrating the binding affinity of ADC3 (DAR of 8) to target antigen+cells. Figure 46C is a graph of experimental data demonstrating the binding affinity of mAb-vcMMAE (DAR of 4) to mAb+ cells.

[0140] Figure 47A is a graph of experimental data showing relative cell viability (%) versus the concentration of mAb-vcMMAE (DAR of 4). Figure 47B is a graph of experimental data showing relative cell viability (%) versus the concentration of ADC3 (DAR of 8).

[0141] Figure 48A is a graph of experimental data showing relative cell viability (%) versus the concentration of mAb-vcMMAE (DAR of 4). Figure 48B is a graph of experimental data showing relative cell viability (%) versus the concentration of ADC3 (DAR of 8).

[0142] Figure 49A are graphs of experimental data demonstrating the cytotoxicity of ADC3 (DAR of 8) in a PC3 colony formation assay. Figure 49B are graphs of experimental data demonstrating the cytotoxicity of the isotype control exatecan in a PC3 colony formation assay. Figure 49C is a graph of experimental data showing colony formation (normalized to untreated colonies) versus concentration (μg / mL) of unconjugated mAb or ADC.

[0143] Figures 50A-50B A non-limiting example of a pharmacokinetic strategy for evaluating the antibody drug conjugates of the present disclosure is shown. Figure 50A Non-limiting examples of LC-MS based analysis of ADC, total mAb, and payload (eg, mAb-vcMMAE) are shown. Figure 50B Non-limiting examples of ELISA-based analysis of ADC, total mAb, and LC-MS of payload (eg, ADC3) are shown.

[0144] Figures 51A-51B Experimental data demonstrating efficacy and tolerability data of ADC3 are shown. Figure 51A Shown is a graph of experimental data demonstrating MDA-MB-231 (TNBC) treatment time versus tumor volume (dosing day 0, day 7, day 14). Copy number = 73,654 (FACS). Figure 51B is a table of experimental data showing a three-dose (Q3W) non-human primate non-GLP toxicity study. Baseline data: HNSTD = 30 mg / kg, NOAEL = 15 mg / kg (cynomolgus monkey); ILD was observed at higher doses in cynomolgus monkeys.

[0145] Figure 52 Experimental data are presented, which show that ADC3 is highly stable in mouse / human / cynomolgus monkey plasma.

[0146] Figures 53A-53C Experimental data are presented showing that trastuzumab-Compound 30 was found to be less potent than T-Dxd ADC in HER2+++ lines in in vitro studies ( Figure 53A and 53C ), but the potency of the free payload is roughly equal ( Figure 53B ). Figure 53A is a table showing the EC50 values ​​of compounds in HER2+++ strains. Figure 53B The percentage of viable cells based on the concentration of Dxd, exitecan, or TOPO1 inhibitor control is shown. Figure 53C The percentage of viable cells based on the concentration of ADC (Trastuzumab-Compound 30, Isotype-Compound 30, or Trastuzumab-Dxd) is shown.

[0147] Figure 54 Experimental data are presented demonstrating that in vivo efficacy shows that the regression achieved with ADC3 is significantly better than with The achieved regressions were more prolonged / sustained. JIMT-1CDX in vivo efficacy (HER2+).

[0148] Figure 55 The proposed cleavage mechanism of the Val-Ala bond in compound 30 is shown.

[0149] Figure 56 Shown is a table of experimental data demonstrating the cleavage rates of various linkers / payloads after 24 hours and 96 hours of reaction with papain.

[0150] Figure 57 The structures of compound 30, compound 33, and AZ-0133 are shown.

[0151] Figure 58 The structures of deruxtecan and compound 50 are shown.

[0152] Figure 59 Trastuzumab-DM1 is shown structure.

[0153] Figure 60 The structures of the metabolites followed in the liver and cynomolgus monkey microsome studies are shown.

[0154] Figures 61A-61Fis a graph of experimental data showing that in the presence of trastuzumab-compound 30 ( Figure 61A ), trastuzumab-compound 33 ( Figure 61B ), trastuzumab-compound 50 ( Figure 61C ), Trastuzumab-AZ-0133 ( Figure 61D ), trastuzumab-drugutecan ( Figure 61E ) and trastuzumab-enmei ( Figure 61F ) when incubated together with ixetine.

[0155] Figures 62A-62F is a graph of experimental data showing that in the presence of trastuzumab-compound 30 ( Figure 62A ) when incubated with Cys-compound 20 (M1); when incubated with trastuzumab-compound 33, Cys-Mal-amino-PEG8-Val-Ala-PABC-isotecan ( Figure 62B ); Formation of Cys-Mal-amino-PEG8-Ixitecan when incubated with trastuzumab-compound 50 ( Figure 62C ); Formation of Cys-Mal-amine-PEG8-Val-Ala-PABC-AZ-1033 when incubated with trastuzumab-AZ-0133 ( Figure 62D ); Formation of Cys-Mc-GGFG-DxD upon incubation with trastuzumab-derutin ( Figure 62E ); and the formation of Lys-MCC-DM1 upon incubation with trastuzumab-enmei ( Figure 62F ). DETAILED DESCRIPTION

[0156] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents and publications mentioned herein are incorporated by reference in their entirety.

[0157] definition

[0158] As used herein, the terms "administer," "administration," or "administering" refer to (1) providing, giving, administering, and / or prescribing by or under the direction of a health practitioner or their authorized agent in accordance with the present disclosure, and / or (2) placing into the body, taking, or consuming by a mammal in accordance with the present disclosure.

[0159] As used herein, the terms "co-administration," "co-administering," "administered in combination with," "administering in combination with," "simultaneous," and "concurrent" encompass the administration of two or more active pharmaceutical ingredients to a subject such that both active pharmaceutical ingredients and / or their metabolites are present in the subject's body at the same time. Co-administration includes simultaneous administration as separate compositions, administration at different times as separate compositions, or administration as a composition in which two or more active pharmaceutical ingredients are present. Simultaneous administration as separate compositions and administration as a composition in which both agents are present are preferred.

[0160] The terms "active pharmaceutical ingredient" and "drug" include the antibodies, conjugates, and compounds described herein. The terms "active pharmaceutical ingredient" and "drug" may also include those compounds described herein that bind to a protein and thereby modulate the activity of the protein.

[0161] The term "isostere" refers to a group or molecule whose chemical and / or physical properties are similar to those of another group or molecule. A "bioisostere" is a type of isostere and refers to a group or molecule whose biological properties are similar to those of another group or molecule. For example, a carboxylic acid can be replaced by one of the following bioisosteres of a carboxylic acid, including but not limited to alkyl esters (COOR), acylsulfonamides (CONR-SO2R), hydroxamic acids (CONR-OH), hydroxamic acids (CONR-OR), tetrazoles, hydroxyisoxazoles, isoxazol-3-ones, and sulfonamides (SO2NR), where each R can independently represent hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocyclylalkyl, heterocyclylalkylalkyl, heteroaryl, or heteroarylalkyl.

[0162] The term "in vivo" refers to events that occur within the body of a subject.

[0163] The term "in vitro" refers to events that occur outside the body of a subject. In vitro assays encompass cell-based assays that employ living or dead cells, and can also encompass cell-free assays that do not employ intact cells.

[0164] The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound or combination of compounds as described herein that is sufficient to achieve the intended application, including but not limited to disease treatment. A therapeutically effective amount can vary depending on the intended application (in vitro or in vivo) or the subject and the disease condition being treated (e.g., the subject's weight, age, and sex), the severity of the disease condition, the mode of administration, etc., which can be easily determined by one of ordinary skill in the art. The term is also applicable to doses that will induce a specific response in the target cell (e.g., platelet adhesion and / or cell migration reduction). The specific dosage will vary depending on the specific compound selected, the dosing regimen to be followed, whether the compound is administered in combination with other compounds, the time of administration, the tissue to which the compound is administered, and the physical delivery system in which the compound is carried.

[0165] As used herein, "therapeutic effect" encompasses therapeutic benefit and / or prophylactic benefit. A prophylactic effect includes delaying or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, stopping, or reversing the progression of a disease or condition, or any combination thereof.

[0166] As used herein, the terms "treat," "treatment," and / or "treating" may refer to the management of a disease, disorder, pathological condition, or its symptoms for the purpose of curing, improving, stabilizing, and / or controlling the disease, disorder, pathological condition, or its symptoms. With respect to control of a disease, disorder, or pathological condition, more specifically, "control" may include the absence of progression of the condition as assessed by response to the methods described herein, wherein such response may be complete (e.g., causing remission of the disease) or partial (e.g., alleviating or ameliorating any symptoms associated with the condition). As used herein, the terms "prevent," "preventing," and / or "prevention" may refer to reducing the risk of developing a disease, disorder, or pathological condition.

[0167] As used herein, the terms "modulate" and "modulation" refer to a change in the biological activity of a biomolecule (e.g., a protein, gene, peptide, antibody, etc.), wherein such change can be associated with an increase in the biological activity of the biomolecule (e.g., increased activity, agonism, activation, expression, upregulation, and / or increased expression) or a decrease in the biological activity (e.g., decreased activity, antagonism, inhibition, inactivation, downregulation, and / or decreased expression).

[0168] The terms “QD,” “qd,” or “qd” mean once a day, quaque die, or once daily. The terms “BID,” “bid,” or “bid” mean twice a day, twice a day, or twice daily. The terms “TID,” “tid,” or “tid” mean three times a day, three times a day, or three times daily. The terms “QID,” “qid,” or “qid” mean four times a day, four times a day, or four times daily.

[0169] The term "pharmaceutically acceptable salt" refers to salts derived from various organic and inorganic counterions known in the art. Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids. Preferred inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Preferred organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins. Specific examples include isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salt is selected from ammonium, potassium, sodium, calcium, and magnesium salts. The term "co-crystal" refers to a molecular complex derived from various co-crystal formers known in the art. Unlike salts, co-crystals generally do not involve hydrogen transfer between the co-crystal and the drug, but rather involve intermolecular interactions such as hydrogen bonding, aromatic ring stacking, or dispersion forces between the co-crystal former and the drug in the crystal structure.

[0170] "Pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" or "physiologically compatible" carrier or carrier medium is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and inert ingredients. The use of such pharmaceutically acceptable carriers or pharmaceutically acceptable excipients for active pharmaceutical ingredients is well known in the art. Except in cases where any conventional pharmaceutically acceptable carrier or pharmaceutically acceptable excipient is incompatible with the active pharmaceutical ingredient, its use in the therapeutic compositions of the present disclosure is contemplated. Additional active pharmaceutical ingredients (such as other drugs) may also be incorporated into the described compositions and methods.

[0171] "Prodrug" refers to a derivative of a compound described herein, the pharmacological action of which is produced by conversion to an active compound in vivo by a chemical or metabolic process. Prodrugs include compounds in which an amino acid residue or a polypeptide chain of two or more (e.g., two, three, or four) amino acid residues is covalently linked to a free amino group, a hydroxyl group, or a carboxylic acid group via an amide bond or an ester bond. Amino acid residues include, but are not limited to, 20 naturally occurring amino acids typically represented by single-letter or three-letter symbols, but also include, for example, 4-hydroxyproline, hydroxylysine, desmosine, isodesmosine, 3-methylhistidine, β-alanine, γ-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine, and methionine sulfone. Other types of prodrugs are also encompassed. For example, a free carboxyl group can be derived into an amide or an alkyl ester (e.g., methyl ester and acetoxymethyl ester). As used herein, prodrug esters include esters and carbonates formed by reacting one or more hydroxyl groups of the compounds of the methods of the present disclosure with alkyl, alkoxy or aryl substituted acylating agents using procedures known to those skilled in the art to produce acetates, pivalates, methyl carbonates, benzoates, etc. As another example, free hydroxyl groups can be derived using groups including but not limited to hemisuccinates, phosphates, dimethylaminoacetates and phosphoryloxymethoxycarbonyl groups, as summarized in Advanced Drug Delivery Reviews, 1996, 19, 115. Also included are carbamate prodrugs of hydroxyl and amino groups, such as carbonate prodrugs, sulfonate prodrugs, sulfonates and sulfates of hydroxyl groups. Free amines can also be derived into amides, sulfonamides or phosphamides. All described prodrug moieties can be incorporated into groups including but not limited to ethers, amines and carboxylic acid functional groups. In addition, any compound that can be converted in vivo to provide a bioactive agent is a prodrug within the scope of the present disclosure. Various forms of prodrugs are well known in the art. A comprehensive description of prodrugs and prodrug derivatives is described in: (a) The Practice of Medicinal Chemistry, Camille G. Wermuth et al., (Academic Press, 1996); (b) Design of Prodrugs, H. Bundgaard, ed., (Elsevier, 1985); (c) A Textbook of Drug Design and Development, P. Krogsgaard-Larson and H. Bundgaard, eds., (Harwood Academic Publishers, 1991).In general, prodrugs can be designed to improve drug penetration across biological membranes to achieve improved drug absorption, extend the duration of drug action (slow release of parent drug from the prodrug, reduced first-pass metabolism of the drug), target drug action (e.g., organ or tumor targeting, lymphocyte targeting), alter or increase the aqueous solubility of the drug (e.g., intravenous formulations and eye drops), improve local drug delivery (e.g., skin and ocular drug delivery), increase the chemical / enzymatic stability of the drug, or reduce off-target drug effects, and more generally, improve the therapeutic efficacy of the compounds used in the present disclosure.

[0172] Unless otherwise stated, chemical structures depicted herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, one or more hydrogen atoms are replaced by deuterium or tritium, or one or more carbon atoms are replaced by isotopically enriched atoms. 13 C or 14 C-enriched carbon-substituted compounds are within the scope of the present disclosure.

[0173] "Alkyl" refers to a group consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to ten carbon atoms (e.g., (C 1-10 ) alkyl or C 1-10 The term "alkyl" refers to a straight or branched hydrocarbon chain group of a group consisting of a plurality of groups. ... Unless stated otherwise specifically in the specification, an alkyl group is optionally substituted with one or more substituents which are independently heteroalkyl, acylsulfonamido, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a)2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0174] "Alkylaryl" refers to an -(alkyl)aryl group where aryl and alkyl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for aryl and alkyl respectively.

[0175] "Alkylheteroaryl" refers to an -(alkyl)heteroaryl group where heteroaryl and alkyl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for aryl and alkyl respectively.

[0176] "Alkylheterocycloalkyl" refers to an -(alkyl)heterocyclyl group where alkyl and heterocycloalkyl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for heterocycloalkyl and alkyl respectively.

[0177] An "alkene" moiety refers to a group consisting of at least two carbon atoms and at least one carbon-carbon double bond, and an "alkyne" moiety refers to a group consisting of at least two carbon atoms and at least one carbon-carbon triple bond. Alkyl moieties, whether saturated or unsaturated, may be branched, linear, or cyclic.

[0178] "Alkenyl" refers to a group consisting solely of carbon and hydrogen atoms, containing at least one double bond and having from two to ten carbon atoms (i.e., (C 2-10 ) alkenyl or C 2-10 The term "alkenyl" refers to a straight or branched hydrocarbon chain group of a group consisting of a alkylene group (e.g., an alkylene group or an alkylene group). Whenever a numerical range such as "2 to 10" appears in this article, it refers to each integer within the given range, for example, "2 to 10 carbon atoms" means that the alkenyl group can be composed of 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms. The alkenyl moiety can be attached to the rest of the molecule by a single bond, such as ethenyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl, and penta-1,4-dienyl. Unless stated otherwise specifically in the specification, an alkenyl group is optionally substituted with one or more substituents which are independently alkyl, heteroalkyl, acylsulfonamido, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R ais independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0179] "Alkenyl-cycloalkyl" refers to an -(alkenyl)cycloalkyl group where alkenyl and cycloalkyl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for alkenyl and cycloalkyl respectively.

[0180] "Alkynyl" refers to a group consisting solely of carbon and hydrogen atoms having two to ten carbon atoms (i.e., (C 2-10 ) alkynyl or C 2-10 Alkynyl) is a straight or branched hydrocarbon chain group. Whenever it occurs in this article, a numerical range such as "2 to 10" refers to each integer in a given range, for example, "2 to 10 carbon atoms" means that the alkynyl group can be composed of 2 carbon atoms, 3 carbon atoms, etc., up to 10 (and including 10) carbon atoms. Alkynyl can be attached to the rest of the molecule by a single bond, such as ethynyl, propynyl, butynyl, pentynyl and hexynyl. Unless otherwise specifically stated in the specification, alkynyl is optionally substituted by one or more substituents independently: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, acylsulfonamido, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxyl, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a(where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0181] "Alkynyl-cycloalkyl" refers to an -(alkynyl)cycloalkyl group where alkynyl and cycloalkyl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for alkynyl and cycloalkyl respectively.

[0182] "Acylsulfonamide" refers to the group -C(=O)NR a -S(=O)2R a , where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0183] "Imidazolecarboxaldehyde" refers to a (C=O)H group.

[0184] "Carbonyl" refers to the group -C(=O)-. The carbonyl group may be substituted with the following exemplary substituents: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, acylsulfonamide, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-NR a -OR a -、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a)C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0185] "Carboxy" refers to a -(C=O)OH group.

[0186] "Cyano" refers to a -CN group.

[0187] "Cycloalkyl" refers to a monocyclic or polycyclic group containing only carbon and hydrogen and may be saturated or partially unsaturated. Cycloalkyl includes groups having 3 to 10 ring atoms (i.e., (C 3-10 ) cycloalkyl or C 3-10 Cycloalkyl). Whenever it appears in this article, a numerical range such as "3 to 10" refers to each integer in the given range, for example, "3 to 10 carbon atoms" means that the cycloalkyl can be composed of 3 carbon atoms, etc., up to 10 (and including 10) carbon atoms. Illustrative examples of cycloalkyl include, but are not limited to, the following moieties: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl, etc. Unless otherwise specifically stated in the specification, cycloalkyl is optionally substituted by one or more substituents independently being: alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, acylsulfonyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxyl, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(where t is 1 or 2), -S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0188] "Cycloalkyl-alkenyl" refers to a -(cycloalkyl)alkenyl group where cycloalkyl and alkenyl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for cycloalkyl and alkenyl respectively.

[0189] "Cycloalkyl-heterocycloalkyl" refers to a -(cycloalkyl)heterocycloalkyl group where cycloalkyl and heterocycloalkyl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for cycloalkyl and heterocycloalkyl respectively.

[0190] "Cycloalkyl-heteroaryl" refers to a -(cycloalkyl)heteroaryl radical where cycloalkyl and heteroaryl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for cycloalkyl and heteroaryl respectively.

[0191] The term "alkoxy" refers to the group -O-alkyl, including one to eight carbon atoms attached to the parent structure through an oxygen in straight chain, branched, cyclic configurations, and combinations thereof. Examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, and cyclohexyloxy. "Lower alkoxy" refers to an alkoxy group containing one to six carbon atoms.

[0192] The term "substituted alkoxy" refers to an alkoxy group in which the alkyl component is substituted (i.e., -O-(substituted alkyl)). Unless stated otherwise specifically in the specification, the alkyl portion of the alkoxy group is optionally substituted with one or more substituents which are independently alkyl, heteroalkyl, alkenyl, acylsulfonamido, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0193] The term "alkoxycarbonyl" refers to a radical of the formula (alkoxy)(C=O)- attached through the carbonyl carbon, wherein the alkoxy has the indicated number of carbon atoms. Thus, (C 1-6 ) Alkoxycarbonyl is an alkoxy group having 1 to 6 carbon atoms attached through its oxygen to a carbonyl linker. "Lower alkoxycarbonyl" refers to an alkoxycarbonyl group in which the alkoxy group is a lower alkoxy group.

[0194] The term "substituted alkoxycarbonyl" refers to the group (substituted alkyl)-OC(O)-, wherein the group is attached to the parent structure through the carbonyl functionality. Unless stated otherwise specifically in the specification, the alkyl portion of the alkoxycarbonyl group is optionally substituted with one or more substituents which are independently alkyl, heteroalkyl, acylsulfonamido, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0195] "Acyl" refers to the groups (alkyl)-C(O)-, (aryl)-C(O)-, (heteroaryl)-C(O)-, (heteroalkyl)-C(O)-, and (heterocycloalkyl)-C(O)-, wherein the groups are attached to the parent structure through a carbonyl functional group. If the R group is heteroaryl or heterocycloalkyl, the heterocyclic or chain atoms contribute to the total number of chain or ring atoms. Unless otherwise specifically stated in the specification, the alkyl, aryl, or heteroaryl portion of the acyl group is optionally substituted with one or more substituents which are independently alkyl, heteroalkyl, acylsulfonamido, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0196] "Acyloxy" refers to an R(C=O)O- group, where R is an alkyl, aryl, heteroaryl, heteroalkyl, or heterocycloalkyl group as described herein. If the R group is heteroaryl or heterocycloalkyl, the heterocyclic or chain atoms contribute to the total number of chain or ring atoms. Unless otherwise specifically stated in the specification, the R of the acyloxy group is optionally substituted with one or more substituents that are independently alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0197] "Amino" or "amine" refers to -N(R a )2 group, wherein each R ais independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl or heteroarylalkyl, unless otherwise specifically stated in the specification. a )2 group has two R a When a substituent is present, it can combine with the nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For example, -N(R a )2 is intended to include, but is not limited to, 1-pyrrolidinyl and 4-morpholinyl. Unless specifically stated otherwise in the specification, an amino group is optionally substituted with one or more substituents which are independently alkyl, acylsulfonamido, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0198] The term "substituted amino" also refers to the groups -NHR, each as described above d and NR d R d N-oxides can be prepared by treating the corresponding amino group with, for example, hydrogen peroxide or m-chloroperoxybenzoic acid.

[0199] "Amide" or "amine" refers to an amino group having the formula C(O)NR a R b or -NR a C(O)R b The chemical part, where R a and R b Selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heteroalicyclic (bonded through a ring carbon), each of which moieties may itself be optionally substituted. -C(O)NR a R b R of amide a and R b It is optionally possible to form a 4-, 5-, 6- or 7-membered ring together with the nitrogen to which it is attached. Unless otherwise specifically stated in the specification, the amido group is optionally independently substituted with one or more substituents in a substituent as described herein of an alkyl, amino, cycloalkyl, aryl, heteroaryl or heterocycloalkyl. Amide can be an amino acid or peptide molecule attached to a compound disclosed herein to form a prodrug. The procedures and specific groups for preparing such amides are known to those skilled in the art and can be easily found in pioneering sources, such as Greene and Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York, NY, 1999, which are incorporated herein by reference in their entirety.

[0200] "Aromatic" or "aryl" or "Ar" refers to a radical having six to ten ring atoms (e.g., C6-C 10 Aromatic compounds or C6-C 10The term "phenyl" refers to an aromatic group (e.g., an aryl group) wherein the ring atoms have at least one ring having a conjugated π electron system that is a carbocyclic ring (e.g., phenyl, fluorenyl, and naphthyl). Divalent groups formed by substituted benzene derivatives and having free valences at the ring atoms are named substituted phenylene groups. Divalent groups derived from monovalent polycyclic hydrocarbon groups whose names end with "-" by removing one hydrogen atom from a carbon atom with a free valence are named by adding "subunit" to the name of the corresponding monovalent group, e.g., naphthyl with two attachment points is called naphthylene. Whenever a numerical range such as "6 to 10" appears in this article, it refers to each integer within the given range, e.g., "6 to 10 ring atoms" means that the aryl group can be composed of 6 ring atoms, 7 ring atoms, etc., up to 10 (and including 10) ring atoms. The term includes monocyclic or fused-ring polycyclic (i.e., rings having adjacent pairs of ring atoms) groups. Unless stated otherwise specifically in the specification, an aryl moiety is optionally substituted with one or more substituents which are independently alkyl, heteroalkyl, acylsulfonamido, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a)2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0201] "Aralkyl" or "arylalkyl" refers to an (aryl)alkyl group where aryl and alkyl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for aryl and alkyl respectively.

[0202] "Ester" refers to a chemical group of the formula -COOR, wherein R is selected from the group consisting of: alkyl, cycloalkyl, aryl, heteroaryl (bonded via a ring carbon), and heteroalicyclic compounds (bonded via a ring carbon). The procedures and specific groups for preparing esters are known to those skilled in the art and can be easily found in seminal sources such as Greene and Wuts, Protective Groups in Organic Synthesis, 3rd ed., John Wiley & Sons, New York, NY, 1999, which are incorporated herein by reference in their entirety. Unless otherwise specifically stated in the specification, an ester group is optionally substituted by one or more substituents independently selected from the group consisting of: alkyl, acylsulfonamide, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxyl, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, trimethylsilyl, -OR. a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t Ra (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0203] "Fluoroalkyl" refers to an alkyl group as defined above substituted with one or more fluoro groups as defined above, for example, trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc. The alkyl portion of the fluoroalkyl group may be optionally substituted as defined above for an alkyl group.

[0204] "Halo," "halide," or alternatively "halogen" is intended to mean fluorine, chlorine, bromine, or iodine. The terms "haloalkyl," "haloalkenyl," "haloalkynyl," and "haloalkoxy" include alkyl, alkenyl, alkynyl, and alkoxy structures substituted with one or more halo groups or combinations thereof. For example, the terms "fluoroalkyl" and "fluoroalkoxy" include haloalkyl and haloalkoxy, respectively, wherein the halo is fluorine.

[0205] "Heteroalkyl", "heteroalkenyl" and "heteroalkynyl" refer to optionally substituted alkyl, alkenyl and alkynyl groups having one or more backbone chain atoms selected from atoms other than carbon, e.g., oxygen, nitrogen, sulfur, phosphorus or combinations thereof. Numerical ranges may be given, e.g., C1-C4 heteroalkyl, which refers to the total chain length, which in this example is 4 atoms long. Heteroalkyl may be substituted with one or more substituents which are independently alkyl, heteroalkyl, alkenyl, alkynyl, acylsulfonamido, cycloalkyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thiooxo, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a)C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0206] "Heteroalkylaryl" refers to a -(heteroalkyl)aryl radical where heteroalkyl and aryl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for heteroalkyl and aryl respectively.

[0207] "Heteroalkylheteroaryl" refers to a -(heteroalkyl)heteroaryl radical where heteroalkyl and heteroaryl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for heteroalkyl and heteroaryl respectively.

[0208] "Heteroalkylheterocycloalkyl" refers to a -(heteroalkyl)heterocycloalkyl group where heteroalkyl and heterocycloalkyl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for heteroalkyl and heterocycloalkyl respectively.

[0209] "Heteroalkylcycloalkyl" refers to a -(heteroalkyl)cycloalkyl group where heteroalkyl and cycloalkyl are as disclosed herein and which are optionally substituted by one or more of the substituents described as suitable substituents for heteroalkyl and cycloalkyl respectively.

[0210] "Heteroaryl" or "heteroaromatic" or "HetAr" refers to a 5- to 18-membered aromatic group (e.g., C5-C8) that includes one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur and that can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system. 13Heteroaryl). Whenever it appears in this article, a numerical range such as "5 to 18" refers to each integer in a given range, for example, "5 to 18 ring atoms" means that the heteroaryl group can be composed of 5 ring atoms, 6 ring atoms, etc., up to 18 (and including 18) ring atoms. A divalent group derived from a monovalent heteroaryl group whose name ends with "-" by removing a hydrogen atom from an atom with a free valence is named by adding "subunit" to the name of the corresponding monovalent group, for example, a pyridyl group with two attachment points is a pyridylene group. The "heteroaromatic" or "heteroaryl" moiety containing N refers to an aromatic group in which at least one of the skeleton atoms of the ring is a nitrogen atom. Polycyclic heteroaryl groups can be fused or non-fused. The heteroatoms in the heteroaryl group are optionally oxidized. One or more nitrogen atoms (if present) are optionally quaternized. The heteroaryl group can be attached to the rest of the molecule through any atom of the ring.Examples of heteroaryl groups include, but are not limited to, azepanyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolanyl, benzofuranyl, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolanyl, benzodioxanyl, benzoxazolyl, benzopyranyl, benzopyrone, benzofuranyl, benzopyrone, benzofurazanyl, benzothiazolyl, benzothiophenyl (benzothiophenyl), benzothiophene[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazole benzo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furazanyl, furanonyl, furano[3,2-c]pyridinyl, 5,6,7,8,9,10-hexahydrocyclooctano[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocyclooctano[d]pyridazinyl, 5,6,7,8,9,10-hexahydrocyclooctano[d]pyridinyl , isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolyl, isoindolyl, isoquinolinyl, indolizinyl, isoxazolyl, isoxazol-3-one, 5,8-methanol-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepine, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d] ]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, thiapyranyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pyridinyl, and phenylthio (i.e., thienyl).Unless stated otherwise specifically in the specification, heteroaryl moieties are optionally substituted with one or more substituents which independently are: alkyl, acylsulfonamido, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo, trimethylsilanyl, -OR. a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、-N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a )2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0211] Substituted heteroaryl groups also include ring systems substituted with one or more oxide (—O—) substituents, such as pyridyl N-oxide.

[0212] "Heteroarylalkyl" refers to a moiety having an aryl moiety as described herein attached to an alkylene moiety as described herein, wherein the connection to the rest of the molecule is through the alkylene.

[0213] "Heterocycloalkyl" refers to a stable 3- to 18-membered non-aromatic ring group comprising two to twelve carbon atoms and one to six heteroatoms selected from nitrogen, oxygen and sulfur. Whenever present in this article, numerical ranges such as "3 to 18" refer to each integer within a given range, for example, "3 to 18 ring atoms" means that heterocycloalkyl can be composed of 3 ring atoms, 4 ring atoms, etc., up to 18 (and including 18) ring atoms. Unless otherwise specifically stated in the specification, heterocycloalkyl is a monocyclic, bicyclic, tricyclic or tetracyclic ring system that can include a fused ring or a bridged ring system. The heteroatoms in heterocycloalkyl can be optionally oxidized. One or more nitrogen atoms (if present) are optionally quaternized. Heterocycloalkyl is partially or completely saturated. Heterocycloalkyl can be attached to the remainder of the molecule through any atom of the ring. Examples of such heterocycloalkyl groups include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuranyl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in the specification, a heterocycloalkyl moiety is optionally substituted with one or more substituents which are independently alkyl, acylsulfonamido, heteroalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, hydroxamic acid, aryl, arylalkyl, heteroaryl, heteroarylalkyl, hydroxy, halo, cyano, nitro, oxo, thioxo, trimethylsilyl, -OR a 、-SR a 、-S(O) t R a -(wherein t is 1 or 2), -OC(O)-R a 、-N(R a )2、-C(O)R a 、-C(O)OR a 、-OC(O)N(R a )2、-C(O)N(R a )2、-N(R a )C(O)OR a 、-N(R a )C(O)R a 、

[0214] -N(R a )C(O)N(R a )2、N(R a )C(NR a )N(R a)2、-N(R a )S(O) t R a (where t is 1 or 2), -S(O) t R a (where t is 1 or 2), -S(O) t OR a (where t is 1 or 2), -S(O) t N(R a )2(where t is 1 or 2) or PO(OR a )2, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0215] “Heterocycloalkyl” also includes bicyclic ring systems in which one non-aromatic ring (typically having 3 to 7 ring atoms) contains at least 2 carbon atoms, in addition to 1-3 heteroatoms independently selected from oxygen, sulfur, and nitrogen, and combinations comprising at least one of the foregoing heteroatoms; and the other ring (typically having 3 to 7 ring atoms) optionally contains 1-3 heteroatoms independently selected from oxygen, sulfur, and nitrogen, and is not aromatic.

[0216] "Hydroxamic acid" refers to -C(O)NR a OR a part, where each R a is independently hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocycloalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0217] "Nitro" refers to a -NO2 group.

[0218] "Oxa" refers to an -O- group.

[0219] "Oxo" refers to a =0 group.

[0220] "Isomers" are different compounds with the same molecular formula. "Stereoisomers" refer to isomers that differ only in the way atoms are arranged in space (i.e., have different stereochemical configurations). "Enantiomers" are a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of an enantiomeric pair is a "racemic" mixture. The term "(±)" is used to represent a racemic mixture where appropriate. "Diastereomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is specified according to the Cahn-Ingold-Prelog RS system. When a compound is a pure enantiomer, the stereochemistry of each chiral carbon can be specified by (R) or (S). A resolved compound whose absolute configuration is unknown can be specified as (+) or (-), depending on the direction (right-handed or left-handed) in which it rotates plane polarized light at the wavelength of the sodium D line. Certain compounds described herein contain one or more asymmetric centers and can thereby produce enantiomers, diastereomers, and other stereoisomeric forms that can be defined in terms of absolute stereochemistry as (R)- or (S). The chemical entities, pharmaceutical compositions, and methods of the present invention are intended to include all such possible isomers, including racemic mixtures, optically pure forms, and mixtures of intermediates. Optically active (R)- and (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. Where the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless otherwise indicated, it is intended that the compounds include both E and Z geometric isomers.

[0221] As used herein, "enantiomeric purity" refers to the relative amount of one particular enantiomer present relative to the other enantiomer, expressed as a percentage. For example, if a compound that may potentially have an (R)- or (S)-isomer configuration is present as a racemic mixture, the enantiomeric purity relative to the (R)- or (S)-isomer is about 50%. If one isomeric form of the compound predominates over the other, for example, 80% (S)-isomer and 20% (R)-isomer, the enantiomeric purity of the compound relative to the (S)-isomer form is 80%. The enantiomeric purity of a compound can be determined in a variety of ways known in the art, including, but not limited to, chromatography using a chiral support, polarization measurement of the rotation of polarized light, nuclear magnetic resonance spectroscopy using chiral shift reagents (including but not limited to chiral complexes containing lanthanides or Pirkle's reagent), or derivatization of the compound with a chiral compound such as Mosher's acid followed by chromatography or nuclear magnetic resonance spectroscopy.

[0222] In some embodiments, in terms of therapeutic utility per unit mass, the efficacy of the enantiomer-enriched composition is higher than that of the racemic mixture of the composition. Enantiomers can be separated from the mixture by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred enantiomers can be prepared by asymmetric synthesis. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions, Wiley Interscience, New York, 1981; E.L. Eliel, Stereochemistry of Carbon Compounds, McGraw-Hill, New York, 1962; and E.L. Eliel and S.H. Wilen, Stereochemistry of Organic Compounds, Wiley-International Science, New York, 1994.

[0223] As used herein, the terms "enantiomerically enriched" and "non-racemic" refer to compositions in which the weight percentage of one enantiomer is greater than the amount of that enantiomer in a control mixture of a racemic composition (e.g., greater than 1:1 by weight). For example, an enantiomerically enriched formulation of the (S)-enantiomer means a formulation of the compound having greater than 50% by weight (e.g., at least 75% by weight or at least 80% by weight) of the (S)-enantiomer relative to the (R)-enantiomer. In some embodiments, the enrichment can be significantly greater than 80% by weight, thereby providing a "substantially enantiomerically enriched" or "substantially non-racemic" formulation, which refers to a formulation of a composition having at least 85% by weight (e.g., at least 90% by weight or at least 95% by weight) of one enantiomer relative to the other. The terms "enantiomerically pure" or "substantially enantiomerically pure" refer to a composition that contains at least 98% of a single enantiomer and less than 2% of the opposite enantiomer.

[0224] "Moiety" refers to a specific segment or functional group of a molecule. A chemical moiety is typically a recognized chemical entity embedded in or appended to a molecule.

[0225] "Tautomers" are structurally different isomers that interconvert via tautomerization. "Tautomerization" is a form of isomerization and includes prototropic tautomerization or proton-shift tautomerization and is considered a subset of acid-base chemistry. "Prototropic tautomerization" or "proton-shift tautomerization" involves the migration of a proton accompanied by a change in bond order, typically the interchange of a single bond with an adjacent double bond. In situations where tautomerization is possible (e.g., in solution), a chemical equilibrium of the tautomers can be achieved. An example of tautomerization is keto-enol tautomerization. A specific example of keto-enol tautomerization is the interconversion of pentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Another example of tautomerization is phenol-keto tautomerization. A specific example of phenol-keto tautomerization is the interconversion of pyridin-4-ol and pyridin-4(1H)-one tautomers.

[0226] A "leaving group or atom" is any group or atom that will cleave from the starting material under the reaction conditions, thereby promoting the reaction at a specific site. Unless otherwise specified, examples of such groups include halogen atoms and methanesulfonyloxy, p-nitrobenzenesulfonyloxy, and toluenesulfonyloxy groups.

[0227] "Protecting group" is intended to mean a group that selectively blocks one or more reactive sites in a multifunctional compound so that a chemical reaction can be selectively carried out at another unprotected reactive site, and the group can then be easily removed or deprotected after the selective reaction is complete. A variety of protecting groups are disclosed, for example, in TH Greene and PGM Wuts, Protecting Groups in Organic Synthesis, 3rd ed., John Wiley & Sons, New York (1999).

[0228] "Solvate" refers to a compound in physical association with one or more molecules of a pharmaceutically acceptable solvent.

[0229] "Substituted" means that the referenced group can be attached with one or more additional groups, radicals, or moieties individually and independently selected from, for example, acyl, alkyl, alkylaryl, cycloalkyl, aralkyl, aryl, carbohydrate, carbonate, heteroaryl, heterocycloalkyl, hydroxamic acid, hydroxy, alkoxy, aryloxy, thiol, alkylthio, arylthio, cyano, halo, carbonyl, ester, thiocarbonyl, isocyanato, thiocyanato, isothiocyanato, nitro, oxo, perhaloalkyl, perfluoroalkyl, phosphate, silyl, sulfinyl, sulfonyl, sulfonamido, sulfonyloxy, sulfonate, urea, and amino (including mono- and di-substituted amino groups), and protected derivatives thereof. A substituent can itself be substituted, for example, a cycloalkyl substituent can itself have a halide substituent on one or more of its ring carbons. The term "optionally substituted" means optional substitution with a specified group, radical, or moiety.

[0230] "Sulfanyl" refers to groups including -S-(optionally substituted alkyl), -S-(optionally substituted aryl), -S-(optionally substituted heteroaryl), and -S-(optionally substituted heterocycloalkyl).

[0231] "Sulfinyl" refers to groups including -S(O)-H, -S(O)-(optionally substituted alkyl), -S(O)-(optionally substituted amino), -S(O)-(optionally substituted aryl), -S(O)-(optionally substituted heteroaryl), and -S(O)-(optionally substituted heterocycloalkyl).

[0232] "Sulfonyl" refers to groups including -S(O2)-H, -S(O2)-(optionally substituted alkyl), -S(O2)-(optionally substituted amino), -S(O2)-(optionally substituted aryl), -S(O2)-(optionally substituted heteroaryl) and -S(O2)-(optionally substituted heterocycloalkyl).

[0233] "Sulfonamido" or "sulfonamido" refers to a -S(=O)2-NRR radical, where each R is independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heteroalicyclic (bonded through a ring carbon). The R groups in the -NRR of the -S(=O)2-NRR radical can be taken together with the nitrogen to which they are attached to form a 4-, 5-, 6-, or 7-membered ring. The sulfonamido group is optionally substituted with one or more of the substituents described for alkyl, cycloalkyl, aryl, and heteroaryl, respectively.

[0234] "Sulfonyloxy" refers to a -S(=O)2OH group.

[0235] "Sulfonate" refers to a S(=O)-OR group, where R is selected from the group consisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ring carbon), and heteroalicyclic (bonded through a ring carbon). The sulfonate group is optionally substituted on R with one or more of the substituents described for alkyl, cycloalkyl, aryl, and heteroaryl, respectively.

[0236] The compounds of the present disclosure also include crystalline and amorphous forms of those compounds, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs and amorphous forms of the compounds, and mixtures thereof. "Crystalline forms" and "polymorphs" are intended to include all crystalline and amorphous forms of the compounds, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs and amorphous forms, and mixtures thereof, unless a specific crystalline or amorphous form is mentioned.

[0237] Antibody

[0238] In one aspect, the present disclosure provides antibodies and antibody fragments that can be used in antibody-drug conjugates (ADCs), linkers and other compounds and / or conjugates as described herein. As used herein, the term "antibody" encompasses the broadest sense and specifically encompasses monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (e.g., bispecific antibodies) and antibody fragments, as long as these antibodies exhibit the desired biological activity. Antibodies can be mouse antibodies, human antibodies, humanized antibodies, chimeric antibodies or derived from other species. Antibodies are proteins produced by the immune system that can recognize and bind to specific antigens. (Janeway, C., Travers, P., Walport, M., Shlomchik (2001) "Immuno Biology", 5th edition, Garland Publishing House, New York). Target antigens typically have many binding sites recognized by CDRs on multiple antibodies, also referred to as epitopes. Each antibody that specifically binds to different epitopes has a different structure. Thus, an antigen can have more than one corresponding antibody. Antibodies include full-length immunoglobulin molecules or immunologically active portions of full-length immunoglobulin molecules, i.e., molecules that contain an antigen binding site that immunospecifically binds to an antigen of a target of interest or portion thereof, such targets including, but not limited to, cancer cells or cells that produce autoimmune antibodies associated with autoimmune diseases.

[0239] The immunoglobulins disclosed herein can be of any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass of immunoglobulin molecules. The immunoglobulins can be derived from any species. However, in one aspect, the immunoglobulins are of human, mouse, or rabbit origin.

[0240] An "antigen-binding fragment" of an antibody refers to a fragment of a full-length antibody that retains the ability to specifically bind to an antigen (preferably with substantially the same binding affinity). Examples of antigen-binding fragments include (i) a Fab fragment, a monovalent fragment consisting of a VL domain, a VH domain, a CL domain, and a CH1 domain; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of a VH domain and a CH1 domain; (iv) a Fv fragment, consisting of a VL domain and a VH domain of a single arm of an antibody; (v) a dAb fragment (Ward et al., 1989 Nature 341:544-546), consisting of a VH domain; and (vi) isolated complementarity-determining regions (CDRs), disulfide-bonded Fvs (dsFvs), and anti-idiotypic (anti-Id) antibodies and intrabodies. In addition, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, the two domains can be linked using recombinant methods (e.g., by a synthetic linker), thereby enabling their production as a single protein chain in which the VL and VH regions pair to form a monovalent molecule (referred to as single-chain Fv (scFv)); see, for example, Bird et al., Science 242:423-426 (1988); and Huston et al., 1988 Proc. Natl. Acad. Sci. USA 85:5879-5883). Other forms of single-chain antibodies, such as diabodies, are also contemplated. Diabodies are bivalent, bispecific antibodies in which the VH and VL domains are expressed on a single polypeptide chain, but using a linker that is too short to allow pairing between the two domains on the same chain, thereby forcing the domains to pair with the complementary domains of another chain and create two antigen-binding sites (see, e.g., Holliger et al., 1993, Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak et al., 1994, Structure 2:1121-1123).

[0241] Antibody "variable domain" refers to the variable region of an antibody light chain (VL) or the variable region of an antibody heavy chain (VH), alone or in combination. As known in the art, the variable regions of the heavy and light chains are each composed of four framework regions (FR) connected by three complementarity determining regions (CDRs) and contribute to the formation of the antigen binding site of the antibody.

[0242] Complementarity determining regions (CDRs) can be identified according to the definitions of Kabat, Chothia, accumulation of both Kabat and Chothia, AbM, contact, North and / or conformational definitions, or any CDR determination method well known in the art. See, for example, Kabat et al., 1991, Protein Sequences of Immunological Significance, 5th edition (hypervariable regions); Chothia et al., 1989, Nature 342:877-883 (structural loop structures). The identity of the amino acid residues constituting the CDRs in a particular antibody can be determined using methods well known in the art. The AbM definition of CDRs is a compromise between Kabat and Chothia, and is determined using the AbM antibody modeling software from Oxford Molecular.

[0243] The "contact" definition of CDR is based on the observed antigen contacts described in MacCallum et al., 1996, J. Mol. Biol. 262:732-745. The "conformation" definition of CDR is based on the residues that make enthalpic contributions to antigen binding (see, for example, Makabe et al., 2008, J. Biol. Chem. 283:1156-1166). North has identified classical CDR conformations using different preferred sets of CDR definitions (North et al., 2011, J. Mol. Biol. 406:228-256). In another method referred to herein as the "conformation definition" of CDR, the position of the CDR can be identified as the residues that make enthalpic contributions to antigen binding (Makabe et al., 2008, J. Biol. Chem. 283:1156-1166).

[0244] Other CDR boundary definitions may still not strictly follow one of the above methods, but will nonetheless overlap with at least a portion of the Kabat CDRs, but the boundary definitions may be shortened or lengthened based on predictions or experimental findings that particular residues or groups of residues, or even the entire CDR, do not significantly affect antigen binding.

[0245] As used herein, CDRs may refer to CDRs defined by any method known in the art (including combinations of methods). The methods used herein can utilize CDRs defined according to any of these methods. For any given embodiment containing more than one CDR, the CDRs (or other residues of an antibody) may be defined according to any of the Kabat, Chothia, North, extension, AbM, contact and / or conformational definitions.

[0246] The residues in the variable domains are numbered according to Kabat, which is a numbering system for the heavy chain variable domains or light chain variable domains compiled for antibodies. Referring to Kabat et al., 1991, " Protein Sequences with Immunological Significance ", the 5th edition, National Institutes of Health Public Health Service, Bethesda, Maryland. Using this numbering system, the actual linear amino acid sequence can contain the shortening corresponding to the FR or CDR of the variable domains or to the less or extra amino acids inserted therein. For example, the heavy chain variable domain can include a single amino acid insert (according to Kabat's residue 52a) after the residue 52 of H2 and the inserted residues (for example, according to Kabat's residues 82a, 82b and 82c) after the heavy chain FR residue 82. By comparing the homology region of the antibody sequence with the "standard" Kabat numbering sequence, the Kabat residue numbering of a given antibody can be determined. Various algorithms for allocating Kabat numbers are available. The algorithm implemented in version 2.3.3 release of Abysis (www.abysis.org) is used herein to assign Kabat numbering to the variable regions CDRL1, CDRL2, CDRL3, CDRH1, CDRH2, and CDRH3.

[0247] "Framework" (FR) residues are antibody variable domain residues other than CDR residues. A VH domain framework or a VL domain framework comprises four framework subregions FR1, FR2, FR3, and FR4 interspersed with CDRs in the following structure: FR1 - CDR1 - FR2 - CDR2 - FR3 - CDR3 - FR4.

[0248] "Epitope" refers to a region or area of ​​an antigen to which an antibody specifically binds, such as a region or area comprising residues that interact with the antibody. An epitope can be linear or conformational.

[0249] The term "paratope" is derived from the above definition of "epitope" by reversing the perspective and refers to the region or region of the antibody molecule that participates in antigen binding, e.g., a region or region comprising residues that interact with the antigen. A paratope can be linear or conformational (e.g., discontinuous residues in a CDR).

[0250] The epitope / paratope of a given antigen / antibody binding pair can be defined and characterized in varying degrees of detail using a variety of experimental and computational epitope mapping methods. Experimental methods include mutagenesis, X-ray crystallography, nuclear magnetic resonance (NMR) spectroscopy, hydrogen / deuterium exchange mass spectrometry (HX-MS), and various competitive binding methods.

[0251] At its most detailed level, the epitope / paratope for the interaction between an antibody (Ab) and an antigen (Ag) can be defined by the spatial coordinates defining the atomic contacts present in the Ag-Ab interaction and information about their relative contributions to binding thermodynamics. At one level, the epitope / paratope residues can be characterized by the spatial coordinates defining the atomic contacts between the Ag and the Ab.

[0252] In one aspect, epitope / paratope residues can be defined by specific criteria, such as the distance between atoms in the Ab and the Ag (e.g., equal to or less than about 100 μg of the distance between the heavy atoms of the cognate antibody and the heavy atoms of the antigen). In another aspect, an epitope / paratope residue can be characterized as participating in hydrogen bonding interactions with a cognate antibody / antigen or a water molecule that also hydrogen bonds to the cognate antibody / antigen (water-mediated hydrogen bonding). In another aspect, an epitope / paratope residue can be characterized as forming a salt bridge with a residue of the cognate antibody / antigen. In yet another aspect, an epitope / paratope residue can be characterized as a residue that has a non-zero change in buried surface area (BSA) due to interaction with the cognate antibody / antigen.

[0253] At a less detailed level, an epitope / paratope can be characterized by function, such as by competitive binding with other Abs. An epitope / paratope can also be defined more generally as comprising an amino acid residue for which substitution by another amino acid alters the properties of the interaction between the Ab and Ag (e.g., alanine scanning).

[0254] An antibody that "preferentially binds" or "specifically binds" (used interchangeably herein) to an epitope is a term well known in the art, and methods for determining such specific or preferential binding are also well known in the art. A molecule is said to exhibit "specific binding" or "preferential binding" if it reacts or associates with a particular cell or substance more frequently, more rapidly, for a longer period of time, and / or with greater affinity than it reacts or associates with an alternative cell or substance. An antibody "specifically binds" or "preferentially binds" to a target if it binds with a greater affinity, avidity, ease, and / or duration than it binds to other substances. It should also be understood from reading this definition that, for example, an antibody (or portion, targeting agent, or epitope) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target. Thus, "specific binding" or "preferential binding" does not necessarily require (although it can include) exclusive binding. Typically, but not necessarily, reference to binding means preferential binding.

[0255] "Specific binding" or "preferential binding" includes recognizing and binding to a specific molecule, but not substantially recognizing other molecules in the sample or compounds that bind thereto, such as proteins, nucleic acids, antibodies, etc. For example, an antibody that recognizes and binds to its cognate antigen in a sample but does not substantially recognize and bind to other molecules in the sample specifically binds to the cognate antigen. Thus, under specified assay conditions, a specified binding moiety (e.g., an antibody or antigen-binding portion thereof) preferentially binds to a specific target molecule and does not bind to other components present in the test sample in significant amounts.

[0256] A variety of assays can be used to select antibodies or peptides that specifically bind to a molecule of interest. For example, solid phase ELISA immunoassays, immunoprecipitation, BIAcore TM (GE Healthcare, Piscataway, NJ), fluorescence-activated cell sorting (FACS), Octet TM (FortéBio, Inc., Menlo Park, CA) and Western blot analysis are a number of assays that can be used to identify antibodies that specifically react with an antigen or receptor or a ligand binding moiety thereof that specifically binds to a cognate ligand or binding partner. Typically, a specific or selective reaction will be at least two times the background signal or noise, and more typically more than 10 times the background, and even more specifically, at an equilibrium dissociation constant (K D) value ≤ 1 μM, such as ≤ 100 nM, ≤ 10 nM, ≤ 100 pM, ≤ 10 pM or ≤ 1 pM. An antibody is said to “specifically bind” to an antigen.

[0257] As used herein with respect to antibodies, the term "competition" means that the binding of a first antibody, or its antigen-binding portion thereof, to an antigen reduces the subsequent binding of a second antibody, or its antigen-binding portion thereof, to the same antigen. Generally, binding to the first antibody results in steric hindrance, conformational change, or binding to a common epitope (or a portion thereof), such that the binding of the second antibody to the same antigen is reduced. Standard competition assays can be used to determine whether two antibodies compete with each other. One suitable assay for antibody competition involves the use of Biacore technology, which can use surface plasmon resonance (SPR) technology, typically using a biosensor system (e.g., The degree of interaction can be measured using a SPR system. For example, SPR can be used in an in vitro competitive binding inhibition assay to determine the ability of one antibody to inhibit the binding of a second antibody. Another assay for measuring antibody competition uses an ELISA-based method.

[0258] In addition, a high-throughput method for "binding" to an antibody based on competition is described in International Patent Application No. WO2003 / 48731, which is incorporated herein by reference in its entirety. If one antibody (or fragment) reduces the binding of another antibody (or fragment) to the target, competition is present. For example, a sequential binding competition assay can be used, in which different antibodies are added sequentially. A first antibody can be added to achieve near-saturated binding. Then, a second antibody is added. In a non-limiting example, if the binding of the second antibody is not detected or significantly reduced (e.g., at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% reduction) compared to a parallel assay in the absence of the first antibody (the value can be set to 100%), it is considered that the two antibodies compete with each other.

[0259] An "antigen-binding portion" (or interchangeably, an "antigen-binding fragment") comprises a portion of a full-length antibody, typically the antigen-binding or variable region. Examples of antigen-binding portions include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; fragments produced by Fab expression libraries, anti-idiotypic (anti-Id) antibodies, CDRs (complementarity-determining regions), and epitope-binding fragments of any of the above that immunospecifically bind to a cancer cell antigen, a viral antigen, or a microbial antigen, a single-chain antibody molecule; and multispecific antibodies formed from antibody fragments. In some embodiments, the antibody, or antigen-binding portion thereof, is selected from a monoclonal antibody, a polyclonal antibody, an antibody fragment, Fab, Fab', Fab'-SH, F(ab')2, Fv, a single-chain Fv, a diabody, a linear antibody, a bispecific antibody, a multispecific antibody, a chimeric antibody, a humanized antibody, a human antibody, and a fusion protein comprising the antigen-binding portion of an antibody.

[0260] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population, that is, except for the possible naturally occurring mutation that may exist in a small amount, the individual antibodies comprising the population are identical. Monoclonal antibodies are highly specific, directed against a single antigenic site. In addition, compared with polyclonal antibody preparations comprising different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to its specificity, the advantage of monoclonal antibodies is that these antibodies can be synthesized without being contaminated by other antibodies. The modifier "monoclonal" represents that the characteristic of an antibody is obtained from a substantially homogeneous antibody population, and should not be construed as needing to produce an antibody by any ad hoc method. For example, the monoclonal antibody to be used according to the present invention can be prepared by the hybridoma method described first by Kohler et al., (1975) Nature 256:495, or can be prepared by a recombinant DNA method.

[0261] "Fv" is the smallest antibody fragment containing a complete antigen recognition site and an antigen binding site. This region is composed of a dimer of a heavy chain variable domain and a light chain variable domain in close non-covalent association. It is in this configuration that the three hypervariable regions of each variable domain interact to limit the antigen binding site to the surface of the VH-VL dimer. In general, the six hypervariable regions confer antibody antigen binding specificity. However, even a single variable domain (or half of an Fv comprising only three hypervariable regions that are specific for an antigen) has the ability to recognize and bind to an antigen, but the binding is carried out with an affinity lower than the affinity of the entire binding site.

[0262] Fab fragments also contain the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. Fab' fragments differ from Fab fragments in that several residues are added to the carboxyl terminus of the heavy chain CH1 domain, including one or more cysteines from the antibody hinge region. Fab'-SH is the designation herein for Fab', in which the cysteine ​​residues of the constant domains carry at least one free thiol group. F(ab')2 antibody fragments were originally produced as pairs of Fab' fragments with hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0263] The light chains of antibodies from any vertebrate species can be assigned to one of two clearly distinct types, called kappa (κ) and lambda (λ), based on the amino acid sequences of their constant domains.

[0264] Single-chain Fv or scFv refers to a single-chain variable region antibody fragment comprising the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. The Fv polypeptide may further comprise a polypeptide linker between the VH and VL domains that enables the scFv to form the desired structure for antigen binding.

[0265] The term "diabody" refers to a small antibody fragment with two antigen-binding sites, which comprises a variable heavy domain (VH) connected to a variable light domain (VL) in the same polypeptide chain (VH-VL). By using a linker short enough to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and create two antigen-binding sites.

[0266] "Humanized" forms of non-human (e.g., rodent) antibodies are chimeric antibodies containing minimal sequence derived from non-human immunoglobulins. For the most part, humanized antibodies are human immunoglobulins (recipient antibodies) in which residues from the hypervariable regions of the recipient are replaced by residues from the hypervariable regions of a non-human species (donor antibody) such as mouse, rat, rabbit, or non-human primate with the desired specificity, affinity, and capacity. In some cases, framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues.

[0267] In addition, humanized antibodies can include residues that do not exist in the recipient antibody or the donor antibody. These modifications are carried out to further improve antibody performance. Generally, humanized antibodies will include at least one of all or substantially all hypervariable loops corresponding to non-human immunoglobulin regions and all or substantially all FRs that are human immunoglobulin sequences and typically substantially all variable domains in two variable domains.

[0268] "Isolated antibodies" are antibodies that have been identified and separated and / or recovered from components of their environment. Contaminant components in their natural environment are materials that interfere with the diagnostic or therapeutic use of the antibody and may include enzymes, hormones, and other proteins or non-protein solutes. In some embodiments, the antibody may be purified (1) to greater than 95% by weight or greater than 99% by weight as determined by the Lowry method, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence using a spinning cup protein sequencer, or (3) to homogeneity by SDS-PAGE performed under reducing or non-reducing conditions using Coomassie blue or silver staining. Isolated antibodies include antibodies that are in situ located within recombinant cells because at least one component of the antibody's natural environment will not be present. However, typically, the isolated antibody will be prepared by at least one purification step.

[0269] In some embodiments, the targeting agent disclosed herein, antibody or its antigen-binding fragment may include one or more conservative amino acid substitutions. It will be appreciated by those skilled in the art that conservative amino acid substitutions are amino acid substitutions in which one amino acid is replaced with another amino acid having similar structure or chemical properties (e.g., similar side chains). Exemplary conservative substitutions are described in the art in, for example, Watson et al., Molecular Biology of the Gene, Benjamin / Cummings Publishing Company, 4th edition (1987).

[0270] In certain embodiments, the targeting agent, antibody or its antigen-binding fragment described herein comprises an Fc domain. The Fc domain can be derived from IgA (e.g., IgA1 or IgA2), IgG, IgE or IgG (e.g., IgG1, IgG2, IgG3 or IgG4). In certain embodiments, the Fc domain comprises the wild-type sequence of the Fc domain. In certain embodiments, the Fc domain comprises one or more mutations that cause a change in biological activity. For example, mutations can be introduced into the Fc domain to increase the homogeneity during the production of the recombinant protein. In certain embodiments, the Fc domain is the Fc domain of human IgG. In certain embodiments, the lysine at the C-terminal position of the Fc domain is deleted to increase the homogeneity during the production of the recombinant protein. In certain embodiments, the lysine at the C-terminal position of the Fc domain is present.

[0271] In certain embodiments, a polypeptide comprising a targeting agent, antibody, or antigen-binding fragment thereof as described herein is encoded by a cDNA polynucleotide sequence. As is well understood in the art, introduction of the cDNA into competent mammalian cells will produce a polypeptide comprising a targeting agent, antibody, or antigen-binding fragment thereof. Exemplary methods for producing antibodies by these means are disclosed in at least U.S. Patent Nos. 8,008,449, 10,934,571, and 11,339,215, which are incorporated herein by reference.

[0272] In one embodiment, the cDNA comprises a polynucleotide encoding a polypeptide comprising an immunoglobulin heavy chain or fragment thereof, wherein the immunoglobulin heavy chain comprises a VH comprising CDR H1, CDR H2, and CDR H3.

[0273] The present disclosure also provides a targeting agent, antibody or its antigen-binding fragment that is combined with the same epitope as any one of the antibodies or their antigen-binding fragments described herein.For example, antibody competition assays (and overlapping epitope analysis) can be assessed by surface plasmon resonance (SPR) or biolayer interferometry (BLI), as described in detail herein.

[0274] The antibodies and antigen-binding fragments provided by the present invention include monoclonal antibodies, polyclonal antibodies, antibody fragments (e.g., Fab, Fab', F(ab')2, Fv, Fc, etc.), chimeric antibodies, bispecific antibodies, heterologous conjugated antibodies (e.g., antibody-drug conjugates), single chains (ScFv), mutants thereof, fusion proteins comprising antibody portions, domain antibodies (dAb), humanized antibodies, and any other modified configurations of immunoglobulin molecules comprising antigen recognition sites with the required specificity, including glycosylation variants of antibodies, amino acid sequence variants of antibodies, and covalently modified antibodies. Antibodies and antigen-binding fragments can be mouse, rat, human, or any other source (including chimeric or humanized antibodies). In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a chimeric, humanized, or human antibody. In certain embodiments, the antibody is an antibody-drug conjugate.

[0275] "Conservative modifications" refer to amino acid modifications that do not significantly affect or change the binding properties of the antibody containing the amino acid sequence. Conservative modifications include amino acid substitutions, additions, and deletions. Conservative substitutions are substitutions in which an amino acid is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains are well defined and include amino acids with acidic side chains (e.g., aspartic acid, glutamic acid), basic side chains (e.g., lysine, arginine, histidine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), uncharged polar side chains (e.g., glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine, tryptophan), aromatic side chains (e.g., phenylalanine, tryptophan, histidine, tyrosine), aliphatic side chains (e.g., glycine, alanine, valine, leucine, isoleucine, serine, threonine), amides (e.g., asparagine, glutamine), beta-branched side chains (e.g., threonine, valine, isoleucine), and sulfur-containing side chains (cysteine, methionine). In addition, any native residue in the polypeptide can be substituted with alanine, as previously described for alanine scanning mutagenesis (MacLennan et al. (1998) Acta Physiol Scand Suppl 643:55-67; ​​Sasaki et al. (1998) Adv Biophys 35:1-24). Amino acid substitutions for the antibodies of the invention can be performed by known methods, for example, by PCR mutagenesis (U.S. Pat. No. 4,683,195).

[0276] The binding affinity of a targeting agent or antibody can be expressed as the equilibrium dissociation constant (K D ) value indicates that the KD refers to the dissociation rate of a specific antigen-antibody interaction. D is the dissociation rate (also known as the “off-rate” (k 脱离 )") and the association rate or "binding rate (k 结合 )”. Therefore, K D Equal to k 脱离 / k 结合 (dissociation / association) and expressed in molar concentration (M), and K D The smaller the K, the stronger the binding affinity. D Values ​​can be determined using methods established in the art. Unless otherwise indicated, "binding affinity" refers to monovalent interactions (intrinsic activity; eg, antibody and antigen binding through a monovalent interaction).

[0277] In certain embodiments, the affinity (K D) value or less than about 350 nM, about 325 nM, about 323.10 nM, about 300 nM, about 286.44 nM, about 275 nM, about 250 nM, about 232.13 nM, about 225 nM, about 219.13 nM, about 200 nM, about 195.54 nM, about 175 nM, about 158 ​​nM, about 150 nM, about 125 nM or about 100 nM.

[0278] In some embodiments, the targeting agent, antibody, or antigen-binding fragment thereof binds to the epitope with a K of or less than about 95 nM, about 90 nM, about 80 nM, about 79.89 nM, about 75 nM, about 70 nM, about 69.50 nM, about 65 nM, about 63.44 nM, about 60 nM, about 55 nM, about 52.88 nM, about 50 nM, about 45 nM, about 44.50 nM, about 41.99 nM, about 40 nM, about 35 nM, about 30 nM, about 25 nM, about 20 nM, about 10 nM, about 5 nM, or about 1 nM. D Value combination.

[0279] In some embodiments, the targeting agent, antibody, or antigen-binding fragment thereof binds to the epitope with a K of or less than about 5 nM, about 4.5 nM, about 4 nM, about 3.5 nM, about 3.12 nM, about 3 nM, about 2.90 nM, about 2.5 nM, about 2 nM, about 1.5 nM, about 1 nM, about 900 pM, about 800 pM, about 700 pM, about 600 pM, about 500 pM, about 400 pM, about 300 pM, about 250 pM, about 200 pM, about 150 pM, about 100 pM, about 50 pM, about 40 pM, about 30 pM, about 25 pM, about 20 pM, about 15 pM, about 10 pM, about 5 pM, or about 1 pM. D Value combination.

[0280] K D The value of K can be determined directly by well-known methods and can even be calculated for complex mixtures by methods such as those shown in Caceci et al. (1984, Byte 9:340-362). For example, K D A dual filter nitrocellulose filter binding assay can be used, such as that disclosed by Wong and Lohman (1993, PNAS 90:5428-5432). Other standard assays for evaluating the binding ability of a ligand (e.g., an antibody) to a target antigen are known in the art and include, for example, ELISA, Western blot, RIA, and flow cytometric analysis, as well as other assays exemplified elsewhere herein.

[0281] To measure binding affinity (K DAn exemplary method for measuring ) is surface plasmon resonance (SPR), typically using a biosensor system (e.g. SPR refers to the SPR that allows The system detects changes in protein concentration within a biosensor matrix to analyze optical phenomena of real-time biospecific interactions. BIAcore kinetic analysis involves analyzing the binding and dissociation of an antigen to a chip with an immobilized molecule (e.g., a molecule comprising an antigen-binding domain) on its surface; or the dissociation of an antibody or antigen-binding fragment thereof from a chip with an immobilized antigen.

[0282] In certain embodiments, SPR measurements are performed using The standard assay conditions for surface plasmon resonance (SPR) are performed using a T100 or T200 instrument. For example, the standard assay conditions for surface plasmon resonance (SPR) can be based on the immobilization of antibodies to approximately 100-500 response units (RU) of IgG on an SPR chip. The purified target protein is diluted to a certain range of final concentrations in a buffer solution and injected with the necessary flow rate (e.g., 10-100 microliters / minute) to allow calculation of Ka. Dissociation is allowed to continue to establish the dissociation rate, followed by 3M MgCl2 (or 20mM NaOH) for chip surface regeneration. The sensorgram is then analyzed using a kinetic evaluation software package. In an exemplary embodiment, SPR is measured according to the conditions described in the examples.

[0283] In certain embodiments, a solution-based kinetic exclusion assay (KinExA TM ) to measure binding affinity (K D In a specific embodiment, KinExA is used TM KinExA measurements were performed using a 3200 instrument (Sapidyne). TM ) is a universal immunoassay platform (essentially, a flow spectrofluorometer) capable of measuring the equilibrium dissociation constant as well as the association and dissociation rate constants of antigen / antibody interactions. TM is performed after equilibrium has been achieved, so it is used to measure the K for high affinity interactions. D In the case of high affinity interactions, the off-rate of the interaction may be very slow. TM Methods may generally be performed as described in Drake et al. (2004) Analytical Biochem. 328, 35-43.

[0284] For determining the K of antibodies DAnother method is by using biolayer interferometry (BLI), typically using the In certain embodiments, BLI measurements are performed using a biosensor system (e.g., Octet QKe system). In certain embodiments, BLI measurements are performed as follows: a sensor tip coated with a proprietary anti-human antibody (ForteBio) is subjected to BLI signal stabilization by immersing it in a running buffer (e.g., 10 mM Hepes-buffered saline (HBS) containing 0.05% tween-20) for 120 seconds. The antibody is then captured by immersing the sensor in a running buffer solution (the buffer may contain 1-10 ug / mL antibody) for 300 seconds. The signal is then stabilized by immersing the sensor tip back into the running buffer for 120 seconds. The tip is then transferred to a solution containing the cognate antigen. Antibody-antigen binding is measured within 180 seconds before the sensor tip is transferred to the running buffer to monitor receptor dissociation within 180 seconds.

[0285] In an embodiment, a 7-point dose response of antigen is measured (which can be in the range of 1-2 nM in serial dilutions). In addition, the sensor tip to which no antibody is captured is exposed to antigen to monitor nonspecific binding of the receptor to the sensor tip. A second reference type also includes a tip on which the antibody is captured but then exposed only to running buffer without antigen. This allows for double referencing to eliminate nonspecific binding as well as system noise and underlying baseline drift due to dissociation of the antibody from the anti-human Fc sensor tip. The original material undergoes double reference subtraction and is then fitted to a 1:1 Langmuir type binding model to determine affinity and kinetic parameters.

[0286] Drug section

[0287] In some embodiments, the drug moiety is a cytotoxic agent, an immunomodulatory agent, an imaging agent, a chemotherapeutic agent, or a therapeutic protein.

[0288] In some embodiments, the drug moiety is a small molecule with a molecular weight of preferably < about 5 kDa, more preferably < about 4 kDa, more preferably < about 3 kDa, most preferably < about 1.5 kDa, or < about 1 kDa.

[0289] In some embodiments, the IC of the drug moiety 50 About less than about 1 nM.

[0290] In some embodiments, the IC of the drug moiety 50 Greater than about 1 nM, e.g., the IC of a therapeutic agent 50 From about 1 nM to about 50 nM.

[0291] IC 50 Some drug moieties greater than about 1 nM (e.g., "low potency drugs") are not suitable for conjugation to antibodies using art-recognized conjugation techniques. Without wishing to be bound by theory, the potency of such drug moieties is insufficient for use in targeted antibody-drug conjugates using conventional techniques because sufficient copies of the drug (i.e., more than 8) cannot be conjugated using art-recognized techniques without causing a decrease in the pharmacokinetic and physicochemical properties of the conjugate. However, sufficiently high loadings of these low potency drugs can be achieved using the conjugation strategies described herein, thereby enabling high loadings of the therapeutic agent while maintaining the desired pharmacokinetic and physicochemical properties. Thus, in some embodiments, the present disclosure also relates to an antibody-drug conjugate comprising an antibody, a linker, and at least eight drug moieties, wherein the IC of the therapeutic agent is less than 100. 50 Greater than about 1 nM.

[0292] In some embodiments, small molecule therapeutics used in the present disclosure (e.g., anti-proliferative (cytotoxic and cytostatic) agents that can be linked to targeting moieties via linkers of the present disclosure) include cytotoxic compounds (e.g., broad spectrum), angiogenesis inhibitors, ceiling cycle progression inhibitors, PI3K / m-TOR / AKT pathway inhibitors, MAPK signaling pathway inhibitors, kinase inhibitors, protein chaperone inhibitors, HDAC inhibitors, PARP inhibitors, nicotinamide phosphoribosyltransferase (NAMPT) inhibitors, Wnt Hedgehog signaling pathway inhibitors, and RNA polymerase inhibitors.

[0293] Broad-spectrum cytotoxins include, but are not limited to, DNA binding agents, intercalating or alkylating agents, microtubule stabilizers and destabilizers, platinum compounds, topoisomerase inhibitors (including topoisomerase I inhibitors and topoisomerase II inhibitors), and protein synthesis inhibitors.

[0294] In some embodiments, the drug moiety comprises one or more cGAS / interferon gene stimulator (STING) pathway agonists. Non-limiting examples of STING agonists include DMXAA, ADUS100 / MIW815, MK-1454, MK-2118, SB11285, GSK3745417, BMS-986301, BI-STING (BI 1387446), E7766, TAK-676, SNX281, SYNB1891. Additional non-limiting examples of STING agonists and combinations with other cytotoxic agents and / or ENPP1 inhibitors can be found in Amouzegar et al., Cancers 13:2695 (2021), which is incorporated herein by reference in its entirety.

[0295] Exemplary DNA binding, intercalating, or alkylating agents include, but are not limited to, CC-1065 and its analogs, anthracyclines (doxorubicin, epirubicin, idarubicin, daunorubicin, nemorubicin and its derivatives, PNU-159682), bis-naphthylimide compounds such as elinafide (LU79553) and its analogs, alkylating agents such as calicheamicin, dactinomycin, mitomycin, pyrrolobenzodiazepines, indolebenzodiazepines, and the like. Exemplary CC-1065 analogs include, but are not limited to, duocarmycin SA, duocarmycin A, duocarmycin CI, duocarmycin C2, duocarmycin B1, duocarmycin B2, duocarmycin D, DU-86, KW-2189, adozelesin, bizelesin, carzelesin, seco-adozelesin, and related analogs and prodrug forms, examples of which are described in U.S. Patent Nos. 5,475,092; 5,595,499; 5,846,545; 6,534,660; 6,586,618; 6,756,397; and 7,049,316. ​​Doxorubicin and its analogs include those described in U.S. Patent No. 6,630,579. Calicinomycins include, for example, enediynes such as esperamicin and enediynes described in U.S. Pat. Nos. 5,714,586 and 5,739,116. Duocarmycins include those described in U.S. Pat. Nos. 5,070,092; 5,101,038; 5,187,186; 6,548,530; 6,660,742; and 7,553,816B2; and Li et al., Tel Letts., 50:2932-2935 (2009), the disclosures of all of which are incorporated herein by reference in their entireties.

[0296] Exemplary topoisomerase inhibitors (e.g., topoisomerase I and / or topoisomerase II) include, but are not limited to, camptothecin, camptothecin derivatives, camptothecin analogs, and unnatural camptothecins, such as irinotecan, Dxd, Sn-38 (7-ethyl-10-hydroxy-camptothecin), CPT-11 (irinotecan), GI-147211C, topotecan, 9-aminocamptothecin, 7-hydroxymethylcamptothecin, 7-aminomethylcamptothecin, 10-hydroxycamptothecin, (20S)-camptothecin, rubitecan, gimatecan, karenitecin, silatecan, lurtotecan, diflomotecan, belotecan, lurtotecan, and S39625, and any analogs thereof. Non-limiting examples of other topoisomerase inhibitors (e.g., topoisomerase I and / or topoisomerase II) that may be used in the present disclosure include topoisomerase inhibitors described in WO 2020 / 00880 and WO 2021 / 148501, the disclosure of each of which is incorporated herein by reference in its entirety. Non-limiting examples of other camptothecin compounds that may be used in the present disclosure include camptothecin compounds described in: Journal of Medicinal Chemistry (J.Med.Chem.), 29: 2358-2363 (1986); Journal of Medicinal Chemistry, 23: 554 (1980); Journal of Medicinal Chemistry, 30: 1774 (1987), the disclosure of each of which is incorporated herein by reference in its entirety. In some embodiments, the drug moiety is exitecan and / or its analogs. In some embodiments, the drug moiety is Dxd and / or its analogs.

[0297] Non-limiting examples of pyrrolobenzodiazepines (PBDs) and their analogs include, but are not limited to, PBDs and their analogs described in Denny, Exp. Opin. Ther. Patents., 10(4): 459-474 (2000); Antonow and Thurston, Chem Rev., 2815-2864 (2010); Min et al., ACS Omega 5: 25798-25809 (2020); and Hartley, Exp. Opin. Biol. Therapy, 7: 931-943 (2020), the disclosures of each of which are incorporated herein by reference in their entirety.

[0298] Exemplary microtubule stabilizers and destabilizers include, but are not limited to, taxane compounds, such as paclitaxel, docetaxel, tesetaxel, and carbazitaxel; maytansinoids, auristatins and their analogs, vinca alkaloid derivatives, epothilones, and cryptophycins.

[0299] Exemplary maytansine compounds or maytansine compounds analogs include but are not limited to maytansinol (maytansinol) and maytansinol analogs, and maytansine or DM-i and DM-4 are U.S. Patent No. 5,208,020;No. 5,416,064;No. 6,333.410;No. 6,441,163;No. 6,716,821;No. RE39,151;And the maytansine or DM-i and DM-4 described in No. 7,276,497.In certain embodiments, cytotoxic agent is maytansine compounds, the anti-tubulin agent (ImmunoGen company (ImmunoGen, Inc.) of another group;See also Chari et al., 1992,《Cancer Research (Cancer Res.)》52:127-131), maytansine compounds or maytansine compounds analogs. Examples of suitable maytansinoids include, but are not limited to, maytansinol and maytansinol analogs. Non-limiting examples of suitable maytansinoids are disclosed in U.S. Pat. Nos. 4,424,219; 4,256,746; 4,294,757; 4,307,016; 4,313,946; 4,315,929; 4,331,598; 4,361,650; 4,362,663; 4,364,866; 4,450,254; 4,322,348; 4,371,533; 6,333,410; 5,475,092; 5,585,499; and 5,846,545, which are incorporated herein by reference in their entireties.

[0300] Exemplary auristatins include, but are not limited to, auristatin E (also known as a derivative of dolastatin-10), auristatin EB (AEB), auristatin EFP (AEFP), monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), auristatin F, auristatin F phenylenediamine (AFP), auristatin F hydroxypropylamide (AF FTP A), monomethyl auristatin F hydroxypropylamide (MMAF HP A), and dolastatin. Non-limiting examples of suitable auristatins are also described in U.S. Publication Nos. 2003 / 0083263; 2011 / 0020343; and 2011 / 0070248; PCX Application Publication Nos. WO 09 / 117531; WO 2005 / 081711; WO 04 / 010957; WO 02 / 088172; and WO No. 01 / 24763; and U.S. Patents Nos. 7,498,298; 6,884,869; 6,323,315; 6,239,104; 6,124,431; 6,034,065; 5,780,588; 5,767,237; 5,665,860; 5,663,149; 5,635,483; 5,599,902; , 554,725; 5,530,097; 5,521,284; 5,504,191; 5,410,024; 5,138,036; 5,076,973; 4,986,988; 4,978,744; 4,879,278; 4,816,444; and 4,486,414, which are incorporated herein by reference in their entireties. In some embodiments, the drug moiety is monomethyl auristatin E (MMAE) and / or its analogs.

[0301] Exemplary vinca alkaloids include, but are not limited to, vincristine, vinblastine, vindesine, and navelbine (vinorelbine). Suitable vinca alkaloids that may be used in the present disclosure are also disclosed in U.S. Patents Nos. 2002 / 0103136 and 2010 / 0305149 and U.S. Patent No. 7,303,749B1, the disclosure of each of which is incorporated herein by reference in its entirety.

[0302] Exemplary epothilone compounds include, but are not limited to, epothilones A, B, C, D, E, and F, and derivatives thereof. Suitable epothilone compounds and derivatives thereof are described, for example, in U.S. Patent Nos. 6,956,036; 6,989,450; 6,121,029; 6,117,659; 6,096,757; 6,043,372; 5,969,145; and 5,886,026; and WO 97 / 19086; WO 98 / 08849; WO 98 / 22461; WO 98 / 25929; WO 98 / 38192; WO 99 / 01124; WO 99 / 02514; WO 99 / 03848; WO 99 / 07692; WO 99 / 27890; and WO No. 99 / 28324, the disclosures of all of the patents in said patent are incorporated herein by reference in their entirety.

[0303] Non-limiting examples of nostoc compounds are described in US Patent Nos. 6,680,311 and 6,747,021, the disclosures of each of which are incorporated herein by reference in their entirety.

[0304] Exemplary platinum compounds include, but are not limited to, cisplatin Carboplatin Oxaliplatin Iproplatin, ormaplatin, and tetraplatin.

[0305] Non-limiting examples of other classes of compounds or compounds having these or other cytotoxic modes of action may be selected, including, for example, mitomycin C, mitomycin A, daunorubicin, doxorubicin, morpholino-doxorubicin, cyanomorpholino-doxorubicin, aminopterin, bleomycin, 1-(chloromethyl)-2,3-dihydro-1H-benz[e]indol-5-ol, pyridobenzodiazepine (PDD), pyrrolobenzodiazepine (PBD), and polyamides and dimers thereof. Non-limiting examples of other suitable cytotoxic agents include puromycin, topotecan, rhizoxin, echinomycin, combretastatin, netropsin, estramustine, cryptophycin, cemadotin, discodermolide, eleutherobin, and mitoxantrone.

[0306] Angiogenesis inhibitors include but are not limited to MetAP2 inhibitors, VEGF inhibitors, PIGF inhibitors, VGFR inhibitors, PDGFR inhibitors, MetAP2 inhibitors. Exemplary VGFR and PDGFR inhibitors include but are not limited to sorafenib (sorafenib) (Nexavar), sunitinib (sunitinib) (Sutent) and vatalanib. Exemplary MetAP2 inhibitors include fumagillol analogs, which means any compound comprising a fumarillin core structure that inhibits the ability of MetAP-2 to remove the NH2-terminal methionine from the protein, including fumarillamine, as described in Rodeschini et al., J. Org. Chem., 69, 357-373, 2004, and Liu et al., Science, 282, 1324-1327, 1998. Non-limiting examples of "fumagillol analogs" are disclosed in J. Org. Chem., 69, 357, 2004; J. Org. Chem., 70, 6870, 2005; European Patent Application No. 0 354 787; J. Med. Chem., 49, 5645, 2006; Bioorg. Med. Chem., 11, 5051, 2003; Bioorg. Med. Chem., 14, 91, 2004; J. Econ. Express, 40, 4797, 1999; WO 99 / 61432; U.S. Pat. Nos. 6,603,812; 5,789,405; 5,767,293; 6,566,541; and 6,207,704, the disclosures of all of which are incorporated herein by reference in their entirety.

[0307] Exemplary ceiling cycle progression inhibitors include, but are not limited to, CDK inhibitors such as BMS-387032 and PD0332991; Rho kinase inhibitors such as GSK429286; checkpoint kinase inhibitors such as AZD7762; Aurora kinase inhibitors such as AZD1 152, MLN8054, and MLN8237; PLK inhibitors such as BI 2536, BI6727 (Volasertib), GSK461364, ON-01910 (Estybon); and KSP inhibitors such as SB 743921, SB 715992 (ispinesib), MK-0731, AZD8477, AZ3146, and ARRY-520.

[0308] Exemplary PBK / m-TOR / AKT signaling pathway inhibitors include, but are not limited to, phosphoinositide 3-kinase (PI3K) inhibitors, GSK-3 inhibitors, ATM inhibitors, DNA-PK inhibitors, and PDK-1 inhibitors.

[0309] Non-limiting examples of exemplary PI3 kinase inhibitors are disclosed in U.S. Pat. No. 6,608,053, the disclosure of which is incorporated herein by reference in its entirety, and include BEZ235, BGT226, BKM120, CAL101, CAL263, demethoxyviridin, GDC-0941, GSK615, IC871 14, LY294002, Pafomid 529, perifosine, PI-103, PF-04691502, PX-866, SAR245408, SAR245409, SF 1 126, Wortmannin, XL 147, and XL765.

[0310] Exemplary AKT inhibitors include, but are not limited to, AT7867.

[0311] Exemplary MAPK signaling pathway inhibitors include, but are not limited to, MEK, Ras, JNK, B-Raf, and p38 MAPK inhibitors.

[0312] Non-limiting exemplary MEK inhibitors are disclosed in U.S. Pat. No. 7,517,994, the disclosure of which is herein incorporated by reference in its entirety, and include GDC-0973, GSK1120212, MSC1936369B, AS703026, R05126766 and R04987655, PD0325901, AZD6244, AZD 8330, and GDC-0973.

[0313] Exemplary B-raf inhibitors include, but are not limited to, CDC-0879, PLX-4032, and SB590885.

[0314] Exemplary B p38 M APK inhibitors include, but are not limited to, BIRB 796, LY2228820, and SB 202190.

[0315] Receptor tyrosine kinases (RTKs) are cell surface receptors that are often associated with signaling pathways that stimulate uncontrolled proliferation and angiogenesis of cancer cells. Many RTKs that are overexpressed or have mutations that result in constitutive activation of the receptor have been identified, including but not limited to VEGFR, EGFR, FGFR, PDGFR, EphR, and RET receptor family receptors. Exemplary specific RTK targets include but are not limited to ErbB2, FLT-3, c-Kit, and c-Met.

[0316] Exemplary inhibitors of the ErbB2 receptor (EGFR family) include, but are not limited to, AEE788 (NVP-AEE 788), BIBW2992, (Afatinib), Lapatinib, Erlotinib (Tarceva), and Gefitinib (Iressa).

[0317] Exemplary RTK inhibitors that target more than one signaling pathway (multi-targeted kinase inhibitors) include, but are not limited to, AP24534 (Ponatinib), which targets FGFR receptors, FLT-3 receptors, VEGFR-PDGFR receptors, and Bcr-Abl receptors; ABT-869 (Linifanib), which targets FLT-3 receptors and VEGFR-PDGFR receptors; AZD2171, which targets VEGFR-PDGFR receptors, Flt-3 receptors, and Bcr-Abl receptors; CHR-258 (Dovitinib), which targets the VEGFR-PDGFR receptor, FGFR receptor, Flt-3 receptor, and c-Kit receptor; Sunitinib (Sutent), which targets VEGFR, PDGFR, KIT, FLT-3, and CSF-IR; Sorafenib (Nexavar) and Vatalanib, which target VEGFR, PDGFR, and intracellular serine / threonine kinases in the Raf / Mek / Erk pathway.

[0318] Exemplary protein chaperone inhibitors include, but are not limited to, HSP90 inhibitors.

[0319] Exemplary HSP90 inhibitors include, but are not limited to, 17AAG derivatives, BIIB021, BIIB028, SX-5422, NVP-AUY-922, and KW-2478.

[0320] Exemplary WD AC inhibitors include, but are not limited to, Belinostat (PXD101), CUDC-101, Droxinostat, ITF2357 (Givinostat, Gavinostat), JNJ-26481585, LAQ824 (NVP-LAQ824, Dacinostat), LBH-589 (Panobinostat), MCI 568, MGCD0103 (Mocetinostat), MS-275 (Entinostat), PCI-24781, Pyroxamide (NSC 696085), SB939, Trichostatin A, and Vorinostat (SAHA).

[0321] Exemplary PARP inhibitors include, but are not limited to, iniparib (BSI 201), olaparib (AZD-2281), ABT-888 (Veliparib), AG014699, CEP 9722, MK 4827, KU-0059436 (AZD2281), LT-673, 3-aminobenzamide, A-966492, and AZD2461.

[0322] Exemplary NAMPT inhibitors include, but are not limited to, FK866 (AP0866) and CHS828, GPP 78, GMX1778 (CHS828), STF-1 18804, STF-31, CB 300919, CB 30865, GNE-617, IS001, TP201565, Nampt-IN-1, P7C3, MPC-9528, CB30865, MPI0479883, and (£)-N-(5-((4-(((2-(1H-indol-3-yl)ethyl)(isopropyl)amino)methyl)phenyl)amino)pentyl)-3-(pyridin-3-yl)acrylamide.

[0323] Exemplary Wnt / Hedgehog signaling pathway inhibitors include, but are not limited to, vismodegib (RG3616 / GDC-0449), cyclopamine (11-deoxojervine) (Hedgehog pathway inhibitor), and XAV-939 (Wnt pathway inhibitor).

[0324] Exemplary RNA polymerase inhibitors include, but are not limited to, amatoxins. Exemplary amatoxins include α-amanitin, β-amanitin, γ-amanitin, ε-amanitin, amanullin, amanullic acid, amaninamide, amanin, and proamanullin.

[0325] Exemplary protein synthesis inhibitors include, but are not limited to, trichothecene compounds.

[0326] In some embodiments, the drug moiety D is a topoisomerase inhibitor (e.g., a non-natural camptothecin compound), a vinca alkaloid, a kinase inhibitor (e.g., a PI3 kinase inhibitor (GDC-0941 and PI-103)), a MEK inhibitor, a KSP inhibitor, an RNA polymerase inhibitor, a protein synthesis inhibitor, a PARP inhibitor, a NAMPT inhibitor, docetaxel, paclitaxel, doxorubicin, a duocarmycin, auristatin, dolastatin, calicheamicin, topotecan, SN38, camptothecin, exitecan, nemorubicin and its derivatives, PNU-1.59682, CC1065, elinefand, a trichothecene, a pyrrolobenzodiazepine, a maytansine, a DNA-binding drug, or a platinum compound and its analogs. In some embodiments, the drug is Sn-38, camptothecin, topotecan, exitecan, calicheamicin, nemorubicin, PNU-159682, anthracycline, maytansinoid, taxane, trichothecene, CC1065, elinefand, vindesine, vinblastine, PI-103, AZD 8330, dolastatin, auristatin E, auristatin F, a duocarmycin compound, ispins, pyrrolobenzodiazepine, ARRY-520, and derivatives of stereoisomers, isosteres, and analogs thereof.

[0327] In some embodiments, the drug moiety D is a topoisomerase inhibitor having the structure of:

[0328]

[0329]

[0330]

[0331] In some embodiments, the drug moiety used in the present disclosure is a combination of two or more drugs, for example, a PI3 kinase inhibitor and a MEK inhibitor, a broad spectrum cytotoxic compound and a platinum compound; 3inhibitors, NAMPT inhibitors and platinum compounds, broad-spectrum cytotoxic compounds and PARP inhibitors.

[0332] In some embodiments, the drug moiety used in the present disclosure is auristatin F-hydroxypropylamide-L-alanine.

[0333] connector

[0334] In one aspect, the drug moiety can be directly or indirectly connected to a targeting agent (e.g., an antibody or its binding fragment) to provide a targeted conjugate. In some embodiments, an antibody drug conjugate (ADC) of the present disclosure (e.g., ADC of formula (I)) contains a linker group, wherein a targeting agent (e.g., an antibody or its binding fragment) is attached to the drug moiety via the linker group. In some embodiments, a compound of the present disclosure (e.g., a compound of formula (III) or its salt, solvate, tautomer, isomer, or mixture) contains a linker group, wherein a targeting agent (e.g., an antibody or its binding fragment) is attached to the drug moiety via the linker group. In some embodiments, the linker is a single bond. In one non-limiting example, when the linker is a single bond, the drug moiety is directly attached to the targeting agent (e.g., an antibody or its binding fragment). In some embodiments, a variety of target conjugates are known in the art and can be used together with a compound of formula (III) and a salt or solvate thereof. In one non-limiting example, a target conjugate is an antibody-drug conjugate wherein one or more compounds of formula (III) are connected to an antibody. In embodiments, the antibody drug conjugates of the present disclosure contain one or more compounds of formula (III) or salts, solvates, tautomers, isomers, or mixtures thereof.

[0335] The present disclosure contemplates any linker suitable for attaching a drug moiety to a targeting agent (eg, an antibody or binding fragment thereof), as would be understood by one of ordinary skill in the art.

[0336] In some embodiments, the linker is a bond or a moiety having 1-200 non-hydrogen atoms selected from C, N, O, S or halogen, and optionally incorporates an alkyl moiety, an ether moiety, an oxo moiety, a carboxyl moiety, a carboxamide moiety, a carboxamido moiety, an ester moiety, a urethane moiety, a branched moiety, a cyclic moiety, an unsaturated moiety, an amino acid moiety, a heterocyclyl moiety, an aryl moiety or a heteroaryl moiety. In embodiments, the linker is unbranched or branched, flexible or rigid, short or long, and optionally incorporates any combination of moieties deemed useful. In some embodiments, at least a portion of the linker comprises a polyalkylene oxide polymer region. In a non-limiting example, the polyalkylene oxide polymer region can enhance the solubility of the drug moiety. In some embodiments, the linker comprises ethylene glycol repeating units.

[0337] In some embodiments, the linker has a number of repeating ethylene glycol units ranging from about 1 to about 25 or between. In some embodiments, the linker comprises about 3 to about 20, about 3 to about 5, about 4 to about 15, about 4 to about 8, about 4 to about 6, about 5 to about 12, about 6 to about 10, or about 7 to about 9 ethylene glycol units. In some embodiments, the linker comprises about 8 ethylene glycol units.

[0338] In certain embodiments, at least a portion of the joint comprises one or more amino acid moieties. In a non-limiting example, one or more amino acid moieties enhance the solubility of the drug moiety and / or provide for enhancing target binding, enhancing compatibility with the targeting agent and / or enhancing the target binding amino acid sequence of recognition. In certain embodiments, the joint comprises one or more amino acid moieties that provide a substrate motif that is applicable to protease. In a non-limiting example, when a group of amino acid moieties is incorporated into a joint that provides a specific substrate motif for selected protease, the drug moiety can be released from the target-bound conjugate to provide a localized cytotoxic effect.

[0339] In some embodiments, the linker comprises an alkylene chain. In some embodiments, the alkylene chain is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbons in length; and suitably, the alkylene chain comprises a -CH2- group. In some embodiments, these substrate motifs are known in the art and are incorporated into the linker as needed to provide selective release from the target-bound conjugate. In a non-limiting example, this selectivity is based on the presence of a desired protease within the local delivery zone of a known conjugate drug. In some embodiments, other polymer types may be incorporated into the linker, including but not limited to polyacids, polysaccharides, or polyamines. In some embodiments, other moieties such as substituted aromatic or heteroaromatic moieties are used to enhance rigidity or provide synthetically accessible sites on the substituents therein for attachment to a reactive moiety or drug moiety.

[0340] In one non-limiting example, the linker comprises ethylene glycol repeating units and / or an amino acid sequence.

[0341] In some embodiments, the linker comprises or consists of the formula:

[0342] -[CH2CH2O] p -X AA -

[0343] where X AA is an amino acid sequence, and p is an integer from 0 to 50. In some embodiments, p is an integer from 1 to 10, from 4 to 10, from 6 to 10, or from 7 to 9. In some embodiments, p is 8.

[0344] In some embodiments, the linker comprises or consists of the formula:

[0345] -[CH2CH2O] p -X AA -

[0346] where X AA is an amino acid sequence, and p is an integer from 0 to 50. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0347] In some embodiments, the linker (e.g., L A ) comprises or consists of the following formula:

[0348] -[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA -

[0349] where X AA is an amino acid sequence, and p is an integer from 0 to 50. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0350] In some embodiments, the linker (e.g., L A ) comprises or consists of the following formula:

[0351] -[CH2CH2O] p -(CH2)1-5 -C(O)-X AA -

[0352] where X AA is an amino acid sequence, and p is an integer from 0 to 50, wherein X AA is not Val-Cit or Phe-Lys. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0353] In some embodiments, the linker (e.g., L A ) comprises or consists of the following formula:

[0354] -[CH2CH2O] p -(CH2) 1-3 -C(O)-X AA -

[0355] where X AA is an amino acid sequence, and p is an integer from 0 to 50. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0356] In some embodiments, the linker (e.g., L A ) comprises or consists of the following formula:

[0357] -[CH2CH2O] p -(CH2) 1-3 -C(O)-X AA -

[0358] where X AA is an amino acid sequence, and p is an integer from 0 to 50, wherein X AA is not Val-Cit or Phe-Lys. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0359] In some embodiments, the linker (e.g., L A ) comprises or consists of the following formula:

[0360] -[CH2CH2O] p -(CH2)2-C(O)-X AA -

[0361] where X AA is an amino acid sequence, and p is an integer from 0 to 50. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0362] In some embodiments, the linker (e.g., L A ) comprises or consists of the following formula:

[0363] -[CH2CH2O] p -(CH2)2-C(O)-X AA -

[0364] where X AA is an amino acid sequence, and p is an integer from 0 to 50, wherein X AAis not Val-Cit or Phe-Lys. In some embodiments, p is an integer from 1 to 40. In some embodiments, p is an integer from 1 to 30. In some embodiments, p is an integer from 6 to 40. In some embodiments, p is an integer from 8 to 30. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 6 to 20. In some embodiments, p is an integer from 8 to 20. In some embodiments, p is an integer from 10 to 30. In some embodiments, p is an integer from 10 to 20. In some embodiments, p is an integer from 1 to 25, 4 to 20, 5 to 15, 6 to 12, or 5 to 10. In some embodiments, p is an integer from 1 to 10, 4 to 10, 6 to 10, or 7 to 9. In some embodiments, p is 8.

[0365] In some embodiments, a suitable number of ethylene glycol units can be used in the joint. In some embodiments, the joint comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 16, 19, 20, 23, 24, 35, 36, 37, 48, 49 or more ethylene glycol units. In some embodiments, the joint comprises 1 to 10, 4 to 10, 6 to 10 or 7 to 9 ethylene glycol units. In some embodiments, the joint comprises 8 ethylene glycol units. Non-limiting examples of commercially available ethylene glycol groups (polyethylene glycol, PEG) suitable for use in joints include H2N- with discrete ("d") polyethylene glycol. 8-C(O)OH, the discrete polyethylene glycol having 8 ethylene glycol repeating units. Non-limiting examples of other discrete PEG units are commercially available and known to those skilled in the art, such as those commercially available and known from Advanced ChemTech. In some embodiments, the linker comprises the formula:

[0366] -HN-PEG-C(O)-X AA -

[0367] wherein PEG has 1-50 ethylene glycol units, and X AA is an amino acid sequence. In some embodiments, PEG has 1-10 ethylene glycol units, about 4-10 ethylene glycol units, or about 7-9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units.

[0368] In some embodiments, the linker comprises the formula:

[0369] -HN-PEG-C(O)-X AA -

[0370] wherein PEG has 1-50 ethylene glycol units, and X AA is an amino acid sequence, provided that X AA Not Val-Cit or Phe-Lys. In some embodiments, PEG has 1-10 ethylene glycol units, about 4-10 ethylene glycol units, or about 7-9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units.

[0371] In some embodiments, the linker (e.g., L A )Inclusion formula:

[0372] -HN-PEG-(CH2) 1-5 -C(O)-X AA -

[0373] wherein PEG has 1-50 ethylene glycol units, and X AA is an amino acid sequence. In some embodiments, PEG has 1-10 ethylene glycol units, about 4-10 ethylene glycol units, or about 7-9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units. In some embodiments, p is 8.

[0374] In some embodiments, the linker (e.g., L A )Inclusion formula:

[0375] -HN-PEG-(CH2) 1-5 -C(O)-X AA -

[0376] wherein PEG has 1-50 ethylene glycol units, and X AA is an amino acid sequence, provided that X AA Not Val-Cit or Phe-Lys. In some embodiments, PEG has 1-10 ethylene glycol units, about 4-10 ethylene glycol units, or about 7-9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units.

[0377] In some embodiments, the linker (e.g., L A )Inclusion formula:

[0378] -HN-PEG-(CH2) 1-3 -C(O)-X AA -

[0379] wherein PEG has 1-50 ethylene glycol units, and X AAis an amino acid sequence. In some embodiments, PEG has 1-10 ethylene glycol units, about 4-10 ethylene glycol units, or about 7-9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units. In some embodiments, p is 8.

[0380] In some embodiments, the linker (e.g., L A )Inclusion formula:

[0381] -HN-PEG-(CH2) 1-3 -C(O)-X AA -

[0382] wherein PEG has 1-50 ethylene glycol units, and X AA is an amino acid sequence, provided that X AA Not Val-Cit or Phe-Lys. In some embodiments, PEG has 1-10 ethylene glycol units, about 4-10 ethylene glycol units, or about 7-9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units.

[0383] In some embodiments, the linker (e.g., L A )Inclusion formula:

[0384] -HN-PEG-(CH2)2-C(O)-X AA -

[0385] wherein PEG has 1-50 ethylene glycol units, and X AA is an amino acid sequence. In some embodiments, PEG has 1-10 ethylene glycol units, about 4-10 ethylene glycol units, or about 7-9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units.

[0386] In some embodiments, the linker (e.g., L A )Inclusion formula:

[0387] -HN-PEG-(CH2)2-C(O)-X AA -

[0388] wherein PEG has 1-50 ethylene glycol units, and X AA is an amino acid sequence, provided that X AA Not Val-Cit or Phe-Lys. In some embodiments, PEG has 1-10 ethylene glycol units, about 4-10 ethylene glycol units, or about 7-9 ethylene glycol units. In some embodiments, PEG has 8 ethylene glycol units.

[0389] In another non-limiting example, the linker comprises an alkylene chain and / or an amino acid sequence. In some embodiments, the linker comprises the formula:

[0390] -[CH2] 0-12 -X AA -

[0391] where X AA is an amino acid sequence; and the linker comprises 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 -CH2- units.

[0392] In another non-limiting example, the linker comprises an alkylene chain and / or an amino acid sequence. In some embodiments, the linker comprises the formula:

[0393] -[CH2] 0-12 -X AA -

[0394] where X AA is an amino acid sequence; and the linker comprises 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 -CH2- units, with the proviso that X AA Not Val-Cit or Phe-Lys.

[0395] In some embodiments, the linker comprises the formula:

[0396] -[CH2] 0-12 -X AA -

[0397] where X AA is an amino acid sequence; and the linker comprises 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 -CH2- units, with the proviso that X AA Not Val-Cit or Phe-Lys.

[0398] In some embodiments, the linker comprises the formula:

[0399] -HN-PEG8-C(O)-Val-Ala-

[0400] PEG has 8 ethylene glycol units.

[0401] In some embodiments, the linker comprises the formula:

[0402] -HN-PEG8-(CH2) 1-5 -C(O)-Val-Ala-

[0403] PEG has 8 ethylene glycol units.

[0404] In some embodiments, the linker comprises the formula:

[0405] -HN-PEG8-(CH2) 1-3 -C(O)-Val-Ala-

[0406] PEG has 8 ethylene glycol units.

[0407] In some embodiments, the linker comprises the formula:

[0408] -HN-PEG8-(CH2)2-C(O)-Val-Ala-

[0409] PEG has 8 ethylene glycol units.

[0410] In some embodiments, the linker also includes various other linking groups that connect the ethylene glycol moiety to the amino acid sequence or connect the ethylene glycol or amino acid sequence to a targeting agent (e.g., an antibody or its binding fragment) or a drug moiety. For example, the amino acid sequence can be connected to the drug moiety through a 4-aminocarboxylic acid benzyl ester group. In some embodiments, the ethylene glycol moiety can be directly connected to the targeting agent (e.g., an antibody or its binding fragment). In some embodiments, the linker comprises or consists of the following formula:

[0411] In some embodiments, the HN group is directly linked to the targeting agent (eg, an antibody or binding fragment thereof).

[0412] In an embodiment, the linker is or comprises:

[0413]

[0414] where X AA is an amino acid sequence; and K2 is -[CH2CH2O] 0-50 -or-[CH2] 0-12 In some embodiments, the linker is attached to the targeting agent (e.g., antibody or binding fragment thereof) and the drug moiety in either direction. In some embodiments, the linker is (i), (ii), (iii), (iv), (vi), (viii), or (ix).

[0415] In an embodiment, the linker is or comprises:

[0416]

[0417] where X AA is an amino acid sequence; and K2 is -[CH2CH2O] 0-50 -[CH2] 0-12-C(O)-. In some embodiments, the linker is attached to the targeting agent (e.g., antibody or binding fragment thereof) and the drug moiety in either orientation. In some embodiments, the linker is (i), (ii), (iii), (iv), (vi), (viii), or (ix).

[0418] In some embodiments, the linker is or comprises:

[0419]

[0420] In some embodiments, the linker further comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 16, 19, 20, 23, 24, 35, 36, 37, 48, 49, or 50 ethylene glycol units. In some embodiments, the linker comprises 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 -CH2- units. In some embodiments, the HN group is directly linked to the targeting agent (e.g., an antibody or binding fragment thereof).

[0421] In some embodiments, the amino acid moiety is an amino acid derivative.

[0422] In some embodiments, the linker comprises an amino acid portion comprising any suitable number of amino acid portions as described above. In one non-limiting example, the amino acid sequence X AA In some embodiments, the linker comprises 1 to 100 amino acid moieties, or 1 to 10 amino acid moieties, or 1 to 5 amino acid moieties. In some embodiments, the linker comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acid moieties. In some embodiments, the linker comprises 2 amino acid moieties. In some embodiments, X AA It's valine-alanine.

[0423] In some embodiments, the amino acid sequence X AA yes:

[0424]

[0425] In some embodiments, the amino acid sequence X AA yes:

[0426]

[0427] In some embodiments, the linker comprises one or more selected from the group consisting of C1-C6 alkyl, C=O, -NH-, ethylene glycol, optionally 2-10 ethylene glycol units, valine-citrulline (val-cit), 6-maleimidocaproyl (mc), 6-succinimidocaproyl, 6-(2,5-dioxo-3λ 3 -pyrrolidin-1-yl) hexanoyl, methoxy-polyethylene glycol maleimide 6 (MalPeg6), p-aminobenzylcarbamate (PABC), dimethylaminoethanol (DMAE), 3-maleimidopropionyl (MP), 3-succinimidylpropionyl, 3-(2,5-dioxo-3λ 3 -pyrrolidin-1-yl) propionyl, hydrolyzed Peg-maleimide, hydrolyzed maleimide, hydrolyzed succinimide, valine-alanine (Val-Ala), alanine-phenylalanine (ala-phe), p-aminobenzyloxycarbonyl (PAB), 4-(2-pyridylthio)pentanoic acid N-succinimidyl ester (SPP), 4-(N-maleimidomethyl)cyclohexane-1 carboxylic acid N-succinimidyl ester (SMCC), (4-iodo -acetyl)aminobenzoic acid N-succinimidyl ester (SIAB), 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (mc-val-cit-PAB) and 6-maleimidocaproyl-valine-citrulline-p-aminobenzylcarbamate (mc-val-cit-PABC) and an amino acid, optionally (D)-valine, (L)-valine, (D)-alanine and / or (L)-alanine and maleimide.

[0428] In some embodiments, the linker comprises a C1-C6 alkyl group, C=O, -NH-, polyethylene glycol (PEG), optionally 2-10 PEG groups, an amino acid, optionally (D)-valine, (L)-valine, (D)-alanine and / or (L)-alanine and maleimide, succinimide or 2,5-dioxo-3λ 3 In some embodiments, the linker comprises and / or consists of valine-citrulline (val-cit). In some embodiments, the linker comprises and / or consists of valine-citrulline (val-cit)-p-aminobenzyloxycarbonyl (PAB).

[0429] In certain embodiments, the linker comprises one or more reactive moieties and / or targeting agents (such as antibodies or antibody fragments) that can react with a targeting agent. The limiting examples of reactive moieties include azide, alkynes, bissulfones, carbohydrazide, hydrazine, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, pyridopyridazine, half hydrazide, succinimidyl ester, sulfodichlorophenol ester, sulfonyl halide, sulfosuccinimidyl ester, 4-sulfo tetrafluorophenyl ester, tetrafluorophenyl ester, thiazole and NHNH . The limiting examples of targeting agents include a part for protein, protein, polypeptide, nucleic acid, hormone, antibody or antibody fragment. In certain embodiments, the targeting agent is an antibody or antibody fragment.

[0430] In some embodiments, the linker comprises R*, wherein R* is a reactive portion capable of reacting with a targeting agent, a linking portion that connects the linker to the targeting agent, or a targeting agent. In some embodiments, the linker comprises or consists of the formula:

[0431] R*-L

[0432] wherein R* is a reactive moiety, a linking moiety, or a targeting agent.

[0433] In some embodiments, R* is a reactive moiety and is capable of reacting with a functional group such as an aldehyde, amine, disulfide, ketone, thiol in a targeting agent or reacting with a targeting agent in a Staudinger reaction, a Pictet-Spengler reaction, and / or click chemistry. For some reactive moieties, a suitable coupling reagent is used to react the reactive moiety with the targeting agent, for example, when R* is a carboxylic acid, a carbodiimide coupling reagent can be used. In some embodiments, R* is selected from azide, alkyne, bissulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, half hydrazide, succinimidyl ester, and sulfonyl halide.

[0434] In some embodiments, R* is or comprises maleimide:

[0435]

[0436] In some embodiments, R* is or comprises a bissulfone. In some embodiments, a bissulfone is or comprises In some embodiments, the bissulfone is or comprises

[0437] In some embodiments, R* is or comprises an azide, an alkyne, a bissulfone, a carbohydrazide, a hydroxylamine, an iodoacetamide, an isothiocyanate, a maleimide, a phosphine, a hemihydrazide, a succinimidyl ester, and a sulfonyl halide.

[0438] Non-limiting examples of other chemicals are known for attaching compounds to antibodies. US 7,595,292 (Brocchini et al.) mentions linkers that form thioesters with sulfur in disulfide bonds of antibodies. US 7,985,783 (Carico et al.) mentions the introduction of aldehyde residues into antibodies for coupling compounds to antibodies, all of which are incorporated herein by reference in their entirety.

[0439] In some embodiments, R* is a targeting agent, wherein the targeting agent is selected from a protein, a portion of a protein, a peptide, a nucleic acid, a hormone, an antibody, or an antibody fragment. In some embodiments, the targeting agent binds to a tumor-associated antigen, a cancer stem cell-associated antigen, or a viral antigen.

[0440] In some embodiments, the targeting agent is selected from a protein, a portion of a protein, a polypeptide, a nucleic acid, an antibody, or an antibody fragment. In some embodiments, the targeting agent is an antibody or an antibody fragment. In some embodiments, the targeting agent is an antibody.

[0441] In various embodiments, the targeting agent can bind to a target selected from the group consisting of acute myeloid leukemia (AML M4) cells, acute promyelocytic leukemia cells, acute lymphoblastic leukemia cells, acute lymphocytic leukemia cells, chronic lymphocytic leukemia cells, chronic myeloid leukemia cells, chronic T-cell lymphocytic leukemia, myelodysplastic syndrome cells, multiple myeloma cells, prostate cancer cells, renal cell adenocarcinoma cells, pancreatic adenocarcinoma cells, lung cancer cells or gastric adenocarcinoma cells, gastric adenocarcinoma cells, breast cancer cells, colon cancer cells, melanoma cells, thyroid cancer cells, ovarian cancer cells, bladder cancer cells, liver cancer cells, head and neck cancer cells, esophageal cancer cells, Hodgkin's lymphoma cells, non-Hodgkin's lymphoma cells, mesothelioma cells, neuroblastoma cells, neuroendocrine tumor cells, neurofibromatosis type 1 (NF1) cells, neurofibromatosis type 2 (NF2) cells, or osteosarcoma cells.

[0442] In some embodiments, the reactive moiety and / or targeting agent further comprises a linking moiety. In some embodiments, the linking moiety is attached to the reactive moiety and / or targeting agent and the linker to connect the reactive moiety and / or targeting agent to the linker. In some embodiments, the linking moiety comprises one or more selected from the group consisting of: -[CH2] 0-12 、-[CH2CH2O] 0-50 -and-[CH2] 0-12 -C(O)NH-.

[0443] In some embodiments, the linker comprises or consists of the formula:

[0444] R*-L1-L A -

[0445] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or a targeting agent. In some embodiments, the linker L A Conjugated with a drug moiety.

[0446] In some embodiments, linker L A Containing or consisting of the following formula:

[0447] -[CH2CH2O] p -X AA -

[0448] where X AA is an amino acid sequence, and p is an integer from 0 to 50. In some embodiments, the linker L A Further includes

[0449] In some embodiments, linker L A Containing or consisting of the following formula:

[0450] -[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA -

[0451] where X AA is an amino acid sequence, and p is an integer from 0 to 50. In some embodiments, the linker L A Further includes

[0452] In some embodiments, linker L A Containing or consisting of the following formula:

[0453] -[CH2CH2O] p -(CH2) 1-3 -C(O)-X AA -

[0454] where X AA is an amino acid sequence, and p is an integer from 0 to 50. In some embodiments, the linker L A Further includes

[0455] In some embodiments, linker L A Containing or consisting of the following formula:

[0456] -[CH2CH2O] p -(CH2)2-C(O)-X AA -

[0457] where XAA is an amino acid sequence, and p is an integer from 0 to 50. In some embodiments, the linker L A Further includes

[0458] In some embodiments, R* is a reactive moiety. In some embodiments, the reactive moiety is maleimide. In some embodiments, the reactive moiety is a bissulfone.

[0459] In some embodiments, L1 is a group comprising one or more selected from: -[CH2] 0-12 、-[CH2CH2O] 0-50 -and-[CH2] 0-12 -C(O)NH-. In some embodiments, L1 is [CH2] 0-12 -C(O)NH-. In some embodiments, L1 is -[CH2]2-C(O)NH-. In some embodiments, -[CH2]5-C(O)NH-.

[0460] In some embodiments, L1 is [CH2] 1-3 -C(O)NH-.

[0461] In some embodiments, L1 further comprises a linking moiety, which is generated by reacting a reactive moiety with a functional group such as an aldehyde, amine, disulfide, ketone, thiol, etc. in a targeting agent or in a Staudinger reaction, a Pickett-Spengler reaction, and / or click chemistry of the targeting agent. In some embodiments, L1 further comprises a linking moiety selected from a triazole, an amide, a thioether, and a succinimide.

[0462] In some embodiments, L1 further comprises a succinimide (ie, a succinimidyl moiety, "2,5-dioxo-3λ 3 -pyrrolidin-1-yl"):

[0463]

[0464] In some embodiments, L1 is or comprises In some embodiments, L1 is or comprises In some embodiments, L1 is or comprises In some embodiments, L1 is or comprises In some embodiments, L1 is or comprises In some embodiments, L1 further comprises

[0465] In some embodiments, R* is a reactive moiety that has been reacted with a functional group such as an aldehyde, amine, disulfide, ketone, thiol, etc. in a targeting agent (e.g., Ab having Formula (I)) or in a Staudinger reaction, a Pickett-Spengler reaction, and / or click chemistry of a targeting agent (e.g., Ab having Formula (I)). In some embodiments, R* is selected from succinimide, triazole, amide, and thioether.

[0466] In some embodiments, L is of the formula -R * -L1-L A In one non-limiting example, R* is a reactive moiety that has reacted with a functional group of a targeting agent (e.g., with a cysteine ​​moiety of an antibody or antibody fragment, such as a cysteine ​​moiety in the form of formula (I)). In some embodiments, R* is selected from succinimide, triazole, amide, and thioether.

[0467] In some embodiments, R* is a succinimide (ie, a succinimidyl moiety, "2,5-dioxo-3λ 3 -pyrrolidin-1-yl"):

[0468]

[0469] In some embodiments, R* is In some embodiments, R* is In some embodiments, R* is In some embodiments, R* is In some embodiments, R* is In some embodiments, R* is

[0470] In some embodiments, the linker comprises or consists of the formula:

[0471]

[0472] In some embodiments, the linker comprises or consists of the formula:

[0473]

[0474] In some embodiments, the linker comprises or consists of the formula:

[0475]

[0476] In some embodiments, the linker comprises or consists of the formula:

[0477]

[0478] In some embodiments, the linker comprises or consists of the formula:

[0479]

[0480] In some embodiments, the linker comprises or consists of the formula:

[0481]

[0482] In some embodiments, the linker comprises or consists of the formula:

[0483]

[0484] In some embodiments, the linker comprises or consists of the formula:

[0485]

[0486] In some embodiments, the linker comprises or consists of the formula:

[0487]

[0488] In some embodiments, the linker comprises or consists of the formula:

[0489]

[0490] In some embodiments, the linker comprises or consists of the formula:

[0491]

[0492] In some embodiments, the linker comprises or consists of the formula:

[0493]

[0494] In some embodiments, the linker comprises or consists of the formula:

[0495]

[0496] In some embodiments, the linker comprises or consists of the formula:

[0497]

[0498] In some embodiments, the linker comprises or consists of the formula:

[0499]

[0500] In some embodiments, the linker comprises or consists of the formula:

[0501]

[0502] In some embodiments, the linker comprises or consists of the formula:

[0503]

[0504] In some embodiments, the linker comprises or consists of the formula:

[0505]

[0506] In some embodiments, the linker comprises or consists of the formula:

[0507]

[0508] In some embodiments, R* is a targeting agent. In some embodiments, the targeting agent is an antibody or antibody fragment. In some embodiments, the targeting agent is an antibody.

[0509] Compound

[0510] In one aspect, the disclosure provides compounds comprising one or more linkers and one or more drug moieties.

[0511] In some embodiments, the compound has Formula (III) or a salt, solvate, tautomer, isomer, or mixture thereof:

[0512] LD

[0513] Formula (III)

[0514] Wherein in formula (III):

[0515] L is a linker; and

[0516] D contains the drug portion.

[0517] In some embodiments, the linker L of formula (III) reacts with a target moiety (e.g., an antibody or a binding fragment thereof, including but not limited to Ab of formula (III)) that has reacted with a targeting moiety to form a covalent bond with the targeting moiety. In one non-limiting example, the linker L of formula (III) comprises a reactive group R* that reacts with the antibody or antibody fragment Ab to provide a conjugate of formula (I), wherein the linker L of formula (I) is a linker of formula (III) comprising the reaction product of R* and the targeting moiety (e.g., R* is maleimide in formula (III) and is succinimide in formula (I), and further, L is equivalent in each of formula (I) and formula (III).

[0518] In some embodiments, the drug moiety D is selected from Exitecan, Dxd, Sn-38, monomethyl auristatin E (MMAE) and pyridobenzodiazepine (PDD). In some embodiments, Exitecan comprises or has the formula: In some embodiments, the PDD comprises or has the formula: In some embodiments, the PDD comprises or has the formula: In some embodiments, the PDD comprises or has the formula:

[0519] In some embodiments, Sn-38 comprises or has the formula:

[0520] In some embodiments, the compound comprises or consists of the formula:

[0521]

[0522] In some embodiments, the compound comprises or consists of the formula:

[0523]

[0524] In some embodiments, the linker comprises or consists of the formula:

[0525]

[0526] In some embodiments, the linker comprises or consists of the formula:

[0527]

[0528] In some embodiments, the linker comprises or consists of the formula:

[0529]

[0530] In some embodiments, the linker comprises or consists of the formula:

[0531]

[0532] In some embodiments, the linker comprises or consists of the formula:

[0533]

[0534] In some embodiments, the linker comprises or consists of the formula:

[0535]

[0536] In some embodiments, the linker comprises or consists of the formula:

[0537]

[0538] In some embodiments, the linker comprises or consists of the formula:

[0539]

[0540] In some embodiments, LD has the formula:

[0541]

[0542] In some embodiments, LD has the formula:

[0543]

[0544] In some embodiments, LD has the formula:

[0545]

[0546] In some embodiments, LD has the formula:

[0547]

[0548] In some embodiments, LD has the formula:

[0549]

[0550] In some embodiments, LD has the formula:

[0551]

[0552] In some embodiments, LD has the formula:

[0553]

[0554] In some embodiments, LD has the formula:

[0555]

[0556] In some embodiments, LD has the formula:

[0557]

[0558] In some embodiments, LD has the formula:

[0559]

[0560] In some embodiments, the conjugates of the present disclosure (e.g., conjugates of formula (I)) comprise a compound of formula (III). In some embodiments, the compound has formula (III), or a salt, solvate, tautomer, isomer, or mixture thereof.

[0561] LD

[0562] Formula (III)

[0563] Wherein in formula (III):

[0564] L is of the formula R*-L1-L A - connector;

[0565] R* is maleimide;

[0566] L1 is -[CH2] 1-3 -C(O)NH-;

[0567] L A is -[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA -, wherein p is an integer from 5 to 10, and X AA is an amino acid sequence having two amino acid portions; and

[0568] D is Dxd or Sn-38.

[0569] In some embodiments, the compound has Formula (III) or a salt, solvate, tautomer, isomer, or mixture thereof:

[0570] LD

[0571] Formula (III)

[0572] Wherein in formula (III):

[0573] L is of the formula R*-L1-L A - connector;

[0574] R* is maleimide;

[0575] L1 is -[CH2] 1-3 -C(O)NH-;

[0576] L A is -[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA -, optionally -[CH2CH2O] p -(CH2) 1-3 -C(O)-X AA -, optionally -[CH2CH2O] p -(CH2)2-C(O)-X AA -, wherein p is an integer from 5 to 10, and X AA is an amino acid sequence having two amino acid portions; and

[0577] D is selected from

[0578]

[0579]

[0580]

[0581] In some embodiments, X AA Selected from Val-Ala, Tyr-Arg, Phe-Arg, Val-Gln, Val-Cit, Tyr-Met, Leu-Gln, Val-Arg, Met-Thr, Phe-Gln , Thr-Thr, Val-Thr, Ala-Ala, Val-Met, Leu-Met, Ala-Asn, D-Val-D-Gln, D-Ala-D-Ala and Phe-Met.

[0582] In some embodiments, X AA It's Val-Ala.

[0583] In some embodiments, the compound has Formula (III-A) or a salt, solvate, tautomer, isomer, or mixture thereof:

[0584] R*-L1-[CH2CH2O] p -(CH2) 1-5 -C(O)-X AA 1 -X AA 2 -D

[0585] Formula (III-A)

[0586] Wherein in formula (III-A):

[0587] R* is

[0588] L1 is -[CH2] 1-3 -C(O)NH-;

[0589] p is an integer from 6 to 20;

[0590] X AA 1 and X AA 2 are independently selected amino acid moieties; and

[0591] D is a topoisomerase inhibitor.

[0592] In some embodiments, formula (III-A) is a compound having the formula -[CH2CH2O] p-(CH2) 1-3 -C(O)-X AA -. In some embodiments, formula (III-A) is a compound having the formula [CH2CH2O] p -(CH2)2-C(O)-X AA -.

[0593] In some embodiments, X AA 1 is selected from Val, Tyr, Phe, Leu, Met, Thr, Ala, D-Val and D-Ala. In some embodiments, X AA 1 It's Val.

[0594] In some embodiments, X AA 2 is selected from the group consisting of Ala, Arg, Gln, Cit, Met, Thr, Asn, D-Gln, and D-Ala. In some embodiments, X AA 2 It's Ala.

[0595] In some embodiments, -X AA 1 -X AA 2 -Selected from Val-Ala, Tyr-Arg, Phe-Arg, Val-Gln, Val-Cit, Tyr-Met, Leu-Gln, Val-Arg, Met-Thr, Phe-Gl n, Thr-Thr, Val-Thr, Ala-Ala, Val-Met, Leu-Met, Ala-Asn, D-Val-D-Gln, D-Ala-D-Ala and Phe-Met.

[0596] In some embodiments, -X AA 1 -X AA 2 -It's Val-Ala.

[0597] In some embodiments, the compound of formula (III) or the compound of formula (III-A) is selected from a compound of any one of formulas 30 or 3031-3064, or a salt, solvate, tautomer, isomer, or mixture thereof:

[0598]

[0599]

[0600]

[0601]

[0602]

[0603]

[0604]

[0605]

[0606] In some embodiments, the compound of formula (III) or the compound of formula (III-A) is selected from a compound of any one of formulas 30 or 3100-3118, or a salt, solvate, tautomer, isomer, or mixture thereof:

[0607]

[0608]

[0609]

[0610] In one embodiment (CI), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0611] LD

[0612] Formula (III)

[0613] Wherein in formula (III):

[0614] L is a linker; and

[0615] D contains the drug portion.

[0616] The connector has the following formula:

[0617] R*-L1-L A -

[0618] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0619] Connector L A Has the following formula:

[0620] -[Alk-O] p -(CH2) 1-5 -C(O)-X AA -

[0621] in

[0622] Alk represents a C2-C4-alkylene group,

[0623] X AA It is a dipeptide.

[0624] p is an integer from 0 to 50.

[0625] R* is a reactive moiety selected from the group consisting of azide, alkyne, bissulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, hemihydrazide, succinimidyl ester, and sulfonyl halide. In addition, R* is or comprises optionally bissulfone, optionally bissulfone is or comprises

[0626] L1 is -[CH2] 0-12 -C(O)NH-.

[0627] The drug moiety is selected from exitecan having the formula:

[0628] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[0629] In one embodiment (CI), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0630] LD

[0631] Formula (III)

[0632] Wherein in formula (III):

[0633] L is a linker; and

[0634] D contains the drug portion.

[0635] The connector has the following formula:

[0636] R*-L1-L A -

[0637] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0638] Connector L A Has the following formula:

[0639] -[Alk-O] p -(CH2) 1-3 -C(O)-X AA -

[0640] in

[0641] Alk represents a C2-C4-alkylene group,

[0642] X AA It is a dipeptide.

[0643] p is an integer from 0 to 50.

[0644] R* is a reactive moiety selected from the group consisting of azide, alkyne, bissulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, hemihydrazide, succinimidyl ester, and sulfonyl halide. In addition, R* is or comprises optionally bissulfone, optionally bissulfone is or comprises

[0645] L1 is -[CH2] 0-12 -C(O)NH-.

[0646] The drug moiety is selected from exitecan having the formula:

[0647] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[0648] In one embodiment (CIb), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0649] LD

[0650] Formula (III)

[0651] Wherein in formula (III):

[0652] L is a linker; and

[0653] D contains the drug portion.

[0654] The connector has the following formula:

[0655] R*-L1-L A -

[0656] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0657] Connector L A Has the following formula:

[0658] -[Alk-O] p -(CH2)2-C(O)-X AA -

[0659] in

[0660] Alk represents a C2-C4-alkylene group,

[0661] XAA It is a dipeptide.

[0662] p is an integer from 0 to 50.

[0663] R* is a reactive moiety selected from the group consisting of azide, alkyne, bissulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, hemihydrazide, succinimidyl ester, and sulfonyl halide. In addition, R* is or comprises optionally bissulfone, optionally bissulfone is or comprises

[0664] L1 is -[CH2] 0-12 -C(O)NH-.

[0665] The drug moiety is selected from exitecan having the formula:

[0666] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[0667] In one embodiment (CII), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0668] LD

[0669] Formula (III)

[0670] Wherein in formula (III):

[0671] L is a linker; and

[0672] D contains the drug portion.

[0673] The connector has the following formula:

[0674] R*-L1-L A -

[0675] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0676] Connector L A Has the following formula:

[0677] -[Alk-O] p -(CH2) 1-5 -C(O)-X AA -

[0678] in

[0679] Alk represents a C2-C4-alkylene group,

[0680] XAA It is a dipeptide.

[0681] p is an integer from 0 to 50.

[0682] R* is a reactive moiety selected from the group consisting of azide, alkyne, bissulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, hemihydrazide, succinimidyl ester, and sulfonyl halide. In addition, R* is or comprises optionally bissulfone, optionally bissulfone is or comprises

[0683] L1 is -[CH2] 0-12 -C(O)NH-.

[0684] The drug portion is selected from:

[0685]

[0686]

[0687]

[0688] In one embodiment (CIIa), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0689] LD

[0690] Formula (III)

[0691] Wherein in formula (III):

[0692] L is a linker; and

[0693] D contains the drug portion.

[0694] The connector has the following formula:

[0695] R*-L1-L A -

[0696] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0697] Connector L A Has the following formula:

[0698] -[Alk-O] p -(CH2) 1-3 -C(O)-X AA -

[0699] in

[0700] Alk represents a C2-C4-alkylene group,

[0701] X AA It is a dipeptide.

[0702] p is an integer from 0 to 50.

[0703] R* is a reactive moiety selected from the group consisting of azide, alkyne, bissulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, hemihydrazide, succinimidyl ester, and sulfonyl halide. In addition, R* is or comprises optionally bissulfone, optionally bissulfone is or comprises

[0704] L1 is -[CH2] 0-12 -C(O)NH-.

[0705] The drug portion is selected from:

[0706]

[0707]

[0708]

[0709] In one embodiment (CIIb), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0710] LD

[0711] Formula (III)

[0712] Wherein in formula (III):

[0713] L is a linker; and

[0714] D contains the drug portion.

[0715] The connector has the following formula:

[0716] R*-L1-L A -

[0717] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0718] Connector L A Has the following formula:

[0719] -[Alk-O] p -(CH2)2-C(O)-X AA -

[0720] in

[0721] Alk represents a C2-C4-alkylene group,

[0722] X AA It is a dipeptide.

[0723] p is an integer from 0 to 50.

[0724] R* is a reactive moiety selected from the group consisting of azide, alkyne, bissulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, hemihydrazide, succinimidyl ester, and sulfonyl halide. In addition, R* is or comprises optionally bissulfone, optionally bissulfone is or comprises

[0725] L1 is -[CH2] 0-12 -C(O)NH-.

[0726] The drug portion is selected from:

[0727]

[0728]

[0729]

[0730] In one embodiment (CIII), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0731] LD

[0732] Formula (III)

[0733] Wherein in formula (III):

[0734] L is a linker; and

[0735] D contains the drug portion.

[0736] The connector has the following formula:

[0737] R*-L1-L A -

[0738] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0739] Connector L A Has the following formula:

[0740] -[Alk-O] p -(CH2)2-C(O)-X AA -

[0741] in

[0742] X AA It is a non-polar dipeptide.

[0743] p is an integer from 5 to 10, for example, 5, 6, 7, 8, 9 or 10.

[0744] R* is a reactive moiety selected from the group consisting of azide, alkyne, bissulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, hemihydrazide, succinimidyl ester, and sulfonyl halide. In addition, R* is or comprises optionally bissulfone, optionally bissulfone is or comprises

[0745] L1 is -[CH2] 0-12 -C(O)NH-.

[0746] The drug moiety is selected from exitecan having the formula:

[0747] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[0748] In one embodiment (CIV), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0749] LD

[0750] Formula (III)

[0751] Wherein in formula (III):

[0752] L is a linker; and

[0753] D contains the drug portion.

[0754] The connector has the following formula:

[0755] R*-L1-L A -

[0756] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0757] Connector L A Has the following formula:

[0758] -[Alk-O] p -(CH2)2-C(O)-X AA -

[0759] in

[0760] Alk represents a C2-C4-alkylene group,

[0761] X AA It is a dipeptide.

[0762] p is an integer from 0 to 50.

[0763] R* is maleimide.

[0764] L1 is -[CH2] 1-3 -C(O)NH-.

[0765] The drug moiety is selected from exitecan having the formula:

[0766] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[0767] In one embodiment (CV), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0768] LD

[0769] Formula (III)

[0770] Wherein in formula (III):

[0771] L is a linker; and

[0772] D contains the drug portion.

[0773] The connector has the following formula:

[0774] R*-L1-L A -

[0775] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0776] Connector L A Has the following formula:

[0777] -[CH2-CH2-O] p -(CH2)2-C(O)-X AA -

[0778] in

[0779] X AA It is a non-polar dipeptide.

[0780] p is an integer from 5 to 10, for example, 5, 6, 7, 8, 9 or 10.

[0781] R* is maleimide.

[0782] L1 is -[CH2] 1-3 -C(O)NH-.

[0783] The drug moiety is selected from exitecan having the formula:

[0784] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[0785] In one embodiment (CVI), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0786] LD

[0787] Formula (III)

[0788] Wherein in formula (III):

[0789] L is a linker; and

[0790] D contains the drug portion.

[0791] The connector has the following formula:

[0792] R*-L1-L A -

[0793] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0794] Connector L A Has the following formula:

[0795] -[Alk-O] p -(CH2)2-C(O)-X AA -

[0796] in

[0797] Alk represents a C2-C4-alkylene group,

[0798] X AA It's Val-Ala,

[0799] p is 7 or 8.

[0800] R* is a reactive moiety selected from the group consisting of azide, alkyne, bissulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, hemihydrazide, succinimidyl ester, and sulfonyl halide. In addition, R* is or comprises optionally bissulfone, optionally bissulfone is or comprises

[0801] L1 is -[CH2] 0-12 -C(O)NH-.

[0802] The drug moiety is selected from exitecan having the formula:

[0803] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[0804] In one embodiment (CVII), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0805] LD

[0806] Formula (III)

[0807] Wherein in formula (III):

[0808] L is a linker; and

[0809] D contains the drug portion.

[0810] The connector has the following formula:

[0811] R*-L1-L A -

[0812] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0813] Connector L A Has the following formula:

[0814] -[CH2-CH2-O] p -(CH2)2-C(O)-X AA -

[0815] in

[0816] X AA It's Val-Ala,

[0817] p is 7 or 8.

[0818] R* is a reactive moiety selected from the group consisting of azide, alkyne, bissulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, hemihydrazide, succinimidyl ester, and sulfonyl halide. In addition, R* is or comprises optionally bissulfone, optionally bissulfone is or comprises

[0819] L1 is -[CH2] 0-12 -C(O)NH-.

[0820] The drug moiety is selected from exitecan having the formula:

[0821] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[0822] In one embodiment (CVIII), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0823] LD

[0824] Formula (III)

[0825] Wherein in formula (III):

[0826] L is a linker; and

[0827] D contains the drug portion.

[0828] The connector has the following formula:

[0829] R*-L1-L A -

[0830] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0831] Connector L A Has the following formula:

[0832] -[Alk-O] p -(CH2)2-C(O)-X AA -

[0833] in

[0834] Alk represents a C2-C4-alkylene group,

[0835] X AA It's Val-Ala,

[0836] p is 7 or 8.

[0837] R* is maleimide.

[0838] L1 is -[CH2] 1-3 -C(O)NH-.

[0839] The drug moiety is selected from exitecan having the formula:

[0840] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[0841] In one embodiment (CIX), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0842] LD

[0843] Formula (III)

[0844] Wherein in formula (III):

[0845] L is a linker; and

[0846] D contains the drug portion.

[0847] The connector has the following formula:

[0848] R*-L1-L A -

[0849] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[0850] Connector L A Has the following formula:

[0851] -[CH2-CH2-O] p -(CH2)2-C(O)-X AA -

[0852] in

[0853] X AA It's Val-Ala,

[0854] p is 7 or 8.

[0855] R* is maleimide.

[0856] L1 is -[CH2] 1-3 -C(O)NH-.

[0857] The drug moiety is selected from exitecan having the formula:

[0858] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[0859] In one embodiment (CX), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0860] LD

[0861] Formula (III)

[0862] As defined in any one of embodiments (CI) to (CX), wherein the exponent p is 8.

[0863] In one embodiment (CXI), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0864] LD

[0865] Formula (III)

[0866] As defined in any one of embodiments (CI) to (CX), wherein L1 is -[CH2]2-C(O)NH-.

[0867] In one embodiment (CXII), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0868] LD

[0869] Formula (III)

[0870] As defined in any one of embodiments (CI) to (CXI), wherein the drug moiety is selected from exitecan having the formula:

[0871] Dxd and SN-38.

[0872] In one embodiment (XIII), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0873] LD

[0874] Formula (III)

[0875] As defined in any one of embodiments (CI) to (CXII), wherein the drug moiety is selected from exitecan having the formula:

[0876] Dxd and SN-38.

[0877] In one embodiment (XIV), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0878] LD

[0879] Formula (III)

[0880] As defined in any one of embodiments (CI) to (CXIII), wherein the drug moiety is exitecan having the formula:

[0881]

[0882] In one embodiment (CXV), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0883] LD

[0884] Formula (III)

[0885] As defined in any one of embodiments (CI) to (CXIV), wherein X AA Not Val-Cit or Phe-Lys.

[0886] In one embodiment (CXVI), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[0887] LD

[0888] Formula (III)

[0889] Some of the LDs have the following structures

[0890]

[0891] In one aspect, the present disclosure provides a conjugate of formula (II):

[0892] Ab-L-

[0893] Formula (II)

[0894] wherein Ab is an antibody or antibody fragment, and L is a linker.

[0895] Any antibody, antibody fragment and / or linker disclosed herein is encompassed within formula (II). In some embodiments, the linker comprises and / or consists of a partial structure that is further conjugated to a drug moiety. In some embodiments, the linker comprises and / or consists of a complete structure that is further conjugated to a drug moiety.

[0896] In some embodiments, the linker comprises or consists of the formula:

[0897]

[0898] In some embodiments, the linker has the formula:

[0899]

[0900] In some embodiments, the linker has the formula:

[0901]

[0902] In some embodiments, the linker comprises or consists of the formula:

[0903]

[0904]

[0905] In some embodiments, the linker comprises or consists of the formula:

[0906]

[0907] In some embodiments, the linker comprises or consists of the formula:

[0908]

[0909] In some embodiments, the linker comprises or consists of the formula:

[0910]

[0911] In some embodiments, the linker comprises or consists of the formula:

[0912]

[0913] In some embodiments, the linker comprises or consists of the formula:

[0914]

[0915] In some embodiments, the linker comprises or consists of the formula:

[0916]

[0917] In some embodiments, the linker comprises or consists of the formula:

[0918]

[0919] In some embodiments, the linker comprises or consists of the formula:

[0920]

[0921] In some embodiments, the linker comprises or consists of the formula:

[0922]

[0923] Antibody-drug conjugates

[0924] In one aspect, the present disclosure provides an antibody-drug conjugate comprising one or more joints and one or more drug moieties. In certain embodiments, the antibody-drug conjugate comprises an antibody, antibody fragment, joint and / or drug moiety as described herein (e.g., Formula (II) and / or Formula (III)). In certain embodiments, the antibody and / or antibody fragment is conjugated to the joint-drug moiety by a sulfur-containing moiety (e.g., thiol) on the antibody and / or antibody fragment. In certain embodiments, the sulfur-containing moiety comprises and / or consists of the sulfur moiety of one or more interchain disulfide bridges of the antibody and / or antibody fragment. In a non-limiting example, a reducing agent is used to destroy the interchain disulfide bridges that hold the arms of the antibody (e.g., mAb) and / or antibody fragment together, and the joint and / or payload is conjugated to the sulfur moiety of the disulfide bridge. In a non-limiting example, all available thiols from interchain disulfides are occupied due to the presence of 4 interchain disulfides in the antibody (e.g., mAb) 8. In certain embodiments, antibodies and / or antibody fragments are conjugated to joint-drug moieties by one or more amino acid residues on antibodies and / or antibody fragments, and the amino acid residues include but are not limited to amino acid residues comprising sulfur-containing side chains (e.g., cysteine ​​and / or methionine). In certain embodiments, antibodies and / or antibody fragments are conjugated to joint-drug moieties by one or more cysteine ​​residues on antibodies and / or antibody fragments. In certain embodiments, antibodies and / or antibody fragments are conjugated to joint-drug moieties by one or more cysteine ​​residues on antibodies. In certain embodiments, antibodies and / or antibody fragments are conjugated to joint-drug moieties by one or more cysteine ​​residues on antibodies. In certain embodiments, amino acid residues are unengineered (e.g., unengineered cysteine ​​residues and / or unengineered methionine residues). In certain embodiments, amino acid residues are engineered (e.g., engineered cysteine ​​residues and / or engineered methionine residues). In certain embodiments, joints are selected from any joints described herein. In certain embodiments, drug moieties are selected from any drug moieties described herein.

[0925] In one aspect, the present disclosure provides an antibody-drug conjugate (ADC) having formula (I):

[0926] Ab-[LD] n

[0927] Formula (I)

[0928] Wherein in formula (I):

[0929] Ab comprises an antibody or a binding fragment thereof;

[0930] L is the connector;

[0931] D comprises a drug moiety; and

[0932] n is an integer from 1 to 20.

[0933] Any of the antibodies, antibody fragments, linkers and / or drug moieties disclosed herein are encompassed by Formula (III).

[0934] In some embodiments, the linker L of formula (I) is a linker of formula (III) that has been reacted with an antibody or binding fragment thereof Ab (eg, the linker comprises a reactive group R* that reacts with the antibody or antibody fragment Ab).

[0935] In some embodiments, the linker comprises or consists of the formula:

[0936]

[0937] In some embodiments, the linker comprises or consists of the formula:

[0938]

[0939] In some embodiments, the linker comprises or consists of the formula:

[0940]

[0941] In some embodiments, the linker comprises or consists of the formula:

[0942]

[0943] In some embodiments, the linker comprises or consists of the formula:

[0944]

[0945] In some embodiments, the linker comprises or consists of the formula:

[0946]

[0947] In some embodiments, the linker comprises or consists of the formula:

[0948]

[0949] In some embodiments, the linker comprises or consists of the formula:

[0950]

[0951] In some embodiments, the linker comprises or consists of the formula:

[0952]

[0953] In some embodiments, the linker comprises or consists of the formula:

[0954]

[0955] In some embodiments, the linker comprises or consists of the formula:

[0956]

[0957] In some embodiments, the linker comprises or consists of valine-citrulline.

[0958] In some embodiments, the drug moiety D is selected from Exitecan, Dxd, Sn-38, monomethyl auristatin E (MMAE) and pyridobenzodiazepine (PDD). In some embodiments, Exitecan comprises or has the formula: In some embodiments, the PDD comprises or has the formula: In some embodiments, the PDD comprises or has the formula: In some embodiments, the PDD comprises or has the formula:

[0959] In some embodiments, Sn-38 comprises or has the formula:

[0960] In some embodiments, LD has the formula:

[0961]

[0962] In some embodiments, LD has the formula:

[0963]

[0964] In some embodiments, LD has the formula:

[0965]

[0966] In some embodiments, LD has the formula:

[0967]

[0968] In some embodiments, LD has the formula:

[0969]

[0970] In some embodiments, LD has the formula:

[0971]

[0972] In some embodiments, LD has the formula:

[0973]

[0974] In some embodiments, LD has the formula:

[0975]

[0976] In some embodiments, LD has the formula:

[0977]

[0978] In some embodiments, LD has the formula:

[0979]

[0980] In some embodiments, LD has the formula:

[0981]

[0982] In some embodiments, LD has the formula:

[0983]

[0984] In some embodiments, LD has the formula -val-cit-MMAE.

[0985] In some embodiments, LD has the formula -val-cit.PAB-MMAE.

[0986] In some embodiments, the antibody-drug conjugate has Formula (IA):

[0987] Ab-[L1-(CH2CH2O) p -X AA 1 -X AA 2 -D] n

[0988] Formula (IA)

[0989] Wherein in formula (IA):

[0990] Ab comprises an antibody or a binding fragment thereof;

[0991] L1 is

[0992] p is an integer from 6 to 20;

[0993] X AA 1 and X AA 2 are independently selected amino acid moieties;

[0994] D is a topoisomerase inhibitor; and

[0995] n is an integer from 1 to 20.

[0996] In some embodiments, the conjugate of formula (I) or the conjugate of formula (IA) is selected from a conjugate of any one of formulas 1030-1064:

[0997]

[0998]

[0999]

[1000]

[1001]

[1002]

[1003]

[1004]

[1005] In some embodiments, the conjugate of formula (I) or the conjugate of formula (IA) is selected from a conjugate of formula 1030 or any one of 1100-1118:

[1006]

[1007]

[1008] Mode <![CDATA[-X AA -or-X AA 1 -X AA 2 -]]> 1030 Val-Ala 1101 Tyr-Arg 1102 Phe-Arg 1103 Val-Gln 1104 Val-Cit 1105 Tyr-Met 1106 Leu-Gln 1107 Val-Arg 1108 Met-Thr 1109 Phe-Gln 1110 Thr-Thr 1111 Val-Thr 1112 Ala-Ala 1113 Val-Met 1114 Leu-Met 1115 Ala-Asn 1116 D-Val-D-Gln 1117 D-Ala-D-Ala 1118 Phe-Met

[1009] In some embodiments, n is an integer from 1 to 10. In some embodiments, n is an integer from 4 to 8. In some embodiments, n is an integer from 2 to 8. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10.

[1010] In one embodiment (I), the present disclosure provides an antibody-drug conjugate having formula (I):

[1011] Ab-[LD] n

[1012] Formula (I)

[1013] Wherein in formula (I):

[1014] Ab is an antibody or its binding fragment;

[1015] L is the connector;

[1016] D is a drug moiety; and

[1017] n is an integer from 1 to 20.

[1018] The connector has the following formula:

[1019] -R*-L1-L A -

[1020] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[1021] Connector L A Has the following formula:

[1022] -[Alk-O] p -(CH2) 1-5 -C(O)-X AA -

[1023] in

[1024] Alk represents a C2-C4-alkylene group,

[1025] X AA It is a dipeptide.

[1026] p is an integer from 0 to 50.

[1027] R* is a reactive moiety selected from the group consisting of succinimide, triazole, amide, and thioether.

[1028] L1 is -[CH2] 0-12 -C(O)NH-.

[1029] The drug moiety is selected from exitecan having the formula:

[1030] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[1031] In one embodiment (Ia), the present disclosure provides an antibody-drug conjugate having formula (I):

[1032] Ab-[LD] n

[1033] Formula (I)

[1034] Wherein in formula (I):

[1035] Ab is an antibody or its binding fragment;

[1036] L is the connector;

[1037] D is a drug moiety; and

[1038] n is an integer from 1 to 20.

[1039] The connector has the following formula:

[1040] -R*-L1-L A -

[1041] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[1042] Connector L A Has the following formula:

[1043] -[Alk-O] p -(CH2)2-C(O)-X AA -

[1044] in

[1045] Alk represents a C2-C4-alkylene group,

[1046] X AA It is a dipeptide.

[1047] p is an integer from 0 to 50.

[1048] R* is a reactive moiety selected from the group consisting of succinimide, triazole, amide, and thioether.

[1049] L1 is -[CH2] 0-12 -C(O)NH-.

[1050] The drug moiety is selected from exitecan having the formula:

[1051] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[1052] In one embodiment (II), the present disclosure provides an antibody-drug conjugate having formula (I):

[1053] Ab-[LD] n

[1054] Formula (I)

[1055] Wherein in formula (I):

[1056] Ab is an antibody or its binding fragment;

[1057] L is the connector;

[1058] D is a drug moiety; and

[1059] n is an integer from 1 to 20.

[1060] The connector has the following formula:

[1061] -R*-L1-L A -

[1062] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent. Linker L A Has the following formula:

[1063] -[Alk-O] p -(CH2) 1-5 -C(O)-X AA -

[1064] in

[1065] Alk represents a C2-C4-alkylene group,

[1066] X AA It is a dipeptide.

[1067] p is an integer from 0 to 50.

[1068] R* is a reactive moiety selected from the group consisting of succinimide, triazole, amide, and thioether.

[1069] L1 is -[CH2] 0-12 -C(O)NH-.

[1070] The drug portion is selected from:

[1071]

[1072]

[1073]

[1074]

[1075] In one embodiment (IIa), the present disclosure provides an antibody-drug conjugate having formula (I):

[1076] Ab-[LD] n

[1077] Formula (I)

[1078] Wherein in formula (I):

[1079] Ab is an antibody or its binding fragment;

[1080] L is the connector;

[1081] D is a drug moiety; and

[1082] n is an integer from 1 to 20.

[1083] The connector has the following formula:

[1084] -R*-L1-L A -

[1085] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[1086] Connector L A Has the following formula:

[1087] -[Alk-O] p -(CH2)2-C(O)-X AA -

[1088] in

[1089] Alk represents a C2-C4-alkylene group,

[1090] X AA It is a dipeptide.

[1091] p is an integer from 0 to 50.

[1092] R* is a reactive moiety selected from the group consisting of succinimide, triazole, amide, and thioether.

[1093] L1 is -[CH2] 0-12 -C(O)NH-. The drug moiety is selected from:

[1094]

[1095]

[1096]

[1097] In one embodiment (III), the present disclosure provides an antibody-drug conjugate having formula (I):

[1098] Ab-[LD] n

[1099] Formula (I)

[1100] Wherein in formula (I):

[1101] Ab is an antibody or its binding fragment;

[1102] L is the connector;

[1103] D is a drug moiety; and

[1104] n is an integer from 1 to 20.

[1105] The connector has the following formula:

[1106] -R*-L1-L A -

[1107] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[1108] Connector L A Has the following formula:

[1109] -[CH2-CH2-O] p -(CH2)2-C(O)-X AA -

[1110] in

[1111] X AA It is a non-polar dipeptide.

[1112] p is an integer from 5 to 10, for example, 5, 6, 7, 8, 9 or 10.

[1113] R* is a reactive moiety selected from the group consisting of succinimide, triazole, amide, and thioether.

[1114] L1 is -[CH2] 0-12 -C(O)NH-.

[1115] The drug moiety is selected from exitecan having the formula:

[1116] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[1117] In one embodiment (IV), the present disclosure provides an antibody-drug conjugate having formula (I):

[1118] Ab-[LD] n

[1119] Formula (I)

[1120] Wherein in formula (I):

[1121] Ab is an antibody or its binding fragment;

[1122] L is the connector;

[1123] D is a drug moiety; and

[1124] n is an integer from 1 to 20.

[1125] The connector has the following formula:

[1126] -R*-L1-L A -

[1127] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[1128] Connector L A Has the following formula:

[1129] -[Alk-O] p -(CH2)2-C(O)-X AA -

[1130] in

[1131] Alk represents a C2-C4-alkylene group,

[1132] X AA It is a dipeptide.

[1133] p is an integer from 0 to 50.

[1134] R* is succinimide.

[1135] L1 is -[CH2] 1-3 -C(O)NH-.

[1136] The drug moiety is selected from exitecan having the formula:

[1137] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[1138] In one embodiment (V), the present disclosure provides an antibody-drug conjugate having formula (I):

[1139] Ab-[LD] n

[1140] Formula (I)

[1141] Wherein in formula (I):

[1142] Ab is an antibody or its binding fragment;

[1143] L is the connector;

[1144] D is a drug moiety; and

[1145] n is an integer from 1 to 20.

[1146] The connector has the following formula:

[1147] -R*-L1-L A -

[1148] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[1149] Connector L A Has the following formula:

[1150] -[CH2-CH2-O] p -(CH2)2-C(O)-X AA -

[1151] in

[1152] X AA It is a non-polar dipeptide.

[1153] p is an integer from 5 to 10, for example, 5, 6, 7, 8, 9 or 10.

[1154] R* is succinimide.

[1155] L1 is -[CH2] 1-3 -C(O)NH-.

[1156] The drug moiety is selected from exitecan having the formula:

[1157] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[1158] In one embodiment (VI), the present disclosure provides an antibody-drug conjugate having formula (I):

[1159] Ab-[LD] n

[1160] Formula (I)

[1161] Wherein in formula (I):

[1162] Ab is an antibody or its binding fragment;

[1163] L is the connector;

[1164] D is a drug moiety; and

[1165] n is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[1166] The connector has the following formula:

[1167] -R*-L1-L A -

[1168] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[1169] Connector L A Has the following formula:

[1170] -[Alk-O] p -(CH2)2-C(O)-X AA -

[1171] in

[1172] Alk represents a C2-C4-alkylene group,

[1173] X AA It's Val-Ala,

[1174] p is 7 or 8.

[1175] R* is a reactive moiety selected from the group consisting of succinimide, triazole, amide, and thioether.

[1176] L1 is -[CH2] 0-12 -C(O)NH-.

[1177] The drug moiety is selected from exitecan having the formula:

[1178] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[1179] In one embodiment (VII), the present disclosure provides an antibody-drug conjugate having formula (I):

[1180] Ab-[LD] n

[1181] Formula (I)

[1182] Wherein in formula (I):

[1183] Ab is an antibody or its binding fragment;

[1184] L is the connector;

[1185] D is a drug moiety; and

[1186] n is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[1187] The connector has the following formula:

[1188] -R*-L1-LA -

[1189] Among them L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[1190] Connector L A Has the following formula:

[1191] -[CH2-CH2-O] p -(CH2)2-C(O)-X AA -

[1192] in

[1193] X AA It's Val-Ala,

[1194] p is 7 or 8.

[1195] R* is a reactive moiety selected from the group consisting of succinimide, triazole, amide, and thioether.

[1196] L1 is -[CH2] 0-12 -C(O)NH-.

[1197] The drug moiety is selected from exitecan having the formula:

[1198] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[1199] In one embodiment (VIII), the present disclosure provides an antibody-drug conjugate having formula (I):

[1200] Ab-[LD] n

[1201] Formula (I)

[1202] Wherein in formula (I):

[1203] Ab is an antibody or its binding fragment;

[1204] L is the connector;

[1205] D is a drug moiety; and

[1206] n is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[1207] The connector has the following formula:

[1208] -R*-L1-L A -

[1209] Among them LA is a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[1210] Connector L A Has the following formula:

[1211] -[Alk-O] p -(CH2)2-C(O)-X AA -

[1212] in

[1213] Alk represents a C2-C4-alkylene group,

[1214] X AA It's Val-Ala,

[1215] p is 7 or 8.

[1216] R* is succinimide.

[1217] L1 is -[CH2] 1-3 -C(O)NH-.

[1218] The drug moiety is selected from exitecan having the formula:

[1219] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[1220] In one embodiment (IX), the present disclosure provides an antibody-drug conjugate having formula (I):

[1221] Ab-[LD] n

[1222] Formula (I)

[1223] Wherein in formula (I):

[1224] Ab is an antibody or its binding fragment;

[1225] L is the connector;

[1226] D is a drug moiety; and

[1227] n is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[1228] The connector has the following formula:

[1229] -R*-L1-L A -

[1230] Among them L Ais a linker, L1 is a connecting moiety, and R* is a reactive moiety or targeting agent.

[1231] Connector L A Has the following formula:

[1232] -[CH2-CH2-O] p -(CH2)2-C(O)-X AA -

[1233] in

[1234] X AA It's Val-Ala,

[1235] p is 7 or 8.

[1236] R* is succinimide.

[1237] L1 is -[CH2] 1-3 -C(O)NH-.

[1238] The drug moiety is selected from exitecan having the formula:

[1239] Dxd, Sn-38, monomethyl auristatin E (MMAE), and pyridobenzodiazepine (PDD).

[1240] In one embodiment (X), the present disclosure provides an antibody-drug conjugate having formula (I):

[1241] Ab-[LD] n

[1242] Formula (I)

[1243] As defined in any one of embodiments (I) to (X), wherein the index p is 8.

[1244] In one embodiment (XI), the present disclosure provides an antibody-drug conjugate having formula (I):

[1245] Ab-[LD] n

[1246] Formula (I)

[1247] As defined in any one of embodiments (I) to (X), wherein L1 is -[CH2]2-C(O)NH-.

[1248] In one embodiment (XII), the present disclosure provides an antibody-drug conjugate having formula (I):

[1249] Ab-[LD] n

[1250] Formula (I)

[1251] As defined in any one of embodiments (I) to (XI), wherein the drug moiety is selected from exitecan having the formula:

[1252] Dxd and SN-38.

[1253] In one embodiment (XIII), the present disclosure provides an antibody-drug conjugate having formula (I):

[1254] Ab-[LD] n

[1255] Formula (I)

[1256] As defined in any one of embodiments (I) to (XII), wherein the drug moiety is selected from exitecan having the formula:

[1257] Dxd and SN-38.

[1258] In one embodiment (XIV), the present disclosure provides an antibody-drug conjugate having formula (I):

[1259] Ab-[LD] n

[1260] Formula (I)

[1261] As defined in any one of embodiments (I) to (XIII), wherein the drug moiety is exitecan having the formula:

[1262]

[1263] In one embodiment (XV), the present disclosure provides an antibody-drug conjugate having formula (I):

[1264] Ab-[LD] n

[1265] Formula (I)

[1266] As defined in any one of embodiments (I) to (XIV), wherein X AA Not Val-Cit or Phe-Lys.

[1267] In one embodiment (XVI), the present disclosure provides an antibody-drug conjugate having formula (I):

[1268] Ab-[LD] n

[1269] Formula (I)

[1270] Some of them – [LD] n With the following structure

[1271]

[1272] wherein the moiety Ab- is as defined in any one of embodiments (I) to (XV), and wherein the moiety Ab- is reacted with a maleimide group.

[1273] In one embodiment (XVII), the present disclosure provides a compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof:

[1274] LD

[1275] Formula (III)

[1276] Wherein in formula (III):

[1277] L is a linker; and

[1278] D contains the drug portion,

[1279] Any one of embodiments (I) to (XVI) of said formula (III) corresponding to formula (I), wherein Ab is absent and n is 1.

[1280] In an embodiment, the present disclosure provides an antibody-drug conjugate (ADC) having formula (I):

[1281] Ab-[LD] n

[1282] Formula (I)

[1283] Wherein in formula (I):

[1284] Ab is an antibody or its binding fragment;

[1285] L is of the formula -R*-L1-L A - connector, where L A is a linker, L1 is a connecting moiety, and R* is a reactive moiety or a targeting agent;

[1286] D is a drug moiety; and

[1287] n is an integer from 1 to 20. In an embodiment, R* is a reactive moiety selected from succinimide, triazole, amide, and thioether. In an embodiment, R* is succinimide. In an embodiment, L1 is [CH2] 0-12 -C(O)NH-. In some embodiments, the linker L A With the formula -[Alk-O] p-(CH2)2-C(O)-X AA -. In some embodiments, the linker L A With the formula -[CH2CH2-O] p -(CH2)2-C(O)-X AA -, wherein p is an integer from 5 to 10. In an embodiment, X AA is selected from the group consisting of Val-Ala, Tyr-Arg, Phe-Arg, Val-Gln, Val-Cit, Tyr-Met, Leu-Gln, Val-Arg, Met-Thr, Phe-Gln, Thr-Thr, Val-Thr, Ala-Ala, Val-Met, Leu-Met, Ala-Asn, D-Val-D-Gln, D-Ala-D-Ala, and Phe-Met. In some embodiments, X AA In some embodiments, L1 is [CH2] 1-3 -C(O)NH-. In some embodiments, L1 is -[CH2]2-C(O)NH-. In some embodiments, p is 7 or 8. In some embodiments, p is 8.

[1288] In some embodiments, the antibody-drug conjugates described herein (eg, Formula (I)) are capable of forming metabolites in vivo. In some embodiments, the antibody-drug conjugates are capable of forming metabolites in vitro.

[1289] In some embodiments, the antibody-drug conjugates described herein (eg, Formula (I)) are capable of forming a metabolite of Formula 300:

[1290]

[1291] In some embodiments, the antibody-drug conjugates described herein (eg, Formula (III)) are capable of forming a metabolite of Formula 301:

[1292]

[1293] In some embodiments, the antibody-drug conjugates described herein (eg, Formula (III)) are capable of forming a metabolite of Formula 302:

[1294]

[1295] In some embodiments, the antibody-drug conjugates described herein (eg, Formula (I)) are capable of forming a metabolite of Formula 303:

[1296]

[1297] In some embodiments, a metabolite of Formula 300 can form a metabolite of Formula 301. In some embodiments, a metabolite of Formula 300 can form a metabolite of Formula 302. In some embodiments, a metabolite of Formula 300 can form a metabolite of Formula 303. In some embodiments, a metabolite of Formula 301 can form a metabolite of Formula 302. In some embodiments, a metabolite of Formula 301 can form a metabolite of Formula 303. In some embodiments, a metabolite of Formula 300 can form a metabolite of Formula 301, and a metabolite of Formula 301 can form a metabolite of Formula 302. In some embodiments, a metabolite of Formula 300 can form a metabolite of Formula 301, a metabolite of Formula 301 can form a metabolite of Formula 302, and a metabolite of Formula 302 can form a metabolite of Formula 303.

[1298] In some embodiments, the antibody drug conjugates as described herein bind to the target antigen at a pH of about 1000 nM, about 500 nM, about 400 nM, about 300 nM, about 200 nM, about 180 nM, about 160 nM, about 140 nM, about 120 nM, or about 100 nM. D Value combination.

[1299] In some embodiments, an ADC having the following: (a) an antibody or antigen-binding fragment thereof, (b) a linker, and (c) a drug moiety can be produced or generated. The drug-antibody ratio (DAR) or drug loading represents the number of drug (D) molecules and / or moieties conjugated to each antibody. In some embodiments, the number of linker-drug moieties attached to the antibody can be any number suitable for developing an ADC. In some embodiments, the number of linker-drug moieties per antibody is in the range of about 1 to about 10. In some embodiments, the number of linker-drug moieties per antibody is about 10. In some embodiments, the number of linker-drug moieties per antibody is about 9. In some embodiments, the number of linker-drug moieties per antibody is about 8. In some embodiments, the number of linker-drug moieties per antibody is about 7. In some embodiments, the number of linker-drug moieties per antibody is about 6. In some embodiments, the number of linker-drug moieties per antibody is about 5. In some embodiments, the number of linker-drug moieties per antibody is about 4. In some embodiments, the number of linker-drug moieties per antibody is about 3. In some embodiments, the number of linker-drug moieties per antibody is about 2. In some embodiments, the number of linker-drug moieties per antibody is about 1. In some embodiments, the number of linker-drug moieties per antibody is greater than 4, such as 5, 6, 7, 8, 9, 10, 11, 12 or more linker-drug moieties per antibody. Non-limiting examples of methods for determining DAR include various conventional means, such as UV spectroscopy, mass spectrometry, ELISA assays, radioactivity measurement methods, hydrophobic interaction chromatography (HIC), electrophoresis, and HPLC. In some embodiments, the DAR of the ADC of the present disclosure is equivalent to "n" as mentioned in Formula (I).

[1300] In some embodiments, the antibody drug conjugates (ADCs) of the present disclosure (e.g., ADCs of the present disclosure comprising an antibody and / or antibody fragment conjugated to a drug moiety via a linker) are capable of internalization. In another embodiment, the antibody drug conjugates (ADCs) of the present disclosure are capable of inducing cell death in cells that endogenously express the target antigen.

[1301] In some embodiments, LD has the formula:

[1302]

[1303] In some embodiments, LD has the formula:

[1304]

[1305] In some embodiments, LD has the formula:

[1306]

[1307] In some embodiments, LD has the formula:

[1308]

[1309] In some embodiments, LD has the formula:

[1310]

[1311] In some embodiments, LD has the formula:

[1312]

[1313] In some embodiments, LD has the formula:

[1314]

[1315] In some embodiments, LD has the formula:

[1316]

[1317] In some embodiments, LD has the formula -val-cit-MMAE.

[1318] In some embodiments, LD has the formula -val-cit-PAB-MMAE.

[1319] Treatment

[1320] In one aspect, the conjugates (e.g., ADCs), compounds, and compositions described herein (e.g., Formula (I), Formula (II), and / or Formula (III)) can be used in a method for treating a disease. In some embodiments, the disease is cancer. In one embodiment, the disease is a hyperproliferative disease. In some embodiments, the hyperproliferative disorder is cancer. In some embodiments, the cancer is pancreatic cancer, breast cancer, prostate cancer, lymphoma, skin cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain cancer, head-neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck carcinoma, breast carcinoma, ovarian carcinoma, lung cancer, small-cell lung carcinoma, Wilms' tumor, cervical cancer, testicular cancer, bladder cancer, pancreatic cancer, stomach cancer, colon cancer, prostatic cancer, carcinoma), genitourinary system cancer, thyroid cancer, esophageal cancer, myeloma, multiple myeloma, adrenal cancer, renal cell carcinoma, endometrial cancer, adrenocortical carcinoma, malignant pancreatic insulinoma, malignant carcinoid, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic myeloid leukemia, acute myeloid leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, polycythemia vera, essential thrombocythemia, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, osteosarcoma, primary macroglobulinemia, or retinoblastoma, etc.In other embodiments, the cancer is an acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, triple negative breast cancer (TNBC), bronchogenic carcinoma, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, embryonal carcinoma, endothelial carcinoma, ependymoma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer, cancer), glioblastoma multiforme, glioma, head and neck cancer, hemangioblastoma, hepatoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelioma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, stomach cancer In some embodiments, the disease is triple-negative breast cancer (TNBC).

[1321] In some embodiments, the antibody drug conjugates described herein reduce the mean tumor volume by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% compared to the mean tumor volume of untreated controls in the MDA-MB-231 model of breast cancer. In some embodiments, the antibody drug conjugates reduce the mean tumor volume by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% compared to the mean tumor volume of untreated controls in a patient-derived xenograft model of breast cancer.

[1322] In some embodiments, a method of treating cancer comprises administering to a subject in need thereof a therapeutically effective amount of an antibody-drug conjugate described herein (eg, Formula (I)) or a pharmaceutical composition thereof.

[1323] In some embodiments, after administration of a therapeutically effective amount of an antibody-drug conjugate to a subject, the antibody-drug conjugate is converted to a metabolite.

[1324] In some embodiments, about 24 hours after administering the therapeutically effective amount of the antibody-drug conjugate to the subject, less than about 75% of the antibody-drug conjugate is converted to metabolites. In some embodiments, about 24 hours after administering the therapeutically effective amount of the antibody-drug conjugate to the subject, less than about 10%, less than about 15%, less than about 20%, less than about 25%, less than about 30%, less than about 35%, less than about 40%, less than about 45%, less than about 50%, less than about 55%, less than about 60%, less than about 65% or less than about 70% of the antibody-drug conjugate is converted to metabolites.

[1325] In some embodiments, about 10% to less than about 50% of the antibody-drug conjugate is converted to metabolites about 24 hours after administering a therapeutically effective amount of an antibody-drug conjugate to a subject. In some embodiments, about 24 hours, about 36 hours, about 48 hours, about 60 hours, about 72 hours, or about 84 hours after administering a therapeutically effective amount of an antibody-drug conjugate to a subject, less than about 50% of the antibody-drug conjugate is converted to metabolites. In some embodiments, about 24 hours after administering the therapeutically effective amount of the antibody-drug conjugate to the subject, less than about 50% of the antibody-drug conjugate is converted to metabolites.

[1326] In some embodiments, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70% or about 75% of the antibody-drug conjugate is converted into metabolites after about 24 hours, about 36 hours, about 48 hours, about 60 hours, about 72 hours, about 84 hours or about 96 hours after a therapeutically effective amount of an antibody-drug conjugate is administered to a subject. In some embodiments, about 10% to about 50% of the antibody-drug conjugate is converted into metabolites after about 36 hours to about 96 hours after a therapeutically effective amount of an antibody-drug conjugate is administered to a subject. In some embodiments, about 50% of the antibody-drug conjugate is converted into metabolites after about 36 hours, about 48 hours, about 60 hours, about 72 hours, about 84 hours or about 96 hours after a therapeutically effective amount of an antibody-drug conjugate is administered to a subject. In some embodiments, about 50% of the antibody-drug conjugate is converted to metabolites about 96 hours after administering the therapeutically effective amount of the antibody-drug conjugate to the subject.

[1327] In some embodiments, after administration of a therapeutically effective amount of an antibody-drug conjugate to a subject, the antibody-drug conjugate is converted to a metabolite of Formula 300:

[1328]

[1329] In some embodiments, after administration of a therapeutically effective amount of an antibody-drug conjugate to a subject, the antibody-drug conjugate is converted to a metabolite of Formula 301:

[1330]

[1331] In some embodiments, after administration of a therapeutically effective amount of an antibody-drug conjugate to a subject, the antibody-drug conjugate is converted to a metabolite of Formula 302:

[1332]

[1333] In some embodiments, a metabolite of Formula 300 is converted to a metabolite of Formula 301. In some embodiments, a metabolite of Formula 300 is converted to a metabolite of Formula 302. In some embodiments, a metabolite of Formula 301 is converted to a metabolite of Formula 302. In some embodiments, a metabolite of Formula 300 is converted to a metabolite of Formula 301, and a metabolite of Formula 301 is converted to a metabolite of Formula 302.

[1334] In some embodiments, the antibody-drug conjugate is converted to a metabolite in vivo. In some embodiments, the antibody-drug conjugate is converted to a metabolite in vitro.

[1335] In a non-limiting example, the percentage of antibody-drug conjugate converted to metabolites is based on the therapeutically effective amount of the antibody-drug conjugate administered to the subject. The conversion of the antibody-drug conjugate to metabolites can be determined using any method known in the art. Non-limiting examples of methods that can be used to determine conversion include mass spectrometry.

[1336] Combination therapy / conjugates

[1337] As described herein, in various embodiments, the present invention is directed to anti-tumor agents that can be part of the conjugates and / or compounds of the present invention or can be used in the context of various combination therapies encompassed by the present invention.

[1338] Combination therapy encompasses the administration of an antibody-drug conjugate and another therapeutic agent as part of a specific treatment regimen, which optionally includes a maintenance phase, wherein the portion is intended to provide a beneficial effect from the co-action of these therapeutic agents. The beneficial effect of the combination includes, but is not limited to, the pharmacokinetic or pharmacodynamic co-action produced by the combination of therapeutic agents. The combined administration of these therapeutic agents is typically carried out within a limited time period (typically minutes, hours, days, or weeks, depending on the selected combination). Combination therapy is generally not intended to encompass the administration of two or more of these therapeutic agents as part of a separate monotherapy regimen, which therapeutic regimen accidentally and arbitrarily produces a combination of the present invention.

[1339] Combination therapy encompasses administering these therapeutic agents in a sequential manner, that is, wherein each therapeutic agent is administered at different times, and at least two of these therapeutic agents or therapeutic agents are administered in a substantially simultaneous manner. The sequential administration of each therapeutic agent or administration substantially simultaneously can be achieved by any appropriate approach, including but not limited to oral route, intravenous route, intramuscular, subcutaneous route, and direct absorption through mucosal tissue. The therapeutic agent can be administered by the same approach or by different approaches. For example, a first therapeutic agent (for example, a chemotherapeutic agent) can be administered orally, and a second agent (for example, ADC) can be administered intravenously. In addition, the first therapeutic agent of the selected combination can be administered by intravenous injection, while other therapeutic agents of the combination can be administered orally. Alternatively, for example, two therapeutic agents can be administered by intravenous injection or subcutaneous injection.

[1340] In the present disclosure, unless otherwise indicated, the term sequential mode is characterized by a conventional order or sequence, for example, in the case where a dosage regimen includes the administration of an ADC and a chemotherapeutic agent, a sequential dosing regimen may include administering the ADC before, simultaneously, substantially simultaneously, or after the administration of the chemotherapeutic agent, but both agents will be administered in a conventional order or sequence. Unless otherwise indicated, the term separate means to keep one agent separate from the other. Unless otherwise indicated, the term simultaneous means to occur or be completed simultaneously, that is, the compounds of the present invention are administered simultaneously. The term substantially simultaneous means that the compounds are administered within a few minutes of each other (e.g., within 10 minutes of each other), and is intended to encompass both combined administration and continuous administration, but if administration is continuous, it is separated in time by only a short period of time (e.g., the time it takes for a medical practitioner to administer the two compounds separately). As used herein, concurrent administration and substantially simultaneous administration are used interchangeably. Sequential administration refers to the administration of an ADC and a chemotherapeutic agent separately in time.

[1341] In some embodiments, the chemotherapeutic agent is selected from the group consisting of alkylating agents such as thiotepa and cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylmelamines including altretamine, triethylenemelamine, triethylenephosphamide, and tritylenamine. ephosphoramide), triethiylenethiophosphoramide, and trimethylolomelamine; acetogenins (e.g., bullatacin and bullatacinone); camptothecins (including the synthetic analogue topotecan); bryostatin; CC-1065 (including its synthetic analogues adolesine, carzelesin, and biszelesin); nostocs (e.g., nostoc 1 and nostoc 8); dolastatin; duocarmycins (including the synthetic analogues KW-2189 and CB-1065); 1-TM1); acanthopanax; pancratistatin; sarcodictyin; spongistatin; nitrogen mustards (e.g., chlorambucil, chlomaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, and uracil mustard);Nitrosoureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics, such as enediyne antibiotics (e.g., calicheamicins, particularly calicheamicin gamma II and calicheamicin omega II) (see, e.g., Agnew, Chem. Intl. Ed. Engl., 33:183-186 (1994)); dynemicins, including dynemicin A; bisphosphonates, such as clodronate; esperamicins; and neocarzinostatin chromophores. chromophores and related pigment proteins (enediyne antibiotic chromophores), aclacinomysin, actinomycin, authramycin, azaserine, bleomycin, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycin omycinis), actinomycin D, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrroline-doxorubicin, and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, acid), nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, and zorubicin;Antimetabolites, such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs, such as denopterin, methotrexate, pteropterin, and trimetrexate; purine analogs, such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine; pyrimidine analogs, such as ancitabine. azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and floxuridine; androgens such as calusterone and dromostanolone propionate propionate, epitiostanol, mepitiostane, testolactone; antiadrenal drugs such as minoglutethimide, mitotane, and trilostane; folic acid supplements such as folinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; demecolcine; diaziquone; elformthine; elliptinium acetate acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansine compounds, such as maytansine and ansamitocin; mitoguazone; mitoxantrone; mopidamol;Nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecenes (e.g., T-2 toxin, verracurin A, roridin A) A) and anguidin); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxanes, for example, TAXOL paclitaxel, ABRAXANE Cremophor-free, albumin-engineered paclitaxel nanoparticle formulations (American Pharmaceutical Partners, Schaumberg, IL), and TAXOTERE docetaxel (Rhone-Poulenc Pharmaceuticals, Antony, France). Rorer, Antony, France); GEMZAR gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin, oxaliplatin, and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; NAVELBINE, vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda;Ibandronate; irinotecan (Camptosar, CPT-11) (including regimens of irinotecan with 5-FU and leucovorin); topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoids, such as retinoic acid; capecitabine; combretastatin; leucovorin (LV); oxaliplatin, including oxaliplatin regimens (FOLFOX), lapatinib (Tykerb); inhibitors of PKC-α, Raf, H-Ras, EGFR (e.g., erlotinib (Tarceva)), and VEGF-A, which reduce cell proliferation; and pharmaceutically acceptable salts, acids, or derivatives of any of the above drugs.

[1342] In some embodiments, the anti-tumor agent is a cytotoxic agent. In some embodiments, the cytotoxic agent is selected from methotrexate, aminopterin, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil; alkylating agents such as methylchlorothiazide, thiotepa, chlorambucil, melphalan, carmustine (BSNU), mitomycin C, lomustine (CCNU), 1-methylnitrosourea, cyclophosphamide, methylchlorothiazide, busulfan, dibromomannitol, streptozotocin, mitomycin C, cis-dichlorodiamine platinum (II) (DDP), cisplatin and carboplatin; anthracyclines include daunorubicin, doxorubicin, detorubicin, carminomycin, idarubicin, epirubicin, mitoxantrone and biguanide; antibiotics include dactinomycin / actinomycin D), bleomycin, calicheamicin, mithramycin, and anthramycin (AMC); and antimitotic agents such as vinca alkaloids, vincristine, and vinblastine, and mixtures thereof.

[1343] In some embodiments, the cytotoxic agent is selected from the group consisting of taxol, ricin, Pseudomonas exotoxin, gemcitabine, cytochalasin B, gramicidin D, ethidium bromide, emetine, etoposide, tenoposide, colchicine, dihydroxyanthracenedione, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, procarbazine, hydroxyurea, and mixtures thereof.

[1344] In some embodiments, the compositions and methods of the present invention can be used in combination with checkpoint inhibitors, for example, for the treatment of various cancers. For example, the compositions and methods of the present invention can be supplemented with checkpoint inhibitor-based cancer therapies, for example, by improving the patient's response thereto (for example, by converting non-responders to responders, and / or increasing the magnitude of the therapeutic response, and / or reducing the dosage or regimen required for the therapeutic response, and / or reducing one or more side effects of checkpoint inhibitor-based cancer therapies).

[1345] In some embodiments, the checkpoint inhibitor is an agent that targets one of TIM-3, BTLA, PD-1, CTLA-4, B7-H4, GITR, Galectin-9, HVEM, PD-L1, PD-L2, B7-H3, CD244, CD160, TIGIT, SIRPα, ICOS, CD172a, and TMIGD2.

[1346] In some embodiments, immune checkpoint immunotherapy agents regulate PD-1. In some embodiments, the agent targeting PD-1 is an antibody or antigen-binding portion thereof that is specific for PD-1, optionally selected from nivolumab, pembrolizumab, and pidilizumab. In some embodiments, the antibody or antigen-binding portion thereof that is specific for PD-1 is nivolumab and can be administered at 240 mg every 2 weeks. In some embodiments, the antibody or antigen-binding portion thereof that is specific for PD-1 is pembrolizumab and can be administered at 200 mg every 3 weeks. In some embodiments, the antibody or antigen-binding portion thereof that is specific for PD-1 is pembrolizumab and can be administered at 200 mg every 3 weeks.

[1347] In some embodiments, the immune checkpoint immunotherapy agent regulates PD-L1. In some embodiments, the agent that regulates PD-L1 is an antibody or its antigen-binding portion thereof that is specific for PD-L1. In some embodiments, the antibody or its antigen-binding portion thereof that is specific for PD-L1 is selected from atezolizumab, avelumab, durvalumab, and BMS-936559. In some embodiments, the antibody or its antigen-binding portion thereof that is specific for PD-L1 is BMS-936559 and can be administered at 0.1 mg / kg every 2 weeks. In some embodiments, the antibody or its antigen-binding portion thereof that is specific for PD-L1 is atezolizumab and can be administered at 1200 mg every 3 weeks. In some embodiments, the antibody or its antigen-binding portion thereof that is specific for PD-L1 is avelumab and can be administered at 10 mg / kg every 2 weeks. In some embodiments, the antibody or its antigen-binding portion thereof that is specific for PD-L1 is durvalumab and can be administered at 10 mg / kg every 2 weeks.

[1348] In some embodiments, the agent targeting CTLA-4 is an antibody or antigen-binding portion thereof specific for CTLA-4, optionally selected from ipilimumab and tremelimumab. In some embodiments, the antibody or antigen-binding portion thereof specific for CTLA-4 is tremelimumab and can be administered at 3 mg / kg, 6 mg / kg, or 10 mg / kg. In some embodiments, the antibody or antigen-binding portion thereof specific for CTLA-4 is ipilimumab and can be administered at 5 mg / mL for 12 weeks.

[1349] In some embodiments, the hyperproliferative disorder (eg, cancer) treated by the compounds and compositions described herein comprises cells having expression of p38α MAPK protein and / or p38α MAPK-related proteins.

[1350] In one embodiment, the present disclosure relates to a method of treating a disease that is alleviated by inhibiting p38α MAPK protein in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a p38α MAPK inhibitor or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal or prodrug thereof and one or more additional therapeutic agents, including chemotherapeutic agents and / or immunotherapeutic agents, wherein the p38α MAPK inhibitor is a compound that is capable of binding to a pocket near the ED substrate docking site of p38α MAPK.

[1351] The compounds and combinations of compounds described herein can be tested for efficacy in treating the indicated diseases or conditions using various models known in the art that provide guidance for the treatment of human disease. Any and all of the described treatment methods may include medical follow-up to determine the therapeutic or preventive effect achieved in subjects undergoing treatment with the compounds and / or compositions described herein.

[1352] Pharmaceutical composition

[1353] In one embodiment, an active pharmaceutical ingredient or combination of active pharmaceutical ingredients (such as any of the conjugates, drug moieties, linkers, compounds, and / or compositions of the present disclosure) is provided in the form of a pharmaceutically acceptable composition.

[1354] In one embodiment, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of one or more conjugates, drug moieties, linkers, compounds and / or compositions of the present disclosure (e.g., Formula (I), Formula (II) and / or Formula (III)) or pharmaceutically acceptable salts, solvates, hydrates, cocrystals or prodrugs thereof; and a physiologically compatible carrier medium, wherein the disease is cancer. In one embodiment, the disease is cancer, such as acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, bronchogenic carcinoma, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, embryonal carcinoma, endothelial carcinoma, ependymoma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer (gastric cancer), cancer), glioblastoma multiforme, glioma, head and neck cancer, hemangioblastoma, hepatoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelioma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, stomach cancer cancer), sweat gland cancer, synovioma, testicular cancer, small cell lung cancer, laryngeal cancer, triple negative breast cancer (TNBC), uterine cancer, Wilms' tumor, blood cancer, acute erythroleukemic leukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute lymphoblastic leukemia, acute megakaryocytic leukemia, acute monocytic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute nonlymphocytic leukemia, acute promyelocytic leukemia, acute undifferentiated leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma, polycythemia vera, or Waldenstrom's macroglobulinemia.

[1355] In some embodiments, each active pharmaceutical ingredient (e.g., any conjugate, drug moiety, linker, compound, and / or composition of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III))) in a pharmaceutical composition of the present disclosure is provided at a concentration of less than, e.g., 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.6%, 0.7%, 0.8%, 0.9%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 10. 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w / w, w / v, or v / v.

[1356] In some embodiments, each active pharmaceutical ingredient in a pharmaceutical composition of the present disclosure, such as any conjugate, drug moiety, linker, compound, and / or composition of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III)), is provided at a concentration greater than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19.75%, 19.50%, 19.25%, 19%, 18.75%, 18.50%, 18.25%, 18%, 17.75%, 17.50%, 17.25%, or more of the pharmaceutical composition. %, 17%, 16.75%, 16.50%, 16.25%, 16%, 15.75%, 15.50%, 15.25%, 15%, 14.75%, 14.50%, 14.25%, 14%, 13.75%, 13.50%, 13.25%, 13%, 12.75%, 12.50%, 12.25%, 12%, 11.75%, 11.50%, 11.25%, 11%, 10.75%, 10.50%, 10.25%, 10%, 9.75%, 9.50%, 9.25% ,9%,8.75%,8.50%,8.25%,8%,7.75%,7.50%,7.25%,7%,6.75%,6.50%,6.25%,6%,5.75%,5.50%,5.25%,5%,4.75%,4.50%,4.25%,4%,3.75%,3.50%,3.25%,3%,2.75%,2.50%,2.25%,2%,1.75%,1.50%,125%,1%,0.5%,0.4%,0.3%,0.2%,0.1%,0. 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w / w, w / v, or v / v.

[1357] In some embodiments, the concentration of each active pharmaceutical ingredient in the pharmaceutical composition of the present disclosure, such as any conjugate, drug moiety, linker, compound and / or composition of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III)), is provided at a concentration of about 0.0001% to about 50%, about 0.001% to about 40%, about 0.01% to about 30%, about 0.02% to about 29%, about 0.03% to about 28%, about 0.04% to about 27%, about 0.05% to about 10%, about 0.06% to about 15%, about 0.07% to about 16%, about 0.08% to about 17%, about 0.10% to about 18%, about 0.11% to about 19%, about 0.12% to about 20%, about 0.13% to about 21%, about 0.14% to about 22%, about 0.15% to about 23%, about 0.16% to about 24%, about 0.17% to about 25%, about 0.18% to about 26%, about 0.19% to about 27%, about 0.20% to about 28%, about 0.21% to about 29%, about 0.22% to about 29%, about 0.23% to about 28%, about 0.24% to about 29%, about 0.25% to about 2 % to about 15%, about 0.8% to about 14%, about 0.9% to about 12%, or about 1% to about 10% w / w, w / v, or v / v.

[1358] In some embodiments, the concentration of each active pharmaceutical ingredient (such as any conjugate, drug moiety, linker, compound and / or composition of the disclosure (e.g., Formula (I), Formula (II), and / or Formula (III))) provided in a pharmaceutical composition of the present disclosure is within the range of about 0.001% to about 10%, about 0.01% to about 5%, about 0.02% to about 4.5%, about 0.03% to about 4%, about 0.04% to about 3.5%, about 0.05% to about 3%, about 0.06% to about 2.5%, about 0.07% to about 2%, about 0.08% to about 1.5%, about 0.09% to about 1%, about 0.1% to about 0.9% w / w, w / v, or v / v of the pharmaceutical composition.

[1359] In some embodiments, the amount of each active pharmaceutical ingredient in the pharmaceutical compositions of the present disclosure, such as any of the foregoing conjugates, drug moieties, linkers, compounds, and / or compositions of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III)), is provided in an amount equal to or less than 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5g, 0.45g, 0.4g, 0.35g, 0.3g, 0.25g, 0.2g, 0.15g, 0.1g, 0.09g, 0.08 g, 0.07g, 0.06g, 0.05g, 0.04g, 0.03g, 0.02g, 0.01g, 0.009g, 0.008g, 0. 007g, 0.006g, 0.005g, 0.004g, 0.003g, 0.002g, 0.001g, 0.0009g, 0.00 08g, 0.0007g, 0.0006g, 0.0005g, 0.0004g, 0.0003g, 0.0002g or 0.0001g.

[1360] In some embodiments, each active pharmaceutical ingredient (e.g., any conjugate, drug moiety, linker, compound, and / or composition of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III))) provided in a pharmaceutical composition of the present disclosure is provided in an amount greater than 0.0001 g, 0.0002 g, 0.0003 g, 0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009 g, 0.0010 g, 0.0011 g, 0.0012 g, 0.0013 g, 0.0014 g, 0.0015 g, 0.0016 g, 0.0017 g, 0.0018 g, 0.0019 g, 0.0020 g, 0008g, 0.0009g, 0.001g, 0.0015g, 0.002g, 0.0025g, 0.003g, 0.0035g, 0.004g, 0.0045g , 0.005g, 0.0055g, 0.006g, 0.0065g, 0.007g, 0.0075g, 0.008g, 0.0085g, 0.009g, 0.009 5g, 0.01g, 0.015g, 0.02g, 0.025g, 0.03g, 0.035g, 0.04g, 0.045g, 0.05g, 0.055g, 0.06g , 0.065g, 0.07g, 0.075g, 0.08g, 0.085g, 0.09g, 0.095g, 0.1g, 0.15g, 0.2g, 0.25g, 0.3g , 0.35g, 0.4g, 0.45g, 0.5g, 0.55g, 0.6g, 0.65g, 0.7g, 0.75g, 0.8g, 0.85g, 0.9g, 0.95g, 1g, 1.5g, 2g, 2.5, 3g, 3.5, 4g, 4.5g, 5g, 5.5g, 6g, 6.5g, 7g, 7.5g, 8g, 8.5g, 9g, 9.5g or 10g.

[1361] It is effective in a wide dosage range according to each active pharmaceutical ingredient of the present disclosure. For example, when treating an adult, the dosage of 0.01mg to 1000mg every day, 0.5mg to 100mg every day, 1mg to 50mg every day and 5mg to 40mg every day independently in the scope is an example of a dosage that can be used. Exact dosage will depend on the form used by route of administration, compound, sex and age of the experimenter to be treated, weight of the experimenter to be treated and preference and experience of the attending physician. If appropriate, the clinically determined dosage of the conjugate of the present disclosure, drug moiety, linker, compound and / or composition (for example, formula (I) formula (II) and / or formula (III) of the present disclosure) can also be used.

[1362] In one embodiment, the molar ratio of the two active pharmaceutical ingredients in the pharmaceutical composition is in the range of 10:1 to 1:10, preferably 2.5:1 to 1:2.5, and more preferably about 1:1. In one embodiment, the weight ratio of the molar ratio of the two active pharmaceutical ingredients in the pharmaceutical composition is selected from the group consisting of: 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19 and 1:20. In one embodiment, the weight ratio of the molar ratio of the two active pharmaceutical ingredients in the pharmaceutical composition is selected from the group consisting of: 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19 and 1:20.

[1363] Non-limiting pharmaceutical compositions and methods for their preparation are described below.

[1364] Pharmaceutical compositions for oral administration

[1365] In one embodiment, the present disclosure provides a pharmaceutical composition for oral administration, comprising an active pharmaceutical ingredient or a combination of active pharmaceutical ingredients described herein (such as a conjugate, drug moiety, linker, compound and / or composition of the present disclosure (e.g., Formula (I), Formula (II) and / or Formula (III))) and a pharmaceutical excipient suitable for oral administration.

[1366] In some embodiments, the present disclosure provides a solid pharmaceutical composition for oral administration, comprising: (i) an effective amount of an active pharmaceutical ingredient or a combination of active pharmaceutical ingredients, and (ii) a pharmaceutical excipient suitable for oral administration. In selected embodiments, the composition further comprises (iii) an effective amount of a third active pharmaceutical ingredient, and optionally (iv) an effective amount of a fourth active pharmaceutical ingredient.

[1367] In certain embodiments, the pharmaceutical composition can be a liquid pharmaceutical composition suitable for oral consumption. The pharmaceutical composition of the present disclosure suitable for oral administration can be presented in discrete dosage forms, such as capsules, pouches or tablets or the liquid or aerosol spray, solution or aqueous or non-aqueous liquid suspension in the active component in the form of powder or granules each containing a predetermined amount, oil-in-water emulsion or water-in-oil liquid emulsion, powder for recovery, powder for oral consumption, bottle (including powder or liquid in bottle), oral dissolving film, lozenge, paste, pipe, glue and packaging. Such dosage forms can be prepared by any pharmaceutical method, but all methods include the step of associating active component with a carrier, and the carrier constitutes one or more essential components. Typically, by making active component and liquid carrier or fine powder solid carrier or both uniform and closely mixed, and then making the product be shaped into the desired presentation form to prepare compositions if necessary. For example, tablets can be prepared optionally with one or more auxiliary ingredients by compression or molding. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in a free-flowing form (such as a powder or granules), optionally mixed with an excipient such as, but not limited to, a binder, a lubricant, an inert diluent, and / or a surfactant or dispersant. Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[1368] The present disclosure further encompasses anhydrous pharmaceutical compositions and dosage forms, because water can promote the degradation of some compounds. For example, in the pharmaceutical field, water (e.g., 5%) can be added as a means of simulating long-term storage to determine the characteristics of the formulation that change over time, such as shelf life or stability. Anhydrous pharmaceutical compositions and dosage forms of the present disclosure can be prepared using anhydrous or low-moisture ingredients and low-moisture or low-humidity conditions. If it is expected that substantial contact will occur with moisture and / or humidity during manufacture, packaging and / or storage, the pharmaceutical compositions and dosage forms of the present disclosure containing lactose can be made anhydrous. Anhydrous pharmaceutical compositions can be prepared and stored in a manner that maintains the anhydrous nature of the anhydrous pharmaceutical composition. Therefore, anhydrous compositions can be packaged using materials known to prevent exposure to water so that the anhydrous compositions can be included in a suitable prescription kit. Examples of suitable packaging include, but are not limited to, airtightly sealed foil, plastics, etc., unit dose containers, blister packs, and strip packs.

[1369] Each active pharmaceutical ingredient can be closely mixed with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier can take a variety of forms, depending on the form of the preparation required for administration. When preparing a composition of an oral dosage form, in the case of an oral liquid preparation (such as a suspension, solution, and elixir) or an aerosol, any conventional pharmaceutical medium can be used as a carrier, such as water, ethylene glycol, oil, alcohol, flavoring, preservative, colorant, etc.; or in the case of an oral solid preparation, carriers such as starch, sugar, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, and disintegrants can be used, and in some embodiments, lactose is not used. For example, suitable carriers include powders, capsules, and tablets with solid oral preparations. If desired, tablets can be coated by standard aqueous or non-aqueous techniques.

[1370] Binders suitable for use in pharmaceutical compositions and dosage forms include, but are not limited to, corn starch, potato starch or other starches, gelatin, natural and synthetic gums (e.g., acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum), cellulose and its derivatives (e.g., ethylcellulose, cellulose acetate, carboxymethylcellulose calcium, sodium carboxymethylcellulose), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropyl methylcellulose, microcrystalline cellulose, and mixtures thereof.

[1371] Examples of fillers suitable for use in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof.

[1372] Disintegrants can be used in the compositions of the present invention to provide tablets that disintegrate when exposed to an aqueous environment. Too much disintegrant can produce tablets that disintegrate in the bottle. Too little disintegrant may not be enough to cause disintegration to occur, thereby changing the rate and extent of release of the active ingredient from the dosage form. Therefore, a sufficient amount of disintegrant (which will not adversely change the release of the active ingredient due to too little or too much) can be used to form a dosage form of the compound disclosed herein. The amount of disintegrant used can vary based on the type of formulation and the mode of administration and can be easily discerned by one of ordinary skill in the art. About 0.5% to about 15% by weight of disintegrant or about 1% to about 5% by weight of disintegrant can be used in the pharmaceutical composition. Disintegrants that can be used to form pharmaceutical compositions and dosage forms of the present disclosure include, but are not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pregelatinized starch, other starches, clays, other algins, other celluloses, gums, or mixtures thereof.

[1373] Lubricants that can be used to form pharmaceutical compositions and dosage forms of the present disclosure include, but are not limited to, calcium stearate, magnesium stearate, sodium stearyl fumarate, mineral oil, light mineral oil, glycerol, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, or a mixture thereof. Additional lubricants include, for example, a condensed aerosol of synthetic silica, synthetic silicon dioxide, silicified microcrystalline cellulose, or a mixture thereof. Lubricants can optionally be added in an amount less than about 0.5% or less than about 1% (by weight) of the pharmaceutical composition.

[1374] When aqueous suspensions and / or elixirs are desired for oral administration, the active pharmaceutical ingredient may be combined with various sweetening or flavoring agents, colorants or dyes and, if desired, emulsifying and / or suspending agents and diluents such as water, ethanol, propylene glycol, glycerin, and various combinations thereof.

[1375] Tablet can be uncoated or can be coated to postpone disintegration and absorption in gastrointestinal tract by known technology, and the sustained action in long period is provided thus.For example, can adopt the time-delay materials such as glyceryl monostearate or glyceryl distearate.Preparation for oral use can also be presented with the form of hard gelatin capsule or soft gelatin capsule, in the case of described hard gelatin capsule, active component and inert solid diluent (for example, calcium carbonate, calcium phosphate or kaolin) mix, in the case of described soft gelatin capsule, active component and water or oil medium (for example, peanut oil, liquid paraffin or olive oil) mix.

[1376] Surfactants that can be used to form the pharmaceutical compositions and dosage forms of the present disclosure include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof. That is, a mixture of hydrophilic surfactants can be employed, a mixture of lipophilic surfactants can be employed, or a mixture of at least one hydrophilic surfactant and at least one lipophilic surfactant can be employed.

[1377] Suitable hydrophilic surfactants generally have an HLB value of at least 10, while suitable lipophilic surfactants generally have an HLB value of about 10 or less. An empirical parameter used to characterize the relative hydrophilicity and hydrophobicity of nonionic amphiphilic compounds is the hydrophilic-lipophilic balance ("HLB" value). Surfactants with lower HLB values ​​are more lipophilic or hydrophobic and have greater solubility in oils, while surfactants with higher HLB values ​​are more hydrophilic and have greater solubility in aqueous solutions. Hydrophilic surfactants are generally considered to be compounds with an HLB value greater than about 10, as well as anionic, cationic, or zwitterionic compounds for which the HLB scale is generally not applicable. Similarly, lipophilic (i.e., hydrophobic) surfactants are compounds with an HLB value equal to or less than about 10. However, the HLB value of a surfactant is only a rough guide that is commonly used to formulate industrial, pharmaceutical, and cosmetic emulsions.

[1378] The hydrophilic surfactant may be ionic or nonionic. Suitable ionic surfactants include, but are not limited to, alkylammonium salts; fusidates; fatty acid derivatives of amino acids, oligopeptides, and polypeptides; glyceride derivatives of amino acids, oligopeptides, and polypeptides; lecithin and hydrogenated lecithin; lysolecithin and hydrogenated lysolecithin; phospholipids and their derivatives; lysolecithin and their derivatives; carnitine fatty acid ester salts; alkyl sulfates; fatty acid salts; sodium docusate; acyl lactylates; monoacetylated and diacetylated tartaric acid esters of monoglycerides and diglycerides; succinylated monoglycerides and diglycerides; citric acid esters of monoglycerides and diglycerides; and mixtures thereof.

[1379] Within the aforementioned group, ionic surfactants include, for example: lecithin, lysolecithin, phospholipids, lysophospholipids and derivatives thereof; carnitine fatty acid ester salts; alkyl sulfates; fatty acid salts; sodium docusate; acyl lactylations; monoacetylated and diacetylated tartaric acid esters of monoglycerides and diglycerides; succinylated monoglycerides and diglycerides; citric acid esters of monoglycerides and diglycerides; and mixtures thereof.

[1380] The ionic surfactant can be an ionized form of lecithin, lysolecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylserine, PEG-phosphatidylethanolamine, PVP-phosphatidylethanolamine, lactic acid esters of fatty acids, stearoyl-2-lactate, stearoyl lactylate, succinylated monoglycerides, monoacetylated / diacetylated tartaric acid esters of monoglycerides / diglycerides, citrate esters of monoglycerides / diglycerides, cholylsarcosine, caproate, caprylate, caprate, laurate, myristate, palmitate, oleate, ricinoleate, linolenate, stearate, lauryl sulfate, tetradecyl sulfate, docusate, lauroylcarnitine, palmitoylcarnitine, myristoylcarnitine, and salts and mixtures thereof.

[1381] The hydrophilic nonionic surfactants may include, but are not limited to, alkyl glucosides; alkyl maltosides; alkyl thioglucosides; polyethylene glycol glycerol laurate; polyoxyalkylene alkyl ethers, such as polyethylene glycol alkyl ethers; polyoxyalkylene alkylphenols, such as polyethylene glycol alkylphenols; polyoxyalkylene alkylphenol fatty acid esters, such as polyethylene glycol fatty acid monoesters and polyethylene glycol fatty acid diesters; polyethylene glycol glycerol fatty acid esters; polyglycerol fatty acid esters; polyoxyalkylene sorbitan fatty acid esters, such as polyethylene glycol sorbitan fatty acid esters; hydrophobic transesterification products of polyols having at least one member selected from the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols; polyoxyethylene sterols, derivatives and analogs thereof; polyoxyethylated vitamins and derivatives thereof; polyoxyethylene-polyoxypropylene block copolymers; and mixtures thereof; polyethylene glycol sorbitan fatty acid esters and hydrophilic transesterification products of polyols having at least one member selected from the group consisting of triglycerides, vegetable oils and hydrogenated vegetable oils. The polyol may be glycerol, ethylene glycol, polyethylene glycol, sorbitol, propylene glycol, pentaerythritol or a sugar.

[1382] Other hydrophilic nonionic surfactants include, but are not limited to, PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32 distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25 triolein, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30 glyceryl laurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate, PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryl laurate, PEG- EG-40 Palm Kernel Oil, PEG-50 Hydrogenated Castor Oil, PEG-40 Castor Oil, PEG-35 Castor Oil, PEG-60 Castor Oil, PEG-40 Hydrogenated Castor Oil, PEG-60 Hydrogenated Castor Oil, PEG-60 Corn Oil, PEG-6 Capric / Caprylic Glycerides, PEG-8 Capric / Caprylic Glycerides, Polyglyceryl-10 Laurate, PEG-30 Cholesterol, PEG-25 Phytosterols, PEG-30 Soy Sterols, PEG -20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitan laurate, polysorbate 20, polysorbate 80, POE-9 lauryl ether, POE-23 lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearyl ether, tocopheryl PEG-100 succinate, PEG-24 cholesterol, polyglyceryl-10 oleate, Tween 40, Tween 60, sucrose monostearate, sucrose monolaurate, sucrose monopalmitate, PEG 10-100 nonylphenol series, PEG 15-100 octylphenol series and poloxamer.

[1383] By way of example only, suitable lipophilic surfactants include: fatty alcohols; glycerol fatty acid esters; acetylated glycerol fatty acid esters; lower alcohol fatty acid esters; propylene glycol fatty acid esters; sorbitan fatty acid esters; polyethylene glycol sorbitan fatty acid esters; sterols and sterol derivatives; polyoxyethylated sterols and sterol derivatives; polyethylene glycol alkyl ethers; sugar esters; sugar ethers; lactic acid derivatives of monoglycerides and diglycerides; hydrophobic transesterification products of polyols having at least one member selected from the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids, and sterols; oil-soluble vitamin / microbial derivatives; and mixtures thereof. Within this group, preferred lipophilic surfactants include glycerol fatty acid esters, propylene glycol fatty acid esters, and mixtures thereof, or hydrophobic transesterification products of polyols having at least one member selected from the group consisting of vegetable oils, hydrogenated vegetable oils, and triglycerides.

[1384] In one embodiment, the composition may include a solubilizing agent to ensure good solubilization and / or dissolution of the disclosed compound and to minimize precipitation of the disclosed compound. This may be particularly important for compositions intended for parenteral use, such as injectable compositions. A solubilizing agent may also be added to increase the solubility of a hydrophilic drug and / or other components such as a surfactant or to maintain the composition in a stable or homogeneous solution or dispersion.

[1385] Examples of suitable solubilizers include, but are not limited to, the following: alcohols and polyols, such as ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediol and isomers thereof, glycerol, pentaerythritol, sorbitol, mannitol, oxyalkyl glycols, dimethyl isosorbide, polyethylene glycol, polypropylene glycol, polyvinyl alcohol, hydroxypropyl methylcellulose and other cellulose derivatives, cyclodextrins and cyclodextrin derivatives; polyethylene glycol ethers having an average molecular weight of about 200 to about 6000, such as tetrahydrofurfuryl alcohol PEG ether (glycogen) or methoxy PEG; amides and other nitrogen-containing compounds, such as 2-pyrrolidone, 2-piperidone, ε-caprolactam, N-alkyl Pyrrolidone, N-hydroxyalkylpyrrolidone, N-alkylpiperidone, N-alkylcaprolactam, dimethylacetamide and polyvinylpyrrolidone; esters such as ethyl propionate, tributyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, triethyl citrate, ethyl oleate, ethyl caprylate, ethyl butyrate, triacetin, propylene glycol monoacetate, propylene glycol diacetate, ε-caprolactone and its isomers, δ-valerolactone and its isomers, β-butyrolactone and its isomers; and other solubilizers known in the art such as dimethylacetamide, dimethyl isosorbide, N-methylpyrrolidone, monocaprylic acid glyceride, diethylene glycol monoethyl ether and water.

[1386] Mixtures of solubilizing agents can also be used. Examples include, but are not limited to, triacetin, triethyl citrate, ethyl oleate, ethyl caprylate, dimethylacetamide, N-methyl pyrrolidone, N-hydroxyethyl pyrrolidone, polyvinyl pyrrolidone, hydroxypropyl methylcellulose, hydroxypropyl cyclodextrin, ethanol, polyethylene glycol 200-100, glycogen, oxyalkyl glycol, propylene glycol, and dimethyl isosorbide. Particularly preferred solubilizing agents include sorbitol, glycerol, triacetin, ethanol, PEG-400, glycogen, and propylene glycol.

[1387] The amount of the solubilizing agent that can be included is not particularly limited.The amount of a given solubilizing agent can be limited to the biologically acceptable amount that those skilled in the art can easily determine.In some cases, it may be advantageous to comprise that amount far exceeds the biologically acceptable amount for example so that the concentration of the medicine is maximized solubilizing agent, wherein excessive solubilizing agent uses conventional techniques such as distillation or evaporation to remove before providing compositions to the patient.Therefore, if present, based on the combined weight of medicine and other excipients, the weight ratio of solubilizing agent can be 10 % by weight, 25 % by weight, 50 % by weight, 100 % by weight or approximately 200 % by weight at the most.If desired, also can use a very small amount of solubilizing agent, as 5%, 2%, 1% or even less.Typically, solubilizing agent can exist with the amount of approximately 1 % by weight to approximately 100 % by weight, more typically approximately 5 % by weight to approximately 25 % by weight.

[1388] The composition may further include one or more pharmaceutically acceptable additives and excipients. Such additives and excipients include, but are not limited to, anti-adherents, defoaming agents, buffers, polymers, antioxidants, preservatives, chelating agents, viscosity modifiers, tonicifiers, flavoring agents, colorants, flavor enhancers, opacifiers, suspending agents, adhesives, fillers, plasticizers, lubricants, and mixtures thereof.

[1389] In addition, acids or bases can be incorporated into the composition to facilitate processing, enhance stability, or achieve other purposes. Examples of pharmaceutically acceptable bases include amino acids, amino acid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide, sodium bicarbonate, aluminum hydroxide, calcium carbonate, magnesium hydroxide, magnesium aluminum silicate, synthetic aluminum silicate, synthetic hydrocalcite, magnesium aluminum hydroxide, diisopropylethylamine, ethanolamine, ethylenediamine, triethanolamine, triethylamine, triisopropanolamine, trimethylamine, tris(hydroxymethyl)aminomethane (TRIS), and the like. Also suitable is the alkali as the salt of pharmaceutically acceptable acid, described pharmaceutically acceptable acid such as acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, amino acid, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acid, formic acid, fumaric acid, gluconic acid, hydroquinone sulfonic acid (hydroquinosulfonic acid), isoascorbic acid, lactic acid, maleic acid, oxalic acid, p-bromobenzenesulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, mercaptoacetic acid, toluenesulfonic acid, uric acid etc..Can also use the salt of polyprotic acid, such as sodium phosphate, disodium hydrogen phosphate and sodium dihydrogen phosphate.When alkali is salt, cation can be any convenient and pharmaceutically acceptable cation, such as ammonium, alkali metal and alkaline earth metal.Example can include but not limited to sodium, potassium, lithium, magnesium, calcium and ammonium.

[1390] Suitable acid is a pharmaceutically acceptable organic acid or inorganic acid. Examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, boric acid, phosphoric acid, etc. Examples of suitable organic acids include acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, amino acids, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroquinonesulfonic acid, isoascorbic acid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid, p-bromobenzenesulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, and uric acid.

[1391] Pharmaceutical compositions for injection

[1392] In some embodiments, a pharmaceutical composition for injection is provided, which contains an active pharmaceutical ingredient or a combination of active pharmaceutical ingredients (such as a conjugate, drug moiety, linker, compound and / or composition of the present disclosure (e.g., Formula (I), Formula (II) and / or Formula (III)) and a pharmaceutical excipient suitable for injection.

[1393] The compositions of the present disclosure may be incorporated into forms for administration by injection including aqueous or oily suspensions or emulsions with sesame oil, corn oil, cottonseed oil or peanut oil as well as elixirs, mannitol, dextrose or sterile aqueous solutions and similar pharmaceutical vehicles.

[1394] Routinely, aqueous solutions in saline are also used for injection. Ethanol, glycerol, propylene glycol and liquid polyethylene glycol (and its applicable mixture), cyclodextrin derivatives and vegetable oils can also be adopted. Suitable fluidity can be maintained, for example, by using coatings such as lecithin to maintain the desired particle size in the case of a dispersant and by using a surfactant. Prevention of the effects of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid and thimerosal.

[1395] Sterile injectable solutions are prepared by incorporating the required amount of the active pharmaceutical ingredient or combination of active pharmaceutical ingredients into a suitable solvent as needed along with the various other ingredients listed above, followed by filtration sterilization. Typically, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle containing a basic dispersion medium and the other ingredients required from the ingredients listed above. In the case of sterile powders for the preparation of sterile injectable solutions, some desirable preparation methods are vacuum drying and freeze drying techniques, which produce a powder of the active ingredient plus any additional desired ingredients from its previously sterile filtered solution.

[1396] Pharmaceutical compositions for local delivery

[1397] In some embodiments, a pharmaceutical composition for transdermal delivery is provided, comprising an active pharmaceutical ingredient or a combination of active pharmaceutical ingredients, such as a conjugate, drug moiety, linker, compound, and / or composition of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III)) and a pharmaceutical excipient suitable for transdermal delivery.

[1398] The compositions of the present disclosure can be formulated into preparations in solid, semisolid or liquid form suitable for local or topical application, such as gels, water-soluble colloids, creams, lotions, suspensions, foams, powders, slurries, ointments, solutions, oils, pastes, suppositories, sprays, emulsions, saline solutions, solutions based on dimethyl sulfoxide (DMSO). In general, carriers with higher density can provide areas exposed to active ingredients for a long time. In contrast, solution formulations can make active ingredients more directly exposed to selected areas.

[1399] The pharmaceutical composition can also include a suitable solid phase or gel phase carrier or excipient, which is a compound that increases the penetration of the stratum corneum permeability barrier of the therapeutic molecule across the skin or helps deliver the therapeutic molecule across the stratum corneum permeability barrier of the skin. For trained technicians in the field of topical formulations, there are many of these penetration enhancing molecules. Examples of such carriers and excipients include, but are not limited to, wetting agents (e.g., urea), glycols (e.g., propylene glycol), alcohols (e.g., ethanol), fatty acids (e.g., oleic acid), surfactants (e.g., isopropyl myristate and sodium lauryl sulfate), pyrrolidone, glyceryl monolaurate, sulfoxides, terpenes (e.g., menthol), amines, amides, alkanes, alkanols, water, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycol.

[1400] Another exemplary formulation for the methods of the present disclosure employs a transdermal delivery device ("patch"). Such transdermal patches can be used to provide continuous or discontinuous infusion of controlled amounts of an active pharmaceutical ingredient or combination of active pharmaceutical ingredients, with or without another active pharmaceutical ingredient.

[1401] The construction and use of transdermal patches for delivering pharmaceutical agents are well known in the art. See, for example, U.S. Patents 5,023,252; 4,992,445; and 5,001,139, the entire contents of which are incorporated herein by reference. Such patches can be configured for continuous, pulsed, or on-demand delivery of pharmaceutical agents.

[1402] Pharmaceutical compositions for inhalation

[1403] Compositions for inhalation or insufflation include solutions and suspensions in the form of pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, as well as powders. Liquid or solid compositions can contain suitable pharmaceutically acceptable excipients and conjugates, drug moieties, joints, compounds and / or compositions of the present disclosure (e.g., Formula (I), Formula (II) and / or Formula (III)) as described above. Preferably, the composition is administered via oral or nasal respiratory route to produce a local or systemic effect. Compositions in the form of preferably pharmaceutically acceptable solvents can be atomized using an inert gas. The atomized solution can be inhaled directly from the atomizing device, or the atomizing device can be attached to a mask tent or intermittent positive pressure breathing machine. Solutions, suspensions or powder compositions can be administered orally or nasally, preferably by a device that delivers the formulation in an appropriate manner. Dry powder inhalers can also be used to provide inhalation delivery of the composition.

[1404] Other pharmaceutical compositions

[1405] Pharmaceutical compositions of the conjugates, drug moieties, linkers, compounds and / or compositions of the present disclosure (e.g., Formula (I), Formula (II) and / or Formula (III)) can also be prepared from the compositions described herein and one or more pharmaceutically acceptable excipients suitable for sublingual, buccal, rectal, intraosseous, intraocular, intranasal, epidural or intraspinal administration. The preparation of such pharmaceutical compositions is well known in the art. See, for example, Anderson, Philip O.; Knoben, James E.; Troutman, William G, eds., Handbook of Clinical Drug Data, 10th ed., McGraw-Hill, 2002; and Pratt and Taylor, eds., Principles of Drug Action, 3rd ed., Churchill Livingston, NY, 1990, each of which is incorporated herein by reference in its entirety.

[1406] The administration of the active pharmaceutical ingredient or combination of active pharmaceutical ingredients or its pharmaceutical composition can be achieved by any method that enables the compound to be delivered to the site of action. These methods include oral routes, intraduodenal routes, parenteral injection (including intravenous, intraarterial, subcutaneous, intramuscular, intravascular, intraperitoneal or infusion), local (e.g., transdermal application), rectal administration, local delivery by catheter or stent, or by inhalation. The active pharmaceutical ingredient or combination of active pharmaceutical ingredients can also be administered intrafatally or intrathecally.

[1407] Exemplary parenteral administration forms include solutions or suspensions of the active compound in sterile aqueous solutions (for example, aqueous propylene glycol or aqueous dextrose). Such dosage forms can be suitably buffered, if necessary.

[1408] Reagent test kit

[1409] The present disclosure also provides a test kit. The test kit includes an active pharmaceutical ingredient or a combination of active pharmaceutical ingredients, alone or in combination, in suitable packaging, and written materials, which may include instructions for use, a discussion of clinical studies, and a list of side effects. Such test kits may also include information indicating or establishing the activity and / or advantages of the composition and / or describing dosage, administration, side effects, drug interactions (such as scientific literature references, package inserts, clinical trial results, and / or summaries of these), or other information useful to health care providers. Such information can be based on the results of various studies, for example, studies using experimental animals involving in vivo models and studies based on human clinical trials. The test kit may further contain another active pharmaceutical ingredient. In selected embodiments, the active pharmaceutical ingredient or combination of active pharmaceutical ingredients is provided in the form of a separate composition in a separate container within the test kit. In selected embodiments, the active pharmaceutical ingredient or combination of active pharmaceutical ingredients is provided in the form of a single composition in a container within the test kit. Suitable packaging and additional use items (e.g., a measuring cup for liquid preparations, foil packaging to minimize exposure to air, etc.) are known in the art and may be included in the test kit. The test kits described herein can be provided, sold, and / or promoted to health providers, including physicians, nurses, pharmacists, prescribing officials, etc. In selected embodiments, the kits may also be sold directly to consumers.

[1410] In some embodiments, the present disclosure provides a kit comprising a composition comprising a therapeutically effective amount of an active pharmaceutical ingredient (e.g., a conjugate, drug moiety, linker, compound, and / or composition of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III))) or a combination of active pharmaceutical ingredients, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof. These compositions are typically pharmaceutical compositions. The kit is used for co-administering the active pharmaceutical ingredients or the combination of active pharmaceutical ingredients simultaneously or separately.

[1411] In some embodiments, the present disclosure provides a kit comprising (1) a composition comprising a therapeutically effective amount of an active pharmaceutical ingredient (e.g., a conjugate, drug moiety, linker, compound, and / or composition of the present disclosure (e.g., Formula (I), Formula (II), and / or Formula (III))) or a combination of active pharmaceutical ingredients, or a pharmaceutically acceptable salt, solvate, hydrate, cocrystal, or prodrug thereof, and (2) a diagnostic test for determining whether a patient's cancer is a particular subtype of cancer. Any of the aforementioned diagnostic methods can be used in the kit.

[1412] The kits described above are preferably used to treat the diseases and conditions described herein.In some embodiments, the kits are used to treat a hyperproliferative disorder, such as cancer.

[1413] In a specific embodiment, the kits described herein are used to treat cancer. In some embodiments, the kits described herein are used to treat a cancer selected from the group consisting of: acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, bronchogenic carcinoma, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, embryonal carcinoma, endothelial carcinoma, ependymoma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer, cancer), glioblastoma multiforme, glioma, head and neck cancer, hemangioblastoma, hepatoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelioma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, stomach cancer In some embodiments, the disease is triple negative breast cancer (TNBC). In some embodiments, the disease is triple negative breast cancer (TNBC). In some embodiments, the disease is triple negative breast cancer (TNBC), including but not limited to: cancer, sweat gland cancer, synovioma, testicular cancer, small cell lung cancer, laryngeal cancer, triple negative breast cancer (TNBC), uterine cancer, Wilms' tumor, blood cancer, acute erythroleukemic leukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute lymphoblastic leukemia, acute megakaryocytic leukemia, acute monocytic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute non-lymphocytic leukemia, acute promyelocytic leukemia, acute undifferentiated leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma, polycythemia vera, or Waldenstrom's macroglobulinemia. In some embodiments, the disease is triple negative breast cancer (TNBC).

[1414] Dosage and administration schedule

[1415] The amount of the pharmaceutical composition administered using the methods herein (e.g., dosage of the conjugates, drug moieties, linkers, compounds and / or compositions of the present disclosure (e.g., Formula (I), Formula (II) and / or Formula (III))) will depend on the human or mammal being treated, the severity of the disorder or condition, the rate of administration, the disposition of the active pharmaceutical ingredient, and the judgment of the prescribing physician. However, in single or divided doses, effective dosages are in the range of about 0.001 mg / kg body weight per day to about 100 mg / kg body weight per day, such as about 1 mg / kg / day to about 35 mg / kg / day. For a 70 kg human, this would be equivalent to about 0.05 g / day to 7 g / day, such as about 0.05 g / day to about 2.5 g / day. In some cases, dosage levels below the lower limit of the aforementioned range may be more than adequate, while in other cases, still larger doses may be employed without causing any adverse side effects, for example, by dividing such larger doses into several smaller doses for administration throughout the day. The dosage of the pharmaceutical composition and active pharmaceutical ingredient may be expressed in mg / kg body mass or mg / m 2 The units are provided in terms of body surface area.

[1416] In some embodiments, the pharmaceutical composition or active pharmaceutical ingredient is administered in a single dose. Such administration can be by injection, for example, intravenous injection, to rapidly introduce the active pharmaceutical ingredient. However, other routes, including the preferred oral route, may be used where appropriate. A single dose of the pharmaceutical composition may also be used to treat acute conditions.

[1417] In some embodiments, the pharmaceutical composition or active pharmaceutical ingredient is administered in multiple doses. In one embodiment, the pharmaceutical composition is administered in multiple doses. Administration can be once a day, twice a day, three times a day, four times a day, five times a day, six times a day, or more than six times a day. Administration can be monthly, biweekly, weekly, or every other day. In other embodiments, the pharmaceutical composition is administered from about once a day to about six times a day. In some embodiments, the pharmaceutical composition is administered once a day, while in other embodiments, the pharmaceutical composition is administered twice a day, and in other embodiments, the pharmaceutical composition is administered three times a day.

[1418] As long as necessary, the administration of active pharmaceutical ingredient can continue. In selected embodiments, the pharmaceutical composition is administered for more than 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 14 days or 28 days. In certain embodiments, the pharmaceutical composition is administered for less than 28 days, 14 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days or 1 day. In certain embodiments, the pharmaceutical composition is administered chronically on a continuous basis, for example, for treating chronic effects. In certain embodiments, the administration of the pharmaceutical composition continues for less than about 7 days. In another embodiment, the administration continues for more than about 6 days, 10 days, 14 days, 28 days, two months, six months or one year. In some cases, continuous administration is achieved and as long as necessary, the continuous administration is maintained.

[1419] In some embodiments, an effective dose of an active pharmaceutical ingredient disclosed herein (e.g., any conjugate, drug moiety, linker, compound, and / or composition disclosed herein (e.g., Formula (I), Formula (II), and / or Formula (III))) is in the range of about 1 mg to about 500 mg, about 10 mg to about 300 mg, about 20 mg to about 250 mg, about 25 mg to about 200 mg, about 10 mg to about 200 mg, about 20 mg to about 150 mg, about 30 mg to about 120 mg, about 10 mg to about 90 mg, about 20 mg to about 80 mg, about 30 mg to about 70 mg, about 40 mg to about 60 mg, about 80 mg to about 90 mg, about 10 mg to about 150 mg, about 150 mg to about 150 mg, about 1 ... In some embodiments, the effective dose...

Claims

1. A compound of formula (III) or a salt, solvate, tautomer, isomer or mixture thereof: LD Formula (III) Wherein in formula (III): L is of the formula R*-L1-L A - connector; R* is maleimide; L1 is -[CH2] 1-3 -C(O)NH-; L A Yes - [CH2CH2O] p -(CH2) 1-5 -C(O)-X AA -, wherein p is an integer from 5 to 10, and X AA is an amino acid sequence having 2 amino acid portions; and D is selected from 2. The compound according to claim 1, wherein L1 is -[CH2]2-C(O)NH-.

3. The compound according to claim 1 or 2, wherein p is 7 or 8.

4. The compound according to any one of claims 1 to 3, wherein p is 8.

5. A compound according to any one of claims 1 to 4, wherein X AA Selected from Val-Ala, Tyr-Arg, Phe-Arg, Val-Gln, Val-Cit, Tyr-Met, Leu-Gln, Val-Arg, Met-Thr, Phe-Gln , Thr-Thr, Val-Thr, Ala-Ala, Val-Met, Leu-Met, Ala-Asn, D-Val-D-Gln, D-Ala-D-Ala and Phe-Met.

6. A compound according to any one of claims 1 to 5, wherein X AA It's valine-alanine.

7. A compound according to any one of claims 1 to 6, wherein L A Yes - [CH2CH2O] p -(CH2) 1-3 -C(O)-X AA -.

8. A compound according to any one of claims 1 to 7, wherein L A Yes - [CH2CH2O] p -(CH2)2-C(O)-X AA -.

9. A compound according to any one of claims 1 to 8, wherein D is 10. The compound according to any one of claims 1 to 9, wherein the linker L has the formula:

11. A compound according to any one of claims 1 to 10, wherein D is 12. A compound according to any one of claims 1 to 11, wherein the compound has the formula:

13. An antibody-drug conjugate (ADC) having formula (I): Ab-[LD] n Formula (I) Wherein in formula (I): Ab comprises an antibody or a binding fragment thereof; L is the formula -R * -L1-L A - connector; R * It is succinimide; L1 is -[CH2] 1-3 -C(O)NH-; L A Yes - [CH2CH2O] p -(CH2) 1-5 -C(O)-X AA -, wherein p is an integer from 5 to 10, and X AA is an amino acid sequence having 2 amino acid portions; D is selected from and n is an integer of 1 to 20.

14. The antibody-drug conjugate of claim 13, wherein L1 is 15. The antibody-drug conjugate according to claim 13 or 14, wherein p is 7 or 8.

16. The antibody-drug conjugate according to any one of claims 13 to 15, wherein p is 8.

17. The antibody-drug conjugate according to any one of claims 13 to 16, wherein X AA Selected from Val-Ala, Tyr-Arg, Phe-Arg, Val-Gln, Val-Cit, Tyr-Met, Leu-Gln, Val-Arg, Met-Thr, Phe-Gln , Thr-Thr, Val-Thr, Ala-Ala, Val-Met, Leu-Met, Ala-Asn, D-Val-D-Gln, D-Ala-D-Ala and Phe-Met.

18. The antibody-drug conjugate according to any one of claims 13 to 17, wherein X AA It's valine-alanine.

19. The antibody-drug conjugate according to any one of claims 13 to 18, wherein L A Yes - [CH2CH2O] p -(CH2) 1-3 -C(O)-X AA -.

20. The antibody-drug conjugate according to any one of claims 13 to 19, wherein L A Yes - [CH2CH2O] p -(CH2)2-C(O)-X AA -.

21. The antibody-drug conjugate according to any one of claims 13 to 20, wherein the linker L has the formula:

22. The antibody-drug conjugate of any one of claims 13 to 21, wherein D is 23. The antibody-drug conjugate according to any one of claims 13 to 22, wherein LD has the formula:

24. The antibody-drug conjugate according to any one of claims 13 to 23, wherein n is an integer from 4 to 8.

25. The antibody-drug conjugate of claim 24, wherein n is 4.

26. The antibody-drug conjugate of claim 24, wherein n is 8.

27. The antibody-drug conjugate of any one of claims 10 to 19, wherein the antibody-drug conjugate has a drug-antibody ratio (DAR) in the range of about 1 to about 10, optionally wherein the DAR is about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9 or about 10, optionally the DAR is about 4, optionally the DAR is about 8.

28. A pharmaceutical composition comprising the antibody drug conjugate according to any one of claims 13 to 29; and a pharmaceutically acceptable carrier.

29. A method of treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of the antibody-drug conjugate according to any one of claims 13 to 28 or the pharmaceutical composition according to claim 28.

30. The method of claim 29, wherein less than about 50% of the antibody-drug conjugate is converted to metabolites about 24 hours after administering the therapeutically effective amount of the antibody-drug conjugate to the subject.

31. The method of claim 29 or 30, wherein about 50% of the antibody-drug conjugate is converted to metabolites about 96 hours after administering the therapeutically effective amount of the antibody-drug conjugate to the subject.

32. The method of any one of claims 29 to 31, wherein the antibody-drug conjugate is converted to a metabolite of formula 300:

33. The method of any one of claims 29 to 32, wherein the antibody-drug conjugate is converted to a metabolite of formula 301:

34. The method of any one of claims 29 to 33, wherein the antibody-drug conjugate is converted to a metabolite of formula 302:

35. A method according to any one of claims 32 to 34, wherein: (a) converting the metabolite of formula 300 into the metabolite of formula 301; (b) converting the metabolite of formula 300 into the metabolite of formula 302; (c) converting the metabolite of formula 301 into the metabolite of formula 302; or (d) converting the metabolite of formula 300 into the metabolite of formula 301, and converting the metabolite of formula 301 into the metabolite of formula 302.

36. The method of any one of claims 29 to 35, wherein the antibody-drug conjugate is converted to a metabolite in vivo.

37. The method of any one of claims 29 to 35, wherein the antibody-drug conjugate is converted to a metabolite in vitro.

38. The method of any one of claims 29 to 37, wherein the cancer is selected from the group consisting of pancreatic cancer, breast cancer, prostate cancer, lymphoma, skin cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain cancer, head-neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck carcinoma, breast carcinoma, ovarian carcinoma, lung cancer, small-cell lung carcinoma, Wilms' tumor, cervical cancer, testicular cancer, bladder carcinoma, pancreatic carcinoma, stomach cancer, colon cancer. carcinoma), prostatic carcinoma, genitourinary cancer, thyroid cancer, esophageal cancer, myeloma, multiple myeloma, adrenal cancer, renal cell carcinoma, endometrial cancer, adrenocortical carcinoma, malignant pancreatic insulinoma, malignant carcinoid, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic granulocytic leukemia, acute myeloid leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, polycythemia vera, essential thrombocythemia, Hodgkin's disease, non-Hodgkin's lymphoma lymphoma), soft tissue sarcoma, osteosarcoma, primary macroglobulinemia or retinoblastoma, etc. In other embodiments,The cancer is acoustic neuroma, adenocarcinoma, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, triple negative breast cancer (TNBC), bronchogenic carcinoma, cervical cancer, chordoma, choriocarcinoma, colon cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, embryonal carcinoma, endothelial carcinoma, ependymoma, epithelial carcinoma, esophageal cancer, Ewing's tumor, fibrosarcoma, gastric cancer cancer), glioblastoma multiforme, glioma, head and neck cancer, hemangioblastoma, hepatoma, kidney cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangioendothelioma, lymphangiosarcoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, myxosarcoma, nasal cancer, neuroblastoma, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, retinoblastoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, squamous cell carcinoma, stomach cancer cancer), sweat gland cancer, synovial tumor, testicular cancer, small cell lung cancer, laryngeal cancer, uterine cancer, Wilm's tumor, blood cancer, acute erythroleukemic leukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monocytic leukemia, acute myeloblastic leukemia, acute myelomonocytic leukemia, acute non-lymphocytic leukemia, acute promyelocytic leukemia, acute undifferentiated leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, multiple myeloma, heavy chain disease, Hodgkin's disease, multiple myeloma, non-Hodgkin's lymphoma, polycythemia vera or Waldenstrom's macroglobulinemia. , 39. An antibody-drug conjugate having formula (I): Ab-[LD] n Formula (I) Wherein in formula (I): Ab comprises an antibody or a binding fragment thereof; n is an integer from 1 to 20; and LD has the formula:

40. The antibody-drug conjugate of claim 39, wherein n is an integer from 1 to 10, 2 to 8, or 4 to 8, optionally n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

41. The antibody-drug conjugate of claim 39, wherein n is 4 or 8.

42. The antibody-drug conjugate of claim 39, wherein n is 4.

43. The antibody-drug conjugate of claim 39, wherein n is 8.

44. The antibody-drug conjugate of claim 39, wherein the antibody-drug conjugate has a drug-antibody ratio (DAR) in the range of about 1 to about 10, optionally wherein the DAR is about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9 or about 10, optionally the DAR is about 4, optionally the DAR is about 8.

45. A compound having formula 30, any one of formulas 3031-3064 or 3101-3118, or a salt, solvate, tautomer, isomer or mixture thereof.

46. ​​A compound having the following formula or a salt, solvate, tautomer, isomer or mixture thereof:

47. An antibody-drug conjugate having any one of formulae 1030-1064 or 1100-1118.

48. A compound according to any one of embodiments (CI) to (CXVI).

49. An antibody-drug conjugate of any one of embodiments (I)-(XVII).

50. An antibody-drug conjugate having formula (I): Ab-[LD] n Formula (I) Wherein in formula (I): Ab comprises an antibody or a binding fragment thereof; n is 1; and LD has the formula:

51. An antibody-drug conjugate having formula (I): Ab-[LD] n Formula (I) Wherein in formula (I): Ab comprises an antibody or a binding fragment thereof; n is 4; and LD has the formula:

52. An antibody-drug conjugate having formula (I): Ab-[LD] n Formula (I) Wherein in formula (I): Ab comprises an antibody or a binding fragment thereof; n is 8; and LD has the formula:

53. A method of treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of an antibody-drug conjugate or compound according to any one of claims 33 to 52.

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