Antibody-drug-conjugates cleavable in a tumor microenvironment

Abstract

This application relates to antibody-drug conjugates cleavable in a tumor microenvironment by proteases such as neutrophil elastase, pharmaceutical compositions comprising the same, processes for preparation thereof, and the use thereof for treating diseases, disorders, or conditions in humans and other mammals, including hyperproliferative disorders such as cancer.

Description

ANTIBODY-DRUG CONJUGATES CLEAVABLE IN A TUMOR MICROENVIRONMENT CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This International Patent Application claims the benefit of European Patent Application No. 22306695.2, filed on November 17, 2022, and of European Patent Application No. 23305485.7, filed on April 3, 2023, each of which is incorporated herein by reference in its entirety. SEQUENCE LISTING

[0002] The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled 59362-725.601.xml, created November 14, 2023, which is 132 kilobytes in size. The information in the electronic format of the Sequence Listing is incorporated by reference in its entirety. BACKGROUND OF THE INVENTION

[0003] This application relates to antibody-drug conjugates, pharmaceutical compositions, processes for preparation thereof, and the use thereof for treating, preventing, or managing diseases and conditions including hyperproliferative disorders, such as, cancer in humans and other mammals. SUMMARY

[0004] In an aspect, provided herein is an antibody-drug conjugate according to Formula I:or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding antibody fragment; B1 and B2 are in each instance, independently, a small molecule target protein binder; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; P1 is a protease-cleavable linker; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1, 2, 3, 4, or 5.

[0005] In some embodiments, provided herein is an antibody-drug conjugate according to Formula I- A, Formula I-B, or Formula I-C:or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding antibody fragment; B1 and B2 are in each instance, independently, a small molecule target protein binder; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; P1 is a protease-cleavable linker; m is an integer from 1 to 20; n’ is an integer from 1 to 20; and p is 1, 2, 3, 4, or 5.

[0006] In another aspect, provided herein is a compound of Formula (VIII) or Formula (IX):or a pharmaceutically acceptable salt thereof, wherein: PR1 and PR2 are in each instance, independently, a protein-reactive group; L1a and L2aare in each instance, independently, a non-cleavable linker; B1 and B2 are in each instance, independently, a small molecule target protein binder; P1 is in each instance, independently, a protease-cleavable linker; D1 is in each instance, independently, a therapeutic payload; and p is 1, 2, 3, 4, or 5. INCORPORATION BY REFERENCE

[0007] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent publications and patents or patent applications incorporated by reference contradict the disclosure contained in the specification, the specification is intended to supersede and / or take precedence over any such contradictory material.P368919WO / 59362-725.601 BRIEF DESCRIPTION OF THE DRAWINGS

[0008] A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings (also “Figure” or “FIG.” herein), of which:

[0009] FIG. 1 shows cytokine release of freshly prepared PBMC from three healthy donors after treatment with the indicated compounds (13i-5657: 37.5 µg / mL; 14i-5657: 24 µg / mL and 15i-5657: 41 µg / mL). DETAILED DESCRIPTION

[0010] While various embodiments of the invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions may occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed.

[0011] The present invention relates to novel pharmaceutical compounds comprising an antibody or antigen-binding fragment thereof capable of binding to an antigen expressed on a tumor cell, or on a cell present in the tumor microenvironment, and which are linked via protease cleavable linkers to one or more payload molecules, the processes for preparation thereof, and to the use thereof for treating, preventing or managing diseases and conditions including hyperproliferative disorders such as cancer in humans and other mammals. Conjugates comprising an antibody or antigen-binding fragment thereof and a payload molecule (e.g., a drug) are often referred to as antibody-drug conjugates (ADCs). ADCs described herein may further comprise one or more binding molecules stably bonded thereto, which may also be capable of bindng to a target protein.

[0012] Examples for binding molecules include binders to target proteins (e.g. cell adhesion proteins) including alpha-v beta-3 integrin, alpha-v beta-6 integrin, prostate specific membrane antigen (PSMA), fibroblast activation protein (FAP), carbonic anhydrase IX (CAIX), a chaperone protein Heat Shock 90 (Hsp 90), Folic Acid Receptor, Glucose transporter 1, Somatostatin receptor, aminopeptidase N (APN), Low density lipoprotein receptor-related protein 1 (LRP1), Bombesin receptor, gonadotropin releasing hormone (GnRH or LHRH) receptor, P32, Membrane type 1 Matrix Metalloprotease (MT1-MMP), Sortilin, or Nectin-4. In some embodiments, the target protein is an alpha-v beta-3 integrin or a fibroblast activation protein. The binding molecules may be bound to the ADC via a linker. In some embodiments, the binding molecule is conjugated to the ADC via the same linker as a payload molecule. In some embodiments, the binding molecule is conjugated to the ADC via a linker that is distinct from a linker to a payload molecule. In some embodiments, the payload molecule is a therapeutic payload (D1), as described herein.P368919WO / 59362-725.601

[0013] Payload molecules employed in ADCs provided herein can be a cytotoxic compound or an immunostimulatory agent. In some preferred embodiments, provided herein are compounds with therapeutic payloads (D1) such as camptothecin derivatives, auristatin derivatives, CDK9 / PTEFb derivatives, kinesin spindle protein inhibitor derivatives, or toll-like receptor 7 and / or 8 (“TLR7 / 8) agonist derivatives.

[0014] ADCs provided herein may comprise one or more protease-cleavable payloads (e.g., therapeutic payloads) which, upon cleavage by a protease, exhibit cytotoxic or immune agonist properties. In order to reduce off-target cytotoxicity or immunostimulatory properties, a payload may be bound to a conjugate (as provided herein) via a protease-cleavable linker, wherein the protease is selectively expressed (e.g., overexpressed) by tumor cells, or by cells present in a tumor microenvironment. The targeted cytotoxic or immunostimulatory effects of a conjugate provided herein (or of a therapeutic payload conjugated thereto) can be further enhanced when the release of the payload takes place in the immediate vicinity of the tumor tissue (i.e., within a tumor microenvironment), or within a tumor cell. Of additional advantage and utility are therapeutic payloads which, upon release from a conjugate, can penetrate a tumor cell. Preferably, a therapeutic payload is not effluxed or transported outside of a cell. In some embodiments, conjugates described herein are selectively cleavable in the microenvironment of cancer cells, with less or no cleavage taking place in healthy cells. In some embodiments, a conjugate described herein or an active metabolite thereof is cytotoxic or immunostimulatory after activation by a tumor-associated enzyme, such as, neutrophil elastase. In some embodiments, a conjugate described herein is non-toxic, in therapeutic concentrations, in the absence of said activation. DEFINITIONS

[0015] Unless defined otherwise, all terms of art, notations and other technical and scientific terms or terminology used herein are intended to have the same meaning as is commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art.

[0016] Throughout this application, various embodiments may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.P368919WO / 59362-725.601

[0017] As used in the specification and claims, the singular forms “a”, “an” and “the” include plural references unless the context clearly dictates otherwise. For example, the term “a sample” includes a plurality of samples, including mixtures thereof.

[0018] The terms “determining,” “measuring,” “evaluating,” “assessing,” “assaying,” and “analyzing” are often used interchangeably herein to refer to forms of measurement. The terms include determining if an element is present or not (for example, detection). These terms can include quantitative, qualitative or quantitative and qualitative determinations. Assessing can be relative or absolute. “Detecting the presence of” can include determining the amount of something present in addition to determining whether it is present or absent depending on the context.

[0019] The terms “subject,” “individual,” or “patient” are often used interchangeably herein. A “subject” can be a biological entity containing expressed genetic materials. The biological entity can be a plant, animal, or microorganism, including, for example, bacteria, viruses, fungi, and protozoa. The subject can be tissues, cells and their progeny of a biological entity obtained in vivo or cultured in vitro. The subject can be a mammal. The mammal can be a human. The subject may be diagnosed or suspected of being at high risk for a disease. In some cases, the subject is not necessarily diagnosed or suspected of being at high risk for the disease.

[0020] As used herein, the term “about” a number refers to that number plus or minus 15% of that number. The term “about” a range refers to that range minus 15% of its lowest value and plus 15% of its greatest value.

[0021] As used herein, the terms “treatment” or “treating” are used in reference to a pharmaceutical or other intervention regimen for obtaining beneficial or desired results in the recipient. Beneficial or desired results include but are not limited to a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit may refer to eradication or amelioration of symptoms or of an underlying disorder being treated. Also, a therapeutic benefit can be achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the subject, notwithstanding that the subject may still be afflicted with the underlying disorder. A prophylactic effect includes delaying, preventing, or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof. For prophylactic benefit, a subject at risk of developing a particular disease, or to a subject reporting one or more of the physiological symptoms of a disease may undergo treatment, even though a diagnosis of this disease may not have been made.

[0022] As used herein, the term “therapeutic payload” generally refers to a chemical group, generally a small molecule (i.e., non-protein) group, having therapeutic activity. Preferably, the therapeutic activity is enhanced (e.g., activated) after separation from a cleavable group. In some embodiments, the cleavable group is an enzymatically-cleavable group. In some embodiments, the therapeutic payload is activated following cleavage by a tumor-associated protein such as neutrophil elastase. A therapeutic payload may be, for example, a drug. In some embodiments, the therapeutic payload is a cytotoxic, cytostatic, orP368919WO / 59362-725.601 immunomodulatory compound. In some embodiments, the therapeutic payload is effective in killing or slowing the growth of cancer cells. In some embodiments, the therapeutic payload is a kinesin spindle protein inhibitor, a camptothecin or a derivative thereof, a CDK9 inhibitor, etc. as described herein.

[0023] As used herein, the term “target protein” generally refers to a protein that is expressed on the surface of a cell (e.g., a cancer cell), which can efficiently bind a small molecule binder. As used herein, the term “target antigen” generally refers to an antigen that is expressed on the surface of a cell (e.g., a cancer cell), which can efficiently bind an antibody or antigen-binding fragment thereof. Efficiently is used herein to generally refer to a compound with micromolar potency or better (e.g., sub-micromolar, nanomolar, sub-nanomolar, etc. as used in the art). Examples of target proteins as defined herein.

[0024] As used herein, the term “non-cleavable linker” refers to a linking unit of atoms (e.g., 1 to 200 atoms selected from C, H, N, O, S, and halogen) that is not known to be chemically or biologically unstable. The term “non-cleavable linker” is intended to differentiate from cleavable linkers (e.g., protease-cleavable linkers, self-immolative linkers, pH-sensitive linkers, etc.). A non-cleavable linker may be an alkyl or heteroalkyl linker, optionally interrupted by one or more cyclyl or heterocyclyl groups (e.g., click partners or artifacts therefrom). A non-cleavable linker may comprise a polymeric section (e.g., PEG, PEI, polysarcosine, etc.) and / or an alkyl section.

[0025] A non-cleavable linker may further comprise another functional group such as a small molecule target protein binder, a pharmacokinetic modulator (e.g., a -COOH group), and / or one or more therapeutic payloads. A non-cleavable may be conjoined with a protease-cleavable linker. In such an instance, the protease-cleavable linker may be cleaved by a protease (e.g., in a tumor microenvironment) to release the therapeutic payload, and leaving the non-cleavable linker stably bound to the antibody. In some embodiments, a non-cleavable linker and / or protease-cleavable linker disclosed herein does not comprise a self-immolative linker group, which may further enhance the stability (e.g., reduce off-target release) of the ADCs disclosed herein.

[0026] Additional elements of stability, including the connecting groups conjoining the antibody to a linker, and / or conjoining a protease-cleavable linker to a payload, are enabled by use of the groups disclosed herein. For example, in some embodiments, provided herein is a protein-coupling group that reacts with a lysine or cysteine residue of an antibody or antigen-binding fragment thereof to form a stable linker. Preferably, such protein-coupling groups and / or protein-coupled linkers are not susceptible to unwanted cleavage (e.g., retro-Michael elimination of a maleimide group). In some embodiments, provided herein is a protease-cleavable linker conjugated to a therapeutic payload via an ester, an amide, or a sulfoximine (R-S(O)(C1-6 alkyl)N=R’) bond. In some embodiments, the protease-cleavable linker is cleaved efficiently and cleanly (i.e., without leaving a post-cleavage artifact bonded to the payload at the point of attachment). In some embodiments, the protease-cleavable linker is cleaved efficiently and cleanly without the use of a self-immolative group, thereby resulting in enhanced stability. In some embodiments, provided herein is a method of delivering a payload disclosed herein to a tumor microenvironment.P368919WO / 59362-725.601

[0001] As used herein, C1-Cx includes C1-C2, C1-C3 . . . C1-Cx. By way of example only, a group designated as "C1-C6" indicates that there are one to six carbon atoms in the moiety, e.g., groups containing 1 carbon atom, 2 carbon atoms, 3 carbon atoms or 4 carbon atoms. Thus, by way of example only, "C1-C4alkyl" indicates that there are one to four carbon atoms in the alkyl group, e.g., the alkyl group is selected from among methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, and t-butyl.

[0002] An “alkyl” group refers to an aliphatic hydrocarbon group. The alkyl group is branched or straight chain. In some embodiments, the “alkyl” group has 1 to 10 carbon atoms, e.g. a C1-C10alkyl. Whenever it appears herein, a numerical range such as “1 to 10” refers to each integer in the given range; e.g., “1 to 10 carbon atoms” means that the alkyl group consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms, although the present definition also covers the occurrence of the term “alkyl” where no numerical range is designated. In some embodiments, an alkyl is a C1-C6alkyl. In one aspect the alkyl is methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, or t-butyl. Typical alkyl groups include, but are in no way limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec- butyl, tertiary butyl, pentyl, neopentyl, or hexyl.

[0003] An “alkylene” group refers to a divalent alkyl radical. Any of the above mentioned monovalent alkyl groups may be an alkylene by abstraction of a second hydrogen atom from the alkyl. In some embodiments, an alkylene is a C1-C6alkylene. In other embodiments, an alkylene is a C1-C4alkylene. Typical alkylene groups include, but not limited to,, -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2- , and the like. In some embodiments, an alkylene is -CH2-.

[0004] An “alkoxy” group refers to a -O(alkyl) group, where alkyl is as defined herein. Examples of alkoxy groups include -OCH3, -OCH2CH3, -OCH2CH2CH3, -OC(CH3)3, and the like.

[0005] An “hydroxyalkyl” refers to an alkyl in which one hydrogen atom is replaced by a hydroxyl. In some embodiments, a hydroxyalkyl is a C1-C4hydroxyalkyl. Typical hydroxyalkyl groups include, but not limited to, -CH2OH, -CH2CH2OH, -CH2CH2CH2OH, -CH2CH2CH2CH2OH, -C(CH3)2OH, and the like.

[0006] The term “alkylamine” refers to the –N(alkyl)xHy group, where x is 0 and y is 2, or where x is 1 and y is 1, or where x is 2 and y is 0.

[0007] An “aminoalkyl” refers to an alkyl in which one hydrogen atom is replaced by an amino. In some embodiments, aminoalkyl is a C1-C4aminoalkyl. Typical aminoalkyl groups include, but not limited to, -CH2NH2, -CH2CH2NH2, -CH2CH2CH2NH2, -CH2CH2CH2CH2NH2, -C(CH3)2NH2, and the like.

[0008] The term, “heteroalkyl” generally refers to a straight-chain and / or branched hydrocarbon chain which has 1 to 30 carbon atoms and may be interrupted once or more than once by one or more of the groups -O-, -S-, -C(=O)-, -S(=O)-, -S(=O)2-, -NRy-, -NRyC(=O)-, -C(=O)-NRy-, -NRyNRy-, -S(=O)2- NRyNRy-, -C(=O)-NRyNRy-, -CRx=N-O-, and where the hydrocarbon chain including the side chains, if present, may be substituted by –NH-C(=O)-NH2, -C(=O)-OH, -OH, -NH2, -NH-C(=NNH2)-, sulfonamide, sulfone, sulfoxide, sulfonic acid, sulfamide, or a combination thereof. In this context, Ry in each case is - H, phenyl, C1-C10-alkyl, C2-C10-alkenyl or C2-C10-alkynyl, which may in turn be substituted in each case by -NHC(O)NH2, -COOH, -OH, -NH2, -NH-C(=NNH2)-, sulfonamide, sulfone, sulfoxide, sulfonic acid,P368919WO / 59362-725.601 sulfamide, or a combnation thereof. In this context, Rx is -H, C1-C3-alkyl or phenyl. The terms “heteroalkyl- aryl” and “heteroalkyl-heteroaryl” as used herein generally refer to a heteroalkyl group (as defined above) substituted with an aromatic carbocycle or an aromatic heterocycle respectively; and wherein each is optionally substituted.

[0009] The term “aromatic” refers to a planar ring having a delocalized ^-electron system containing 4n+2 ^ electrons, where n is an integer. The term “aromatic” includes both carbocyclic aryl (“aryl”, e.g., phenyl) and heterocyclic aryl (or “heteroaryl” or “heteroaromatic”) groups (e.g., pyridine). The term includes monocyclic or fused-ring polycyclic (or rings which share adjacent pairs of carbon atoms) groups.

[0010] The term “carbocyclic” or “carbocycle” refers to a ring or ring system where the atoms forming the backbone of the ring are all carbon atoms. The term thus distinguishes carbocyclic from “heterocyclic” rings or “heterocycles” in which the ring backbone contains at least one atom which is different from carbon. In some embodiments, at least one of the two rings of a bicyclic carbocycle is aromatic. In some embodiments, both rings of a bicyclic carbocycle are aromatic. Carbocycles include aryls and cycloalkyls.

[0011] As used herein, the term “aryl” refers to an aromatic ring wherein each of the atoms forming the ring is a carbon atom. In one aspect, aryl is phenyl or a naphthyl. In some embodiments, an aryl is a phenyl. In some embodiments, an aryl is a phenyl, naphthyl, indanyl, indenyl, or tetrahydronaphthyl. In some embodiments, an aryl is a C6-C10aryl. Depending on the structure, an aryl group is a monoradical or a diradical (or an arylene group). As used herein, the term “aralkyl” generally refers to a monocyclic aromatic carbocycle (e.g., phenyl), to which a C1-4-alkyl group is bonded. Illustrative aralkyl groups include benzyl and ethylphenyl.

[0012] The term “cycloalkyl” refers to a monocyclic or polycyclic aliphatic, non-aromatic radical, wherein each of the atoms forming the ring (or skeletal atoms) is a carbon atom. In some embodiments, cycloalkyls are spirocyclic or bridged compounds. In some embodiments, cycloalkyls are optionally fused with an aromatic ring, and the point of attachment is at a carbon that is not an aromatic ring carbon atom. Cycloalkyl groups include groups having from 3 to 10 ring atoms. In some embodiments, cycloalkyl groups are selected from among cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, spiro[2.2]pentyl, norbornyl and bicycle[1.1.1]pentyl. In some embodiments, a cycloalkyl is a C3-C6cycloalkyl. In some embodiments, a cycloalkyl is a C3-C4cycloalkyl.

[0013] The term “halo” or, alternatively, “halogen” or “halide” means fluoro, chloro, bromo or iodo. In some embodiments, halo is fluoro, chloro, or bromo.

[0014] The term “fluoroalkyl” refers to an alkyl in which one or more hydrogen atoms are replaced by a fluorine atom. In one aspect, a fluoroalkyl is a C1-C6fluoroalkyl.

[0015] The term “optionally substituted” or “substituted” means that the referenced group is optionally substituted with one or more additional group(s) individually and independently selected from halogen, -CN, -NH2, -NH(alkyl), -N(alkyl)2, -OH, -CO2H, -CO2alkyl, -C(=O)NH2, -C(=O)NH(alkyl), - C(=O)N(alkyl)2, -S(=O)2NH2, -S(=O)2NH(alkyl), -S(=O)2N(alkyl)2, alkyl, cycloalkyl, fluoroalkyl, heteroalkyl, alkoxy, fluoroalkoxy, heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio,P368919WO / 59362-725.601 alkylsulfoxide, arylsulfoxide, alkylsulfone, and arylsulfone. In some other embodiments, optional substituents are independently selected from halogen, -CN, -NH2, -NH(CH3), -N(CH3)2, -OH, -CO2H, - CO2(C1-C4alkyl), -C(=O)NH2, -C(=O)NH(C1-C4alkyl), -C(=O)N(C1-C4alkyl)2, -S(=O)2NH2, - S(=O)2NH(C1-C4alkyl), -S(=O)2N(C1-C4alkyl)2, C1-C4alkyl, C3-C6cycloalkyl, C1-C4fluoroalkyl, C1- C4heteroalkyl, C1-C4alkoxy, C1-C4fluoroalkoxy, -SC1-C4alkyl, -S(=O)C1-C4alkyl, and -S(=O)2C1-C4alkyl. In some embodiments, optional substituents are independently selected from halogen, -CN, -NH2, -OH, - NH(CH3), -N(CH3)2, -CH3, -CH2CH3, -CHF2, -CF3, -OCH3, -OCHF2, and -OCF3. In some embodiments, substituted groups are substituted with one or two of the preceding groups. In some embodiments, an optional substituent on an aliphatic carbon atom (acyclic or cyclic) includes oxo (=O).

[0016] “Small molecule,” as used herein, generally refers to any molecule having a molecular weight of about 1000 atomic mass units (Daltons) or less. In some embodiments, a moiety within a compound described herein is refered to as a small molecule, meaning that moiety has a molecular weight of about 1000 Da or less. Small molecules, as used herein, excludes proteins or antibodies, but may comprise peptides or amino acids. In some instances, a compound described herein is a conjugate of two small molecule moieties. Therefore, compounds described herein may be referred to as “small molecule drug conjugates” (SMDCs) or “small molecule prodrug conjugates” (SMPCs). For example, a SMPC may contain a cleavable group (e.g., enzymatically or chemically cleavable) such as an ester which, upon cleavage, results in a SMDC. In some examples, an SMPC (sometimes referred to as a prodrug) described herein can first be cleaved (e.g., in plasma) before an enzyme can efficiently recognize and cleave the SMDC, thus liberating the active agent(s) in two steps. In some embodiments, a SMPC enables slow conversion to the SMDC, which is cleaved more rapidly (i.e., in the presence of a suitable enzyme (e.g., a tumor associated enzyme such as elastase, legumain, or cathepsin)), thus enhancing the therapeutic window or reducing side effects.

[0017] “Pharmaceutically acceptable,” as used herein, refers a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively nontoxic at the concentration or amount used, e.g., the material is administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0018] The term “pharmaceutically acceptable salt” refers to a form of a therapeutically active agent that consists of a cationic form of the therapeutically active agent in combination with a suitable anion, or in alternative embodiments, an anionic form of the therapeutically active agent in combination with a suitable cation. Handbook of Pharmaceutical Salts: Properties, Selection and Use. International Union of Pure and Applied Chemistry, Wiley-VCH 2002. S.M. Berge, L.D. Bighley, D.C. Monkhouse, J. Pharm. Sci. 1977, 66, 1-19. P. H. Stahl and C. G. Wermuth, editors, Handbook of Pharmaceutical Salts: Properties, Selection and Use, Weinheim / Zürich:Wiley-VCH / VHCA, 2002. Pharmaceutical salts typically are more soluble and more rapidly soluble in stomach and intestinal juices than non-ionic species and so are useful in solid dosage forms. Furthermore, because their solubility often is a function of pH, selective dissolution in one or another part of the digestive tract is possible and this capability can be manipulatedP368919WO / 59362-725.601 as one aspect of delayed and sustained release behaviors. Also, because the salt-forming molecule can be in equilibrium with a neutral form, passage through biological membranes can be adjusted.

[0019] The term “acceptable” with respect to a formulation, composition or ingredient, as used herein, means having no persistent detrimental effect on the general health of the subject being treated.

[0020] The terms "administer," "administering", "administration," and the like, as used herein, refer to the methods that may be used to enable delivery of compounds or compositions to the desired site of biological action. These methods include, but not limited to, oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular or infusion), topical and rectal administration. Those of skill in the art are familiar with administration techniques that can be employed with the compounds and methods described herein. In some embodiments, the compounds and compositions described herein are administered orally.

[0021] The terms “effective amount” or “therapeutically effective amount,” as used herein, refer to a sufficient amount of an agent or a compound being administered, which will relieve to some extent one or more of the symptoms of the disease or condition being treated. The result includes reduction or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. For example, an “effective amount” for therapeutic uses is the amount of the composition comprising a compound as disclosed herein required to provide a clinically significant decrease in disease symptoms. An appropriate “effective” amount in any individual case is optionally determined using techniques, such as a dose escalation study.

[0027] The terms “enhance” or “enhancing,” as used herein, means to increase or prolong either in potency or duration a desired effect. Thus, in regard to enhancing the effect of therapeutic agents, the term “enhancing” refers to the ability to increase or prolong, either in potency or duration, the effect of other therapeutic agents on a system. An “enhancing-effective amount,” as used herein, refers to an amount adequate to enhance the effect of another therapeutic agent in a desired system.

[0028] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. ANTIBODY-DRUG CONJUGATES

[0029] Antibody-drug conjugates (ADCs) are an emerging drug class that utilizes antibodies to improve targeting of cytotoxic or immune-oncology payloads to tumor cells. In most cases of current ADCs which are on the market or in development, the active metabolite is formed in the lysosomal compartment of cancer cells upon internalization and trafficking of the ADC to this compartment. The active metabolite is formed either by enzymatic cleavage of the linker or by antibody degradation under the specific conditions (pH, presence of degrading enzymes) in the lysosomes. The approval of meanwhile 13 ADCs demonstrates the potential of ADCs to improve cancer treatment. Many antibodies are currently not suitable for delivering payloads to cancer cells, because they are not efficiently internalized upon binding and / or are not trafficking to lysosomal compartments, where the payload can be released.P368919WO / 59362-725.601

[0030] The current invention discloses antibody-drug conjugates which have been designed for payload release and activation in the tumor microenvironment (TME). The linkers disclosed herein have been optimized to be substrate sequences for enzymes which are upregulated in the tumor microenvironment. Many of these enzymes are part of the protease family of enzymes which may be involved in the modulation of the tumor stroma and the motility of tumor cells. Some of these tumor microenvironment shaping proteases are, for example, serine proteases like plasmin activator, seprase, hepsin or kallikreins. Another family of protease in the TME are cysteine proteases such as cathepsin B and cathepsin K or aspartyl proteases such as cathepsin D and cathepsin E, but also other proteases such as heparanase, endoglycosidase and hyaluronidase were shown to be upregulated and activated within the TME. Such enzymes can be proteases such as matrix metalloproteases cathepsins and neutrophil elastase.

[0031] Another key aspect for ADC performance is the physicochemical profile of the payload which is released. Payloads which are extracellularly cleaved in TME from the ADCs disclosed in the current invention should be membrane permeable and efficiently penetrate into tumor tissue. The release of cell- permeable payloads is associated with a bystander killing effect, which is considered to be particularly beneficial for the treatment of tumors with heterogenous target expression.

[0032] Disclosed herein are conjugates comprising an antibody, optionally one or more small molecule target protein binders linked via a linker, and one or more therapeutic payloads linked via a protease- cleavable linker. The linker may further comprise a non-cleavable linker. Non-cleavable linkers described herein may be bivalent or multivalent. For example, in some embodiments, provided herein is a conjugate comprising a non-cleavable linker which conjoins an antibody to multiple therapeutic payloads, wherein each therapeutic payload is linked to the non-cleavable linker via a protease-cleavable linker. In other embodiments, the non-cleavable linker conjoins a single antibody to a single therapeutic payload, though a given conjugate may comprise multiple non-cleavable linkers each conjoined to a therapeutic payload via a protease-cleavable linker. Non-cleavable linkers may comprise functional elements, including physicochemical-modulating elements (e.g., solubility enhancers), pharmacokinetic-modulating elements (e.g., tumor-targeting or tumor-retained groups), proximity-modulating elements (e.g., spacers), or any combination thereof. In some embodiments, a non-cleavable linker disclosed herein (e.g., L1, L2), is conjoined to a protease-cleavable linker (e.g., P1, P1a).

[0033] Therapeutic payloads for use in accordance with the present invention may be, for example, a drug. Thus, conjugates are generally referred to herein as antibody-drug conjugates (ADCs). However, the term ADC as used herein may refer to any conjugate comprising an antibody and a therapeutic payload, and is not limited specifically to therapeutic payload that is necessarily defined as a “drug.” A therapeutic payload can be, for example, a cytotoxic agent or an immunostimulatory agent. Preferably, the therapeutic payload is permeable to cell membranes (e.g., to tumor cell membranes). More preferably, the therapeutic payload is penetrant to tumor cells and produces cytotoxic or antiproliferative effects in a cell. A therapeutic payload may be configured to be released extracellularly in a tumor microenvironment (e.g., by cleavage of a protease-cleavable linker by an extracellular tumor-associated protein (e.g., neutrophilP368919WO / 59362-725.601 elastase)). An extracellularly released therapeutic payload, e.g., a tumor-penetrant therapeutic payload, may enter a tumor cell and produce potent cytotoxic or immunostimulatory effects.

[0034] In some embodiments, the therapeutic payload is a microtubule toxin (e.g., maytansinoids maytansin, DM1, DM4, DM21, DM23), auristatin (MMAE, MMAF, auristatin F, dolastatin, PF-06380101, amberstatin269, auristatin F-HPA , auristatin W analog, duostatin 5.2, duostatin5, MMAD, SHR152852, Combretastatin A (CBA)), epothilone (epothilone B, epothilone D, ixampra), taxoid (paclitaxel, docetaxel), tubulysin (tubulysin, Tub196, Tub114, Tub201, Tub255, AZ13599185), vinca alkaloid (vinorelbine, vinflunine, vinblastine silanol), cytolysine (TAM470), hemiasterlin (SC209, E7974), or eribulin.

[0035] In some embodiments, the therapeutic payload is a DNA toxin. In some embodiments, the therapeutic payload is an anthracycline (e.g., doxorubicin, daunorubicin, epiburicin, PNU-159682, panobinostat), a topoisomerase I inhibitor (e.g., AZ’0133, camptothecin, belotecan, irinotecan, topotecan, DXd / DX8951, exatecan, FL-118, SN-38, VIP126), a duocarmycin or an analog thereof (e.g., duocarmycin, duocarmycin-hydroxy benzamide azaindole (DUBA), MED-A / DNAMGBA toxin), a calicheamicin, a DNA cross linking agent (e.g., PBD-dimers -FGX20-75, SC-DR003, SG2000, SG3199, SG1882, FGX2- 62, indolino-benzodiazepine dimer -DGN462, DGN549, IGN-P1- , cyclopropylpyrroloindole, isoquinolidinobenzodiazepine –D211) bleomycin A2, dactinomycin, or mitomycin C.

[0036] In some embodiments, the therapeutic payload is a transcription toxin. In some embodiments, the therapeutic payload is an amatoxin (e.g., targeting RNA polymerase II, e.g., amanitin), thailanstatin A (e.g., targeting spliceosome), an oxidative phosphorylation inhibitor (e.g., oligomycin), a protein kinase inhibitor (e.g., an inhibitor of protein kinase B (Akt) such as ipatasertib), an EGFR inhibitor (e.g., erlotinib), a VEGFR inhibitor, (e.g., sorafenib, sunitinib, bevacizumab, Lenvatinib, vandetanib, pazopanib, axitinib, cabozantinib, regorafenib, nintedanib, apatinib), a PDGFR inhibitor, a dihydrofolate reductase (DHFR) inhibitor, e.g., methotrexate, aminopterin, a histone deacetylase inhibitor (e.g., HST746AA1), or a kinesin spindle protein inhibitor (KSPi).

[0037] In some instances, the therapeutic payload is a microtubule toxin, DNA toxin, transcription toxin, or an immune stimulator. In some instances, the microtubule toxin is a maytansinoid, auristatin, epithilone, taxoid, tubulysin, eribulin alkaloid, vinca alkaloid, eribulin, or any combination thereof. In some instances, the DNA toxin is an anthracycline, topoisomerase I inhibitor, duacarmycin or analogs thereof, calichearmicins, DNA cross linking agents, bleomycin A2, dactinomucin, mitomycin C, or any combination thereof. In some instances, the transcription toxin is an amatoxin, thailanstatin A, oxidating phosphorylation inhibitor, protein kinase inhibitor, dihydrofolate reductase (DHFR) inhibitor, or histone deactylase inhibitor.

[0038] ADCs according to the present invention may further comprise one or more small molecule binders (SMBs) of a target protein. Small molecule binders refer to non-antibody chemical groups capable of binding a target protein. The target protein is, in some instances, a target protein on a tumor cell. In some embodiments, the target protein is selected from alpha-v beta-3 (“αvß3” or “avß3”) integrin, alpha-v beta- 6 (“αvß6” or “avß6”) integrin, carbonic anhydrase IX (“CA9” or “CAIX”), fibroblast activating proteinP368919WO / 59362-725.601 (“FAP”), prostate specific membrane antigen (“PSMA”), heat shock protein 90 (“Hsp 90”), folic acid receptor, glucose transporter 1 (GLUT1), somatostatin receptor, aminopeptidase N (APN), low density lipoprotein receptor-related protein 1 (LRP1), bombesin receptor, gonadotropin releasing hormone (GnRH) receptor, luteinizing hormone-releasing hormone (LHRH) receptor, p32, membrane type 1 matrix metalloprotease (MT1-MMP), Sortilin, or Nectin-4. In some instances, a conjugate may comprise a SMB of a target protein on a tumor cell, and the antibody attached thereto is a non-binding antibody. In other instances, a conjugate may comprise a SMB of a target protein on a tumor cell, and the antibody attached thereto is configured to bind to a target antigen (e.g., a tumor antigen). In such cases, the conjugate is configured to bind to one or both of the target protein and the target antigen. Other target proteins are B- lymphocyte antigen CD20 (“CD20”), complement receptor type 2 (“CD21”), Lyb-2 (“CD72”), programmed cell death ligand 1 (“PD-L1”), carcinoembryonic antigen cell adhesion molecule (“CEACAM5”), galectin-3-binding protein (“Gal-3-BP"), leucine-rich alpha-2-glycoprotein 1 (“LRG1”), matrix metallopeptidase 9 (“MMP9”), tumour-associated glycoprotein 72 (“TAG72”), fibronectin 1 (“FN1”), tenascin-C (“TN-C”), collagen type XI alpha 1 chain (“COL11A1”), Collagen type XII alpha 1 chain (“COL12A1”), collagen type I alpha 1 chain (“COL1A1”), collagen type I alpha 2 chain (“COL1A2”), collagen type III alpha 1 chain (“COL3A1”), collagen type V alpha 1 chain (“COL5A1”), collagen type V alpha chain 2 (“COL5A2”), collagen type VI alpha 3 chain (“COL6A3”), collagen type VIII alpha 1 chain (“COL8A1”), inhibin subunit beta A (“INHBA”), periostin (“POSTN”), thrombospondin 2 (“THBS2”), zinc-alpha-2-glxcoprotein (“AZGP1”), AE binding protein 1 (“AEBP1”), anterior gradient 3 (“AGR3”), asporin (“ASPN”), BMP / retinoic acid inducible neural specific 3 (“BRINP3”), chitinase 3 like 1 (“CHI3L1”), cartilage intermediate layer protein (“CILP”), collagen type X alpha 1 chain (“COL10A1”), cartilage oligomeric matrix protein (“COMP”), cystatin SN (“CST1”), collagen triple helix repeat containing 1 (“CTHRC1”), cellular fibronectins containing the extracellular domain A (“ED-A”), cellular fibronectins containing the extracellular domain B (“ED-B”), epiphycan (“EPYC”), follicular dendritic cell secreted protein (“FDCSP”), gastrin releasing peptide (“GRP”), integrin binding sialoprotein (“IBSP”), interleukin 4 induced 1 (“IL4I1”), lumican (“LUM”), matrilin 3 (“MATN3”), midkine (“MDK”), microfibril associated protein 2 (“MFAP2”), matrix Gla protein (“MGP”), matrix metallopeptidase 1 (“MMP1”), matrix metallopeptidase 11 (“MMP11”), matrix metallopeptidase 12 (“MMP12”), matrix metallopeptidase 13 (”MMP13”), matrix metallopeptidase 3 (“MMP3”), matrix metallopeptidase 7 (“MMP7”), mucin like 1 (“MUCL1”), matrix remodeling associated 5 (“MXRA5”), signal peptide CUB domain and EGF like domain containing 2 (“SCUBE2”), secreted frizzled related protein 4 (“SFRP4”), stanniocalcin 2 (“STC2”) and zymogen granule protein 16B (“ZG16B”). Further examples of target proteins compatible with the present invention include, but are not limited to, those disclosed in F. Fauteux, Oncotarget, 2015, 7, 3, 2555 and in N. Ashman, Chem Soc Rev, 2022, Advance Article. In still other instances, the conjugate or ADC does not comprise a SMB, and the antibody attached thereto is configured to bind to a target antigen on a tumor cell or in a tumor microenvironment. Antibodies for use in accordance with the present invention do not need to be efficiently internalized by a cell in order to produce the desired effect In fact non-internalized or poorly-internalizedantibodies (e.g., PD-L1; Tenascin-C, CEACAM5, ED-A, ED-B, CAIX, αvβ6 integrin) are fully compatible with the conjugates disclosed herein.

[0039] In an aspect, provided herein is an antibody-drug conjugate according to Formula I:or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; P1 is a protease-cleavable linker; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1, 2, 3, 4, or 5.

[0040] In some embodiments, provided herein is an antibody-drug conjugate according to Formula I- A, Formula I-B, or Formula I-C:or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; P1 is a protease-cleavable linker; m is an integer from 1 to 20; n’ is an integer from 1 to 20; and p is 1, 2, 3, 4, or 5.

[0041] In some embodiments, provided herein is an antibody-drug conjugate of Formula I-A:P368919WO / 59362-725.601 Formula I-A or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; D1 is a therape'utic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; P1 is a protease-cleavable linker; m is an integer from 1 to 20; n’ is an integer from 1 to 20; and p is 1, 2, 3, 4, or 5.

[0042] In some embodiments, provided herein is an antibody-drug conjugate of Formula I-A’:Formula I-A’ or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B2 is a small molecule target protein binder; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; P1 is a protease-cleavable linker; m is an integer from 1 to 20; n’ is an integer from 1 to 20; and p is 1, 2, 3, 4, or 5.

[0043] In some embodiments, p is 1, 2, 3, 4, or 5. In some embodiments, p is 1 or 2. In some embodiments, p is 1. In some embodiments, p is 2.

[0044] In some embodiments, provided herein is an antibody-drug conjugate of Formula I-A’’:Formula I-A’’ or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B2 is a small molecule target protein binder; is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; P1 is a protease-cleavable linker; is an integer from 1 to 20; and is an integer from 1 to 20.P368919WO / 59362-725.601

[0045] In some embodiments, n’ is an integer from 1 to 10. In some embodiments, n’ is an integer from 1 to 8. In some embodiments, n’ is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, n’ is an integer from 1 to 6. In some embodiments, n’ is an integer from 2 to 6. In some embodiments, n’ is 2, 3, 4, 5, or 6. 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.

[0046] In some embodiments, provided herein is an antibody-drug conjugate of Formula I-B:Formula I-B or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 is a small molecule target protein binder; D1 is a therapeutic payload; L1 is a non-cleavable linker; P1 is a protease-cleavable linker; m is an integer from 1 to 20; p is 1, 2, 3, 4, or 5.

[0047] In some embodiments, provided herein is an antibody-drug conjugate of Formula I-B’:Formula I-B’ or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 is a small molecule target protein binder; D1 is a therapeutic payload; L1 is a non-cleavable linker; P1 is a protease-cleavable linker; and m is an integer from 1 to 20.

[0048] In some embodiments, provided herein is an antibody-drug conjugate of Formula I-C:Formula I-C or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; L1 is a non-cleavable linker; P1 is a protease-cleavable linker; andP368919WO / 59362-725.601 D1 is a therapeutic payload; P is 1, 2, 3, 4, or 5; and m is an integer from 1 to 20.

[0049] In some embodiments, p is 1 or 2. In some embodiments, p is 2. In some embodiments, provided herein is an antibody-drug conjugate of Formula I-C’:Formula I-C’ or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; L1 is a non-cleavable linker; P1 is in each instance a protease-cleavable linker; D1 is in each instance a therapeutic payload; m is an integer from 1 to 20.

[0050] In some embodiments, p is 1. In some embodiments, provided herein is an antibody-drug conjugate of Formula I-C’’:Formula I-C’’ or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; L1 is a non-cleavable linker; P1 is in each instance a protease-cleavable linker; D1 is in each instance a therapeutic payload; m is an integer from 1 to 20.

[0051] In some embodiments, P1 is a neutrophil elastase-cleavable linker. In some embodiments, P1 comprises a neutrophil-elastase cleavable peptide selected from L-Asp-L-Pro-L-Val, L-Asn-L-Pro-L-Val, -Gly-L-Pro-L-Val-, L-Ala-L-Pro-L-Val-, L-Nva-L-Pro-L-Val-, L-His-L-Pro-L-Val-, L-Asp-L-Pro-L-Ala, L-Asn-L-Pro-L-Ala, L-Asp-L-Pro-L-Ile, L-Asn-L-Pro-L-Ile, -Gly-L-Pro-L-Ile-, L-Ala-L-Pro-L-Ile-, L- Nva-L-Pro-L-Ile-, L-His-L-Pro-L-Ile-, L-Asp-L-Pro-L-Leu, L-Asn-L-Pro-L-Leu, -Gly-L-Pro-L-Leu-, L- Ala-L-Pro-L-Leu-, L-Nva-L-Pro-L-Leu-, and L-His-L-Pro-L-Leu-. In some embodiments, P1 comprises the sequence Asn-Pro-Val-§, Asn-Pro-Ala-§, Asp-Pro-Val-§, Asp-Pro-Ala-§, Gly-Pro-Val-§, Gly-Pro- Ala-§, Gly-Pro-Leu-§, Gly-Pro-Ile-§, Asn-Pro-Leu-§, Asn-Pro-Ile-§, Asp-Pro-Leu-§ or Asp-Pro-Ile-§; wherein § represents a bond to a therapeutic payload (D1). In some embodiments, P1 comprises the sequence:P368919WO / 59362-725.601 -(L-Asn)(L-Pro)(L-Val)-§, -(L-Asn)(L-Pro)(L-Ala)-§, -(L-Asp)(L-Pro)(L-Val)-§, -(L-Asp)(L-Pro)(L-Ala)-§, -(Gly)(L-Pro)(L-Val)-§, or -(Gly)(L-Pro)(L-Ala)-§, wherein § represents a bond to a therapeutic payload (D1).

[0052] In some embodiments, P1 is -(L-Asn)(L-Pro)(L-Val)-§ or -(L-Asp)(L-Pro)(L-Val)-§. In some embodiments, P1 is -(L-Asp)(L-Pro)(L-Val)-§. In some embodiments, P1 is -(L-Asn)(L-Pro)(L-Val)-§.

[0053] In some embodiments, D1 of Formula I (e.g., Formula I-A, I-A’, I-A’’, I-B, I-B’, I-C, I-C’, or I- C’’) is a cytotoxic drug or immunostimulatory agent. In some embodiments, D1 of Formula I (e.g., Formula I-A, I-A’, I-A’’, I-B, I-B’, I-C, I-C’, or I-C’’) is a cytotoxic compound. In some embodiments, D1 of Formula I (e.g., Formula I-A, I-A’, I-A’’, I-B, I-B’, I-C, I-C’, or I-C’’) is an immunostimulatory agent. In some embodiments, D1 is a cell-permeable topoisomerase inhibitor, a cell-permeable kinesin spindle protein inhibitor, a cell-permeable cyclin dependent kinase 9 inhibitor, a cell-permeable tubulin inhibitor, or a cell-permeable agonist of toll like receptor 7 and / or 8. In some embodiments, D1 is a topoisomerase inhibitor provided herein. In some embodiments, D1 is a CDK9 (e.g., PTEFb) inhibitor provided herein. In some embodiments, D1 is an auristatin. In some embodiments, D1 is a kinesin spindle protein inhibitor. In some embodiments, D1 is a toll-like receptor (e.g., TLR 7 and / or TLR8) agonist. In some embodiments, D1 is biologically active (i.e., cytotoxic or immunostimulatory) following activation by a protease such as neutrophil elastase. In some embodiments, D1 is not biologically active until activation via cleavage by neutrophil elastase.

[0054] In some embodiments, provided herein is an antibody-drug conjugate according to Formula II:Formula II or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, -CH2COOH, or -CH2COOR3; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R3 is C1-12alkyl substituted with 0-6 R4; R4 is hydrogen, halogen, -C(O)OR5, -C(O)N(R5) , - 5 5 + 5 5 2 N(R )2, -N(R )3 , -OR , -SR , -S(O)R5, -S(O) R5, -S(O) N(R5), -S(O) N(R5)C(O) R5, - 5 5 2 2 2 S(O)2N(R )C(O)OR , -N(R5)S(O) N(R5) , -N(R5)S(O) N(R5)C(O)R5, or -N(R5)S( 5 5 2 2 2 O)2N(R )C(O)OR ; R5 is in each instance, independently, hydrogen or C1-6 alkyl;P368919WO / 59362-725.601 m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1, 2, 3, 4, or 5.

[0055] In some embodiments, the target protein of B1 is a protein that is over-expressed by a tumor cell or in a tumor microenvironment. In some embodiments, the target protein of B1 is a protein that is expressed on the surface of a tumor cell. In some embodiments, the target protein of binder B (e.g., B1 or B2) is a protein selected from alpha-v beta-3 (“αvß3” or “avß3”) integrin, alpha-v beta-6 (“αvß6” or “avß6”) integrin, carbonic anhydrase IX (“CA9” or “CA IX”), fibroblast activating protein (“FAP”), prostate specific membrane antigen (“PSMA”), heat shock protein 90 (“Hsp 90”), folic acid receptor, glucose transporter 1, somatostatin receptor, aminopeptidase N (APN), low density lipoprotein receptor- related protein 1 (LRP1), bombesin receptor, gonadotropin releasing hormone (GnRH or LHRH) receptor, p32, membrane type 1 matrix metalloprotease (MT1-MMP), Sortilin, or Nectin-4. In some embodiments, B1 is absent. In some embodiments, B2 is absent. In some embodiments, both B1 and B2 are absent. In some embodiments, either B1 or B2 is absent. In some embodiments, B1 and B2 are both present.

[0056] In some embodiments, the antibody-drug conjugate of Formula II has the structure of Formula (II-1), (II-2), (II-3), (II-4), (II-5), or (II-6):P368919WO / 59362-725.601or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof comprising cysteine and lysine; B1 and B2 are in each instance, independently, a small molecule target protein binder; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; wherein: L1' denotes L1, bonded via a sulfur atom of a cysteine side-chain of AK; and L1'' denotes L1, bonded via a nitrogen atom of a lysine side-chain of AK; R1 is hydrogen, -CHCONH, -CHCOOH, or -CHCOOR3 2 2 2 2 ; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R3 is C alkyl substituted w 4 1-12 ith 0-6 R; R4 is hydrogen, halogen, -C(O)OR5, -C(O)N(R5), -N(R5), -N(R5)+, -OR5, -SR5, 5 2 2 3 -S(O)R, -S(O)R5, -S(O)N(R5), -S(O)N(R5)C(O) R5, -S 5 5 5 5 2 2 2 (O)2N(R)C(O)OR, -N(R)S(O)2N(R)2, -N(R5)S(O)N(R5) 5 5 5 5 2 C(O)R, or -N(R)S(O)2N(R)C(O)OR; R5 is in each instance, independently, hydrogen or C1-6 alkyl; m is an integer from 1 to 20;m' is an integer from 1 to 10; m'' is an integer from 1 to 10; and n is an integer from 1 to 20.

[0057] In some embodiments, n is an integer from 1 to 20. In some embodiments, n is 0.

[0058] In some embodiments, provided herein is an antibody-drug conjugate of Formula II-A:or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; D1 is a therapeutic payload; L1 is a non-cleavable linker; B1 is a small molecule target protein binder; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); m is an integer from 1 to 20; and p is 1, 2, 3, 4, or 5.

[0059] In some embodiments, R1 is -CH 1 2CONH2, or -CH2COOH. In some embodiments, R is - CH CONH . In some embodiment 1 1 2 2 s, R is -CH2COOH. In some embodiments, R is hydrogen. In some embodiments, R2 is -CH 2 3 or -CH(CH3)2. In some embodiments, R is -CH(CH3)2.

[0060] In some embodiments, provided herein is an antibody-drug conjugate of Formula II-A’:or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; D1 is a therapeutic payload; L1 is a non-cleavable linker; B1 is a small molecule target protein binder; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); m is an integer from 1 to 20; andP368919WO / 59362-725.601 p is 1, 2, 3, 4, or 5.

[0061] In some embodiments, provided herein is an antibody-drug conjugate of Formula II-A’’:Formula II-A’’ or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; D1 is a therapeutic payload; L1 is a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); m is an integer from 1 to 20; and p is 1, 2, 3, 4, or 5.

[0062] In some embodiments, p is 1 or 2. In some embodiments, p is 2. In some embodiments, p is 1.

[0063] In some embodiments, provided herein is an antibody-drug conjugate of Formula II-A’’’:Formula II-A’’’ or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; D1 is a therapeutic payload; L1 is a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); and m is an integer from 1 to 20.

[0064] In some embodiments, provided herein is an antibody drug conjugate of Formula (II-1):(II-1) or a pharmaceutically acceptable salt thereof: AK is an antibody or antigen binding fragment thereof;D1 is a therapeutic payload; L2 is a non-cleavable linker; B2 is a small molecule target protein binder; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); m is an integer from 1 to 20; and n is an integer from 1 to 20.

[0065] In some embodiments, provided herein is an antibody drug conjugate of Formula (II-2):or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; D1 is a therapeutic payload; L2 is a non-cleavable linker; B2 is a small molecule target protein binder; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); m is an integer from 1 to 20; and n is an integer from 1 to 20.

[0066] In some embodiments, provided herein is an antibody-drug conjugate of Formula II-3:or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; L1 is a non-cleavable linker; B1 is a small molecule target protein binder;P368919WO / 59362-725.601 D1 is a therapeutic payload; R1 is hydrogen, -CH CONH 3 2 2, -CH2COOH, or -CH2COOR ; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); m is an integer from 1 to 20; and p is 1, 2, 3, 4, or 5.

[0067] In some embodiments, provided herein is an antibody-drug conjugate of Formula II-4:Formula II-4 or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; D1 is a therapeutic payload; L1 is a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); and m is an integer from 1 to 20.

[0068] In some embodmients, provided herein is an antibody-drug conjugate of Formula (II-5):or a pharmaceutically acceptable salt thereof.

[0069] In some embodmients, provided herein is an antibody-drug conjugate of Formula (II-6):P368919WO / 59362-725.601 (II-6) or a pharmaceutically acceptable salt thereof; wherein: AK is an antibody or antigen binding fragment thereof comprising cysteine and lysine; L1' is a non-cleavable linker, bonded via a sulfur of the cysteine of AK; L1'' is a non-cleavable linker, bonded via a nitrogen of the lysine of AK; m' is an integer from 1 to 10; and m'' is an integer from 1 to 10.

[0070] In some embodiments, provided herein is an antibody-drug conjugate of Formula II’:Formula II’ or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B2 is a small molecule target protein binder; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, -CH2COOH, or -CH2COOR3; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R3 is C1-12alkyl substituted with 0-6 R4; R4 is hydrogen, halogen, -C(O)OR5, -C(O)N(R5) 5 5 + 5 5 2, -N(R)2, -N(R)3, -OR, -SR, -S(O)R5, -S(O)R5, -S(O)N(R5), -S(O)N(R5)C 5 5 5 2 2 2 (O) R, -S(O)2N(R)C(O)OR, -N(R5)S(O) 5 5 5 5 5 5 5 2N(R)2, -N(R)S(O)2N(R)C(O)R, or -N(R)S(O)2N(R)C(O)OR; R5 is in each instance, independently, hydrogen or C1-6alkyl; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1, 2, 3, 4, or 5.

[0071] In some embodiments, R1 is hydrogen, -CH2CONH2, or -CH2COOH. In some embodiments, B2 is a small molecule target protein binder having the structure:or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0072] In some embodiments, provided herein is an antibody-drug conjugate of Formula II-B:Formula II-B or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH CONH , -CH C 3 2 2 2 OOH, or CH2COOR ; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R4 is hydrogen, halogen, -C(O)OR5, -C(O)N(R5) 5 5 + 5 5 2, -N(R )2, -N(R )3 , -OR , -SR , -S(O)R5, -S(O)2R5, -S(O)2N(R5), -S(O)2N(R5)C(O) R5, -S(O)2N(R5)C(O)OR5, -N(R5)S(O)2N(R5)2, -N(R5)S(O)2N(R5)C(O)R5, or -N(R5)S(O)2N(R5)C(O)OR5; R5 is in each instance, independently, hydrogen or C1-6 alkyl; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1, 2, 3, 4, or 5.

[0073] In some embodiments, provided herein is an antibody-drug conjugate of Formula II-B’:P368919WO / 59362-725.601Formula II-B’ or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CHCONH, -CHCOOH, or C 3 2 2 2 H2COOR; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R4 is hydrogen, halogen, -C(O)OR5, -C(O)N(R5), -N(R5), -N(R5)+, -OR5, - 5 2 2 3 SR, -S(O)R5, -S(O)R5, -S(O)N(R5), -S(O)N(R5)C(O) R5, -S(O)N(R5)C 5 2 2 2 2 (O)OR, -N(R5)S(O)N(R5), -N(R5)S(O)N(R5)C(O)R5, or -N(R5)S(O) 5 5 2 2 2 2N(R)C(O)OR; R5 is in each instance, independently, hydrogen or C1-6 alkyl; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1, 2, 3, 4, or 5.

[0074] In some embodiments, p of Formulua II-B or II-B’ is 1 or 2. In some embodiments, p of Formulua II-B or II-B’ is 2. In some embodiments, p of Formulua II-B or II-B’ is 1.

[0075] In some embodiments, provided herein is an antibody-drug conjugate of Formula II-C:Formula II-C or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B2 is a small molecule target protein binder;P368919WO / 59362-725.601 D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CHCON 3 2 H2, -CH2COOH, or -CH2COOR; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R3 is C alky 4 1-12 l substituted with 0-6 R; R4 is hydrogen, halogen, -C(O)OR5, -C(O)N(R5) 5 5 + 5 5 2, -N(R)2, -N(R)3, -OR, -SR, -S(O)R5, -S(O)R5, -S(O)N(R5), -S( 5 5 5 5 2 2 O)2N(R)C(O) R, -S(O)2N(R)C(O)OR, -N(R5)S(O)N(R5), -N(R5)S(O)N(R5)C(O)R5, or -N(R5)S(O)N 5 5 2 2 2 2 (R)C(O)OR; R5 is in each instance, independently, hydrogen or C1-6 alkyl; m is an integer from 1 to 20; and n is an integer from 0 to 20.

[0076] In some embodiments, n is 0. In some embodiments, provided herein is an antibody-drug conjugate of Formula II-C’:Formula II-C’ or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; D1 is a therapeutic payload; L1 is a non-cleavable linker; R1 is hydrogen, -CH 3 2CONH2, -CH2COOH, or -CH2COOR; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R3 is C alkyl substitu 4 1-12 ted with 0-6 R; R4 is hydrogen, halogen, -C(O)OR5, -C(O)N(R5), -N 5 5 + 5 5 2 (R)2, -N(R)3, -OR, -SR, -S(O)R5, -S(O)R5, - 5 5 5 5 5 2 S(O)2N(R), -S(O)2N(R)C(O) R, -S(O)2N(R)C(O)OR, -N(R5)S(O)N(R5), -N(R5)S(O)N(R5)C(O)R5, 5 5 5 2 2 2 or -N(R)S(O)2N(R)C(O)OR; R5 is in each instance, independently, hydrogen or C1-6 alkyl; m is an integer from 1 to 20.

[0077] In some embodiments, D1 is a membrane-permeable therapeutic payload. In some embodiments, D1 is a cytotoxic agent or an immunostimulatory agent. In some embodiments, D1 is atopoisomerase inhibitor, a kinesin spindle protein inhibitor, a tubulin polymerization inhibitor, a cyclin dependent kinase 9 inhibitor, or an agonist of toll like receptor 7 and / or 8.

[0078] In some embodiments, each D1 is independently:or a pharmaceutically acceptable salt thereof, wherein: each R6 and R7 is independently hydrogen, halogen, CN, -C1-6 alkyl, or C1-6 haloalkyl; R8 is hydrogen, halogen, CN, -C1-6 alkyl, C1-6 haloalkyl, C3-6 cycloalkyl, or 5- to 7-membered heterocycloalkyl; R9 is hydrogen, halogen, CN, C1-6 alkyl, -C(O)NH2, -C(O)NHC1-6 alkyl, -C(O)N(C1-6 alkyl)2, -C(O)NHC1-6 alkyl-C(O)NHC1-6 alkyl, -C(O)NHC1-6 alkyl-NHC(O)C1-6 alkyl, - NH2, -NHC1-6 alkyl, -N(C1-6 alkyl)2, -NHC(O)C1-6 alkyl, -OH, or -OC1-6 alkyl; wherein each C alkyl is substitut 10 1-6 ed with 0-5 R ; R10 is in each instance independently selected from the group consisting of hydrogen, halogen, CN, -COOH, -CONH2, -NH2, -NHCH3, -N(CH3)2, -OH, and -OCH3; R11 and R12 are each independently hydrogen, halogen, C1-6alkyl, C1-6haloalkyl, or -OH; or R11 and R12, taken together, form a 5- or 6-membered heterocycle; R13 and R14 are each independently hydrogen, C1-6alkyl, or C1-6alkylamine; or R13 and R14, taken together, form a C carbocycle subst 15 6 ituted with -N(R )2; each R15 is independently hydrogen, C1-6alkyl, -C(O)C1-6alkyl, -C(O)NHC1-6alkyl or -C(O)OC1-6alkyl ; wherein the C1-6alkyl of R15 is optionally substituted with halogen, hydroxy, phenyl, or heteroaryl; or R15 is a cleavable prodrug group; R16 is hydrogen, halogen, C1-6alkyl, C1-6haloalkyl, -OH, -OCH3, or -OCF3; R17 is hydrogen, halogen, C1-6alkyl, C1-6haloalkyl, -OH, -OCH3, or -OCF3; or R16 and R17, taken together, form a heteroalkylene group of the formula:-O-C2-10 alkylene-O-, -NH-C2-10 alkylene-O-, or -NH-C2-10 alkylene-NH-; wherein the heteroalkylene group is optionally substituted with R21; R18 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, -OH, -OCH3, or -OCF3; or R16 and R18, taken together, form a heteroalkylene group of the formula: -O-C2-10 alkylene-O-, -NH-C2-10 alkylene-O-, or -NH-C2-10 alkylene-NH-; wherein the heteroalkylene group is optionally substituted with R21; R19 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, -OH, -OCH3, or -OCF3; R20 is hydrogen or methyl; R21 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, or OH; R22 is hydrogen or C1-6alkyl, wherein the C1-6alkyl is unsubstituted or substituted with R24; R23 is hydrogen, C1-6alkyl, or benzyl, wherein the C1-6alkyl or benzyl is unsubstituted or substituted with one, two, or three R25 groups; R24 is -OH, -O(C1-6alkyl), -NH2, -NH(C1-6alkyl), -N(C1-6alkyl)2, -SH, or -S(C1-6alkyl); R25 is -OH, -O(C1-6alkyl), -NH2, -NH(C1-6alkyl), -N(C1-6alkyl)2, -NHS(O)2(C1-6alkyl), C1-6alkyl, C1-6aminoalkyl, or OCH2CH2NHC(O)(C1-6aminoalkyl); each Y1, Y2, Y3, and Y4 is independently -CH, -CF, or N; Y5 is CH2, NH, or O; q is 0, 1, 2, 3, 4, or 5; and r is 0, 1, 2, 3, 4, or 5.

[0079] In some embodiments, D1 is:or a pharmaceutically acceptable salt thereof, wherein: each R6 and R7 is independently hydrogen, halogen, CN, -C1-6 alkyl, or C1-6 haloalkyl; R8 is hydrogen, halogen, CN, -C1-6 alkyl, C1-6 haloalkyl, C3-6 cycloalkyl, or 5- to 7-membered heterocycloalkyl; R9 is hydrogen, halogen, CN, C1-6 alkyl, -C(O)NH2, -C(O)NHC1-6 alkyl, -C(O)N(C1-6 alkyl)2, -C(O)NHC1-6 alkyl-C(O)NHC1-6 alkyl, -C(O)NHC1-6 alkyl-NHC(O)C1-6 alkyl, - NH2, -NHC1-6 alkyl, -N(C1-6 alkyl)2, -NHC(O)C1-6 alkyl, -OH, or -OC1-6 alkyl; wherein each C alkyl i 10 1-6 s substituted with 0-5 R ;P368919WO / 59362-725.601 R10 is in each instance independently selected from the group consisting of hydrogen, halogen, CN, -COOH, -CONH2, -NH2, -NHCH3, -N(CH3)2, -OH, and -OCH3; q is 0, 1, 2, 3, 4, or 5; and r is 0, 1, 2, 3, 4, or 5.

[0080] In some embodiments, D1 is:or a pharmaceutically acceptable salt thereof, wherein: R11 and R12 are each independently hydrogen, halogen, C1-6alkyl, C1-6haloalkyl, or -OH; or R11 and R12, taken together, form a 5- or 6-membered heterocycle; R13 and R14 are each independently hydrogen, C1-6alkyl, or C1-6alkylamine; or R13 and R14, taken together, form a C6carbocycle substituted with -N(R15)2; each R15 is independently hydrogen, C1-6alkyl, -C(O)C1-6alkyl, -C(O)NHC1-6alkyl or -C(O)OC1-6alkyl ; wherein the C1-6alkyl of R15 is optionally substituted with halogen, hydroxy, phenyl, or heteroaryl; or R15 is a cleavable prodrug group (e.g, a benzyl ester).

[0081] In some embodiments, D1 is:or a pharmaceutically acceptable salt thereof, wherein: R16 is hydrogen, halogen, C1-6alkyl, C1-6haloalkyl, -OH, -OCH3, or -OCF3; R17 is hydrogen, halogen, C1-6alkyl, C1-6haloalkyl, -OH, -OCH3, or -OCF3; or R16 and R17, taken together, form a heteroalkylene group of the formula: -O-C2-10alkylene-O-, -NH-C2-10alkylene-O-, or -NH-C2-10alkylene-NH-; wherein the heteroalkylene group is optionally substituted with R21; R18 is hydrogen, halogen, C1-6alkyl, C1-6haloalkyl, -OH, -OCH3, or -OCF3; or R16 and R18, taken together, form a heteroalkylene group of the formula: -O-C2-10alkylene-O-, -NH-C2-10alkylene-O-, or -NH-C2-10alkylene-NH-; wherein the heteroalkylene group is optionally substituted with R21;P368919WO / 59362-725.601 R19 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, -OH, -OCH3, or -OCF3; R21 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, or OH; each Y1, Y2, Y3, and Y4 is independently -CH, -CF, or N; and Y5 is CH2, NH, or O.

[0082] In some embodiments, D1 is:or a pharmaceutically acceptable salt thereof, wherein R20is hydrogen or methyl.

[0083] In some embodiments, D1 is:or a pharmaceutically acceptable salt thereof, wherein: R22 is hydrogen or C alkyl, wherei 24 1-6 n the C1-6 alkyl is unsubstituted or substituted with R ; R23 is hydrogen, C1-6 alkyl, or benzyl, wherein the C1-6 alkyl or benzyl is unsubstituted or substituted with one, two, or three R25 groups; R24 is -OH, -O(C1-6 alkyl), -NH2, -NH(C1-6 alkyl), -N(C1-6 alkyl)2, -SH, or -S(C1-6 alkyl); R25 is -OH, -O(C1-6 alkyl), -NH2, -NH(C1-6 alkyl), -N(C1-6 alkyl)2, -NHS(O)2(C1-6 alkyl), C1-6 alkyl, C1-6 aminoalkyl, or OCH2CH2NHC(O)(C1-6 aminoalkyl).

[0084] In some embodiments, D1 is:,

[0085] In some embodiments, provided herein is an antibody-drug conjugate having the structure of Formula (II-D) or Formula (II-E):P368919WO / 59362-725.601 Formula II-DFormula II-E or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; D1 is a therapeutic payload; L2 is a non-cleavable linker; L3 is a protein-coupled group; L4 is a bond or a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); Ra is hydrogen or -CH3; Rb is in each instance, independently, hydrogen or CH3; or two Rb, together with the carbon to which they are attached, form a carbonyl; Rc is hydrogen or -CH3; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1 or 2.

[0086] In some embodiments, provided herein is an antibody-drug conjugate having the structure of Formula III:Formula (III) or a pharmaceutically acceptable salt thereof, wherein:AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); each R6 and R7 is independently hydrogen, halogen, CN, -C1-6 alkyl, or C1-6 haloalkyl; R8 is hydrogen, halogen, CN, -C1-6 alkyl, C1-6 haloalkyl, C3-6 cycloalkyl, or 5- to 7-membered heterocycloalkyl; R9 is hydrogen, halogen, CN, C1-6 alkyl, -C(O)NH2, -C(O)NHC1-6 alkyl, -C(O)N(C1-6 alkyl)2, -C(O)NH-C1-6alkyl-C(O)NH-C1-6alkyl, -C(O)NHC1-6alkyl-NHC(O)-C1-6alkyl, -NH2, - NHC1-6alkyl, -N(C1-6alkyl)2, -NHC(O)C1-6alkyl, -OH, or -OC1-6alkyl; wherein each C1-6alkyl is substituted with 0-5 R10; R10 is in each instance independently selected from the group consisting of hydrogen, halogen, CN, -COOH, -CONH2, -NH2, -NHCH3, -N(CH3)2, -OH, and -OCH3; m is an integer from 1 to 20; n is an integer from 0 to 20; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and r is 0, 1, 2, 3, 4, or 5.

[0087] In some embodiments, R6 and R7 are hydrogen or halogen; q is 0, 1, or 2; and r is 0, 1, or 2. In some embodiments, R6 is hydrogen. In some embodiments, q is 0. In some embodiments, R7 is halogen and r is 2. In some embodiments, R7 is fluoro, q is 0, and r is 2.. In some embodiments, R8 is -C1-6 alkyl. In some embodiments, R8 is t-butyl. In some embodiments, provided herein is an antibody-drug conjugate having the structure of Formula III-A:or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof;B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R9 is hydrogen, C1-6 alkyl, -(CH2)0-2C(O)NH2, -(CH2)0-2C(O)NHC1-6 alkyl, or -(CH2)0- 2C(O)N(C1-6 alkyl)2; wherein each C1-6 alkyl is optionally substituted with 0-5 halogen; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1 or 2.

[0088] In some embodiments, R9 is H, -CH3, -CH2F, -CHF2, -CF3, -C(O)NH2, -C(O)NHCH3, or - C(O)N(CH3)2. In some embodiments, R9 is H, -CHF2, or -C(O)NHCH3. In some embodiments, R9 is hydrogen. In some embodiments, R9 is methyl, ethyl, isopropyl, or t-butyl. In some embodiments, R9 is - CH2F, CHF2, -CF3, -CH2CH2F, -CH2CHF2, or -CH2CF3. In some embodiments, R9 is -(CH2)0-2C(O)NH2, -(CH2)0-2C(O)NHCH3, or -(CH2)0-2C(O)N(CH3)2. In some embodiments, R9 is -CH2C(O)NH2, - CH2C(O)NHCH3, or -CH2C(O)N(CH3)2. In some embodiments, R9 is -CH2C(O)NHCH3. In some embodiments, R9 is -CH2C(O)N(CH3)2. In some embodiments, R9 is -CH2C(O)NHCH2CF3or - CH2C(O)NHCHF2. In some embodiments, R9 is -C(O)NH2, -C(O)NHCH3, or -C(O)N(CH3)2. In some embodiments, R9 is -C(O)NHCH3. In some embodiments, R9 is -C(O)N(CH3)2. In some embodiments, R1 is -CH2CONH2, or -CH2COOH. In some embodiments, R1 is -CH2CONH2. In some embodiments, R1 is - CH2COOH. In some embodiments, R2 is -CH3 or -CH(CH3)2. In some embodiments, R2 is -CH(CH3)2. In some embodiments, R2 is -CH3.

[0089] In some embodiments, provided herein is an antibody-drug conjugate having the structure of Formula IV:or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance independently a small molecule target protein binder;L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R11 and R12 are each independently hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, or -OH; or R11 and R12, taken together, form a 5- or 6-membered heterocycle; R13 and R14 are each independently hydrogen, C1-6 alkyl, or C1-6 alkylamine; or R13 and R14, taken together, form a C carbocycle substitute 15 6 d with -N(R )2; each R15 is independently hydrogen, C1-6 alkyl, -C(O)C1-6 alkyl, -C(O)NHC1-6 alkyl, or -C(O)OC al 15 1-6 kyl; wherein the C1-6 alkyl of R is optionally substituted with halogen, hydroxy, phenyl, or heteroaryl; or R15 is a cleavable prodrug group; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1, 2, 3, 4, or 5.

[0090] In some embodiments, provided herein is an antibody-drug conjugate having the structure of Formula IV-A, IV-B, IV-C, or IV-D:P368919WO / 59362-725.601 (IV-B)Formula IV-D or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); each R15 is independently hydrogen, C1-6alkyl, or -C(O)C1-6alkyl; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1, 2, 3, 4, or 5.

[0091] In some embodiments, R11 is hydrogen, halogen, C1-6alkyl, or -OH. In some embodiments, R12 is hydrogen, halogen, C1-6alkyl, or -OH. In some embodiments, R11 is halogen and R12 is C1-6alkyl. In some embodiments R11 is fluoro and R12 is CH3In some embodiments R11 is fluoro R12 is CH3and R13P368919WO / 59362-725.601 and R14, taken together, form a C6 carbocycle substituted with -N(R15)2; wherein each R15 is independently hydrogen, C1-6 alkyl, -C(O)C1-6 alkyl, -C(O)NHC1-6 alkyl, or -C(O)OC1-6 alkyl; wherein the C1-6 alkyl of R15 is optionally substituted with halogen. In some embodiments, R11 and R12, taken together, form a 5- or 6-membered cyclic ether. In some embodiments, R11 and R12, taken together, form a 5- or 6-membered cyclic ether; R13 is hydrogen; and R14 is hydrogen or C 11 12 1-6 alkyl. In some embodiments, R and R , taken together, form a 5- or 6-membered cyclic ether; and R13and R14are hydrogen. In some embodiments, R11, R12, and R13are each hydrogen; and R14 is hydrogen or C 11 12 13 1-6 alkyl. In some embodiments, R , R , and R are each hydrogen; and R14is ethyl.

[0092] In some embodiments, provided herein is an antibody-drug conjugate having the structure of Formula V:Formula V or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R16 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, -OH, -OCH3, or -OCF3; R17 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, -OH, -OCH3, or -OCF3; or R16 and R17, taken together, form a heteroalkylene group of the formula: -O-C2-10 alkylene-O-, -NH-C2-10 alkylene-O-, or -NH-C2-10 alkylene-NH-; R18 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, -OH, -OCH3, or -OCF3; or R16 and R18, taken together, form a heteroalkylene group of the formula: -O-C2-10 alkylene-O-, -NH-C2-10 alkylene-O-, or -NH-C2-10 alkylene-NH-; R19 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, -OH, -OCH3, or -OCF3; each Y1, Y2, Y3, and Y4 is independently -CH, -CF, or N; Y5 is CH2, NH, or O; m is an integer from 1 to 20; n is an integer from 0 to 20; andP368919WO / 59362-725.601 p is 12, 3, 4, or 5.

[0093] In some embodiments, R16 and R18 are each hydrogen; and R17 is -OCH3. In some embodiments, provided herein is an antibody-drug conjugate having the structure of Formula V-A or Formula V-B:Formula V-B or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R16 and R17 taken together form the group -O-C2-10 alkylene-O- or -NH-C2-10 alkylene-O-; or R16 and R18 taken together form the group -NH-C2-10 alkylene-O-; Y3 is CH or N; Y4 is CH or N; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1 or 2.

[0094] In some embodiments, provided herein is an antibody-drug conjugate having the structure of Formula VI:P368919WO / 59362-725.601Formula VI or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R20 is hydrogen or -CH3; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1 or 2.

[0095] In some embodiments, provided herein is an antibody-drug conjugate having the structure of Formula VII:Formula VII or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R22 is hydrogen or C1-6alkyl, wherein the C1-6alkyl is unsubstituted or substituted with R24; R23 is hydrogen, C1-6alkyl, or benzyl, wherein the C1-6alkyl or benzyl is unsubstituted or substituted with one, two, or three R25 groups;R24 is -OH, -O(C1-6 alkyl), -NH2, -NH(C1-6 alkyl), -N(C1-6 alkyl)2, -SH, or -S(C1-6 alkyl); R25 is -OH, -O(C1-6 alkyl), -NH2, -NH(C1-6 alkyl), -N(C1-6 alkyl)2, -NHS(O)2(C1-6 alkyl), C1-6 alkyl, C1-6 aminoalkyl, or OCH2CH2NHC(O)(C1-6 aminoalkyl); m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1 or 2.

[0096] In another aspect, provided herein is an antibody-drug conjugate of Formula I, I-A, I-B, I-C, II, II-A, II-B, II-C, II-D, II-E, II-F, II-G III, III-A, IV, IV-A, IV-B, IV-C, V, V-A, V-B, or VI, for use in the manufacture of a medicament for treating a disease. In another aspect, provided herein is a method of treating a disease or disorder in a subject in need thereof, the method comprising administering a therapetuically effective amount of an antibody-drug conjugate disclosed in any one or more of Formulae I, I-A, I-B, I-C, II, II-A, II-B, II-C, II-D, II-E, II-F, II-G III, III-A, IV, IV-A, IV-B, IV-C, V, V-A, V-B, and / or VI, or a pharmaceutically acceptable salt or solvate thereof, to the subject. In some embodiments, the disease or disorder is a hyperproliferative disease or disorder. In some embodiments, provided herein is an antibody-drug conjugate for use as a medicament. In some embodiments, the medicament is for use in the treatment of a disease (e.g., a hyperproliferative disease or disorder). In some embodiments, the hyperproliferative disease or disorder is a cancer. COMPOUNDS

[0097] In another aspect, provided herein is a precursor molecule (i.e., a compound) comprising a therapeutic payload, and further comprising a protein-reactive group (alternatively, a protein-coupling group). Precursor molecules provided herein, may be useful in the synthesis or manufacture of a medicament for treating diseases or disorders (e.g., cancers) in a subject. In some embodiments, provided herein is a compound for use in the manufacture of a medicament, according to Formula I or Formula II:

[0098] The antibody-drug conjugates of the current invention may be obtained by coupling of payload- linker precursor compounds of Formula VIII, and optionally also binder-linker precursor compounds of Formula IX to the respective antibody:or a pharmaceutically acceptable salt thereof, wherein: PR1 and PR2 are in each instance, independently, a protein-reactive group; L1a and L2aare in each instance, independently, a non-cleavable linker; B1 and B2 are in each instance, independently, a small molecule target protein binder; P1 is in each instance, independently, a protease-cleavable linker; D1 is in each instance, independently, a therapeutic payload; and p is 1, 2, 3, 4, or 5.P368919WO / 59362-725.601

[0099] In some embodiments, each PR (PR1, PR2) is independently selected from a protein reactive group enabling either cysteine conjugation or lysine conjugation. In some embodiments, PR is a substrate for enzymes capable to fuse a payload linker precursor compounds or a binder-linker precursor compound with the antibody. For example, PR1 or PR2 may be a protein-reactive group such as a lysine residue which, in the presence of a bacterial transglutaminase, may react the lysine residue with a glutamine side-chain of the antibody. In some embodiments, wherein p is 2 or more, each P1 and / or D1 is the same. In some embodiments, wherein p is 2 or more, each D1 is different. In some embodiments, a first protease-cleavable linker P1 is efficiently cleaved by a first protease, and the second protease-cleavable linker P2 is slowly cleaved by a second protease, wherein the first and second proteases may be the same or different. [000100]Antibody-drug conjugates which are composed of a payload and an additional binder molecule attached to the antibody are obtained by subsequent coupling of the two different precursors depending on the protein reactive group (PR1, PR2).The resulting ADCs comprise a payload linker and a binder- linker motif, both attached to lysine residues, or both attached to cysteine residues or one motif attached to lysine residues and the other motif attached to cysteine residues. The cysteine residues suitable for payload linker or binder linker attachment can be obtained by reduction of interchain disulfide bridges or by engineering of cysteine residues into the antibody. [000101]In some embodiments, provided herein is a payload-linker precursor compound of Formula VIII, having the structure: , ,,P368919WO / 59362-725.601or a pharmaceutically acceptable salt thereof wherein:P368919WO / 59362-725.601 Ra is hydrogen or -CH3; R1 is hydrogen, -CH2C(O)NH2, or -CH2C(O)OH; R2 is -CH3 or -CH(CH3)2; and D1 is a therapeutic payload. [000102]In some embodiments, provided herein is a binder-linker precursor compound of Formula IX, having the structure:,; or a pharmaceutically acceptable salt thereof. LINKERS [000103]Antibody-drug conjugates and compounds described herein may comprise a non-cleavable linker. According to Formula I and subformulae thereof, the linker conjoining AK to P1 is denoted as L1. In some embodiments, L1 is a bivalent non-cleavable linker. In other embodiments, L1 is a trivalent non- cleavable linker. In some embodiments, L1 conjoins AK and P1 in a linear heteroalkyl chain. In some embodiments, L1 conjoins AK and two or more P1 in a branched structure. In some embodiments, L1 is a trivalent non-cleavable linker. In some embodiments, L1 conjoins AK, P1, and a third group selected from P1, B1, and -COOH, wherein each P1 is a protease-cleavable linker and B1 is a small molecule target protein binder. In some embodiments, L1 is a C heteroalkyl linker 1 1-60 . In some embodiments, L is linear. In some embodiments, L1 is branched. [000104]As illustrated in Formula I, L2 is a non-cleavable linker conjoining AK to B2, wherein B2 is a small-molecule target protein binder. In some embodiments, L2 is a C1-30 heteroalkyl group, wherein one or more methylene units of the alkyl group is replaced with -O- -NH- or -S- and which may be furthersubstituted (i.e., with -COOH, -NH , -SH, or monocycli 2 2 c heterocycloalkyl). In some embodiments, L is a heteroalkyl polyamide, polyamine, or polyethylene glycol polymeric section, comprising 1 to 12 monomers of PEG, PEI, or PSAR. In some embodiments, the PEI linker is partially or entirely N-methylated, and comprises 1 to 12 PEI monomers.In some embodiments, L2 is represented by the structure:wherein: * denotes a bond to a sulfur atom of a cysteine side-chain of AK; and ** denotes a bond to a nitrogen atom of a lysine side-chain of AK; and which may further comprise a polymeric group of the formula (CH2CH2O)1-8CH2CH2X, wherein X is NH, N(CH3), CO, NHCO, N(CH3)CO, CONH, or CON(CH3), and X forms a bond with B2. [000105]The non-cleavable linker of L1 may, in some embodiments, be further defined as comprising one or more of L3, L4, and / or a polymeric linker, wherein L3 is a protein-coupled group, and L4 is a bond or non-cleavable linker. [000106]In some embodiments, the polymeric linker is a polyethylene glycol (PEG) linker. In some embodiments, the PEG linker comprises 1 to 12 PEG monomers. In some embodiments, the polymeric linker is a polyamine linker. In some embodiments, the polyamine linker is a polyethyleneimine glycol (PEI) linker. In some embodiments, the PEI linker is partially or entirely N-methylated, and comprises 1 to 12 PEI monomers. In some embodiments, the polymeric linker is a polyamide linker. In some embodiments, the polyamide linker is a polysarcosine (PSAR) linker. In some embodiments, the PSAR linker comprises 1 to 12 PSAR monomers. [000107]In some embodiments, L3 is a lysine-conjugated group, or a cysteine-conjugated group. In some embodiments, L3 is a heteroalkyl or alkyl adduct of an amino acid, wherein the amino acid is a part of AK. In some embodiments L3 is:; wherein: * denotes a bond to a sulfur atom of a cysteine side-chain of AK; ** denotes a bond to a nitrogen atom of a lysine side-chain of AK; and denotes a bond to L4 or to the polymeric linker. [000108]In some embodiments, L4 is a bond. In some embodiments, L4 is a divalent linker, conjoining the polymeric linker to the protein-coupled group (L3). In some embodiments, L4 is a trivalent linker conjoining the polymeric linker, and the protein-coupled group (L3), and a third group selected from P1, B1, and -COOH. In some embodiments, L4 is a bond, and the protein-coupled group is therefore conjugated directly to the polymeric linker. In other embodiments, L4 is a trivalent linker, conjoining the protein- coupled group to the polymeric linker via an amino-acid-containing heteroalkyl linker. In some embodiments, the amino acid is a lysine or a glutamic acid. In some embodiments, the amino acid linker is a trivalent amino acid, and the linker conjoins a small-molecule protein binder, a second protease- cleavable linker (or polymeric linker), or a PK-modifying group (e.g., COOH) to the protein-coupled group and first polymeric linker. In some embodiments, the amino acid is extended or adapted via C1-12 alkyl or heteroalkyl bridging group. In some embodiments, the heteroalkyl bridging group is a C1-12 alkyl group, wherein 1 or more methylene units are replaced with -O-, -NH-, or -N(CH3)-, and wherein one or more of the remaining methylene groups are optionally substituted with an oxo (=O) group (i.e., forming a carbonyl). Examples of bridging groups include, but are not limited to: -CO-(CH2)1-12-CO-, NH-(CH2)1-12- NH, -NH-(CH2)1-12-CO-, and -CONH-(CH2)1-12-CO-. [000109]In some embodiments, L4 is,wherein: *** denotes a bond to L3; and denotes a bond to the polymeric linker of L1; or to the protease-cleavable linker (P1). ANTIBODIES [000110]In accordance with the present invention, the term "antibody" is to be understood in its broadest meaning and comprises immunoglobulin molecules, for example intact or modified monoclonal antibodies, polyclonal antibodies or multispecific antibodies (e.g. bispecific antibodies). An immunoglobulin molecule preferably comprises a molecule having four polypeptide chains, two heavy chains (H chains) and two light chains (L chains) which are typically linked by disulfide bridges. Each heavy chain comprises a variable domain of the heavy chain (abbreviated VH) and a constant domain of the heavy chain. The constant domain of the heavy chain may, for example, comprise three domains CH1, CH2 and CH3. Each light chain comprises a variable domain (abbreviated VL) and a constant domain. The constant domain of the light chain comprises a domain (abbreviated CL). The VH and VL domains may be subdivided further into regions having hypervariability, also referred to as complementarity determining regions (abbreviated CDR) and regions having low sequence variability (framework region, abbreviated FR). Typically, each VH and VL region is composed of three CDRs and up to four FRs. For example from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. An antibody may be obtained from any suitable species, e.g. rabbit, llama, camel, mouse or rat. In one embodiment, the antibody is of human or murine origin. An antibody may, for example, be human, humanized or chimeric. [000111]In some embodiments, the antibody or antigen-binding fragment thereof (“AK”) is conjugated to, or is configured to be conjugated to, a compound or conjugate as described herein. In some embodiments, AK is a target antigen binder. In some embodiments, AK is a non-binding antibody. In some embodiments, AK is an antibody or an antigen-binding antibody fragment. [000112]In some embodiments, the antibody is conjugated to a compound or conjugate (e.g., via a bond or a linker) by means of a heteroatom of the antibody Heteroatoms according to the invention which canP368919WO / 59362-725.601 be used for attachment include, but are not limited to, sulfur (in one embodiment via a sulfhydryl group of a cysteine side-chain of the antibody), oxygen (according to the invention by means of a carboxyl or hydroxyl group of the antibody) and nitrogen (in one embodiment via a primary or secondary amine group or amide group of the antibody). These heteroatoms may be present in the natural antibody or are introduced by chemical methods or methods of molecular biology. According to the invention, the attachment of the antibody to a small molecule target protein binder or therapeutic payload (e.g., via a linker such as L1-P1 or L2, as defined herein) has only a minor effect on the binding activity of the antibody with respect to the target antigen. In a preferred embodiment, the linkage has no effect on the binding activity of the antibody with respect to the target antigen. In some embodiments, the antibody is conjugated to one or more linkers further comprising a small molecule target protein binder or a therapeutic payload via a sulfhydryl group of a cysteine side-chain of the antibody, via an amine group of a lysine side-chain of the antibody, or via an amide group of a glutamine side-chain of the antibody. [000113]The term “monoclonal” antibody refers to antibodies obtained from a population of substantially homogeneous antibodies, i.e. individual antibodies of the population are identical except for naturally occurring mutations, of which there may be a small number. Monoclonal antibodies recognize a single antigenic binding site with high specificity. The term monoclonal antibody does not refer to a particular preparation process. [000114]The term “intact” antibody refers to antibodies comprising both an antigen-binding domain and the constant domain of the light and heavy chain. The constant domain may be a naturally occurring domain or a variant thereof having a number of modified amino acid positions, and may also be aglycosylated. [000115]The term “modified intact” antibody refers to intact antibodies fused via their amino terminus or carboxy terminus by means of a covalent bond (e.g. a peptide bond) with a further polypeptide or protein not originating from an antibody. Furthermore, antibodies may be modified such that, at defined positions, reactive cysteines are introduced to facilitate coupling to a toxophore. [000116]"Amino acid modification" or "mutation" as used herein generally refers to an amino acid substitution, insertion and / or deletion in a polypeptide sequence. The preferred amino acid modification here is a substitution. "Amino acid substitution" or "substitution" here means an exchange of an amino acid at a given position in a protein sequence for another amino acid. For example, the substitution Y50W describes a variant of a parent polypeptide in which the tyrosine at position 50 has been exchanged for a tryptophan. A "variant" of a polypeptide describes a polypeptide having an amino acid sequence substantially identical to a reference polypeptide, typically a native or "parent" polypeptide. The polypeptide variant may have one or more amino acid exchanges, deletions and / or insertions at particular positions in the native amino acid sequence. [000117]The term “human” antibody refers to antibodies which can be obtained from a human or which are synthetic human antibodies. A "synthetic" human antibody is an antibody which is partially or entirely obtainable in silico from synthetic sequences based on the analysis of human antibody sequences. A human antibody can be encoded, for example, by a nucleic acid isolated from a library of antibody sequences ofP368919WO / 59362-725.601 human origin. Such "human" and "synthetic" antibodies also include aglycosylated variants which have been produced either by deglycosylation by PNGaseF or by mutation of N297 (Kabat numbering) of the heavy chain to any other amino acid. [000118]The term “humanized” or “chimeric” antibody describes antibodies consisting of a non-human and a human portion of the sequence. In these antibodies, part of the sequences of the human immunoglobulin (recipient) is replaced by sequence portions of a non-human immunoglobulin (donor). In many cases, the donor is a murine immunoglobulin. In the case of humanized antibodies, amino acids of the CDR of the recipient are replaced by amino acids of the donor. Sometimes, amino acids of the framework, too, are replaced by corresponding amino acids of the donor. In some cases the humanized antibody contains amino acids present neither in the recepient nor in the donor, which were introduced during the optimization of the antibody. In the case of chimeric antibodies, the variable domains of the donor immunoglobulin are fused with the constant regions of a human antibody. Such "humanized" and "chimeric" antibodies also include aglycosylated variants which have been produced either by deglycosylation by PNGaseF or by mutation of N297 (Kabat numbering) of the heavy chain to any other amino acid. [000119]The term complementarity determining region (CDR) as used herein refers to those amino acids of a variable antibody domain which are required for binding to the antigen. Typically, each variable region has three CDR regions referred to as CDR1, CDR2 and CDR3. Each CDR region may embrace amino acids according to the definition of Kabat and / or amino acids of a hypervariable loop defined according to Chotia. The definition according to Kabat comprises, for example, the region from about amino acid position 24 – 34 (CDR1), 50 – 56 (CDR2) and 89 – 97 (CDR3) of the variable light chain / domain (VL) and 31 – 35 (CDR1), 50 – 65 (CDR2) and 95 – 102 (CDR3) of the variable heavy chain / domain (VH) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)). The definition according to Chotia comprises, for example, the region from about amino acid position 26 – 32 (CDR1), 50 – 52 (CDR2) and 91 –96 (CDR3) of the variable light chain (VL) and 26 – 32 (CDR1), 53 – 55 (CDR2) and 96 – 101 (CDR3) of the variable heavy chain (VH) (Chothia and Lesk; J Mol Biol 196: 901-917 (1987)). In some cases, a CDR may comprise amino acids from a CDR region defined according to Kabat and Chotia. [000120]Depending on the amino acid sequence of the constant domain of the heavy chain, antibodies may be categorized into different classes. There are five main classes of intact antibodies: IgA, IgD, IgE, IgG and IgM, and several of these can be divided into further subclasses. (Isotypes), e.g. IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The constant domains of the heavy chain, which correspond to the different classes, are referred to as [alpha / ^], [delta / ^], [epsilon / ^], [gamma / ^] and [my / ^]. Both the three- dimensional structure and the subunit structure of antibodies are known. [000121]The term “functional fragment” or “antigen-binding antibody fragment” of an antibody / immunoglobulin is defined as a fragment of an antibody / immunoglobulin (e.g. the variable domains of an IgG) which still comprise the antigen binding domains of the antibody / immunoglobulin.P368919WO / 59362-725.601 The “antigen binding domain” of an antibody typically comprises one or more hypervariable regions of an antibody, for example the CDR, CDR2 and / or CDR3 region. However, the “framework” or “skeleton” region of an antibody may also play a role during binding of the antibody to the antigen. The framework region forms the skeleton of the CDRs. Preferably, the antigen binding domain comprises at least amino acids 4 to 103 of the variable light chain and amino acids 5 to 109 of the variable heavy chain, more preferably amino acids 3 to 107 of the variable light chain and 4 to 111 of the variable heavy chain, especially preferably the complete variable light and heavy chains, i.e. amino acids 1 – 109 of the VL and 1 to 113 of the VH (numbering according to WO97 / 08320). [000122]“Functional fragments” or “antigen-binding antibody fragments” of the invention encompass, non-conclusively, Fab, Fab’, F(ab’)2and Fv fragments, diabodies, Single Domain Antibodies (DAbs), linear antibodies, individual chains of antibodies (single-chain Fv, abbreviated to scFv); and multispecific antibodies, such as bi- and tri-specific antibodies, for example, formed from antibody fragments. Antibodies other than “multispecific” or “multifunctional” antibodies are those having identical binding sites. Multispecific antibodies may be specific for different epitopes of an antigen or may be specific for epitopes of more than one antigen. A F(ab')2or Fab molecule may be constructed such that the number of intermolecular disulfide interactions occurring between the Ch1 and the CL domains can be reduced or else completely prevented. [000123]“Epitopes” refer to protein determinants capable of binding specifically to an immunoglobulin or T cell receptors. Epitopic determinants usually consist of chemically active surface groups of molecules such as amino acids or sugar side chains or combinations thereof, and usually have specific 3-dimensional structural properties and also specific charge properties. [000124]“Functional fragments” or “antigen-binding antibody fragments” may be fused with another polypeptide or protein, not originating from an antibody, via the amino terminus or carboxyl terminus thereof, by means of a covalent bond (e.g. a peptide linkage). Furthermore, antibodies and antigen-binding fragments may be modified by introducing reactive cysteines at defined locations, in order to facilitate coupling to a toxophore (see Junutula et al. Nat Biotechnol.2008 Aug; 26(8):925-32). [000125]Antibodies of the invention may be obtained from recombinant antibody libraries consisting for example of the amino acid sequences of a multiplicity of antibodies compiled from a large number of healthy volunteers. Antibodies may also be produced by means of known recombinant DNA technologies. The nucleic acid sequence of an antibody can be obtained by routine sequencing or is available from publically accessible databases. [000126]An “isolated” antibody or binder has been purified to remove other constituents of the cell. Contaminating constituents of a cell which may interfere with a diagnostic or therapeutic use are, for example, enzymes, hormones, or other peptidic or non-peptidic constituents of a cell. A preferred antibody or binder is one which has been purified to an extent of more than 95% by weight, relative to the antibody or binder (determined for example by Lowry method, UV-Vis spectroscopy or by SDS capillary gel electrophoresis). Moreover an antibody which has been purified to such an extent that it is possible toP368919WO / 59362-725.601 determine at least 15 amino acids of the amino terminus or of an internal amino acid sequence, or which has been purified to homogeneity, the homogeneity being determined by SDS-PAGE under reducing or non-reducing conditions (detection may be determined by means of Coomassie Blue staining or preferably by silver coloration). However, an antibody is normally prepared by one or more purification steps. [000127]The term “specific binding” or “binds specifically” refers to an antibody or binder which binds to a predetermined antigen / target molecule. Specific binding of an antibody or binder typically describes an antibody or binder having an affinity of at least 10-7 M (as Kd value; i.e. preferably those with Kd values smaller than 10-7 M), with the antibody or binder having an at least two times higher affinity for the predetermined antigen / target molecule than for a non-specific antigen / target molecule (e.g. bovine serum albumin, or casein) which is not the predetermined antigen / target molecule or a closely related antigen / target molecule. Specific binding of an antibody or binder does not exclude the antibody or binder binding to a plurality of antigens / target molecules (e.g. orthologs of different species). The antibodies preferably have an affinity of at least 10-7 M (as Kd value; in other words preferably those with smaller Kd values than 10-7 M), preferably of at least 10-8 M, more preferably in the range from 10-9 M to 10-11 M. The Kd values may be determined, for example, by means of surface plasmon resonance spectroscopy. [000128]The antibody-drug conjugates of the invention likewise exhibit affinities in these ranges. The affinity is preferably not substantially affected by the conjugation of the drugs (in general, the affinity is reduced by less than one order of magnitude, in other words, for example, at most from 10-8 M to 10-7 M). [000129]The antibodies used in accordance with the invention are also notable preferably for a high selectivity. A high selectivity exists when the antibody of the invention exhibits an affinity for the target protein which is better by a factor of at least 2, preferably by a factor of 5 or more preferably by a factor of 10, than for an independent other antigen, e.g. human serum albumin (the affinity may be determined, for example, by means of surface plasmon resonance spectroscopy). Antibodies directed against a cancer target antigen [000130] The target antigen towards which the antibody or antigen-binding fragment thereof, is directed is preferably a cancer target antigen. The term “cancer target antigen” describes a target antigen which is more abundantly present on one or more cancer cell species than on non-cancer cells of the same tissue type. Preferably, the cancer target antigen is selectively present on one or more cancer cell species compared with non-cancer cells of the same tissue type, where selectively describes an at least two-fold enrichment on cancer cells compared to non-cancer cells of the same tissue type (a “selective cancer target antigen”). In some embodiments, the selective cancer target antigen is at least three-fold, four-fold, five- fold, ten-fold, twenty-fold, or more, enriched in a cancer cell compared to a non-cancer cell of the same tissue type. The use of a cancer target antigen allows the selective therapy of cancer cells using the conjugates according to the invention. [000131] Antibodies which are specific against an antigen, for example cancer cell antigen, can be prepared by a person of ordinary skill in the art by means of methods with which he or she is familiar (such as recombinant expression, for example) or may be acquired commercially (as for example from MerckP368919WO / 59362-725.601 KGaA, Germany). Examples of known commercially available antibodies in cancer therapy are Erbitux® (cetuximab, Merck KGaA), Avastin® (bevacizumab, Roche), Bavencio® (avelumab, Merck KGaA / Pfizer), and Herceptin® (trastuzumab, Genentech). Trastuzumab is a recombinant humanized monoclonal antibody of the IgG1kappa type which in a cell-based assay (Kd = 5 nM) binds the extracellular domains of the human epidermal growth receptor with high affinity. The antibody is produced recombinantly in CHO cells. All these antibodies can also be produced as aglycosylated variants of these antibodies, either by deglycosylation by means of PNGase F or by mutation of N297 (Kabat numbering) of the heavy chain to any amino acid. [000132] Examples of cancer target antigens include, but are not limited to, the following: 1) Carboanhydrase IX (CA9, SwissProt Reference Q16790), NCBI Gene ID: 768) 2) C4.4a (NCBI Reference Sequence NP_055215.2; synonym LYPD3, NCBI Gene ID: 27076) 3) CD52 (NCBI Reference Sequence NP_001794.2) 4) Her2 (ERBB2; NCBI Reference Sequence NP_004439.2; NCBI Gene ID: 2064) 5) CD20 (NCBI Reference Sequence NP_068769.2) 6) the lymphocyte activation antigen CD30 (SwissProt ID P28908) 7) the lymphocyte adhesion molecule CD22 (SwissProt ID P20273; NCBI Gene ID: 933) 8) the myloid cell surface antigen CD33 (SwissProt ID P20138; NCBI Gene ID: 945) 9) the transmembrane glycoprotein NMB (GPNMB, SwissProt ID Q14956, NCBI Gene ID: 10457) 10) the adhesion molecule CD56 (SwissProt ID P13591) 11) the surface molecule CD70 (SwissProt ID P32970, NCBI Gene ID: 970) 12) the surface molecule CD74 (SwissProt ID P04233, NCBI Gene ID: 972) 13) the B-lymphocyte antigen CD19 (SwissProt ID P15391, NCBI Gene ID: 930) 14) the surface protein Mucin-1 (MUC1, SwissProt ID P15941, NCBI Gene ID: 4582) 15) the surface protein CD138 (SwissProt ID P18827) 16) the integrin alphaV (NCBI reference sequence: NP_002201.1, NCBI Gene ID: 3685) 17) the teratocarcinoma-derived growth factor 1 protein TDGF1 (NCBI Reference Sequence: NP_003203.1, NCBI Gene ID: 6997) 18) the prostate-specific membrane antigen PSMA (Swiss Prot ID: Q04609; NCBI Gene ID: 2346) 19) the tyrosine protein kinase EPHA2 (Swiss Prot ID: P29317, NCBI Gene ID: 1969) 20) the surface protein SLC44A4 (NCBI Reference Sequence: NP_001171515.1, NCBI Gene ID: 80736) 21) the surface protein BMPR1B (SwissProt: O00238) 22) the transport protein SLC7A5 (SwissProt: Q01650) 23) the epithelial prostate antigen STEAP1 (SwissProt: Q9UHE8, Gene ID: 26872) 24) the ovarian carcinoma antigen MUC16 (SwissProt: Q8WXI7, Gene ID: 94025) 25) the transport protein SLC34A2 (SwissProt: O95436, Gene ID: 10568) 26) the surface protein SEMA5b (SwissProt: Q9P283) 27) the surface protein LYPD1 (SwissProt: Q8N2G4)P368919WO / 59362-725.601 28) the endothelin receptor type B EDNRB (SwissProt: P24530, NCBI Gene ID: 1910) 29) the ring finger protein RNF43 (SwissProt: Q68DV7) 30) the prostate carcinoma-associated protein STEAP2 (SwissProt: Q8NFT2) 31) the cation channel TRPM4 (SwissProt: Q8TD43) 32) the complement receptor CD21 (SwissProt: P20023) 33) the B-cell antigen receptor complex-associated protein CD79b (SwissProt: P40259, NCBI Gene ID: 974) 34) the cell adhesion antigen CEACAM6 (SwissProt: P40199) 35) the dipeptidase DPEP1 (SwissProt: P16444) 36) the interleukin receptor IL20Ralpha (SwissProt: Q9UHF4, NCBI Gene ID: 3559) 37) the proteoglycan BCAN (SwissProt: Q96GW7) 38) the ephrin receptor EPHB2 (SwissProt: P29323) 39) the prostate stem cell-associated protein PSCA (NCBI Reference Sequence: NP_005663.2 ) 40) the surface protein LHFPL3 (SwissProt: Q86UP9) 41) the receptor protein TNFRSF13C (SwissProt: Q96RJ3) 42) the B-cell antigen receptor complex-associated protein CD79a (SwissProt: P11912) 43) the receptor protein CXCR5 (CD185; SwissProt: P32302; NCBI Gene ID 643, NCBI Reference Sequence: NP_001707.1) 44) the ion channel P2X5 (SwissProt: Q93086) 45) the lymphocyte antigen CD180 (SwissProt: Q99467) 46) the receptor protein FCRL1 (SwissProt: Q96LA6) 47) the receptor protein FCRL5 (SwissProt: Q96RD9) 48) the MHC class II molecule Ia antigen HLA-DOB (NCBI Reference Sequence: NP_002111.1) 49) the T-cell protein VTCN1 (SwissProt: Q7Z7D3) 50) TWEAKR (FN14, TNFRSF12A, NCBI Reference Sequence: NP_057723.1, NCBI Gene ID: 51330) 51) the lymphocyte antigen CD37 (Swiss Prot: P11049, NCBI Gene ID: 951) 52) the FGF receptor 2; FGFR2 (NCBI Gene ID: 2263; Official Symbol: FGFR2) 53) the transmembrane glycoprotein B7H3 (CD276; NCBI Gene ID: 80381 NCBI Reference Sequence: NP_001019907.1, Swiss Prot: Q5ZPR3-1) 54) the B cell receptor BAFFR (CD268; NCBI Gene ID:115650) 55) the receptor protein ROR 1 (NCBI Gene ID: 4919) 56) the surface receptor CD123 (IL3RA; NCBI Gene ID: 3563; NCBI Reference Sequence: NP_002174.1; Swiss-Prot: P26951) 57) the receptor protein syncytin ( NCBI Gene ID 30816) 58) aspartate beta-hydroxylase (ASPH; NCBI Gene ID 444) 59) the cell surface glycoprotein CD44 (NCBI Gene ID: 960) 60) CDH15 (Cadherin 15 NCBI Gene ID: 1013)P368919WO / 59362-725.601 61) the cell surface glycoprotein CEACAM5 (NCBI Gene ID: 1048) 62) the cell adhesion molecule L1-like (CHL1, NCBI Gene ID: 10752) 63) the receptor tyrosine kinase c-Met (NCBI Gene ID: 4233) 64) the notch ligand DLL3 (NCBI Gene ID: 10683) 65) the ephrin A4 (EFNA4, NCBI Gene ID: 1945) 66) ectonucleotide pyrophosphatase / phosphodiesterase 3 (ENPP3, NCBI Gene ID: 5169) 67) coagulation factor III (F3, NCBI Gene ID: 2152) 68) FGF receptor 3 (FGFR3, NCBI Gene ID: 2261) 69) the folate hydrolase FOLH1 (NCBI Gene ID: 2346) 70) the folate receptor 1 (FOLR1; NCBI Gene ID: 2348) 71) the guanylate cyclase 2C (GUCY2C, NCBI Gene ID: 2984) 72) the KIT proto-oncogen receptor tyrosine kinase (NCBI Gene ID: 3815) 73) lysosomal-associated membrane protein 1 (LAMP1, NCBI Gene ID: 3916) 74) lymphocyte antigen 6 complex, locus E (LY6E, NCBI Gene ID: 4061) 75) the protein NOTCH3 (NCBI Gene ID: 4854) 76) protein tyrosine kinase 7 (PTK7, NCBI Gene ID: 5754) 77) nectin cell adhesion molecule 4 (PVRL4, NECTIN4, NCBI Gene ID: 81607) 78) the transmembrane protein syndecan 1 (SDC1, NCBI Gene ID: 6382) 79) SLAM family member 7 (SLAMF7, NCBI Gene ID: 57823) 80) the transport protein SLC39A6 (NCBI Gene ID: 25800) 81) SLIT- and NTRK-like family member 6 (SLITRK6, NCBI Gene ID: 84189) 82) the cell surface receptor TACSTD2 (NCBI Gene ID: 4070) 83) the receptor protein TNFRSF8 (NCBI Gene ID: 943) 84) the receptor protein TNFSF13B (NCBI Gene ID: 10673) 85) the glycoprotein TPBG (NCBI Gene ID: 7162) 86) the cell surface receptor TROP2 (TACSTD2, NCBI Gene ID: 4070) 87) the galanin-like G protein-coupled receptor KISS1R (GPR54, NCBI Gene ID: 84634) 88) the transport protein SLAMF6 (NCBI Gene ID: 114836) 89) the fibroblast activation protein alpha FAP (NCBI Ref Seq: NP_004451) 90) the fibronectin glycoprotein CEACAM5 (NCBI Gene ID: 1048; RefSeq: NP_004354) 91) the collagen 11 alpha 1 protein COL11A1 (Ref Seq: AAI17698) 92) the galectin-3-binding protein (Uniprot Ident. Q08380) 93) the anthrax toxin receptor 1 (tumor endothelial marker 8; Ref Seq: Q9H6X2) 94) Tenascin-C (RefSeq: NP_002151.2) 95) the surface receptor IL2RA (CD25; Gene ID: 3559) 96) the programmed cell death-ligand 1 PD-L1 (NCBI Reference Sequence: NP_054862.1); and 97) the integrin beta6 protein ITGB6 (NCBI Reference Sequence: NP_000879).P368919WO / 59362-725.601 [000133]Preferred examples of cancer target antigens include, but are not limited to, the following: (1) Her2 (NCBI reference sequence NP_004439.2), SEQ ID NO: 115 (MELAALCRWG LLLALLPPGA ASTQVCTGTD MKLRLPASPE THLDMLRHLY QGCQVVQGNL ELTYLPTNAS LSFLQDIQEV QGYVLIAHNQ VRQVPLQRLR IVRGTQLFED NYALAVLDNG DPLNNTTPVT GASPGGLREL QLRSLTEILK GGVLIQRNPQ LCYQDTILWK DIFHKNNQLA LTLIDTNRSR ACHPCSPMCK GSRCWGESSE DCQSLTRTVC AGGCARCKGP LPTDCCHEQC AAGCTGPKHS DCLACLHFNH SGICELHCPA LVTYNTDTFE SMPNPEGRYT FGASCVTACP YNYLSTDVGS CTLVCPLHNQ EVTAEDGTQR CEKCSKPCAR VCYGLGMEHL REVRAVTSAN IQEFAGCKKI FGSLAFLPES FDGDPASNTA PLQPEQLQVF ETLEEITGYL YISAWPDSLP DLSVFQNLQV IRGRILHNGA YSLTLQGLGI SWLGLRSLRE LGSGLALIHH NTHLCFVHTV PWDQLFRNPH QALLHTANRP EDECVGEGLA CHQLCARGHC WGPGPTQCVN CSQFLRGQEC VEECRVLQGL PREYVNARHC LPCHPECQPQ NGSVTCFGPE ADQCVACAHY KDPPFCVARC PSGVKPDLSY MPIWKFPDEE GACQPCPINC THSCVDLDDK GCPAEQRASP LTSIISAVVG ILLVVVLGVV FGILIKRRQQ KIRKYTMRRL LQETELVEPL TPSGAMPNQA QMRILKETEL RKVKVLGSGA FGTVYKGIWI PDGENVKIPV AIKVLRENTS PKANKEILDE AYVMAGVGSP YVSRLLGICL TSTVQLVTQL MPYGCLLDHV RENRGRLGSQ DLLNWCMQIA KGMSYLEDVR LVHRDLAARN VLVKSPNHVK ITDFGLARLL DIDETEYHAD GGKVPIKWMA LESILRRRFT HQSDVWSYGV TVWELMTFGA KPYDGIPARE IPDLLEKGER LPQPPICTID VYMIMVKCWM IDSECRPRFR ELVSEFSRMA RDPQRFVVIQ NEDLGPASPL DSTFYRSLLE DDDMGDLVDA EEYLVPQQGF FCPDPAPGAG GMVHHRHRSS STRSGGGDLT LGLEPSEEEA PRSPLAPSEG AGSDVFDGDL GMGAAKGLQS LPTHDPSPLQ RYSEDPTVPL PSETDGYVAP LTCSPQPEYV NQPDVRPQPP SPREGPLPAA RPAGATLERP KTLSPGKNGV VKDVFAFGGA VENPEYLTPQ GGAAPQPHPP PAFSPAFDNL YYWDQDPPER GAPPSTFKGT PTAENPEYLG LDVPV) (2) TWEAKR (FN14, TNFRSF12A, NCBI Reference Sequence: NP_057723.1, NCBI Gene ID: 51330) (SEQ ID NO: 114 (MARGSLRRLL RLLVLGLWLA LLRSVAGEQA PGTAPCSRGS SWSADLDKCM DCASCRARPH SDFCLGCAAA PPAPFRLLWP ILGGALSLTF VLGLLSGFLV WRRCRRREKF TTPIEETGGE GCPAVALIQ)) (3) The CXC chemokine receptor CXCR5 (CD185; Gene ID 643) (SwissProt: P32302) (4) The surface receptor IL3RA (CD123; Gene ID: 3561) (5) The programmed cell death-ligand 1 PD-L1 (NCBI Reference Sequence: NP_054862.1); (6) Tenascin-C TNC (NCBI Reference Sequence: NP_002151.2) (7) the integrin beta6 protein ITGB6 (NCBI Reference Sequence: NP_000879) [000134] In some embodiments, the cancer target antigen is selected from the group consisting of PD- L1 Her2 TWEAKR Tenascin C alphavbeta6 (“α ß6” or “avß6”) integrin and CD123 In someP368919WO / 59362-725.601 embodiments, AK binds to an extracellular cancer target antigen selected from the group consisting of PD- L1, Her2, TWEAKR, Tenascin-C, integrin alphavbeta6 and CD123. [000135] In some embodiments, AK binds specifically to an extracellular cancer target antigen selected from the group consisting of PD-L1, Her2, TWEAKR, Tenascin-C, integrin alphavbeta6, and CD123. In a preferred embodiment, AK, after binding to its extracellular target antigen on the target tumor cell, is not efficiently internalized by the target tumor cell through the binding. This causes the conjugate, which may be an antibody-drug conjugate (ADC) or an antibody-binder conjugate (ABC), not to be taken up by the target cell. The antibody or antigen binding fragment thereof is processed (including possible cleavage by a protease, e.g., neutrophil elastase) extracellularly. Thus, in certain embodiments of the present disclosure, provided herein is an ADC or ABC comprising a non-internalizing antibody or antigen-binding fragment thereof, wherein a payload (e.g., D1) is released extracellularly in the presence of a protease (e.g., neutrophil elastase), preferably in a tumor microenvironment. [000136] In a preferred embodiment, AK is an isolated antibody or an isolated antigen-binding antibody fragment. Preferred antigen-binding antibody fragments are Fab, Fab', F(ab')2 and Fv fragments, diabodies, DAbs, linear antibodies and scFv. Particularly preferred are Fab, diabodies and scFv. Particularly preferred are monoclonal antibodies or antigen-binding antibody fragments thereof. Further particularly preferred are human, humanized or chimeric antibodies or antigen-binding antibody fragments thereof. [000137] Antibodies or antigen-binding antibody fragments which bind cancer target antigens may be prepared by a person of ordinary skill in the art using known processes, such as, for example, chemical synthesis or recombinant expression. Binders for cancer target molecules may be acquired commercially or may be prepared by a person of ordinary skill in the art using known processes, such as, for example, chemical synthesis or recombinant expression. Further processes for preparing antibodies or antigen- binding antibody fragments are described in WO 2007 / 070538 (see page 22 “Antibodies”). [000138]In another preferred embodiment, the antibody employed in the ADCs is a non-binding antibody. This antibody is further modified with binding molecules, which bind to a cancer target proteins. In some embodiments, the cancer target antigen is an extracellular cancer target antigen. [000139]Examples of cancer target molecules are but are not limited to an alpha-v beta-3 (“αvß3” or “avß3”) integrin, alpha-v beta-6 (“αvß6” or “avß6”) integrin, carbonic anhydrase IX (“CA9” or “CA IX”), fibroblast activating protein (“FAP”), prostate specific membrane antigen (“PSMA”), heat shock protein 90 (“Hsp 90”), folic acid receptor, glucose transporter 1, somatostatin receptor, aminopeptidase N (APN), low density lipoprotein receptor-related protein 1 (LRP1), bombesin receptor, gonadotropin releasing hormone (GnRH or LHRH) receptor, p32, membrane type 1 matrix metalloprotease (MT1-MMP), Sortilin, Nectin-4, PD-L1, Her2, Tenascin-C (TNSC), and integrin alphavbeta6. [000140]In some embodiments, the antibody or antigen binding fragment thereof (AK, e.g., AK1, AK2, AK3) comprises an antibody or antibody region (e.g., CDR, variable domain) shown in the table below:P368919WO / 59362-725.601P368919WO / 59362-725.601P368919WO / 59362-725.601P368919WO / 59362-725.601P368919WO / 59362-725.601P368919WO / 59362-725.601P368919WO / 59362-725.601[000141] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth inP368919WO / 59362-725.601 SEQ ID NO: 2; b) a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 3; c) a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 4; d) a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 6; e) a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 7; and / or f) a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 8. [000142] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 12; b) a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 13; c) a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 14; d) a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 16; e) a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 17; and / or f) a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 18. [000143] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 22; b) a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 23; c) a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 24; d) a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 26; e) a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 27; and / or f) a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 28. [000144] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 32; b) a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 33; c) a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 34; d) a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 36; e) a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 37; and / or f) a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 38. [000145] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 42; b) a heavy chain complementarity determining region 2 (HCDR2) comprising the aminoP368919WO / 59362-725.601 acid sequence set forth in SEQ ID NO: 43; c) a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 44; d) a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 46; e) a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 47; and / or f) a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 48. [000146] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 52; b) a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 53; c) a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 54; d) a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 56; e) a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 57; and / or f) a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 58. [000147] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 62; b) a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 63; c) a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 64; d) a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 66; e) a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 67; and / or f) a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 68. [000148] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 72; b) a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 73; c) a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 74; d) a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 76; e) a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 77; and / or f) a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 78. [000149] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 1; and / or b) an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO:P368919WO / 59362-725.601 5. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 1; and / or b) an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 5. [000150] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 9; and / or b) an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 10. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 9; and / or b) an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 10. [000151] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 11; and / or b) an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 15. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 11; and / or b) an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 15. [000152] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 19; and / or b) an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 20. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 19; and / or b) an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 20. [000153] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 21; and / or b) an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 25. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 21; and / or b) an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 25. [000154] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 29; and / or b) an immunoglobulin light chain comprising an amino acidP368919WO / 59362-725.601 sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 30. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 29; and / or b) an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 30. [000155] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 31; and / or b) an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 35. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 31; and / or b) an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 35. [000156] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 39; and / or b) an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 40. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 39; and / or b) an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 40. [000157] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 41; and / or b) an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 45. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 41; and / or b) an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 45. [000158] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 49; and / or b) an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 50. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 49; and / or b) an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 50. [000159] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 51; and / or b) an immunoglobulin light chain variable regionP368919WO / 59362-725.601 comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 55. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 51; and / or b) an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 55. [000160] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 59; and / or b) an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 60. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 59; and / or b) an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 60. [000161] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 61; and / or b) an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 65. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 61; and / or b) an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 65. [000162] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 69; and / or b) an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 70. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 69; and / or b) an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 70. [000163] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 71; and / or b) an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 75. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 71; and / or b) an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 75. [000164] In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, orP368919WO / 59362-725.601 99% identical to SEQ ID NO: 79; and / or b) an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 80. In some embodiments, the antibody or antigen binding fragment thereof comprises: a) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 79; and / or b) an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 80. [000165] The term “identity” refers to a relationship between the sequences of two or more polypeptide molecules or two or more nucleic acid molecules, as determined by aligning and comparing the sequences. Percent (%) amino acid sequence identity with respect to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALIGN (DNAStar, Inc.) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. Bacterial Expression [000166] The person skilled in the art is aware of the way in which antibodies, antigen-binding fragments thereof or variants thereof can be produced with the aid of bacterial expression. [000167] Suitable expression vectors for bacterial expression of desired proteins are constructed by insertion of a DNA sequence which encodes the desired protein within the functional reading frame together with suitable translation initiation and translation termination signals and with a functional promoter. The vector comprises one or more phenotypically selectable markers and a replication origin in order to enable the retention of the vector and, if desired, the amplification thereof within the host. Suitable prokaryotic hosts for transformation include but are not limited to E. coli, Bacillus subtilis, Salmonella typhimurium and various species from the genus Pseudomonas, Streptomyces, and Staphylococcus. Bacterial vectors may be based, for example, on bacteriophages, plasmids, or phagemids. These vectors may contain selectable markers and a bacterial replication origin, which are derived from commercially available plasmids. Many commercially available plasmids typically contain elements of the well-known cloning vector pBR322 (ATCC 37017). In bacterial systems, a number of advantageous expression vectors can be selected on the basis of the intended use of the protein to be expressed. [000168] After transformation of a suitable host strain and growth of the host strain to an appropriate cell density, the selected promoter is de-reprimed / induced by suitable means (for example a change in temperature or chemical induction), and the cells are cultivated for an additional period. The cells are typically harvested by centrifugation and if necessary digested in a physical manner or by chemical means, and the resulting raw extract is retained for further purification.P368919WO / 59362-725.601 [000169] Therefore, a further embodiment of the present invention is an expression vector comprising a nucleic acid which encodes a novel antibody of the present invention. [000170] Antibodies of the present invention or antigen-binding fragments thereof include naturally purified products, products which originate from chemical syntheses, and products which are produced by recombinant technologies in prokaryotic hosts, for example E. coli, Bacillus subtilis, Salmonella typhimurium and various species from the genus Pseudomonas, Streptomyces, and Staphylococcus, preferably E. coli. Mammalian Cell Expression [000171] The person skilled in the art is aware of the way in which antibodies, antigen-binding fragments thereof or variants thereof can be produced with the aid of mammalian cell expression. [000172] Preferred regulatory sequences for expression in mammalian cell hosts include viral elements which lead to high expression in mammalian cells, such as promoters and / or expression amplifiers derived from cytomegalovirus (CMV) (such as the CMV promoter / enhancer), simian virus 40 (SV40) (such as the SV40 promoter / enhancer), from adenovirus, (for example the adenovirus major late promoter (AdMLP)) and from polyoma. The expression of the antibodies may be constitutive or regulated (for example induced by addition or removal of small molecule inductors such as tetracycline in combination with the Tet system). [000173] For further description of viral regulatory elements and sequences thereof, reference is made, for example, to U.S.5,168,062 by Stinski, U.S.4,510,245 by Bell et al. and U.S.4,968,615 by Schaffner et al. The recombinant expression vectors may likewise include a replication origin and selectable markers (see, for example, U.S. 4,399,216, 4,634,665 and U.S. 5,179,017). Suitable selectable markers include genes which impart resistance to substances such as G418, puromycin, hygromycin, blasticidin, zeocin / bleomycin, or methotrexate, or selectable markers which lead to auxotrophy of a host cell, such as glutamine synthetase (Bebbington et al., Biotechnology (N Y). 1992 Feb;10(2):169-75), when the vector has been introduced into the cell. [000174] For example, the dihydrofolate reductase (DHFR) gene imparts resistance to methotrexate, the neo gene imparts resistance to G418, the bsd gene from Aspergillus terreus imparts resistance to blasticidin, puromycin N-acetyltransferase imparts resistance to puromycin, the Sh ble gene product imparts resistance to zeocin, and resistance to hygromycin is imparted by the E. coli hygromycin resistance gene (hyg or hph). Selectable markers such as DHFR or glutamine synthetase are also helpful for amplification techniques in conjunction with MTX and MSX. [000175] The transfection of an expression vector into a host cell can be executed with the aid of standard techniques, including by electroporation, nucleofection, calcium phosphate precipitation, lipofection, polycation-based transfection such as polyethyleneimine (PEI)-based transfection and DEAE- dextran transfection. [000176] Suitable mammalian host cells for the expression of antibodies, antigen-binding fragments thereof or variants thereof include Chinese hamster ovary (CHO) cells such as CHO-K1 CHO-S CHO-P368919WO / 59362-725.601 K1SV [including DHFR-CHO cells, described in Urlaub and Chasin, (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220 and Urlaub et al., Cell. 1983 Jun;33(2):405-12, used with a DHFR-selectable marker, as described in R. J. Kaufman and P. A. Sharp (1982) Mol. Biol. 159:601-621, and other knockout cells, as detailed in Fan et al., Biotechnol Bioeng. 2012 Apr;109(4):1007-15), NS0 myeloma cells, COS cells, HEK293 cells, HKB11 cells, BHK21 cells, CAP cells, EB66 cells, and SP2 cells. [000177] The expression of antibodies, antigen-binding fragments thereof, or variants thereof can also be effected in a transient or semi-stable manner in expression systems such as HEK293, HEK293T, HEK293-EBNA, HEK293E, HEK293-6E, HEK293 Freestyle, HKB11, Expi293F, 293EBNALT75, CHO Freestyle, CHO-S, CHO-K1, CHO-K1SV, CHOEBNALT85, CHOS-XE, CHO-3E7 or CAP-T cells (for example like Durocher et al., Nucleic Acids Res.2002 Jan 15;30(2):E9) [000178] In some embodiments, the expression vector is constructed in such a way that the protein to be expressed is secreted into the cell culture medium in which the host cells are growing. The antibodies, the antigen-binding fragments thereof, or the variants thereof can be obtained from the cell culture medium with the aid of protein purification methods known to those skilled in the art. Purification [000179] The antibodies, the antigen-binding fragments thereof, or the variants thereof can be obtained and purified from recombinant cell cultures with the aid of well-known methods, examples of which include ammonium sulfate or ethanol precipitation, acid extraction, protein A chromatography, protein G chromatography, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography (HIC), affinity chromatography, hydroxyapatite chromatography and lectin chromatography. High-performance liquid chromatography ("HPLC") can likewise be employed for purification. See, for example, Colligan, Current Protocols in Immunology, or Current Protocols in Protein Science, John Wiley & Sons, NY, N.Y., (1997-2001), e.g., Chapters 1, 4, 6, 8, 9, 10. [000180] Antibodies of the present invention or antigen-binding fragments thereof, or variants thereof include naturally purified products, products from chemical synthesis methods and products which are produced with the aid of recombinant techniques in prokaryotic or eukaryotic host cells. Eukaryotic hosts include, for example, yeast cells, higher plant cells, insect cells and mammalian cells. Depending on the host cell chosen for the recombinant expression, the protein expressed may be in glycosylated or non- glycosylated form. [000181] In a preferred embodiment, the antibody is purified (1) to an extent of more than 95% by weight, measured, for example, by the Lowry method, by UV-vis spectroscopy or by SDS capillary gel electrophoresis (for example with a Caliper LabChip GXII, GX 90 or Biorad Bioanalyzer instrument), and in more preferred embodiments more than 99% by weight, (2) to a degree suitable for determination of at least 15 residues of the N-terminal or internal amino acid sequence, or (3) to homogeneity determined by SDS-PAGE under reducing or non-reducing conditions with the aid of Coomassie blue or preferably silver staining. [000182] Usually, an isolated antibody is obtained with the aid of at least one protein purification step.P368919WO / 59362-725.601 Anti-CD123 antibodies [000183] According to the invention, it is possible to use anti-CD123 antibodies. [000184] The expression “anti-CD123 antibody” or “an antibody which binds specifically to CD123” relates to an antibody which binds the cancer target molecule CD123 (IL3RA; NCBI-Gene ID: 3563; NCBI Reference sequence: NP_002174.1; Swiss-Prot: P26951; SEQ ID NO:111 (MVLLWLTLLL IALPCLLQTK EDPNPPITNL RMKAKAQQLT WDLNRNVTDI ECVKDADYSM PAVNNSYCQF GAISLCEVTN YTVRVANPPF STWILFPENS GKPWAGAENL TCWIHDVDFL SCSWAVGPGA PADVQYDLYL NVANRRQQYE CLHYKTDAQG TRIGCRFDDI SRLSSGSQSS HILVRGRSAA FGIPCTDKFV VFSQIEILTP PNMTAKCNKT HSFMHWKMRS HFNRKFRYEL QIQKRMQPVI TEQVRDRTSF QLLNPGTYTV QIRARERVYE FLSAWSTPQR FECDQEEGAN TRAWRTSLLI ALGTLLALVC VFVICRRYLV MQRLFPRIPH MKDPIGDSFQ NDKLVVWEAG KAGLEECLVT EVQVVQKT)), preferably with an affinity sufficient for a diagnostic and / or therapeutic application. In particular embodiments, the antibody binds CD123 with a dissociation constant (KD) of ≤ 1µM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM. [000185] Sun et al. (Sun et al., 1996, Blood 87(1)83-92) describe the generation and properties of the monoclonal antibody 7G3, which binds the N-terminal domain of IL-3Rα, CD123. US Patent Number 6,177,078 (Lopez) relates to the anti-CD123 antibody 7G3. A chimeric variant of this antibody (CSL360) is described in WO 2009 / 070844, and a humanized version (CSL362) in WO 2012 / 021934. The sequence of the 7G3 antibody is disclosed in EP2426148. This sequence constitutes the starting point for the humanized antibodies obtained by CDR grafting. [000186] An antibody which, after cell surface antigen binding, is internalized particularly well is the anti-CD123 antibody 12F1 disclosed by Kuo et al. (Kuo et al., 2009, Bioconjug Chem. 20(10):1975-82). The antibody 12F1 binds with higher affinity to CD123 than the antibody 7G3 and, after cell surface antigen binding, is internalized markedly faster than 7G3. Bispecific scFv immunofusion proteins based on 12F1 are disclosed in WO 2013 / 173820. Antibody TPP-6013 is a chimeric variant of 12F1. [000187] The invention relates in particular to conjugates with antibodies or antigen-binding antibody fragments thereof or variants thereof derived from the antibodies 7G3 (Sun et al., 1996, Blood 87(1):83- 92) and 12F1 (Kuo et al., 2009, Bioconjug Chem. 20(10):1975-82) originating from the mouse, or to conjugates with antibodies or antigen-binding antibody fragments thereof or variants thereof derived from the antibody 12F1 (Kuo et al., 2009, Bioconjug Chem.20(10):1975-82) originating from the mouse. [000188] Humanized variants of the murine 7G3 antibody and the murine 12F1 antibody were generated by CDR grafting into a human framework and subsequent optimization and are preferred examples in the context of the present invention. [000189] Particular preference is given in the context of the present invention to the anti-CD123 antibody TPP-9476. Anti-TWEAKR antibodies [000190] According to the invention it is possible to use anti-TWEAKR antibodiesP368919WO / 59362-725.601 [000191] The expression “anti-TWEAKR antibody” or “an antibody which binds specifically to TWEAKR” relates to an antibody which binds the cancer target molecule TWEAKR (NCBI Reference Sequence: NP_057723.1 SEQ ID NO: 114 (MARGSLRRLL RLLVLGLWLA LLRSVAGEQA PGTAPCSRGS SWSADLDKCM DCASCRARPH SDFCLGCAAA PPAPFRLLWP ILGGALSLTF VLGLLSGFLV WRRCRRREKF TTPIEETGGE GCPAVALIQ)), preferably with an affinity sufficient for a diagnostic and / or therapeutic application. In particular embodiments, the antibody binds TWEAKR with a dissociation constant (KD) of ≤ 1µM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM. [000192] Examples of antibodies which bind to TWEAKR are disclosed, for example, in WO2009 / 020933(A2), WO2009 / 140177 (A2), WO 2014 / 198817 (A1) and WO 2015 / 189143 (A1). These antibodies and antigen-binding fragments can be used in the context of this invention. [000193] ITEM-4 is an anti-TWEAKR antibody which was described by Nakayama et al. (Nakayama, et al., 2003, Biochem Biophy Res Comm, 306:819-825). Humanized variants of this antibody based on CDR grafting are described by Zhou et al. (Zhou et al., 2013, J Invest Dermatol.133(4):1052-62) and in WO 2009 / 020933. These antibodies and antigen-binding fragments can be used in the context of this invention. [000194] Particular preference is given in the context of the present invention to the anti-TWEAKR antibodies TPP-7006 and TPP-7007. These are humanized variants of the antibody ITEM-4. These antibodies and antigen-binding fragments can be used with preference in the context of this invention. Anti-HER2 antibodies [000195] According to the invention, it is possible to use anti-HER2 antibodies. [000196] The expression “anti-HER2 antibody” or “an antibody which binds specifically to HER2” relates to an antibody which binds the cancer target molecule HER2 (NCBI Reference Sequence: NP_004439.2 SEQ ID NO: 115 (MELAALCRWG LLLALLPPGA ASTQVCTGTD MKLRLPASPE THLDMLRHLY QGCQVVQGNL ELTYLPTNAS LSFLQDIQEV QGYVLIAHNQ VRQVPLQRLR IVRGTQLFED NYALAVLDNG DPLNNTTPVT GASPGGLREL QLRSLTEILK GGVLIQRNPQ LCYQDTILWK DIFHKNNQLA LTLIDTNRSR ACHPCSPMCK GSRCWGESSE DCQSLTRTVC AGGCARCKGP LPTDCCHEQC AAGCTGPKHS DCLACLHFNH SGICELHCPA LVTYNTDTFE SMPNPEGRYT FGASCVTACP YNYLSTDVGS CTLVCPLHNQ EVTAEDGTQR CEKCSKPCAR VCYGLGMEHL REVRAVTSAN IQEFAGCKKI FGSLAFLPES FDGDPASNTA PLQPEQLQVF ETLEEITGYL YISAWPDSLP DLSVFQNLQV IRGRILHNGA YSLTLQGLGI SWLGLRSLRE LGSGLALIHH NTHLCFVHTV PWDQLFRNPH QALLHTANRP EDECVGEGLA CHQLCARGHC WGPGPTQCVN CSQFLRGQEC VEECRVLQGL PREYVNARHC LPCHPECQPQ NGSVTCFGPE ADQCVACAHY KDPPFCVARC PSGVKPDLSY MPIWKFPDEE GACQPCPINC THSCVDLDDK GCPAEQRASP LTSIISAVVG ILLVVVLGVV FGILIKRRQQ KIRKYTMRRL LQETELVEPL TPSGAMPNQA QMRILKETEL RKVKVLGSGA FGTVYKGIWI PDGENVKIPV AIKVLRENTS PKANKEILDE AYVMAGVGSP YVSRLLGICL TSTVQLVTQLP368919WO / 59362-725.601 MPYGCLLDHV RENRGRLGSQ DLLNWCMQIA KGMSYLEDVR LVHRDLAARN VLVKSPNHVK ITDFGLARLL DIDETEYHAD GGKVPIKWMA LESILRRRFT HQSDVWSYGV TVWELMTFGA KPYDGIPARE IPDLLEKGER LPQPPICTID VYMIMVKCWM IDSECRPRFR ELVSEFSRMA RDPQRFVVIQ NEDLGPASPL DSTFYRSLLE DDDMGDLVDA EEYLVPQQGF FCPDPAPGAG GMVHHRHRSS STRSGGGDLT LGLEPSEEEA PRSPLAPSEG AGSDVFDGDL GMGAAKGLQS LPTHDPSPLQ RYSEDPTVPL PSETDGYVAP LTCSPQPEYV NQPDVRPQPP SPREGPLPAA RPAGATLERP KTLSPGKNGV VKDVFAFGGA VENPEYLTPQ GGAAPQPHPP PAFSPAFDNL YYWDQDPPER GAPPSTFKGT PTAENPEYLG LDVPV)), preferably with an affinity sufficient for a diagnostic and / or therapeutic application. In particular embodiments, the antibody binds HER2 with a dissociation constant (KD) of ≤ 1µM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM. [000197] An example of an antibody binding to the cancer target molecule Her2 is trastuzumab (Genentech). Trastuzumab is a humanized antibody used inter alia for the treatment of breast cancer. In a particularly preferred embodiment, the anti-HER2 antibody is TPP-1015 (trastuzumab analogue). [000198] Further examples of antibodies binding to HER2 are, in addition to trastuzumab (INN 7637, CAS No.: RN: 180288-69-1) and pertuzumab (CAS No.: 380610-27-5), the antibodies disclosed in WO 2009 / 123894-A2, WO 200 / 8140603-A2 or in WO 2011 / 044368-A2. An example of an anti-HER2 conjugate is trastuzumab-emtansine (INN-No.9295). These antibodies and antigen-binding fragments can be used in the context of this invention. [000199] Particular preference is given in the context of this invention to the anti-HER2 antibody TPP- 1015 (analogous to trastuzumab). Anti-Tenascin-C antibodies [000200] According to the invention, it is possible to use anti-Tenascin-C antibodies. [000201] The expression “anti-Tenascin-C antibody” or “an antibody which binds specifically to Tenascin-C” relates to an antibody which binds the cancer target molecule TNC (NCBI Reference Sequence: NP_002151.2), preferably with an affinity sufficient for a diagnostic and / or therapeutic application. In particular embodiments, the antibody binds TNC with a dissociation constant (KD) of ≤ 1µM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM. [000202] An example of an antibody binding to the cancer target molecule TNC is the anti-human TNC recombinant antibody (F16-131l) specific to F16 (specific to the A1 domain of human Tenascin-C). Anti-PD-L1 antibodies [000203] According to the invention, it is possible to use anti-PD-L1 antibodies. [000204] The expression “anti-PD-L1 antibody” or “an antibody which binds specifically to PD-L1” relates to an antibody which binds the cancer target molecule PD-L1 (CD274; NCBI Reference: Gene ID: 29126) preferably with an affinity sufficient for a diagnostic and / or therapeutic application. InP368919WO / 59362-725.601 particular embodiments, the antibody binds PD-L1 with a dissociation constant (KD) of ≤ 1µM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, ≤ 0.1 nM, ≤ 0.01 nM, or ≤ 0.001 nM. Examples of antibodies which bind to PD-L1 are disclosed, for example, in KEGG DRUG Database [000205] These antibodies and antigen-binding fragments can be used in the context of this invention. [000206] Particular preference is given in the context of the present invention to the anti-PD-L1 antibody avelumab. This antibody and antigen-binding fragments thereof can be used with preference in the context of this invention. EXAMPLES Abbreviations and Acronyms A431NS human tumour cell line A549 human tumour cell line ABCBl ATP-binding cassette sub-family B member I (synonym for P-gp and MDRl) abs. absolute Ac acetyl ACN acetonitrile ADC antibody drug conjugate A2DC antibody 2 drug conjugate AMAS N-α-maleimidoacetoxysuccinimide ester aq. aqueous, aqueous solution ATP adenosine triphosphate BCRP breast cancer resistance protein, an efflux transporter BEP 2-bromo-l-ethylpyridinium tetrafluoroborate Boc tert-butoxycarbonyl br. broad (in NMR) Ex. Example BxPC3 human tumour cell line ca. circa, about C-DAR cysteine drug to antibody ratio (linker attached to cystein residues) Cl chemical ionization (in MS) D doublet (in NMR) D day(s) TLC thin-layer chromatography DCI direct chemical ionization (in MS) DCM dichloromethane Dd doublet of doublets (in NMR) DMAP 4-N,N-dimethylaminopyridine DME 12-dimethoxyethaneP368919WO / 59362-725.601 DMEM Dulbecco’s Modified Eagle Medium (standardized nutrient medium for cell culture) DMF N,N-dimethylformamide DMSO dimethyl sulphoxide DAR drug to antibody ratio DPBS, D-PBS, PBS Dulbecco’s phosphate-buffered salt solution PBS=DPBS=D-PBS, pEl 7.4, from Sigma, No D8537. Composition: 0.2 g KCl; 0.2 g KH2PO4 (anhyd); 8.0 g NaCl; 1.15 g Na2HPO4 (anhyd); made up ad I I with H2O Dt doublet of triplets (in NMR) DTT DL-dithiothreitol d. Th. of theory (in chemical yield) EDC N'-(3-dimethylaminopropyl)-N-ethylcarbodi- imide hydrochloride EI electron impact ionization (in MS) ELISA enzyme-linked immunosorbent assay eq. equivalent(s) ESI electrospray ionization (in MS) ESI-MicroTofq ESI-MicroTofq (name of the mass spectrometer with Tof=time of flight and q=quadrupol) FCS foetal calf serum Fmoc (9H-fluoren-9-ylmethoxy)carbonyl sat. saturated GTP guanosine-5'-triphosphate h hour(s) HATU O-(7-azabenzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluoro phosphate HCT-116 human tumour cell line HEPES 4-(2-hydroxyethyl)piperazine-l-ethane- sulphonic acid HOAc acetic acid HOAt 1-hydroxy-7-azabenzotriazole HOBt 1-hydroxy-1 H-benzotriazole hydrate HOSu N-hydroxysuccinimide HPLC high-pressure, high-performance liquid chromatography HT29 human tumour cell line IC50 half-maximal inhibitory concentration i.m. intramuscularly, administration into the muscle i.v. intravenously, administration into the vein K-DAR lysine drug to antibody ratio (linker attached to lysine residues) KPL-4 human tumour cell linesP368919WO / 59362-725.601 LC-MS liquid chromatography-coupled mass spectrometry LLC-PKl cells Lewis lung carcinoma pork kidney cell line L-MDR human MDRl transfected LLC-PKl cells m multiplet (in NMR) Me methyl MDRl Multidrug resistance protein I MeCN acetonitrile min minute(s) MOLM 13 human cancer cell line MS mass spectrometry MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl- 2H-tetrazolium bromide NCI-H292 human tumour cell line NCI-H69 human tumour cell line NMM N-methylmorpholine NMP N-methyl-2-pyrrolidinone NMR nuclear magnetic resonance spectrometry NMRI mouse strain originating from the Naval Medical Research Institute Nude mice experimental animals NSCLC non small cell lung cancer PBS phosphate-buffered salt solution Pd / C palladium on activated carbon P-gp P-glycoprotein, a transporter protein PNGaseF enzyme for cleaving sugar PSAR polysarcosine Quant quantitative (in yield) quart quartet (in NMR) quint quintet (in NMR) RT room temperature Rt retention time (in HPLC) s singlet (in NMR) s.c. subcutaneously, administration under the skin SCC-4 human tumour cell line SCC-9 human tumour cell line SCID mice test mice with severe combined immunodeficiency SK-HEP-I human tumour cell line t triplet (in NMR) TBAF tetra-n-butylammonium fluoride TCEP tris(2-carboxyethyl)phosphineP368919WO / 59362-725.601 TEMPO (2,2,6,6-tetramethylpiperidin-l-yl)oxyl tert tertiary TFA trifluoroacetic acid THF tetrahydrofuran T3P® 2,4,6-tripropyl-l,3,5,2,4,6-trioxatriphosphi- nane 2,4,6-trioxide UV ultraviolet spectrometry v / v volume to volume ratio (of a solution) Z benzyloxycarbonyl 786-0 human tumour cell line Amino Acid abbreviations Ala = Alanine Lys = Lysine Arg = Arginine Met = Methionine Asn = Asparagine Nva = Norvaline Asp = Aspartic acid Phe = Phenylalanine Cys = Cysteine Pro = Proline Glu = Glutamic acid Ser = Serine Gln = Glutamine Thr = Threonine Gly = Glycine Trp = Tryptophane His = Histidine Tyr = Tyrosine Ile = Isoleucine Val = Valine Leu = LeucineP368919WO / 59362-725.601 SYNTHESIS EXAMPLES Analytical Methods (LC-MS) [000207]Method 1 (LC-MS): System MS: Waters TOF instrument; System UPLC: Waters Acquity I- CLASS; Column: Waters, HSST3, 2.1 x 50 mm, C181.8 µm; Eluent A: 1 l Water + 0.01% Formic acid; Eluent B: 1 l Acetonitrile + 0.01% Formic acid; Gradient: 0.0 min 2% B → 0.5 min 2% B → 7.5 min 95% B → 10.0 min 95% B; Oven: 50°C; Flow: 1.00 ml / min; UV-Detection: 210 nm. [000208]Method 2 (LC-MS): System MS: Thermo Scientific FT-MS; System UHPLC+: Thermo Scientific UltiMate 3000; Column: Waters, HSST3, 2.1 x 75 mm, C18 1.8 µm; Eluent A: 1 l Water + 0.01% Formic acid; Eluent B: 1 l Acetonitrile + 0.01% Formic acid; Gradient: 0.0 min 10% B → 2.5 min 95% B → 3.5 min 95% B; Oven: 50°C; Flow: 0.90 ml / min; UV-Detection: 210 nm / Optimum Integration Path 210-300 nm. [000209]Method 3 (LC-MS): Instrument: Waters ACQUITY SQD UPLC System; Column: Waters Acquity UPLC HSS T31.8 µm 50 x 1 mm; Eluent A: 1 l Water + 0.25 ml 99% Formic acid , Eluent B: 1 l Acetonitrile + 0.25 ml 99% Formic acid; Gradient: 0.0 min 95% A → 6.0 min 5% A → 7.5 min 5% A Oven: 50°C; Flow: 0.35 ml / min; UV-Detection: 210 nm. [000210]Method 4 (LC-MS): System MS: Waters TOF instrument; System UPLC: Waters Acquity I- CLASS; Column: Waters Acquity UPLC HSS T31.8 µm 50 x 1 mm; Eluent A: 1 l Water + 0.100 ml 99% Formic acid, Eluent B: 1 l Acetonitrile + 0.100 ml 99% Formic acid; Gradient: 0.0 min 90% A →1.2 min 5% A → 2.0 min 5% A Oven: 50°C; Flow: 0.40 ml / min; UV-Detection: 210 nm. [000211]Method 5 (LC-MS): System MS: Waters TOF instrument; System UPLC: Waters Acquity I- CLASS; Column: Waters Acquity UPLC HSS T31.8 µm 50 x 1 mm; Eluent A: 1 l Water + 0.100 ml 99% Formic acid, Eluent B: 1 l Acetonitrile + 0.100 ml 99% Formic acid; Gradient: 0.0 min 95% A →6.0 min 5% A → 7.5 min 5% A Oven: 50°C; Flow: 0.35 ml / min; UV-Detection: 210 nm. [000212]Method 6 (LC-MS): System MS: Thermo Scientific FT-MS; System UHPLC+: Thermo Scientific Vanquish; Column: Waters, HSST3, 2.1 x 75 mm, C181.8 µm; Eluent A: 1 l Water + 0.01% Formic acid; Eluent B: 1 l Acetonitrile + 0.01% Formic acid; Gradient: 0.0 min 10% B → 2.5 min 95% B → 3.5 min 95% B; Oven: 50°C; Flow: 0.90 ml / min; UV-Detection: 210 nm. ADC Precursor molecules applied to the synthesis of A2DCs Precursors for ADCs Intermediate Q1 [000213](4S)-4,11-Diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinolin-4-yl N-{18-[(2,5-dioxopyrrolidin-1-yl)oxy]-14,18-dioxo-4,7,10-trioxa-13-azaoctadecan-1- oyl}-L-alpha-aspartyl-L-prolyl-L-valinateP368919WO / 59362-725.601[000214]Intermediate Q11 (360.0 mg, 358.2 µmol) was dissolved in DMF (30.0 ml). Disuccinimidyl glutarate (350.6 mg, 1074.6 µmol) was added. The reaction was stirred at RT for 1h and was then concentrated in vacuo. The residue was separated by prep. HPLC to give Intermediate Q1 (251 mg, 98% purity, 63% yield) as a light yellow foam. LC-MS (Method 1): Rt = 3.62 min; MS (ESIpos): m / z = 1102 [M+H]+. Intermediate Q2 [000215](4S)-4,11-Diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinolin-4-yl N-[19-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-14-oxo-4,7,10-trioxa-13-azanonadecan-1- oyl]-L-alpha-aspartyl-L-prolyl-L-valinateP368919WO / 59362-725.601[000216]To a solution of Intermediate Q11 (10mg, 9.95 µmol) in DMF (5.0 ml) were added N- succinimidyl 6-maleimidocaproate (6.14 mg, 19.9 µmol) and DIEA (6.93 µl, 39.8 µmol). The mixture was stirred at RT for 1h and concentrated in vacuo. The residue was purified by prep. HPLC and lyophilized to give Intermediate Q2 (9.0 mg, 93% purity, 78% yield) as a yellow amorphous residue. LC-MS (Method 2): R + t = 1.61 min; MS (ESIpos): m / z = 1084 [M+H] . Intermediate Q3 [000217](1S,9S)-1-Acetamido-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-{18-[(2,5-dioxopyrrolidin-1-yl)oxy]- 14,18-dioxo-4,7,10-trioxa-13-azaoctadecan-1-oyl}-L-alpha-aspartyl-L-prolyl-L-valinateP368919WO / 59362-725.601[000218]Intermediate Q3 was prepared in analogy as described for the synthesis of Intermediate Q1 starting with Intermediate Q14. Intermediate Q3 (2.36g, 95% purity, 21% yield) was obtained as a yellow foam. LC-MS (Method 3): R + t = 2.54 min; MS (ESIpos): m / z = 1203 [M+H] . Intermediate Q4 [000219](1S,9S)-1-Amino-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H- benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-[19-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)- 14-oxo-4,7,10-trioxa-13-azanonadecan-1-oyl]-L-alpha-aspartyl-L-prolyl-L-valinate trifluoroacetate (1:1)P368919WO / 59362-725.601[000220]Intermediate Q4 was prepared in analogy as described for the synthesis of Intermediate Q2 starting with Intermediate Q21. Intermediate Q4 (3.15mg, 100% purity, 27% yield) was obtained as a yellow foam. LC-MS (Method 1): R + t = 2.77 min; MS (ESIpos): m / z = 1143 [M+H] . Intermediate Q5 [000221](1S,9S)-1-Amino-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H- benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-{18-[(2,5-dioxopyrrolidin-1-yl)oxy]-14,18- dioxo-4,7,10-trioxa-13-azaoctadecan-1-oyl}-L-alpha-aspartyl-L-prolyl-L-valinateP368919WO / 59362-725.601[000222]Intermediate Q5 was prepared in analogy as described for the synthesis of Intermediate Q1 starting with Intermediate Q21. Intermediate Q5 (6.20mg, 100% purity, 52% yield) was obtained as a light yellow foam. LC-MS (Method 1): R + t = 2.63 min; MS (ESIpos): m / z = 1161 [M+H] . Intermediate Q6 [000223](1S,9S)-1-(Dimethylamino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-{18-[(2,5-dioxopyrrolidin-1-yl)oxy]- 14,18-dioxo-4,7,10-trioxa-13-azaoctadecan-1-oyl}-L-alpha-aspartyl-L-prolyl-L-valinate[000224]Intermediate Q6 was prepared in analogy as described for the synthesis of Intermediate Q1 starting with Intermediate Q27. Intermediate Q6 (4.45mg, 94% purity, 38% yield) was obtained as a yellow foam. LC-MS (Method 1): R = 2 - t .92 min; MS (ESIneg): m / z = 1187 [M-H] . Intermediate Q7 [000225](1S,9S)-1-(Diethylamino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-{18-[(2,5-dioxopyrrolidin-1-yl)oxy]- 14,18-dioxo-4,7,10-trioxa-13-azaoctadecan-1-oyl}-L-alpha-aspartyl-L-prolyl-L-valinate[000226]Intermediate Q7 was prepared in analogy as described for the synthesis of Intermediate Q1 starting with Intermediate Q22. Intermediate Q7 (3.81mg, 99% purity, 35% yield) was obtained as a yellow foam. LC-MS (Method 3): R - t = 2.33 min; MS (ESIneg): m / z = 1216 [M-H] . Intermediate Q8 [000227](7S)-7-Ethyl-8,11-dioxo-7,8,11,13-tetrahydro-10H-[1,3]dioxolo[4,5- g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl N-[19-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-14-oxo- 4,7,10-trioxa-13-azanonadecan-1-oyl]-L-alpha-aspartyl-L-prolyl-L-valinate[000228]Intermediate Q8 was prepared in analogy as described for the synthesis of Intermediate Q2 starting with Intermediate Q32. Intermediate Q8 (3.60mg, 92% purity, 31% yield) was obtained as an amorphous residue. LC-MS (Method 4): Rt= 0.83 min; MS (ESIpos): m / z = 1100 [M+H]+. Intermediate Q9 [000229](3R)-3-({[4-({[2-({5-[(2,5-dioxopyrrolidin-1-yl)oxy]-5- oxopentanoyl}amino)ethyl]carbamoyl}amino)phenyl]carbamoyl}amino)-3-{3-[({3- [(propylcarbamoyl)amino]phenyl}sulfonyl)amino]phenyl}propanoic acid H H[000230]Intermediate Q9 was prepared in analogy as described for the synthesis of Intermediate Q1 starting with Intermediate Q36. Intermediate Q9 (6.73mg, 96% purity, 57% yield) was obtained as a colorless foam LC-MS (Method 3): Rt= 201 min; MS (ESIpos): m / z = 852 [M+H]+P368919WO / 59362-725.601 Intermediate Q10 [000231](1S,9S)-1-Acetamido-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-[19-(2,5-dioxo-2,5-dihydro-1H- pyrrol-1-yl)-14-oxo-4,7,10-trioxa-13-azanonadecan-1-oyl]-L-alpha-aspartyl-L-prolyl-L-valinate[000232]Intermediate Q10 was prepared in analogy as described for the synthesis of Intermediate Q2 starting with Intermediate Q14. Intermediate Q10 (5.0mg, 89% purity, 44% yield) was obtained as an amorphous residue. LC-MS (Method 1): R + t = 3.58 min; MS (ESIpos): m / z = 1185 [M+H] . Intermediate Q11 [000233](4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinolin-4-yl N-(3-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}propanoyl)-L-alpha-aspartyl-L-prolyl-L- valinate trifluoroacetate (1:1)[000234]Step 1: (2S)-1-[(19S)-19-(2-tert-butoxy-2-oxoethyl)-2,2-dimethyl-4,17,20-trioxo-3,8,11,14- tetraoxa-5,18-diazaicosan-20-yl]pyrrolidine-2-carboxylic acid (Intermediate Q39) (50.0 g, 84.8 mmol) was dissolved in 900 ml DMF and the solution was cooled down to 0°C.1.15 eq 1-(3- Dimethylaminopropyl)-3-ethylcarbodiimidhydrochloride (17.0 g, 88.8 mmol) and 1.3 eq Ethyl cyanohydroxyiminoacetate (14.9 g, 105 mmol) and trifluoroacetic acid—(4S)-4,11- diethyl-3,14-dioxo- 3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indolizino[1,2-b]quinolin-4-yl L-valinate (1 / 1) (Intermediate Q40) (47.6 g, 80.8 mmol) were added and subsequently 3.0 eq (42 ml, 240 mmol) N,N- diisopropylethylamine was added dropwise. The mixture was stirred 10 overnight at 0°C. It was diluted with 4L EtOAc and the organic layer was washed with 10% aqueous citric acid (2 x 2l), with 10% aqueous NaHCO3-solution (2 x 2l) and with saturated aqueous NaCl-solution (2 x 3l). Subsequently it was dried over Mg2SO4, filtered and concentrated. The residue was dissolved in 250ml DCM and purified using flash chromatography (DCM:MeOH 100:1). Relevant fractions were collected and evaporated in vacuo to yield OtBu-Intermediate Q11 as a yellow foam (72.8 g, 98 % purity, 84 % yield). [000235]Step 2: 12 g (11.5 mmol) of tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(4S)-4,11-diethyl-3,14- dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl]oxy}-3- methyl-1- oxobutan-2-yl]carbamoyl} pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo-3,8,11,14- tetraoxa-5,18- diazahenicosan-21-oate (OtBu-Intermediate Q11) was dissolved in 100 ml of 20 dichloromethane and 40 ml of anhydrous trifluoroacetic acid was added and the solution was stirred for 2 days at rt. After concentrating in vacuo 50 ml toluene were added and again evaporated. The residue was dissolved in 50 ml DCM / MeOH and subsequently poured into 600 - 184 - ml of diethyl ether. The precipitating product(Intermediate Q11) was filtered, washed with diethyl ether and dried in vacuo to give the title compound (11.7 g, 95% purity, 97 % yield). Intermediate Q14 [000236](1S,9S)-1-Acetamido-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-(3-{2-[2-(2-aminoethoxy) ethoxy] ethoxy}propanoyl)-L-alpha-aspartyl-L-prolyl-L-valinate—trifluoroacetic acid (1 / 1)[000237]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(1S,9S)-1-acetamido-9-ethyl-5-fluoro-4-methyl-10,13- dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl]oxy}- 3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo-3,8,11,14- tetraoxa-5,18-diazahenicosan-21-oate (Intermediate Q15) (45.0 mg, 39.2 µmol) was dissolved in DCM (4.7 mL), then TFA (940 µl) was added. The reaction was stirred for 4h at rt and then concentrated in vacuo. The residue was dissolved in ACN / water and freeze-dried to give a yellow oil, which was purified by prep. HPLC to give Intermediate Q14 as two rotamers as a yellow foam (14.2 mg, 100% purity, 32% yield). LC-MS(Method 2): Rt= 1.98 min; MS (ESIpos): m / z = 992 [M+H]+. Intermediate Q15 [000238]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(1S,9S)-1-acetamido-9-ethyl-5-fluoro-4-methyl-10,13- dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl]oxy}- 3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo-3,8,11,14- tetraoxa-5,18-diazahenicosan-21-oateP368919WO / 59362-725.601[000239]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(1S,9S)-1-amino-9-ethyl-5-fluoro-4-methyl-10,13- dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl]oxy}- 3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo-3,8,11,14- tetraoxa-5,18-diazahenicosan-21-oate (Intermediate Q16) (59.9mg, 0.053 mmol) was dissolved in DMF (5.0 ml). Ac-OSu (16.8 mg, 0.107 mmol) and DIEA (27.6 mg, 213 µmol) were added. The reaction was stirred at RT for 20 h and then concentrated in vacuo. The residue was separated by prep. HPLC to give Intermediate Q15 (35.0mg, 100% purity, 57% yield) as a light yellow foam. LC-MS(Method 3): Rt = 3.64 min; MS (ESIpos): m / z = 1148 [M+H]+. Intermediate Q16 [000240]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(1S,9S)-1-amino-9-ethyl-5-fluoro-4-methyl-10,13- dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl]oxy}- 3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo-3,8,11,14- tetraoxa-5,18-diazahenicosan-21-oate[000241]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(1S,9S)-1-{[(benzyloxy)carbonyl]amino}-9-ethyl-5- fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7] indolizino[1,2-b]quinolin-9-yl]oxy}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2- dimethyl-4,17-dioxo-3,8,11,14-tetraoxa-5,18-diazahenicosan-21-oate (350 mg, 282 µmol) (Intermediate Q17) was dissolved in ethanol (50 ml). The catalyst Pd / C 10% (50.0 mg) was added. The reaction was hydrogenated for 2 hour at standard pressure. The catalyst was filtered off and the filtrate was concentrated on a rotary evaporator. The residue was dissolved in ACN / H2O and freeze-dried to give Intermediate Q16 (287 mg, 86 % purity, 79 % yield) as a colorless foam. LC-MS (Method 1): Rt = 3.59 min; MS (ESIpos): m / z = 1107 [M+H]+. Intermediate Q17 [000242]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(1S,9S)-1-{[(benzyloxy)carbonyl]amino}-9-ethyl-5- fluoro-4- methyl- 10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano [3',4':6,7] indolizino [1,2-b]quinolin-9-yl]oxy}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl- 4,17-dioxo-3,8,11,14-tetraoxa-5,18-diazahenicosan-21-oateP368919WO / 59362-725.601[000243]To a solution of trifluoroacetic acid—(1S,9S)-1-{[(benzyloxy)carbonyl]amino}-9-ethyl-5- fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano [3',4':6,7]indolizino [1,2-b]quinolin-9-yl L-valinate (1 / 1) (Intermediate Q18) (44.0 mg, 98 % purity, 54.9 µmol) and (2S)-1- [(19S)-19-(2-tert-butoxy-2-oxoethyl)-2,2-dimethyl-4,17,20-trioxo-3,8,11,14-tetraoxa-5,18-diazaicosan- 20-yl]pyrrolidine-2-carboxylic acid (Intermediate Q39) (38.9 mg, 65.9 µmol) in DMF (8.0 mL) were added HATU ((27.1 mg, 71.4 µmol) and DIEA (29 µl, 160 µmol). The solution was stirred at rt for 30 min and then concentrated in vacuo. The residue was purified by prep. HPLC, then lyophilized to give Intermediate Q17 (50.0 mg, 100% purity, 73% yield) as a yellow foam. LC-MS(Method 3): Rt = 4.28 min; MS (ESIpos): m / z = 1239 [M+H]+. Intermediate Q18 [000244]Trifluoroacetic acid—(1S,9S)-1-{[(benzyloxy)carbonyl] amino}-9-ethyl-5-fluoro-4-methyl- 10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de] pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl L-valinate (1 / 1)P368919WO / 59362-725.601[000245](1S,9S)-1-{[(Benzyloxy)carbonyl]amino}-9-ethyl-5-fluoro-4-methyl-10,13-dioxo- 2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-(tert- butoxycarbonyl)-L-valinate (Intermediate Q19) (44.0 mg, 98 % purity, 55.9 µmol) was dissolved in DCM (6.3 mL), TFA (1.3 mL) was added and the reaction was stirred at RT for 30min. The reaction was concentrated in vacuo, the residue was dissolved in ACN / H2O and lyophilized to give Intermediate Q18 (44.0 mg, 98% purity, 98% yield). LC-MS(Method 2): R + t = 2.56 min; MS (ESIpos): m / z = 669 [M+H] . Intermediate Q19 [000246](1S,9S)-1-{[(Benzyloxy)carbonyl]amino}-9-ethyl-5-fluoro-4-methyl-10,13-dioxo- 2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7] indolizino[1,2-b]quinolin-9-yl N-(tert- butoxycarbonyl)-L-valinateP368919WO / 59362-725.601 [000247]Benzyl [(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl]carbamate (Intermediate Q20) (50.0 mg, 87.8 µmol) was added to DCM (20.0 mL), then tert-butyl (4S)-2,5-dioxo-4-(propan-2-yl)-1,3- oxazolidine-3-carboxylate (51.2 mg, 211 µmol) and DMAP (19.3 mg, 158 µmol; CAS: 1122-58-3) were added. The mixture was refluxed while stirring for 7h. The reaction was concentrated in vacuo. Water was added, upon which the product precipitated. The mixture was filtered, the filter residue was very poorly soluble, but was soluble in DMSO + ACN or MeOH / DCM. The compound was purified by prep. HPLC to give Intermediate Q19 (54.0 g, 98% purity, 78% yield). LC-MS(Method 2): Rt = 2.48 min; MS (ESIpos): m / z = 769 [M+H]+. Intermediate Q20 [000248]Benzyl [(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl]carbamate[000249]Methanesulfonic acid—(1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15- hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (1 / 1) (exatecan mesylate) (50.0 mg, 94.1 µmol) was dissolved in DMF (10.0 mL), then 1- {[(benzyloxy)carbonyl]oxy}pyrrolidine-2,5-dione (28.1 mg, 113 µmol) and DIEA (49 µl, 280 µmol) were added. The reaction was stirred overnight at rt. The reaction was concentrated in vacuo and the residue was mixed with ACN / H2O / DMF. A solid precipitated, which was filtered off, dried and purified by prep. HPLC to give Intermediate Q20 (50.0 mg, 92% purity, 87% yield) as a white foam. LC- MS(Method 1): Rt= 3.21 min; MS (ESIpos): m / z = 570 [M+H]+. Intermediate Q21 [000250](1S,9S)-1-Amino-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H- benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-(3-{2-[2-(2-aminoethoxy) ethoxy]ethoxy}propanoyl)-L-alpha-aspartyl-L-prolyl-L-valinate trifluoroacetic acid (1 / 1)P368919WO / 59362-725.601[000251]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(1S,9S)-1-amino-9-ethyl-5-fluoro-4-methyl-10,13- dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl]oxy}- 3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo-3,8,11,14- tetraoxa-5,18-diazahenicosan-21-oate (287 mg, 86 % purity, 223 µmol) (Intermediate Q16) was dissolved in DCM (30 ml), then TFA (5.0 ml) was added. The reaction was stirred for 1 hour at RT, then concentrated in vacuo. The residue was purified by prep. HPLC to give Intermediate Q21 (216 mg, 95 % purity, 87 % yield) as a yellow foam. LC-MS (Method 1): Rt = 2.04 min; MS (ESIpos): m / z = 950 [M+H]+. Intermediate Q22 [000252](1S,9S)-1-(diethylamino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-(3-{2-[2-(2- aminoethoxy)ethoxy]ethoxy}propanoyl)-L-alpha-aspartyl-L-prolyl-L-valinate trifluoroacetic acid (1 / 1)P368919WO / 59362-725.601[000253]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(1S,9S)-1-(diethylamino)-9-ethyl-5-fluoro-4-methyl- 10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9- yl]oxy}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo- 3,8,11,14-tetraoxa-5,18-diazahenicosan-21-oate (18.1 mg, 15.6 µmol) (Intermediate Q23) was dissolved in DCM (5.0 ml), then TFA (2.0 ml) was added. The reaction was stirred for 1 hour at RT and concentrated in vacuo. The residue was dissolved in ACN / H2O and freeze-dried to give Intermediate Q22 (17.0 mg, 95 % purity, 92 % yield) as a light yellow foam. LC-MS (Method 1): Rt= 2.65 min; MS (ESIpos): m / z = 1006 [M+H]+. Intermediate Q23 [000254]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(1S,9S)-1-(diethylamino)-9-ethyl-5-fluoro-4-methyl- 10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9- yl]oxy}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo- 3,8,11,14-tetraoxa-5,18-diazahenicosan-21-oateP368919WO / 59362-725.601[000255]Trifluoroacetic acid (1S,9S)-1-(diethylamino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo- 2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl L-valinate (1 / 1) (14.0 mg, 19.9 µmol) (Intermediate Q24) was dissolved in DMF (5.0 ml). (2S)-1-[(19S)-19-(2- Tert-butoxy-2-oxoethyl)-2,2-dimethyl-4,17,20-trioxo-3,8,11,14-tetraoxa-5,18-diazaicosan-20- yl]pyrrolidine-2-carboxylic acid (14.1 mg, 23.8 µmol) (Intermediate Q39), HATU (9.82 mg, 25.8 µmol; CAS-RN:[148893-10-1]) and N,N-diisopropylethylamine (10 µl, 60 µmol; CAS-RN:[7087-68-5]) were added. The reaction was stirred at RT for 1 hour and concentrated in vacuo. The residue was purified by prep. HPLC to give Intermediate Q23 (18.1 mg, 91 % purity, 71 % yield) as a light yellow foam. LC-MS (Method 1): R + t = 4.69 min; MS (ESIpos): m / z = 1163 [M+H] . Intermediate Q24 [000256]Trifluoroacetic acid (1S,9S)-1-(diethylamino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo- 2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl L-valinate (1 / 1)P368919WO / 59362-725.601[000257](1S,9S)-1-(Diethylamino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-(tert-butoxycarbonyl)-L-valinate (15.0 mg, 21.7 µmol) (Intermediate Q25) was dissolved in DCM (7.5 ml), then TFA (1.5 ml) was added. The reaction was stirred for 1 hour at RT and concentrated in vacuo. The residue was dissolved in ACN / H2O and freeze-dried to give Intermediate Q24 (14.0 mg, 100 % purity, 91 % yield) as a beige foam. LC-MS (Method 1): R + t = 2.54 min; MS (ESIpos): m / z = 591 [M+H] . Intermediate Q25 [000258](1S,9S)-1-(Diethylamino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-(tert-butoxycarbonyl)-L-valinate[000259](1S,9S)-1-(Diethylamino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro- 10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (20.0 mg, 40.7 µmol) (Intermediate Q26) was dissolved in DCM (5.0 ml). Boc-Val-NCA: tert-butyl (4S)-2,5-dioxo-4- (propan-2-yl)-13-oxazolidine-3-carboxylate (238 mg 976 µmol) and DMAP (895 mg 732 µmol;CAS-RN:[1122-58-3]) were added. The reaction was stirred for 22h at reflux and then concentrated in vacuo. The residue was separated by prep. HPLC to give Intermediate Q25 (15.0 mg, 100 % purity, 53 % yield) as a beige foam. LC-MS (Method 1): R = 4.74 min; MS (ESIpos): m / z = 691 [M+H + t ] . Intermediate Q26 [000260](1S,9S)-1-(Diethylamino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro- 10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione[000261]Methanesulfonic acid (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15- hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (1 / 1) (exatecan mesylate) (25.0 mg, 47.0 µmol) was dissolved in DCM (8.0 mL). Sodium hydrogen carbonate (7.90 mg, 94.1 µmol) was added. Acetaldehyde (21 µl, 380 µmol) was added and the reaction was stirred for 1 h at reflux. Acetaldehyde (21 µl, 380 µmol) was added twice more and the reaction was stirred for 1h at reflux. The reaction was cooled to 0°C, then sodium triacetoxyborohydride (39.9 mg, 188 µmol) was added. The reaction was stirred at RT for 20h. The reaction was concentrated in vacuo and purified by prep. HPLC to give Intermediate Q26 (20.5 mg, 100% purity, 89% yield) as a colorless foam. LC-MS (Method 1): Rt = 1.22 min; MS (ESIpos): m / z = 492 [M+H]+.1H-NMR (600 MHz, DMSO-d6): δ [ppm] = 9.35-9.28 (m, 2H), 7.89 (s, 1H), 7.88 (s, 1H), 7.77-7.67 (m, 1H), 7.35 (s, 2H), 7.31-7.28 (m, 1H), 6.62- 6.49 (m, 2H), 5.54-5.48 (m, 2H), 5.46-5.44 (m, 1H), 5.48-5.39 (m, 5H), 5.31 (br d, 2H), 5.36-5.23 (m, 1H), 5.07 (br d, 2H), 4.52-4.46 (m, 1H), 3.43-3.27 (m, 7H), 3.24-3.11 (m, 5H), 3.07-2.98 (m, 2H), 2.98- 2.89 (m, 1H), 2.83-2.79 (m, 1H), 2.65-2.56 (m, 2H), 2.40 (br s, 5H), 2.44-2.28 (m, 5H), 2.23-2.15 (m, 2H), 2.07-1.95 (m, 1H), 1.93-1.81 (m, 6H), 1.34 (br t, 6H), 1.18-1.03 (m, 11H), 0.89 (br t, 8H). Intermediate Q27 [000262](1S,9S)-1-(Dimethylamino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-(3-{2-[2-(2- aminoethoxy)ethoxy]ethoxy}propanoyl)-L-alpha-aspartyl-L-prolyl-L-valinate trifluoroacetic acidP368919WO / 59362-725.601[000263]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(1S,9S)-1-(dimethylamino)-9-ethyl-5-fluoro-4-methyl- 10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9- yl]oxy}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo- 3,8,11,14-tetraoxa-5,18-diazahenicosan-21-oate (36.0 mg, 31.7 µmol) (Intermediate Q28) was dissolved in DCM (5.0 ml), then TFA (2.5 ml) was added. The reaction was stirred for 3 hour at RT and concentrated in vacuo. The residue was dissolved in ACN / H2O and freeze-dried to give Intermediate Q27 (33.0 mg, 100 % purity, 95 % yield) as a light yellow foam. LC-MS (Method 2): Rt = 0.91 min; MS (ESIneg): m / z = 976 [M-H]- . Intermediate Q28 [000264]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(1S,9S)-1-(dimethylamino)-9-ethyl-5-fluoro-4-methyl- 10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9- yl]oxy}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo- 3,8,11,14-tetraoxa-5,18-diazahenicosan-21-oateP368919WO / 59362-725.601[000265]Trifluoroacetic acid (1S,9S)-1-(dimethylamino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo- 2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl L-valinate (1 / 1) (23.0 mg, 34.0 µmol) (Intermediate Q29) was dissolved in DMF (8.6 ml). (2S)-1-[(19S)-19-(2- Tert-butoxy-2-oxoethyl)-2,2-dimethyl-4,17,20-trioxo-3,8,11,14-tetraoxa-5,18-diazaicosan-20- yl]pyrrolidine-2-carboxylic acid (24.1 mg, 40.8 µmol) (Intermediate Q39), HATU (16.8 mg, 44.2 µmol; CAS-RN:[148893-10-1]) and N,N-diisopropylethylamine (18 µl, 100 µmol; CAS-RN:[7087-68-5]) were added. The reaction was stirred at RT for 1 hour and then concentrated in vacuo. The residue was separated by prep. HPLC to give Intermediate Q28 (36.0 mg, 99 % purity, 92 % yield) as a light yellow foam. LC-MS (Method 3): Rt= 3.03 min; MS (ESIpos): m / z = 1134 [M+H]+. Intermediate Q29 [000266]Trifluoroacetic acid (1S,9S)-1-(dimethylamino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo- 2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl L-valinate (1 / 1)P368919WO / 59362-725.601[000267](1S,9S)-1-(Dimethylamino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-(tert-butoxycarbonyl)-L-valinate (23.0 mg, 34.7 µmol) (Intermediate Q30) was dissolved in DCM (5.2 ml), then TFA (1.0 ml) was added. The reaction was stirred for 1 hour at RT and concentrated in vacuo. The residue was dissolved in ACN / H2O and freeze-dried to give Intermediate Q29 (23.0 mg, 97 % purity, 95 % yield) as a beige foam. LC-MS (Method 1): R = 2.06 min; MS (ESIpos): m + t / z = 563 [M+H] . Intermediate Q30 [000268](1S,9S)-1-(Dimethylamino)-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro- 1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-(tert-butoxycarbonyl)-L-valinate[000269](1S,9S)-1-(Dimethylamino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro- 10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (40.0 mg, 86.3 µmol) (Intermediate Q31) was dissolved in DCM (33 ml). Boc-Val-NCA: tert-butyl (4S)-2,5-dioxo-4-(propan- 2-yl)-1,3-oxazolidine-3-carboxylate (50.4 mg, 207 µmol) and 4-(Dimethylamino) pyridine (DMAP): (19.0 mg, 155 µmol; CAS-RN:[1122-58-3]) were added. The reaction was stirred for 22 hours at reflux and then concentrated in vacuo. The residue was separated by prep. HPLC to give Intermediate Q30 (23.0 mg, 100 % purity, 40 % yield)) as a beige foam. LC-MS (Method 1): Rt = 3.96 min; MS (ESIpos): m / z = 663 [M+H]+.P368919WO / 59362-725.601 Intermediate Q31 [000270](1S,9S)-1-(Dimethylamino)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro- 10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione[000271]Methanesulfonic acid (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15- hexahydro-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (1 / 1) (Exatecan mesylate) (50.0 mg, 94.1 µmol) was dissolved in methanol (10 ml). Sodium hydrogen carbonate (15.8 mg, 188 µmol; CAS-RN:[144-55-8]) was added. Formaldehyde (61.1 mg, 37 % purity, 752 µmol) was added and the reaction was stirred for 1 hour at 50°C.2X more formaldehyde was added and stirring was continued for 1h at 50°C. The reaction was then cooled to 0 ° C and sodium triacetoxyborohydride (79.7 mg, 376 µmol; CAS-RN:[56553-60-7]) was added. The reaction was stirred at RT for 2h and then concentrated in vacuo. The residue was dissolved in ACN / H2O, the precipitate was filtered off with suction and the filtrate was purified by prep. HPLC. The precipitate was dissolved in ACN / H2O and freeze-dried to give Intermediate Q31 (15.0 mg, 87 % purity, 30 % yield) as a colourless foam. LC-MS (Method 2): Rt= 0.94 min; MS (ESIpos): m / z = 464 [M+H]+. Intermediate Q32 [000272](7S)-7-Ethyl-8,11-dioxo-7,8,11,13-tetrahydro-2H,10H-[1,3]dioxolo[4,5-g]pyrano[3',4':6,7] indolizino[1,2-b]quinolin-7-yl N-(3-{2-[2-(2-aminoethoxy) ethoxy] ethoxy}propanoyl)-L-alpha-aspartyl- L-prolyl-L-valinate—trifluoroacetic acid (1 / 1)P368919WO / 59362-725.601[000273]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(7S)-7-ethyl-8,11-dioxo-7,8,11,13-tetrahydro-2H,10H- [1,3]dioxolo[4,5-g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl]oxy}-3-methyl-1-oxobutan-2- yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa-5,18-diazahenicosan- 21-oate (Intermediate Q33) (37.0 mg, 34.8 µmol) was dissolved in DCM (5.0 mL) and TFA (1.0 mL) was added. The reaction was stirred for 30min at rt and then concentrated in vacuo. The residue was dissolved in ACN / H2O and freeze-dried to give Intermediate Q32 (36.0 mg, 100% purity, 101% yield) as a colourless foam. LC-MS(Method 3): R + t = 2.66 min; MS (ESIpos): m / z = 907 [M+H] . Intermediate Q33 [000274]Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-{[(7S)-7-ethyl-8,11-dioxo-7,8,11,13-tetrahydro-2H,10H- [1,3]dioxolo[4,5-g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl]oxy}-3-methyl-1-oxobutan-2- yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa-5,18-diazahenicosan- 21-oateP368919WO / 59362-725.601[000275]Trifluoroacetic acid—(7S)-7-ethyl-8,11-dioxo-7,8,11,13-tetrahydro-2H,10H-[1,3]dioxolo[4,5- g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl L-valinate (1 / 1) (Intermediate Q34) (25.0 mg, 41.3 µmol) was dissolved in DMF (5.0 mL). (2S)-1-[(19S)-19-(2-tert-butoxy-2-oxoethyl)-2,2-dimethyl- 4,17,20-trioxo-3,8,11,14-tetraoxa-5,18-diazaicosan-20-yl]pyrrolidine-2-carboxylic acid (Intermediate Q39) (29.2 mg, 49.5 µmol), HATU (23.5 mg, 61.9 µmol) and DIEA (22 µl, 120 µmol) were added. The reaction was stirred at rt for 1h and then concentrated in vacuo. The residue was purified by prep. HPLC to give Intermediate Q33 (20.9 mg, 100% purity, 48% yield) as a light, yellow foam. LC-MS(Method 3): R + t = 1.98 min; MS (ESIpos): m / z = 1063 [M+H] . Intermediate Q34 [000276]Trifluoroacetic acid—(7S)-7-ethyl-8,11-dioxo-7,8,11,13-tetrahydro-2H,10H-[1,3]dioxolo[4,5- g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl L-valinate (1 / 1)P368919WO / 59362-725.601[000277](7S)-7-Ethyl-8,11-dioxo-7,8,11,13-tetrahydro-2H,10H-[1,3]dioxolo[4,5- g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl N-(tert-butoxycarbonyl)-L-valinate (Intermediate Q35) (50.0 mg, 84.5 µmol) was dissolved in DCM (15.0 mL), then TFA (3.0 mL) was added. The reaction was stirred for 30min at rt. The reaction was concentrated in vacuo and the residue was dissolved in ACN / H2O and freeze-dried to give Intermediate Q34 (10.2 mg, 82% purity, 16% yield) as a colourless foam. LC-MS(Method 1): R = 1.69 min; MS (ES + t Ipos): m / z = 492 [M+H] . ¹H-NMR (500 MHz, DMSO-d6) δ [ppm]: 0.935 (1.23), 0.949 (2.16), 0.964 (1.23), 1.029 (1.70), 1.043 (1.71), 1.085 (1.20), 1.097 (2.50), 1.111 (1.69), 1.141 (1.17), 1.155 (1.18), 1.232 (2.32), 1.248 (12.26), 1.262 (16.00), 1.276 (8.18), 2.217 (0.48), 2.232 (0.62), 2.248 (0.53), 2.731 (0.46), 2.891 (0.49), 3.125 (0.41), 3.131 (1.08), 3.140 (1.11), 3.146 (1.10), 3.154 (1.06), 3.594 (0.54), 3.602 (0.60), 3.607 (0.95), 3.615 (1.00), 3.620 (1.20), 3.628 (1.20), 3.633 (1.00), 3.641 (0.96), 3.646 (0.61), 3.654 (0.57), 5.243 (1.37), 5.260 (0.58), 5.520 (1.14), 5.527 (1.87), 6.293 (2.87), 7.126 (1.11), 7.231 (1.55), 7.431 (1.66), 7.531 (1.62), 7.537 (1.18), 8.387 (0.53), 8.481 (1.09), 8.492 (0.80), 8.574 (0.50). Intermediate Q35 [000278](7S)-7-Ethyl-8,11-dioxo-7,8,11,13-tetrahydro-2H,10H-[1,3]dioxolo[4,5- g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl N-(tert-butoxycarbonyl)-L-valinateP368919WO / 59362-725.601 [000279](7S)-7-ethyl-7-hydroxy-2H,10H-[1,3]dioxolo[4,5-g]pyrano[3',4':6,7]indolizino[1,2- b]quinoline-8,11(7H,13H)-dione (10,11-methylenedioxy-camptothecin) (20 mg, 0.051 mmol) was dissolved in DCM (5 mL), then tert-butyl (4S)-2,5-dioxo-4-(propan-2-yl)-1,3-oxazolidine-3-carboxylate (24.8 mg, 0.102 mmol) and DMAP (4.80 mg, 0.039 mmol) were added. The reaction was stirred for 12h at rt. The same amounts of tert-butyl (4S)-2,5-dioxo-4-(propan-2-yl)-1,3-oxazolidine-3-carboxylate and DMAP were added again to the reaction and the reaction was stirred for a further 4h. The reaction was concentrated in vacuo and purified by prep. HPLC to give Intermediate Q35 (7.20 mg, 80% purity, 19% yield) as an almost colourless foam. LC-MS(Method 3): R = 3.47 mi + t n; MS (ESIpos): m / z = 592 [M+H] . Intermediate Q36 [000280](3R)-3-{[(4-{[(2-Aminoethyl)carbamoyl]amino}phenyl)carbamoyl]amino}-3-{3-[({3- [(propylcarbamoyl)amino]phenyl}sulfonyl)amino]phenyl}propanoic acid trifluoroacetate (1:1)[000281](3R)-3-[[4-[2-(tert-butoxycarbonylamino)ethylcarbamoylamino]phenyl]carbamoylamino]-3- [3-[[3-(propylcarbamoylamino)phenyl]sulfonylamino]phenyl]propanoic acid (Intermediate Q37) was dissolved in DCM (5.0 ml), then TFA (1.0 ml) was added. The reaction was stirred for 1h at RT. The reaction was concentrated at RT on the oil pump. The resulting residue was dissolved in ACN / water and freeze-dried to give Intermediate Q36 (29.0 mg, 100% purity, 92% yield) as a colorless foam. LC- MS(Method 2): R = 0.93 min; MS (ESIpos): m / z = 641 [ + t M+H] . Intermediate Q37 [000282](3R)-3-[({4-[({2-[(tert- butoxycarbonyl)amino]ethyl}carbamoyl)amino]phenyl}carbamoyl)amino]-3-{3-[({3- [(propylcarbamoyl)amino]phenyl}sulfonyl)amino]phenyl}propanoic acidP368919WO / 59362-725.601[000283]Intermediate Q38 was dissolved in DMF (10.0 ml). N-Boc-ethylenediamine (17.8mg, 111.2 µmol) and DIEA (96.8 µl, 555.8 µmol) were added. The reaction was stirred for 15 minutes at RT and then concentrated in vacuo. The residue was separated by prep. HPLC to give Intermediate Q37 (31.0mg, 97% purity, 73% yield) as a beige foam. LC-MS(Method 2): Rt = 1.45 min; MS (ESIpos): m / z = 741 [M+H]+. Intermediate Q38 [000284](3R)-3-{[(4-{[(4-Nitrophenoxy)carbonyl]amino} phenyl)carbamoyl]amino}-3-{3-[({3- [(propylcarbamoyl)amino]phenyl} sulfonyl)amino]phenyl}propanoic acid[000285]The synthesis of Intermediate Q38 has been described in WO2020 / 094471. Intermediate Q39: [000286](2S)-1-[(19S)-19-(2-tert-butoxy-2-oxoethyl)-2,2-dimethyl-4,17,20-trioxo-3,8,11,14-tetraoxa- 5,18-diazaicosan-20-yl]pyrrolidine-2-carboxylic acidP368919WO / 59362-725.601[000287]Intermediate Q39 was synthesized using classical methods of peptide synthesis starting with the coupling of Z-Asp(OtBu)-OH with benzyl L-prolinate hydrochloride (1:1) in THF in the presence of T3P and DIPEA and subsequent removal of the Z-protecting group as well as the benzyl ester by hydrogenolysis over Pd / C to give (2S)-1-[(2S)-2-amino-4-tert-butoxy-4-oxobutanoyl] pyrrolidine-2- carboxylic acid. This partially protected dipeptide was acylated with tert-butyl{2-[2-(2-{3-[(2,5- dioxopyrrolidin-1-yl)oxy]-3-oxopropoxy}ethoxy)ethoxy]ethyl} carbamate to give the title compound. Tert-butyl{2-[2-(2-{3-[(2,5-dioxopyrrolidin-1-yl)oxy]-3-oxopropoxy}ethoxy) ethoxy]ethyl}carbamate was previously prepared by reacting 2,2-dimethyl-4-oxo-3,8,11,14-tetraoxa-5-azaheptadecan-17-oic acid with N-Hydroxysuccinimide in dioxane in the presence of EDCI. LC-MS: Rt = 0.81 min; MS (ESIpos): m / z = 590 [M+H]+. Intermediate Q40: [000288](4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indolizino[1,2- b]quinolin-4-yl L-valinate • trifluoroacetic acid (1:1)[000289]2.59 g (10.6 mmol) of N-(tert-butoxycarbonyl)-valine-N-carboxyanhydride and 0.5 g of 4- (N,N-dimethylamino)-pyridine were added to a stirred suspension of 2 g (5.3 mmol) of (4S)-4,11-diethyl- 4-hydroxy-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dione in 150 ml of absolute dichloromethane. The mixture was stirred at rt for 20 h and subsequently concentrated in vacuo.8 ml ACN were added to the residue and subsequently 5 mL diethyl ether. The mixture was filtrated and the remaining residue was dried in vacuo.2964 mg (92%) of the protected intermediate were obtained. LC- MS: Rt = 119 min; MS (ESIpos): m / z = 576 (M+H)+ Next 2964 mg (515 mmol) of this Boc protectedintermediate compound in 6 ml of dichloromethane and 60 ml of anhydrous trifluoroacetic acid was stirred for 30 min. at rt and subsequently sonicated for 1 h. After concentrating in vacuo the product was lyophilized from a mixture of acetonitrile / water.3.622 g (quant) of Intermediate Q40 was obtained. LC- MS: R + t = 0.68 min; MS (ESIpos): m / z = 476 [M+H] . Intermediate Q41: [000290](3R)-3-{[(4-{[(2-{[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1- yl)hexanoyl]amino}ethyl)carbamoyl]amino}phenyl)carbamoyl]amino}-3-{3-[({3- [(propylcarbamoyl)amino]phenyl}sulfonyl)amino]phenyl}propanoic acid[000291]Intermediate Q36 (20.0 mg, 26.5 µmol) was dissolved in DMF (6.67 ml). N-Succinimidyl 6- maleimidohexanoate (12.3 mg, 39.7 µmol) and DIEA (13.8 µl, 79.5 µmol) were added. The reaction was stirred at RT for 1 h and then concentrated in vacuo. The residue was separated by prep. HPLC to give Intermediate Q41 as a colorless foam (3.96mg, 95% purity, 17% yield). LC-MS (Method 3): Rt = 2.16 min; MS (ESIpos): m / z = 834 [M+H]+. Intermediate Q42: [000292]N-{18-[(2,5-dioxopyrrolidin-1-yl)oxy]-14,18-dioxo-4,7,10-trioxa-13-azaoctadecan-1-oyl}-L- alpha-aspartyl-L-prolyl-N-[(R*)-{[(4R*)-15,19-difluoro-4-methyl-3,4-dihydro-2H,11H-12,16-(azeno)- 10,6-(metheno)-1,5,11,13-benzodioxadiazacyclooctadecin-8-yl]methyl}(methyl)oxido-λ6-sulfanylidene]- L-valinamide[000293]N-(3-{2-[2-(2-Aminoethoxy)ethoxy]ethoxy}propanoyl)-L-alpha-aspartyl-L-prolyl-N-[(R*)- {[(4R*)-15,19-difluoro-4-methyl-3,4-dihydro-2H,11H-12,16-(azeno)-10,6-(metheno)-1,5,11,13- benzodioxadiazacyclooctadecin-8-yl]methyl}(methyl)oxido-λ6-sulfanylidene]-L-valinamide trifluoroacetate (1:1) (Intermediate Q43) was dissolved in DMF (4.0 ml). Di(N-succinimidyl)glutarate (19.2mg, 0.059 mmol) and DIEA (15.4 ul, 0.088 mmol) were added. The reaction was stirred for 1h at RT and then concentrated in vacuo. The residue was separated by prep. HPLC to give Intermediate Q42 (10.0 mg, 96% purity, 55% yield) as a colorless foam. LC-MS (Method 1): Rt= 4.16 min; MS (ESIpos): m / z = 1186 [M+H]+. Intermediate Q43: [000294]N-(3-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}propanoyl)-L-alpha-aspartyl-L-prolyl-N-[(R*)- {[(4R*)-15,19-difluoro-4-methyl-3,4-dihydro-2H,11H-12,16-(azeno)-10,6-(metheno)-1,5,11,13- benzodioxadiazacyclooctadecin-8-yl]methyl}(methyl)oxido-λ6-sulfanylidene]-L-valinamide trifluoroacetate (1:1)P368919WO / 59362-725.601[000295]Tert-butyl (19S)-19-{[(2S)-2-{[(2S)-1-{[(R*)-{[(11R*)-5,22-difluoro-11-methyl-8,12-dioxa- 18,20,23-triazatetracyclo[17.3.1.113,17.02,7]tetracosa-1(23),2,4,6,13(24),14,16,19,21-nonaen-15- yl]methyl}(methyl)oxido-λ6-sulfanylidene]amino}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidin-1- yl]carbonyl}-2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa-5,18-diazahenicosan-21-oate (Intermediate Q44) (223.8 mg, 0.198 mmol) was dissolved in DCM (15.0 ml), then TFA (5.0 ml) was added. The reaction was stirred for 3 hour at RT and then concentrated in vacuo. The residue was dissolved in ACN / water and freeze-dried to give Intermediate Q43 (209.7 mg, 96% purity, 93% yield) as a light yellow foam. LC-MS (Method 2): Rt= 1.43 min; MS (ESIpos): m / z = 975 [M+H]+. Intermediate Q44: [000296] tert-butyl (19S)-19-{[(2S)-2-{[(2S)-1-{[(R*)-{[(11R*)-5,22-difluoro-11-methyl-8,12-dioxa- 18,20,23-triazatetracyclo[17.3.1.113,17.02,7]tetracosa-1(23),2,4,6,13(24),14,16,19,21-nonaen-15- yl]methyl}(methyl)oxido-λ6-sulfanylidene]amino}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidin-1- yl]carbonyl}-2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa-5,18-diazahenicosan-21-oateP368919WO / 59362-725.601[000297]N-[(R*)-{[(4R*)-15,19-difluoro-4-methyl-3,4-dihydro-2H,11H-12,16-(azeno)-10,6-(metheno)- 1,5,11,13-benzodioxadiazacyclooctadecin-8-yl]methyl}(methyl)oxido-λ6-sulfanylidene]-L-valinamide trifluoroacetate (1:1) (Intermediate Q45) (139.5 mg, 0.207 mmol) was dissolved in DMF (14 ml). Intermediate Q39 (134 mg, 0.228 mmol), HATU (126 mg, 0.331 mmol) and DIEA (108 ul, 0.621 mmol) were added. The reaction was stirred at RT for 30min and then concentrated in vacuo. The residue was separated by prep. HPLC to give Intermediate Q44 (224 mg, 100% purity, 96% yield). LC-MS (Method 2): R + t = 2.34 min; MS (ESIpos): m / z = 1131 [M+H] . Intermediate Q45: [000298]N-[(R*)-{[(4R*)-15,19-difluoro-4-methyl-3,4-dihydro-2H,11H-12,16-(azeno)-10,6-(metheno)- 1,5,11,13-benzodioxadiazacyclooctadecin-8-yl]methyl}(methyl)oxido-λ6-sulfanylidene]-L-valinamide trifluoroacetate (1:1)[000299]Tert-butyl [(2S)-1-{[(R*)-{[(4R*)-15,19-difluoro-4-methyl-3,4-dihydro-2H,11H-12,16- (azeno)-10,6-(metheno)-1,5,11,13-benzodioxadiazacyclooctadecin-8-yl]methyl}(methyl)oxido- lambda6sulfanylidene]amino}-3-methyl-1-oxobutan-2-yl]carbamate (Intermediate Q46) (1391 mgP368919WO / 59362-725.601 210.8 umol) was dissolved in DCM (14.0 ml), then TFA (2.34 ml) was added. The reaction was stirred for 1 hour at RT and then concentrated in vacuo. The residue was dissolved in ACN / water and freeze- dried to give Intermediate Q45 (139.7 mg, 100% purity, 98% yield). LC-MS (Method 2): Rt = 1.45 min; MS (ESIpos): m / z = 560 [M+H]+. Intermediate Q46: [000300] tert-butyl [(2S)-1-{[(R*)-{[(4R*)-15,19-difluoro-4-methyl-3,4-dihydro-2H,11H-12,16-(azeno)- 10,6-(metheno)-1,5,11,13-benzodioxadiazacyclooctadecin-8-yl]methyl}(methyl)oxido-λ6- sulfanylidene]amino}-3-methyl-1-oxobutan-2-yl]carbamate[000301](4R*)-15,19-Difluoro-4-methyl-8-[(S-methylsulfonimidoyl)methyl]-3,4-dihydro-2H,11H- 12,16-(azeno)-10,6-(metheno)-1,5,11,13-benzodioxadiazacyclooctadecine (Intermediate Q47) (130 mg, 0.282 mmol) was dissolved in DMF (15.0 ml). Boc-Val-OH (73.6 mg, 0.339 mmol), HATU (161.0 mg, 0.423 mmol) and DIEA (147.5 ul, 0.847 mmol) were added. The reaction was stirred at RT for 24 hour and then concentrated in vacuo to give Intermediate Q46 (139.1 mg, 100% purity, 75% yield). LC-MS (Method 2): Rt= 2.43 min; MS (ESIpos): m / z = 660 [M+H]+. Intermediate Q47: [000302](4R*)-15,19-difluoro-4-methyl-8-[(S-methylsulfonimidoyl)methyl]-3,4-dihydro-2H,11H- 12,16-(azeno)-10,6-(metheno)-1,5,11,13-benzodioxadiazacyclooctadecineP368919WO / 59362-725.601 [000303]Intermediate Q47 was prepared from commercially available intermediates according to the following steps: Step 1: Synthesis of 4-[tert-butyl(diphenyl)silyl]oxybutan-2-ol (B16-1)[000304]To a solution of butane-1,3-diol (20 g, 221.9 mmol, 1 eq) in DCM (200 mL) was added imidazole (30.22 g, 443.8 mmol, 2 eq) and TBDPSCl (61.0 g, 221.9 mmol, 57.01 mL, 1 eq) at 0°C. The mixture was stirred at 0°C for 1 hour. The reaction mixture was diluted with DCM (300 mL). The organic layer was washed with water 600 mL (2x300 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10:1 to 2:1) to give B16-1 (58.2 g, 177.16 mmol, 79.8% yield) as a colorless oil.1H NMR (400 MHz, CDCl3-d) δ 7.73 - 7.66 (m, 4H), 7.50 - 7.36 (m, 6H), 4.11 (dt, J = 2.8, 6.0 Hz, 1H), 3.88 (s, 2H), 3.28 (d, J = 2.6 Hz, 1H), 1.81 - 1.60 (m, 2H), 1.22 (d, J = 6.0 Hz, 3H), 1.06 (s, 9H). Step 2: Synthesis of tert-butyl-[3-[3-(methylsulfinylmethyl)-5-nitro-phenoxy]butoxy]-diphenyl- silane (B16-2)[000305]To a solution of 3-(methylsulfinylmethyl)-5-nitro-phenol (28 g, 130.10 mmol, 1 eq), B16-1 (42.74 g, 130.10 mmol, 1 eq) and PPh3 (85.31 g, 325.24 mmol, 2.5 eq) in THF (500 mL) was added DIAD (65.77 g, 325.24 mmol, 63.24 mL, 2.5 eq) at 0°C under N2. The mixture was stirred at 20°C for 16 hours. The reaction mixture was diluted with EtOAc (500 mL). The organic layer was washed with NaHCO3 (200 ml), NH4Cl (200 mL), H2O (200 mL) and brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 5:1 to 0:1) to give B16-2 (62 g, 117.93 mmol, 90.6% yield) as a light-yellow oil.1H NMR (400 MHz, CDCl3-d) δ 7.74 - 7.53 (m, 6H), 7.50 - 7.28 (m, 6H), 7.18 - 7.11 (m, 1H), 4.86 - 4.73 (m, 1H), 4.01 - 3.90 (m, 2H), 3.89 - 3.73 (m, 2H), 2.51 (d, J = 5.6 Hz, 3H), 2.07 - 1.78 (m, 2H), 1.34 (d, J = 6.0 Hz, 3H), 1.04 (s, 9H). Step 3: Synthesis of tert-butyl N-[[3-[3-[tert-butyl(diphenyl)silyl]oxy-1-methyl-propoxy]-5-nitro- phenyl]methyl-methyl-oxo-λ⁶-sulfanylidene]carbamate (B16-3)P368919WO / 59362-725.601[000306]To a mixture of B16-2 (23.5 g, 44.70 mmol, 1 eq), tert-butyl carbamate (7.85 g, 67.05 mmol, 1.5 eq), MgO (7.21 g, 178.80 mmol, 2.01 mL, 4 eq) and PhI(OAc)2 (21.60 g, 67.05 mmol, 1.5 eq) in DCM (350 mL) was added Rh2(OAc)4 (493.92 mg, 1.12 mmol, 0.025 eq) at 20°C under N2. Then mixture was stirred at 45°C for 16 hours. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1:1 to 0:1) to give B16-3 (62 g, 91.91 mmol, 68.5% yield, 95% purity) as a light- yellow oil.1H NMR (400 MHz, CDCl3-d) δ 7.82 (d, J = 1.4 Hz, 1H), 7.75 (t, J = 2.0 Hz, 1H), 7.61 (dd, J = 6.6, 17.6 Hz, 4H), 7.48 - 7.28 (m, 6H), 7.24 (s, 1H), 4.89 - 4.69 (m, 3H), 3.91 - 3.73 (m, 2H), 2.97 (s, 3H), 2.04 - 1.82 (m, 2H), 1.52 (s, 9H), 1.34 (d, J = 6.0 Hz, 3H), 1.04 (s, 9H). Step 4: Synthesis of tert-butyl N-[[3-(3-hydroxy-1-methyl-propoxy)-5-nitro-phenyl]methyl-methyl- oxo-λ⁶-sulfanylidene]carbamate (B16-4)[000307]Two reactions were carried out in parallel. To a solution of B16-3 (30 g, 46.81 mmol, 1 eq) in THF (40 mL) was added TBAF (1 M, 46.81 mL, 1 eq) at 20°C. The mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated under reduced pressure to give a residue. The two reactions mixture were combined and diluted with EtOAc (200 mL). The organic layer was washed with NH4Cl (50 mL), NaHCO3(50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1:1 to 0:1) to give B16-4 (32 g, 79.51 mmol, 84.9% yield) as a light-yellow oil.1H NMR (400 MHz, CDCl3) δ 7.83 - 7.79 (m, 2H), 7.42 (d, J = 1.6 Hz, 1H), 4.82 - 4.72 (m, 3H), 3.84 - 3.72 (m, 2H), 3.07 (d, J = 4.4 Hz, 3H), 2.04 - 1.96 (m, 1H), 1.93 - 1.81 (m, 1H), 1.51 (s, 9H), 1.39 (dd, J = 1.6, 6.0 Hz, 3H). Step 5: Synthesis of tert-butyl N-[[3-[3-[2-(2-chloro-5-fluoro-pyrimidin-4-yl)-5-fluoro-phenoxy]-1- methyl-propoxy]-5-nitro-phenyl]methyl-methyl-oxo-λ⁶-sulfanylidene]carbamate (B16-5)P368919WO / 59362-725.601[000308]To a solution of B16-4 (18 g, 44.72 mmol, 1 eq) and 2-(2-chloro-5-fluoro-pyrimidin-4-yl)-5- fluoro-phenol (10.85 g, 44.72 mmol, 1 eq) in toluene (180 mL) was added 2-(tributyl-λ5- phosphanylidene)acetonitrile (16.19 g, 67.09 mmol, 1.5 eq) at 20°C under N2. The mixture was stirred at 80°C for 16 hours. The reaction mixture was diluted with EtOAc (150 mL). The organic layer was washed with H2O (3x100 mL), NaHCO3 (3x100 mL), brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 3:1 to 1:1) to give B16-5 (24 g, 38.27 mmol, 85.5% yield) as a light-yellow oil.1H NMR (400 MHz, CDCl3) δ 8.55 (dd, J = 1.5, 3.6 Hz, 1H), 7.85 - 7.80 (m, 1H), 7.76 (t, J = 2.0 Hz, 1H), 7.51 (dd, J = 6.5, 8.5 Hz, 1H), 7.34 - 7.28 (m, 1H), 6.81 (dt, J = 2.3, 8.3 Hz, 1H), 6.72 (dd, J = 2.0, 10.5 Hz, 1H), 4.84 - 4.70 (m, 3H), 4.23 - 4.15 (m, 2H), 2.99 (d, J = 4.9 Hz, 3H), 2.19 - 2.07 (m, 2H), 1.54 - 1.50 (m, 9H), 1.42 - 1.35 (m, 3H). Step 6: Synthesis of tert-butyl N-[[3-amino-5-[3-[2-(2-chloro-5-fluoro-pyrimidin-4-yl)-5-fluoro- phenoxy]-1-methyl-propoxy]phenyl]methyl-methyl-oxo-λ⁶-sulfanylidene]carbamate (B16-6)[000309]To a solution of B16-5 (22 g, 35.08 mmol, 1 eq) in EtOH (50 mL) and H2O (10 mL) was added Fe (9.80 g, 175.42 mmol, 5 eq) and NH4Cl (9.38 g, 175.42 mmol, 5 eq) at 20°C. The mixture was stirred at 80°C for 2 hours. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was diluted with EtOAc (200 mL). The organic layer was washed with H2O (2x50 mL), brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=2:1 toP368919WO / 59362-725.601 1:1) to give B16-6 (18.5 g, 30.98 mmol, 88.3% yield) as a light-yellow oil.1H NMR (400 MHz, CDCl3) δ 8.50 (dd, J = 1.4, 4.6 Hz, 1H), 7.49 (dd, J = 6.6, 8.6 Hz, 1H), 6.86 - 6.67 (m, 2H), 6.39 - 6.22 (m, 2H), 6.16 (s, 1H), 4.67 - 4.39 (m, 3H), 4.23 - 4.04 (m, 2H), 2.91 (d, J = 1.4 Hz, 3H), 2.15 - 1.96 (m, 2H), 1.56 - 1.47 (m, 9H), 1.28 (d, J = 5.8 Hz, 3H). Step 7: Synthesis of tert-butyl N-[(5,22-difluoro-11-methyl-8,12-dioxa-18,20,23- triazatetracyclo[17.3.1.113,17.02,7]tetracosa-1(22),2,4,6,13,15,17(24),19(23),20-nonaen-15- yl)methyl-methyl-oxo-λ⁶-sulfanylidene]carbamate (B16-7)[000310]To a solution of B16-6 (13 g, 21.77 mmol, 1 eq) in toluene (90 mL) and NMP (9 mL) were added K3PO4 (23.11 g, 108.86 mmol, 5 eq), XPhos (1.04 g, 2.18 mmol, 0.1 eq) and Xhos Pd G1 (1.61 g, 2.18 mmol, 0.1 eq) at 20°C under N2. The mixture was stirred at 110°C for 5 hours. The reaction mixture was diluted with EtOAc (200mL). The organic layer was washed with H2O (2x50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO2, petroleum ether / ethyl acetate = 1:1.5) to give B16-7 (8.2 g, 14.33 mmol, 65.8% yield, 98% purity) as a light-yellow gum.1H NMR (400 MHz, CDCl3) δ 8.81 (d, J = 2.0 Hz, 1H), 8.39 (d, J = 3.0 Hz, 1H), 7.67 - 7.59 (m, 1H), 7.30 (br s, 1H), 6.92 - 6.72 (m, 2H), 6.59 - 6.46 (m, 2H), 4.78 - 4.52 (m, 3H), 4.25 - 4.14 (m, 2H), 2.98 (d, J = 3.8 Hz, 3H), 2.57 (br t, J = 12.8 Hz, 1H), 1.86 - 1.76 (m, 1H), 1.54 (d, J = 3.4 Hz, 9H), 1.47 (d, J = 6.0 Hz, 3H). Step 8: Synthesis of (5,22-difluoro-11-methyl-8,12-dioxa-18,20,23- triazatetracyclo[17.3.1.113,17.02,7]tetracosa-1(22),2,4,6,13,15,17(24),19(23),20-nonaen-15- yl)methyl-imino-methyl-oxo-λ⁶-sulfane (B16-8)P368919WO / 59362-725.601[000311]To a solution of B16-7 (6 g, 10.70 mmol, 1 eq) in DCM (60 mL) was added TFA (6 mL) at 20°C. The mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated under reduced pressure to give crude product B16-8, which was used in the next step. Step 9: Chiral separation of B16-8 to give building block 1, building block 2, Intermediate Q47 and building block 4was separated by prep-SFC (column: DAICEL C10um); mobile phase: [0.1%NH3H2O ETOH]; B%: 60%-60%, 8.4min) to give P1 (2 isomers), P2 (Building block 16), P3 (Building block 17). Then, the P1 residue was repurified by prep-SFC (column: DAICEL CHIRALPAK AD (250mm*30mm, 10um); mobile phase: [0.1%NH3H2O ETOH]; B%: 55%- 55%, 7min) to give P4 (Intermediate Q47) and P5 (Building block 4). P2 (Building block 1) (1.1 g, 2.34 mmol, 21.8% yield, 98% purity) was obtained as a light-yellow solid. P3 (Building block 2) (1 g, 2.13 mmol, 19.8% yield, 98% purity) was obtained as a light yellow solid. P4 (Intermediate Q47) (1.2 g, 2.61 mmol, 24.3% yield) and P5 (Building block 4) (1.3 g, 2.82 mmol, 26.3% yield) was obtained as a light- yellow solid. [000313]Note: The R / S configuration of these four compounds were not confirmed. [000314]Building block 1: 1H NMR (400 MHz, DMSO-d6) δ = 9.80 (s, 1H), 8.67 (d, J = 2.7 Hz, 2H), 7.66 - 7.57 (m, 1H), 7.35 (dd, J = 2.1, 12.1 Hz, 1H), 6.92 (dt, J = 2.1, 8.3 Hz, 1H), 6.74 (s, 1H), 6.47 (s, 1H), 4.47 (br t, J = 11.0 Hz, 1H), 4.37 (br dd, J = 6.1, 9.8 Hz, 1H), 4.22 (q, J = 13.4 Hz, 2H), 4.08 (br d, J = 10.5 Hz, 1H), 3.55 (s, 1H), 2.82 (s, 3H), 2.48 - 2.27 (m, 1H), 1.77 - 1.64 (m, 1H), 1.43 (d, J = 6.1 Hz, 3H). The desired enantiomer (optical rotation -112.52˚ ± 0.00˚, 20C, 589 nm) was obtained with 96.52% ee at R 1.45- + t 1.64 min. LC-MS: Rt = 2.31 min; MS (ESIpos): m / z = 461 [M+H] . Single (-) isomer, absolute stereochemistry unknown.P368919WO / 59362-725.601 [000315]Building block 2: 1H NMR (400 MHz, DMSO-d6) δ = 9.83 (s, 1H), 8.71 - 8.66 (m, 2H), 7.62 (ddd, J = 4.5, 7.1, 8.5 Hz, 1H), 7.36 (dd, J = 2.2, 12.1 Hz, 1H), 6.92 (dt, J = 2.2, 8.3 Hz, 1H), 6.75 (s, 1H), 6.49 (s, 1H), 4.47 (br t, J = 10.9 Hz, 1H), 4.41 - 4.31 (m, 3H), 4.13 - 4.04 (m, 1H), 2.95 (s, 3H), 2.52 - 2.52 (m, 1H), 2.41 - 2.29 (m, 2H), 1.77 - 1.66 (m, 1H), 1.44 (d, J = 6.0 Hz, 3H). The desired enantiomer (optical rotation -129.68˚ ± 0.70˚, 20C, 589 nm) was obtained with 98.04% ee at Rt 1.60-1.83 min. LC- MS: R = 2.32 min + t ; MS (ESIpos): m / z = 461 [M+H] . Single (-) isomer, absolute stereochemistry unknown. [000316]Intermediate Q47: 1H NMR (400 MHz, DMSO-d6) δ 9.81 (s, 1H), 8.67 (d, J = 3.0 Hz, 2H), 7.62 (ddd, J = 4.6, 7.0, 8.4 Hz, 1H), 7.35 (dd, J = 2.0, 12.0 Hz, 1H), 6.92 (dt, J = 2.2, 8.2 Hz, 1H), 6.73 (s, 1H), 6.47 (s, 1H), 4.47 (br t, J = 10.8 Hz, 1H), 4.40 - 4.28 (m, 1H), 4.25 - 4.17 (m, 2H), 4.08 (br d, J = 10.4 Hz, 1H), 3.56 (s, 1H), 2.81 (s, 3H), 2.41 - 2.27 (m, 1H), 1.81 - 1.63 (m, 1H), 1.43 (d, J = 6.0 Hz, 3H). The desired enantiomer (optical rotation 133.00˚ ± 0.00˚, 20C, 589 nm) was obtained with 98.70% ee at Rt2.10-2.80 min. LC-MS: Rt= 2.33 min; MS (ESIpos): m / z = 461 [M+H]+. Single (+) isomer, absolute stereochemistry unknown. [000317]Building block 4: 1H NMR (400 MHz, DMSO-d6) δ 9.82 (s, 1H), 8.67 (d, J = 2.8 Hz, 2H), 7.62 (ddd, J = 4.6, 7.0, 8.4 Hz, 1H), 7.36 (dd, J = 2.0, 12.0 Hz, 1H), 6.92 (dt, J = 2.2, 8.2 Hz, 1H), 6.73 (s, 1H), 6.50 - 6.38 (m, 1H), 4.47 (br t, J = 11.0 Hz, 1H), 4.41 - 4.32 (m, 1H), 4.28 - 4.15 (m, 2H), 4.15 - 4.02 (m, 1H), 3.56 (s, 1H), 2.82 (s, 3H), 2.35 (br t, J = 12.6 Hz, 1H), 1.78 - 1.63 (m, 1H), 1.43 (d, J = 6.0 Hz, 3H). The desired enantiomer (optical rotation 120.80˚ ± 0.00˚, 20C, 589 nm) was obtained with 94.14% ee at R + t 2.50-3.05 min. LC-MS: Rt = 2.33 min; MS (ESIpos): m / z = 461 [M+H] . Single (+) isomer, absolute stereochemistry unknown. Intermediate Q48: [000318]N-{18-[(2,5-dioxopyrrolidin-1-yl)oxy]-14,18-dioxo-4,7,10-trioxa-13-azaoctadecan-1-oyl}-L- alpha-aspartyl-L-prolyl-N-[2-(ethoxymethyl)-1-(2-hydroxy-2-methylpropyl)-1H-imidazo[4,5-c]quinolin- 4-yl]-L-valinamideP368919WO / 59362-725.601[000319]Intermediate Q49 (13.0 mg, 0.014 mmol) was dissolved in DMF (5.0 ml). Di(N- succinimidyl)glutarate (11.2mg, 0.034 mmol) and DIEA (7.20 uL, 0.041 mmol) were added. The reaction was stirred at RT for 30min, concentrated in vacuo and purified by prep HPLC to give Intermediate Q48 (3.32mg, 98% purity, 23% yield) as a colorless foam. LC-MS (Method 5): Rt= 2.07 min; MS (ESIpos): m / z = 1041 [M+H]+. Intermediate Q49: [000320]N-(3-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}propanoyl)-L-alpha-aspartyl-L-prolyl-N-[2- (ethoxymethyl)-1-(2-hydroxy-2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-yl]-L-valinamide trifluoroacetate (1:1)[000321]Intermediate Q50 (65.0 mg, 70.0 umol) was dissolved in DCM (5.0 ml), then TFA (2.0 mL) was added. The reaction was stirred for 2 hour at RT, then it was concentrated on an oil pump. Theresidue was dissolved in ACN / water and freeze-dried to give Intermediate Q49 (55.2 mg, 100% purity, 84% yield) as a colorless foam. LC-MS (Method 2): R - t = 0.98 min; MS (ESIneg): m / z = 827 [M-H] . Intermediate Q50: [000322]N-(2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa-5-azaheptadecan-17-yl)-L-alpha-aspartyl-L- prolyl-N-[2-(ethoxymethyl)-1-(2-hydroxy-2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-yl]-L- valinamide[000323]Intermediate Q51 (93.0 mg, 0.126 mmol) was dissolved in DMF (5.0 ml). Commercially available t-Boc-N-amido-PEG3-NHS ester (63.1 mg, 0.15 mmol) and DIEA (65.7 ul, 377.2 umol) were added. The reaction was stirred at RT for 3 hours, concentrated in vacuo and purified by prep HPLC to give Intermediate Q50 (65.0 mg, 91% purity, 51% yield) as a colorless oil. LC-MS (Method 5): Rt = 2.48 min; MS (ESIpos): m / z = 930 [M+H]+. Intermediate Q51: [000324]L-alpha-aspartyl-L-prolyl-N-[2-(ethoxymethyl)-1-(2-hydroxy-2-methylpropyl)-1H- imidazo[4,5-c]quinolin-4-yl]-L-valinamide trifluoroacetate (1:1)P368919WO / 59362-725.601 [000325]Intermediate Q52 (99.0 mg, 126 umol) was dissolved in DCM (5.0 ml), then TFA (2.0 ml) was added. The reaction was stirred for 2 hour at RT and concentrated on an oil pump. The residue was dissolved in ACN / water and freeze-dried to give Intermediate Q51 (93.0 mg, 94% purity, 94% yield) as a colorless foam. LC-MS (Method 2): Rt= 0.95 min; MS (ESIneg): m / z = 624 [M-H]- . Intermediate Q52: [000326] tert-butyl (3S)-3-[(tert-butoxycarbonyl)amino]-4-[(2S)-2-{[(2S)-1-{[2-(ethoxymethyl)-1-(2- hydroxy-2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-yl]amino}-3-methyl-1-oxobutan-2- yl]carbamoyl}pyrrolidin-1-yl]-4-oxobutanoate[000327]Intermediate Q53 (75.0 mg, 117 umol) was dissolved in DMF (6.0 ml). Boc-Asp(OtBu)-Pro- OH (54.2 mg, 140.3 umol), HATU (57.8 mg, 152.0 umol) and DIEA (61.1 ul, 350.7 umol) were added. The reaction was stirred at RT for 1 hour, concentrated on an oilo pump and purified by prep HPLC to give Intermediate Q52 (99.0 mg, 82% purity, 89% yield) as a colorless foam. LC-MS (Method 2): Rt = 1.89 min; MS (ESIneg): m / z = 780 [M-H]- . Intermediate Q53: [000328]N-[2-(ethoxymethyl)-1-(2-hydroxy-2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-yl]-L- valinamide trifluoroacetate (1:2)[000329]Intermediate Q54 (565 mg, 1.10 mmol) was dissolved in DCM (40 ml), then TFA (10 ml) was added. The reaction was stirred for 2 hour at RT and concentrated on an oil pump. The residue was dissolved in ACN / water and freeze-dried to give Intermediate Q53 (608mg, 100% purity, 86% yield) as a colorless foam. LC-MS (Method 2): R + t = 0.97 min; MS (ESIpos): m / z = 414 [M+H] .P368919WO / 59362-725.601 Intermediate Q54: [000330] tert-butyl [(2S)-1-{[2-(ethoxymethyl)-1-(2-hydroxy-2-methylpropyl)-1H-imidazo[4,5- c]quinolin-4-yl]amino}-3-methyl-1-oxobutan-2-yl]carbamate[000331]Resiquimod (240 mg, 0.763 mmol) was suspended in DCM (50 ml), then Boc-Val-NCA (371 mg, 1.53 mmol) and DMAP (93.3 mg, 763 umol) were added. The clear reaction was stirred for 2 h and then concentrated in vacuo. The residue was purified by prep. HPLC to give Intermediate Q54 (565 mg, 100% purity, quantitative yield) as a colorless foam. LC-MS (Method 2): Rt= 1.64 min; MS (ESIpos): m / z = 514 [M+H]+. Intermediate Q56: [000332]N-[1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2,15-dioxo-6,9,12-trioxa-3-azapentadecan-15- yl]-L-alpha-aspartyl-L-prolyl-N-[2-(ethoxymethyl)-1-(2-hydroxy-2-methylpropyl)-1H-imidazo[4,5- c]quinolin-4-yl]-L-valinamide[000333] Intermediate Q49 (10.0 mg, 10.6 umol), AMAS (5.35mg, 21.2 umol) and DIEA (7.39 ul, 42.4 umol) were dissolved in DMF (4.0 ml) and the reaction was stirred at RT for 30 min. DMF was evaporated in vacuo and the residue was purified by preparative HPLC, then lyophilized to give Intermediate Q56 (4.0 mg, 73% purity, 39% yield) as a white foam. LC-MS (Method 5): Rt= 1.95 min; MS (ESIpos): m / z = 966 [M+H]+.P368919WO / 59362-725.601 Intermediate Q57: [000334](7S)-7-Ethyl-8,11-dioxo-7,8,11,13-tetrahydro-2H,10H-[1,3]dioxolo[4,5- g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl N-[1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2,15- dioxo-6,9,12-trioxa-3-azapentadecan-15-yl]-L-alpha-aspartyl-L-prolyl-L-valinate[000335] Intermediate Q32 (20.0 mg, 92 % purity, 18.0 µmol), AMAS (9.09 mg, 36.0 µmol) and DIEA (9.4 µl, 54 µmol) were dissolved in DMF (3.0 ml) and the reaction was stirred overnight at RT. DMF was evaporated in vacuo and the residue was purified by preparative HPLC, then lyophilized to give Intermediate Q57 (11.0 mg, 94% purity, 55% yield). LC-MS (Method 1): Rt = 3.29 min; MS (ESIpos): m / z = 1044 [M+H]+. Intermediate Q58: [000336](4S)-4,11-Diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinolin-4-yl N-[1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2,15-dioxo-6,9,12-trioxa-3-azapentadecan- 15-yl]-L-alpha-aspartyl-L-prolyl-L-valinate[000337]Intermediate Q11 (20.0 mg, 98 % purity, 19.5 µmol), AMAS (9.84 mg, 39.0 µmol) and DIEA (10 µl, 59 µmol) were dissolved in DMF (3.0 ml) and the reaction was stirred overnight at RT. DMF was evaporated in vacuo and the residue was purified by preparative HPLC, then lyophilized to give Intermediate Q58 (10.4 mg, 92% purity, 48% yield). LC-MS (Method 1): Rt = 3.62 min; MS (ESIpos): m / z = 1028 [M+H]+. Intermediate Q59: [000338] (4S)-4,11-Diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indolizino[1,2- b]quinolin-4-yl L-asparaginyl-L-prolyl-L-valinate • trifluoroacetic acid (1:1)[000339]Step 1: Trifluoroacetic acid—(4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H- pyrano[3',4':6,7] indolizino[1,2-b]quinolin-4-yl L-valinate (1 / 1) (150 mg, 254 µmol) (Intermediate Q40) (150 mg, 254 µmol) and N2-(tert-butoxycarbonyl)-L-asparaginyl-L-proline (101 mg, 305 µmol) weredissolved in DMF (10 mL) and 1.3 eq EDCI (63.4 mg, 331 µmol), 1.5 eq HOBT hydrate (58.4 mg, 382 µmol) as well as 3 eq N,N-diisopropylethylamine (130 µl, 760 µmol) were added. After stirring for 1h at rt, the mixture was concentrated in vacuo. The residual was purified by prep. HPLC, then concentrated and lyophilized to yield Boc-Intermediate Q59 (185 mg, 93% purity, 86%). LC-MS: Rt = 3.08 min; MS (ESIpos): m / z = 787 [M+H]+. [000340]Step 2: (4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinolin-4-yl N2-(tert-butoxycarbonyl)-L-asparaginyl-L-prolyl-L-valinate (185 mg, 235 µmol) was dissolved in DCM (15 mL), TFA (3 mL) was added and the reaction mixture was stirred for 30 min and concentrated in vacuo. The residual was dissolved in ACN / water and lyophilized to yield Intermediate Q59 (188 mg, 100% purity, quant.). LC-MS: Rt= 1.13 min; MS (ESIpos): m / z = 687 [M+H]+ Intermediate Q60: [000341](4S)-4,11-Diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinolin-4-yl N-{2-[{2-[(2-aminoethyl)(methyl) amino]ethyl}(methyl)amino]ethyl}-N-methylglycyl- L-asparaginyl-L-prolyl-L-valinate • trifluoroacetic acid (1:1)[000342]Step 1: Trifluoroacetic acid—(4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H- pyrano[3',4':6,7] indolizino[1,2-b]quinolin-4-yl L-asparaginyl-L-prolyl-L-valinate (1 / 1) (Intermediate Q57) (150 mg, 187 µmol) was dissolved in DMF (12 mL) and 1.5 eq HATU (107 mg, 281 µmol), 1 eq N-methyl-N-(2,2,8,11-tetramethyl-4-oxo-3-oxa-5,8,11-triazatridecan-13-yl)glycine (64.9 mg, 187 µmol) as well as 3.0 eq N,N-diisopropylethylamine (98 µl, 560 µmol) were added. After stirring for 1 h at rt, the mixture was concentrated in vacuo and the residual was purified by prep. HPLC, then concentrated in vacuo to yield Boc-Intermediate 4 as a light yellow foam (128 mg, 55% purity, 37%). LC-MS: Rt = 2.83 min; MS (ESIpos): m / z = 1015 [M+H]+. [000343]Step 2: (4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7] indolizino[1,2- b] quinolin-4-yl N-methyl-N-(2,2,8,11-tetramethyl-4-oxo-3-oxa-5,8,11-triazatridecan-13-yl) glycyl-L- asparaginyl-L-prolyl-L-valinate (128 mg, 55 % purity, 69.3 µmol) was dissolved in DCM (15 mL), TFA (3 mL) was added and the reaction mixture was stirred for 30 min. It was concentrated in vacuo, and theP368919WO / 59362-725.601 residual was purified by prep. HPLC, then concentrated in vacuo to yield Intermediate 4 as a yellow foam (88 mg, 100% purity, quant.). LC-MS: R + t = 2.46 min; MS (ESIpos): m / z = 915 [M+H] . Intermediate Q61: [000344](4S)-4,11-Diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinolin-4-yl N-{2-[{2-[{2-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1- yl)acetamido]ethyl}(methyl)amino]ethyl}(methyl)amino]ethyl}-N-methylglycyl-L-asparaginyl-L-prolyl- L-valinate[000345]Intermediate Q60 (21.5 mg, 96 % purity, 20.1 µmol), AMAS (10.1 mg, 40.2 µmol) and DIEA (11 µl, 60 µmol) were dissolved in DMF (3.0 ml) and the reaction was stirred overnight at RT. DMF was evaporated in vacuo and the residue was purified by preparative HPLC, then lyophilized to give Intermediate Q61 (14.4 mg, 100% purity, 68% yield). LC-MS (Method 1): Rt= 2.66 min; MS (ESIpos): m / z = 1052 [M+H]+. Intermediate Q62: [000346](4S)-4,11-Diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinolin-4-yl N-{2-[(2-{[2-({5-[(2,5-dioxopyrrolidin-1-yl)oxy]-5- oxopentanoyl}amino)ethyl](methyl)amino}ethyl)(methyl)amino]ethyl}-N-methylglycyl-L-asparaginyl- L-prolyl-L-valinateP368919WO / 59362-725.601[000347]Intermediate Q60 (23.2 mg, 88 % purity, 19.9 µmol), 1,1'-[(1,5-dioxopentane-1,5- diyl)bis(oxy)]di(pyrrolidine-2,5-dione) (19.5 mg, 59.8 µmol) and DIEA (14 µl, 80 µmol) were dissolved in DMF (3.0 ml) and the reaction was stirred at RT for 1h. DMF was evaporated in vacuo and the residue was purified by preparative HPLC, then lyophilized to give Intermediate Q62 (15 mg, 96% purity, 64% yield). LC-MS (Method 1): Rt= 2.72 min; MS (ESIpos): m / z = 1126 [M+H]+. Intermediate Q63: [000348](1S,9S)-1-Amino-9-ethyl-5-fluoro-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H- benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-9-yl N-[1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)- 2,15-dioxo-6,9,12-trioxa-3-azapentadecan-15-yl]-L-alpha-aspartyl-L-prolyl-L-valinate.trifluoroacetic acid (1 / 1)[000349]Intermediate Q21 (20.0 mg, 18.8 µmol), AMAS (5.69 mg, 22.6 µmol) and DIEA (9.8 µl, 56 µmol) were dissolved in DMF (2.0 ml) and the reaction was stirred at RT for 1h30. DMF was evaporated in vacuo and the residue was purified by preparative HPLC, then lyophilized to give Intermediate Q63 (9.2 mg, 91% purity, 37% yield) as a yellow foam. LC-MS (Method 1): Rt= 2.69 min; MS (ESIpos): m / z = 1087 [M+H]+. Intermediate Q64: [000350](3R)-3-{[(4-{[(2-{2-[2-(2- aminoethoxy)ethoxy]ethoxy}ethyl)carbamoyl]amino}phenyl)carbamoyl]amino}-3-[3-({3- [(propylcarbamoyl)amino]benzene-1-sulfonyl}amino)phenyl]propanoic acid.trifluoroacetic acid (1 / 1)[000351]Step 1: Intermediate Q38 (60.0 mg, 90 % purity, 74.9 µmol) was dissolved in DMF (10.0 ml). tert-butyl (2-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}ethyl)carbamate (43.8 mg, 150 µmol) and DIEA (130 µl, 750 µmol) were added. The reaction was stirred for 15 minutes at RT and then concentrated inP368919WO / 59362-725.601 vacuo. The residue was separated by prep. HPLC to give (3R)-3-{[(4-{[(2,2-dimethyl-4-oxo-3,8,11,14- tetraoxa-5-azahexadecan-16-yl)carbamoyl]amino}phenyl)carbamoyl]amino}-3-[3-({3- [(propylcarbamoyl)amino]benzene-1-sulfonyl}amino)phenyl]propanoic acid (54.8 mg, 100 % purity, 84% yield). LC-MS (Method 2): R + t = 1.52 min; MS (ESIpos): m / z = 873 [M+H] . [000352]Step 2: (3R)-3-{[(4-{[(2,2-dimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadecan-16- yl)carbamoyl]amino}phenyl)carbamoyl]amino}-3-[3-({3-[(propylcarbamoyl)amino]benzene-1- sulfonyl}amino)phenyl]propanoic acid (54.3 mg, 100 % purity, 62.2 µmol) was dissolved in DCM (5.0 ml), then TFA (1.0 ml) was added. The reaction was stirred for 1h at RT. The reaction was concentrated at RT on the oil pump. The resulting residue was dissolved in ACN / water and freeze-dried to give Intermediate Q64 (53.7 mg, 100% purity, 97% yield). LC-MS(Method 2): Rt= 0.94 min; MS (ESIpos): m / z = 774 [M+H]+. Intermediate Q65: [000353](3R)-3-{[(4-{[(2-{2-[2-(2-{[N6-(tert-butoxycarbonyl)-L- lysyl]amino}ethoxy)ethoxy]ethoxy}ethyl)carbamoyl]amino}phenyl)carbamoyl]amino}-3-[3-({3- [(propylcarbamoyl)amino]benzene-1-sulfonyl}amino)phenyl]propanoic acid[000354]Step 1: Intermediate Q64 (53.1 mg, 100 % purity, 59.9 µmol) was dissolved in DMF (3.0 ml). 2,5-dioxopyrrolidin-1-yl N2-[(benzyloxy)carbonyl]-N6-(tert-butoxycarbonyl)-L-lysinate (37.2 mg, 77.8 µmol) and DIEA (31 µl, 180 µmol) were added. The reaction was stirred for 1h at RT and then concentrated in vacuo. The residue was separated by prep. HPLC to give (3R)-3-({[4-({[2-(2-{2-[2- ({N2-[(benzyloxy)carbonyl]-N6-(tert-butoxycarbonyl)-L- lysyl}amino)ethoxy]ethoxy}ethoxy)ethyl]carbamoyl}amino)phenyl]carbamoyl}amino)-3-[3-({3- [(propylcarbamoyl)amino]benzene-1-sulfonyl}amino)phenyl]propanoic acid (61.2 mg, 100 % purity, 90% yield). LC-MS (Method 2): R + t = 1.72 min; MS (ESIpos): m / z = 1136 [M+H] . [000355]Step 2: (3R)-3-({[4-({[2-(2-{2-[2-({N2-[(benzyloxy)carbonyl]-N6-(tert-butoxycarbonyl)-L- lysyl}amino)ethoxy]ethoxy}ethoxy)ethyl]carbamoyl}amino)phenyl]carbamoyl}amino)-3-[3-({3-P368919WO / 59362-725.601 [(propylcarbamoyl)amino]benzene-1-sulfonyl}amino)phenyl]propanoic acid (60.7 mg, 53.5 µmol) was dissolved in DCM (10 ml) and Methanol (10 ml), then Pd / C (10 mg) was added. The reaction was hydrogenated for 2h at RT. The reaction was filtered and concentrated under reduced pressure. The resulting residue was dissolved in ACN / water and freeze-dried to give Intermediate Q65 (52.1 mg, 100% purity, 97% yield). LC-MS (Method 2): R + t = 1.19 min; MS (ESIpos): m / z = 1001 [M+H] . Intermediate Q66: [000356](7S)-7-ethyl-8,11-dioxo-7,8,11,13-tetrahydro-2H,10H-[1,3]dioxolo[4,5- g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl N-{18-[(2,5-dioxopyrrolidin-1-yl)oxy]-14,18-dioxo- 4,7,10-trioxa-13-azaoctadecanan-1-oyl}-L-alpha-aspartyl-L-prolyl-L-valinate[000357] Intermediate Q32 (30.0 mg, 92 % purity, 27.0 µmol), 1,1'-[(1,5-dioxopentane-1,5- diyl)bis(oxy)]di(pyrrolidine-2,5-dione) (26.5 mg, 81.1 µmol) and DIEA (14 µl, 81 µmol) were dissolved in DMF (3.0 ml) and the reaction was stirred at RT for 1h. DMF was evaporated in vacuo and the residue was purified by preparative HPLC, then lyophilized to give Intermediate Q66 (12.3 mg, 97% purity, 40% yield). LC-MS (Method 1): R + t = 3.37 min; MS (ESIpos): m / z = 1118 [M+H] . Intermediate Q67: [000358](7S)-7-ethyl-8,11-dioxo-7,8,11,13-tetrahydro-2H,10H-[1,3]dioxolo[4,5- g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl N-[(20S)-20-(4-aminobutyl)-35-{4-[({(1R)-2-carboxy- 1-[3-({3-[(propylcarbamoyl)amino]benzene-1-sulfonyl}amino)phenyl]ethyl}carbamoyl)amino]anilino}- 14,18,21,35-tetraoxo-4,7,10,25,28,31-hexaoxa-13,19,22,34-tetraazapentatriacontanan-1-oyl]-L-alpha- aspartyl-L-prolyl-L-valinate.trifluoroacetic acid (1 / 1)P368919WO / 59362-725.601[000359]Step 1: Intermediate Q65 (20.0 mg, 20.0 µmol) was dissolved in DMF (2.0 ml). Intermediate Q66 (26.8 mg, 24.0 µmol) and DIEA (14 µl, 80 µmol) were added. The reaction was stirred overnight at RT and then concentrated in vacuo. The residue was separated by prep. HPLC to give ((7S)-7-ethyl-8,11- dioxo-7,8,11,13-tetrahydro-2H,10H-[1,3]dioxolo[4,5-g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl N-[(20S)-20-{4-[(tert-butoxycarbonyl)amino]butyl}-35-{4-[({(1R)-2-carboxy-1-[3-({3- [(propylcarbamoyl)amino]benzene-1-sulfonyl}amino)phenyl]ethyl}carbamoyl)amino]anilino}- 14,18,21,35-tetraoxo-4,7,10,25,28,31-hexaoxa-13,19,22,34-tetraazapentatriacontanan-1-oyl]-L-alpha- aspartyl-L-prolyl-L-valinate (23.2 mg, 98 % purity, 57% yield). LC-MS (Method 1): Rt = 4.02 min; MS (ESIpos): m / z = 2004 [M+H]+. [000360]Step 2: (7S)-7-ethyl-8,11-dioxo-7,8,11,13-tetrahydro-2H,10H-[1,3]dioxolo[4,5- g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl N-[(20S)-20-{4-[(tert-butoxycarbonyl)amino]butyl}-35- {4-[({(1R)-2-carboxy-1-[3-({3-[(propylcarbamoyl)amino]benzene-1- sulfonyl}amino)phenyl]ethyl}carbamoyl)amino]anilino}-14,18,21,35-tetraoxo-4,7,10,25,28,31-hexaoxa- 13,19,22,34-tetraazapentatriacontanan-1-oyl]-L-alpha-aspartyl-L-prolyl-L-valinate (23.0 mg, 100 % purity, 11.5 µmol) was dissolved in DCM (2.5 ml), then TFA (250 µl) was added. The reaction was stirred overnight at RT. The reaction was concentrated at RT on the oil pump. The resulting residue was dissolved in ACN / water and freeze-dried to give Intermediate Q67 (22.2 mg, 100% purity, 96% yield) as am amorphous residue. LC-MS (Method 1): Rt= 3.28 min; MS (ESIpos): m / z = 1904 [M+H]+. Intermediate Q68: [000361](7S)-7-ethyl-8,11-dioxo-7,8,11,13-tetrahydro-2H,10H-[1,3]dioxolo[4,5- g]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-7-yl N-{(20S)-35-{4-[({(1R)-2-carboxy-1-[3-({3- [(propylcarbamoyl)amino]benzene-1-sulfonyl}amino)phenyl]ethyl}carbamoyl)amino]anilino}-20-[4- ({5-[(2,5-dioxopyrrolidin-1-yl)oxy]-5-oxopentanoyl}amino)butyl]-14,18,21,35-tetraoxo-4,7,10,25,28,31- hexaoxa-13,19,22,34-tetraazapentatriacontanan-1-oyl}-L-alpha-aspartyl-L-prolyl-L-valinateP368919WO / 59362-725.601[000362] Intermediate Q67 (21.6 mg, 100 % purity, 10.7 µmol), 1,1'-[(1,5-dioxopentane-1,5- diyl)bis(oxy)]di(pyrrolidine-2,5-dione) (10.5 mg, 32.1 µmol) and DIEA (5.6 µl, 32 µmol) were dissolved in DMF (2.5 ml) and the reaction was stirred at RT for 1h. DMF was evaporated in vacuo and the residue was purified by preparative HPLC, then lyophilized to give Intermediate Q68 (15.9 mg, 95% purity, 67% yield). LC-MS (Method 1): R = 3.74 m 2+ t in; MS (ESIpos): m / z = 1058 [M+2H] . Intermediate Q69: [000363](4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinolin-4-yl N-[(20S)-20-(4-aminobutyl)-35-{4-[({(1R)-2-carboxy-1-[3-({3- [(propylcarbamoyl)amino]benzene-1-sulfonyl}amino)phenyl]ethyl}carbamoyl)amino]anilino}- 14,18,21,35-tetraoxo-4,7,10,25,28,31-hexaoxa-13,19,22,34-tetraazapentatriacontanan-1-oyl]-L-alpha- aspartyl-L-prolyl-L-valinate.trifluoroacetic acid (1 / 1)[000364]Step 1: Intermediate Q65 (20.0 mg, 20.0 µmol)) was dissolved in DMF (2.0 ml). Intermediate Q1 (26.9 mg, 24.0 µmol) and DIEA (14 µl, 80 µmol) were added. The reaction was stirred overnight at RT and then concentrated in vacuo. The residue was separated by prep. HPLC to give 4S)-4,11-diethyl- 3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl N-[(20S)-20-{4- [(tert-butoxycarbonyl)amino]butyl}-35-{4-[({(1R)-2-carboxy-1-[3-({3-P368919WO / 59362-725.601 [(propylcarbamoyl)amino]benzene-1-sulfonyl}amino)phenyl]ethyl}carbamoyl)amino]anilino}- 14,18,21,35-tetraoxo-4,7,10,25,28,31-hexaoxa-13,19,22,34-tetraazapentatriacontanan-1-oyl]-L-alpha- aspartyl-L-prolyl-L-valinate (28.4 mg, 99 % purity, 71% yield). LC-MS (Method 1): Rt = 4.24 min; MS (ESIpos): m / z = 1988 [M+H]+. [000365]Step 2: (4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinolin-4-yl N-[(20S)-20-{4-[(tert-butoxycarbonyl)amino]butyl}-35-{4-[({(1R)-2-carboxy-1-[3-({3- [(propylcarbamoyl)amino]benzene-1-sulfonyl}amino)phenyl]ethyl}carbamoyl)amino]anilino}- 14,18,21,35-tetraoxo-4,7,10,25,28,31-hexaoxa-13,19,22,34-tetraazapentatriacontanan-1-oyl]-L-alpha- aspartyl-L-prolyl-L-valinate (28.2 mg, 100 % purity, 14.2 µmol) was dissolved in DCM (3.0 ml), then TFA (300 µl) was added. The reaction was stirred overnight at RT. The reaction was concentrated at RT on the oil pump. The resulting residue was dissolved in ACN / water and freeze-dried to give Intermediate Q69 (28.1 mg, 100% purity, 99% yield) as am amorphous residue. LC-MS (Method 1): Rt= 3.47 min; MS (ESIpos): m / z = 1888 [M+H]+. Intermediate Q70: [000366](4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinolin-4-yl N-{(20S)-35-{4-[({(1R)-2-carboxy-1-[3-({3-[(propylcarbamoyl)amino]benzene- 1-sulfonyl}amino)phenyl]ethyl}carbamoyl)amino]anilino}-20-[4-({5-[(2,5-dioxopyrrolidin-1- yl)oxy]-5-oxopentanoyl}amino)butyl]-14,18,21,35-tetraoxo-4,7,10,25,28,31-hexaoxa- 13,19,22,34-tetraazapentatriacontanan-1-oyl}-L-alpha-aspartyl-L-prolyl-L-valinate[000367] Intermediate Q69 (27.7 mg, 100 % purity, 13.8 µmol), 1,1'-[(1,5-dioxopentane-1,5- diyl)bis(oxy)]di(pyrrolidine-2,5-dione) (13.5 mg, 41.5 µmol) and DIEA (7.2 µl, 42 µmol) were dissolved in DMF (2.5 ml) and the reaction was stirred at RT for 1h. DMF was evaporated in vacuo and the residue was purified by preparative HPLC, then lyophilized to give Intermediate Q70 (18.5 mg, 96% purity, 61% yield). LC-MS (Method 1): Rt = 3.95 min; MS (ESIpos): m / z = 1050 [M+2H]2+.P368919WO / 59362-725.601 Intermediate Q71 [000368](3R)-3-[[4-[2-[[2-(2,5-dioxopyrrol-1- yl)acetyl]amino]ethylcarbamoylamino]phenyl]carbamoylamino]-3-[3-[[3- (propylcarbamoylamino)phenyl]sulfonylamino]phenyl]propanoic acid[000369]Intermediate Q71 was prepared in analogy as described for the synthesis of Intermediate Q57 starting with Intermediate Q36. Intermediate Q71 (11.3 mg, 94% purity, 52% yield) was obtained. LC-MS (Method 6): R + t = 1.24 min; MS (ESIpos): m / z = 778 [M+H] . Intermediate Q72 [000370](3R)-3-[[4-[2-[2-[2-[2-[[2-(2,5-dioxopyrrol-1- yl)acetyl]amino]ethoxy]ethoxy]ethoxy]ethylcarbamoylamino]phenyl]carbamoylamino]-3-[3-[[3- (propylcarbamoylamino)phenyl]sulfonylamino]phenyl]propanoic acid[000371]Intermediate Q72 was prepared in analogy as described for the synthesis of Intermediate Q57 starting with Intermediate Q64. Intermediate Q72 (9.0 mg, 98% purity, 48% yield) was obtained. LC-MS (Method 1): Rt= 2.96 min; MS (ESIpos): m / z = 910 [M+H]+. Intermediate Q73P368919WO / 59362-725.601 [000372](3R)-3-[[4-[2-[2-[2-[2-[[5-(2,5-dioxopyrrolidin-1-yl)oxy-5-oxo- pentanoyl]amino]ethoxy]ethoxy]ethoxy]ethylcarbamoylamino]phenyl]carbamoylamino]-3-[3-[[3- (propylcarbamoylamino)phenyl]sulfonylamino]phenyl]propanoic acid[000373]Intermediate Q73 was prepared in analogy as described for the synthesis of Intermediate Q1 starting with Intermediate Q64. Intermediate Q73 (10.0 mg, 88% purity, 44% yield) was obtained as a colorless foam. LC-MS (Method 1): R = + t 3.06 min; MS (ESIpos): m / z = 984 [M+H] . Intermediate Q74 [000374]Benzyl L-prolyl-L-valinate - trifluoroacetic acid (2 / 1)[000375]Intermediate Q74 was synthesized using classical peptide synthesis methods starting with the coupling of 1-(tert-butoxycarbonyl)-L-proline with 4-methylbenzene-1-sulfonic acid.benzyl L-valinate (1 / 1) in DMF in the presence of HATU and N,N-Diisopropylethylamine and subsequent removal of the Boc-protecting group with TFA in DCM. LC-MS (Method 2): Rt= 0.94 min; MS (ESIpos): m / z = 305 [M+H]+. Intermediate Q75 [000376]N2-(2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa-5-azaheptadecan-17-yl)-L-asparaginyl-L- prolyl-L-valineP368919WO / 59362-725.601[000377]Intermediate Q75 was synthesized using classical peptide synthesis methods starting with the coupling of Intermediate Q74 with 2,5-dioxopyrrolidin-1-yl N2-(tert-butoxycarbonyl)-L-asparaginate in DMF in the presence of N,N-Diisopropylethylamine and subsequent removal of the Boc-protecting group with TFA in DCM. This partially protected tripeptide was acylated with tert-butyl {2-[2-(2-{3-[(2,5- dioxopyrrolidin-1-yl)oxy]-3-oxopropoxy}ethoxy)ethoxy]ethyl}carbamate in DMF in the presence of N, N-Diisopropylethylamine. In the final step the benzylester was removed by hydrogenolysis over 10% Pd / charcoal. LC-MS (Method 2): R = 1.10 min; MS (E + t SIpos): m / z = 632 [M+H] . Intermediate Q76 [000378]N2-(3-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}propanoyl)-L-asparaginyl-L-prolyl-N-[(2S)-4- [{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2-yl]-2,2-dimethylpropyl}(hydroxyacetyl)amino]- 1-(methylamino)-1-oxobutan-2-yl]-L-valinamideP368919WO / 59362-725.601 [000379]Step 1: To a solution of (2S)-2-amino-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol- 2-yl]-2,2-dimethylpropyl}(hydroxyacetyl)amino]-N-methylbutanamide (63.0 mg, 120 µmol, CAS Nr: 1800460-13-2, as described in WO2015096982) in DMF (5 ml) were added N2-(2,2-dimethyl-4,17- dioxo-3,8,11,14-tetraoxa-5-azaheptadecan-17-yl)-L-asparaginyl-L-prolyl-L-valine (90.7 mg, 144 µmol) (Intermediate Q75), HATU (59.1 mg, 156 µmol) and DIEA (63 µl, 360 µmol). The mixture was stirred at rt for 1 h and then concentrated under reduced pressure. The residue was purified by preparative HPLC and lyophilized to afford N2-(2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa-5-azaheptadecan-17-yl)-L- asparaginyl-L-prolyl-N-[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2-yl]-2,2- dimethylpropyl}(hydroxyacetyl)amino]-1-(methylamino)-1-oxobutan-2-yl]-L-valinamide (92 mg, 98 % purity, 66 % yield) as an amorphous residue. LC-MS (Method 1): Rt= 4.93 min; MS (ESIpos): m / z = 1141 [M+H]+ [000380]Step 2: To a solution of N2-(2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa-5-azaheptadecan-17- yl)-L-asparaginyl-L-prolyl-N-[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2-yl]-2,2- dimethylpropyl}(hydroxyacetyl)amino]-1-(methylamino)-1-oxobutan-2-yl]-L-valinamide (92.0 mg, 98 % purity, 79.2 µmol) in Trifluoroethanol (4 ml), was added zinc chloride (86.3 mg, 633 µmol). The mixture was stirred at 50°C for 5 h. The mixture was freezed at -20°C for 4 days and then stirred again at 50°C for 3 h. EDTA (185 mg, 633 µmol) and water 0.1% TFA (3 ml) were then added and the resulting mixture was purified over preparative HPLC and lyophilized to afford Intermediate Q76 (84 mg, 96 % purity, 97 % yield) as an amorphous residue. LC-MS (Method 1): Rt = 3.44 min; MS (ESIpos): m / z = 1041 [M+H]+ Intermediate Q77: [000381]N2-{18-[(2,5-dioxopyrrolidin-1-yl)oxy]-14,18-dioxo-4,7,10-trioxa-13-azaoctadecanan-1-oyl}- L-asparaginyl-L-prolyl-N-[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2-yl]-2,2- dimethylpropyl}(hydroxyacetyl)amino]-1-(methylamino)-1-oxobutan-2-yl]-L-valinamideP368919WO / 59362-725.601 [000382] Intermediate Q76 (21.6 mg, 100 % purity, 10.7 µmol), 1,1'-[(1,5-dioxopentane-1,5- diyl)bis(oxy)]di(pyrrolidine-2,5-dione) (20.0 mg, 96 % purity, 18.4 µmol) and DIEA (9.6 µl, 55 µmol) were dissolved in DMF (3.0 ml) and the reaction was stirred at RT for 1h. DMF was evaporated in vacuo and the residue was purified by preparative HPLC, then lyophilized to give Intermediate Q77 (11.1 mg, 77% purity, 37% yield). LC-MS (Method 1): R = 4.45 m + t in; MS (ESIpos): m / z = 1252 [M+H] . Intermediate Q78: [000383](3,20-difluoro-13-oxa-5,7,18,25-tetrazatetracyclo[17.3.1.12,6.18,12]pentacosa- 1(22),2,4,6(25),8,10,12(24),19(23),20-nonaen-10-yl)methyl-imino-methyl-oxo-λ⁶-sulfane (single enantiomer)[000384]Intermediate Q78 was prepared from commercially available intermediates according to the following steps: [000385]Step 1: Synthesis of tert-butyl N-[[3-amino-5-(4-hydroxybutoxy)phenyl]methyl-methyl-oxo-λ⁶- sulfanylidene]carbamate[000386]To a mixture of tert-butyl N-[[3-(4-hydroxybutoxy)-5-nitro-phenyl]methyl-methyl-oxo-λ⁶- sulfanylidene]carbamate (4 g, 9.94 mmol, 1 eq) in EtOH (60 mL) and H2O (12 mL) was added Fe (2.78 g, 49.69 mmol, 5 eq) and NH4Cl (2.66 g, 49.69 mmol, 5 eq) at 20°C under N2. The mixture was stirred at 80°C for 1 hour. The reaction mixture was filtered and concentrated under reduced pressure to remove EtOH. The mixture was diluted with H2O (50 mL) and extracted with EtOAc (3x80 mL). The combined organic layers were washed with brine (2x80 mL), dried over anhydrous Na2SO4,filtered, and concentrated under reduced pressure to give Q78-1 (3.8 g, crude) as a yellow solid.1H NMR (400 MHz,P368919WO / 59362-725.601 CDCl3) δ 6.32 - 6.29 (m, 2H), 6.24 (t, J = 2.0 Hz, 1H), 4.59 (s, 2H), 3.94 (t, J = 6.2 Hz, 2H), 3.70 (t, J = 6.2 Hz, 2H), 2.94 (s, 3H), 1.90 - 1.78 (m, 2H), 1.78 - 1.65 (m, 2H), 1.51 (s, 9H). [000387]Step 2: Synthesis of 2-chloro-5-fluoro-4-(4-fluoro-3-nitro-phenyl)pyrimidine (Q78-2)[000388]To a solution of 2,4-dichloro-5-fluoro-pyrimidine (9.48 g, 56.78 mmol, 1 eq) and (4-fluoro-3- nitro-phenyl)boronic acid (10.5 g, 56.78 mmol, 1 eq) in DME (150 mL) were added K2CO3 (2 M, 85.17 mL, 3 eq) and Pd(dppf)Cl2.CH2Cl2 (4.64 g, 5.68 mmol, 0.1 eq) at 20°C under N2. The mixture was stirred at 80°C for 2.5 hours. TLC showed that the starting material had disappeared and that a new main spot was formed. The mixture was extracted with EtOAc (2x100 mL) and H2O (100 mL). The combined organic phase was washed with brine (3x50 mL), dried with anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 20:1 to 8:1) to give Q78-2 (27 g, 99.41 mmol, 87.5% yield) as a light-yellow solid.1H NMR (400 MHz, DMSO-d6) δ 9.06 (d, J = 3.0 Hz, 1H), 8.73 (dd, J = 2.2, 7.2 Hz, 1H), 8.47 - 8.39 (m, 1H), 7.83 (dd, J = 8.8, 11.2 Hz, 1H). [000389]Step 3: Synthesis of 5-(2-chloro-5-fluoro-pyrimidin-4-yl)-2-fluoro-aniline (Q78-3)[000390]To a solution of Q78-2 (19.7 g, 72.53 mmol, 1 eq) in EtOH (200 mL) and H2O (40 mL) were added Fe (20.25 g, 362.66 mmol, 5 eq) and NH4Cl (19.40 g, 362.66 mmol, 5 eq) at 20°C. The mixture was stirred at 80°C for 2 hours. The mixture was filtered through a pad of Celite, and the pad cake was washed with EtOH (5x15 mL). The mixture was concentrated under reduced pressure to remove EtOH. The mixture was diluted with H2O (100 mL) and extracted with ethyl acetate (3x100 mL). The combined organic layers were washed with brine (2x50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 20:1 to 8:1) to give Q78-3 (12.4 g, 51.32 mmol, 70.75% yield) as a yellow solid.1H NMR (400 MHz, DMSO-d6) δ 8.89 (d, J = 3.6 Hz, 1H), 7.56 (dd, J = 1.8, 8.8 Hz, 1H), 7.28 - 7.14 (m, 2H), 5.53 (s, 2H). [000391]Step 4: Synthesis of N-[5-(2-chloro-5-fluoro-pyrimidin-4-yl)-2-fluoro-phenyl]-2-nitro- benzenesulfonamide (Q78-4)[000392]To a mixture of Q78-3 (7.35 g, 30.42 mmol, 1 eq) and 2-nitrobenzenesulfonyl chloride (10.79 g, 48.67 mmol, 1.6 eq) in DCM (80 mL) was added DMAP (260.14 mg, 2.13 mmol, 0.07 eq) and pyridine (3.85 g, 48.67 mmol, 3.93 mL, 1.6 eq) at 20°C under N2. The mixture was stirred at 20°C for 16 hours. The reaction mixture was poured into water (50 mL). The aqueous phase was extracted with ethyl acetate (3x100 mL). The combined organic phase was washed with brine (2x100 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude product was washed with DCM (2x8 mL) to give Q78-4 (21 g, 49.21 mmol, 80.88% yield) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 8.58 (d, J = 2.8 Hz, 1H), 8.49 - 8.41 (m, 1H), 8.05 - 7.93 (m, 3H), 7.73 - 7.62 (m, 3H), 7.18 (t, J = 9.2 Hz, 1H). [000393]Step 5: Synthesis of N-[5-(2-chloro-5-fluoro-pyrimidin-4-yl)-2-fluoro-phenyl]-2-nitro- benzenesulfonamide (Q78-5)[000394]To a mixture of Q78-1 (3.8 g, 10.20 mmol, 1 eq) and Q78-4 (4.35 g, 10.20 mmol, 1 eq) in NMP (6 mL) and toluene (60 mL) was added Xphos Pd G1 (376.84 mg, 510.10 umol, 0.05 eq), Xphos (486.35 mg, 1.02 mmol, 0.1 eq) and K3PO4(10.83 g, 51.01 mmol, 5 eq) at 20°C under N2. The mixture was stirred at 110°C for 4 hours. TLC (petroleum ether:ethyl acetate = 0:1, Rf= 0.4) indicated that the starting material was consumed completely and that one new spot formed. The reaction mixture was poured into water (50 mL). The aqueous phase was extracted with ethyl acetate (2x100 mL). The combined organic phase was washed with brine (2x100 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 20:1 to 1:1) to give Q78-5 (6.5 g, 8.52 mmol, 83.5% yield) as a yellow solid.1H NMR (400 MHz CDCl3) δ 843 (dd J = 22 76 Hz 1H) 839 - 833 (m 1H) 801 - 786 (m 3H) 779 - 768 (mP368919WO / 59362-725.601 1H), 7.67 - 7.57 (m, 1H), 7.50 (s, 1H), 7.38 - 7.31 (m, 2H), 7.13 (t, J = 9.2 Hz, 1H), 6.64 (s, 1H), 4.81 - 4.63 (m, 2H), 4.03 (t, J = 6.2 Hz, 2H), 3.72 (t, J = 6.4 Hz, 2H), 3.02 (s, 3H), 1.94 - 1.84 (m, 2H), 1.79 - 1.71 (m, 2H), 1.48 (s, 9H) [000395]Step 6: Synthesis of tert-butyl N-[[3,20-difluoro-18-(2-nitrophenyl)sulfonyl-13-oxa-5,7,18,25- tetrazatetracyclo[17.3.1.12,6.18,12]pentacosa-1(22),2,4,6(25),8,10,12(24),19(23),20-nonaen-10- yl]methyl-methyl-oxo-λ⁶-sulfanylidene]carbamate (Q78-6)[000396]To a mixture of Q78-5 (5.4 g, 7.08 mmol, 1 eq) in toluene (110 mL) was added CMBP (3.42 g, 14.16 mmol, 2 eq) at 20°C under N2. The mixture was stirred at 110°C for 16 hours. The reaction mixture was diluted with EtOAc (30 mL) and filtered, the solid was dried in vacuo to give Q78-6 (4.6 g, 6.18 mmol, 87.2% yield) as a white solid.1H NMR (400 MHz, DMSO-d6) δ 10.03 (s, 1H), 8.74 (d, J = 3.6 Hz, 1H), 8.32 (br d, J = 5.8 Hz, 1H), 8.15 (br d, J = 4.4 Hz, 1H), 8.07 (s, 1H), 7.98 - 7.95 (m, 1H), 7.89 (t, J = 7.6 Hz, 1H), 7.84 - 7.81 (m, 1H), 7.79 - 7.74 (m, 1H), 7.49 (t, J = 9.4 Hz, 1H), 6.85 (s, 1H), 6.60 (s, 1H), 4.78 - 4.70 (m, 2H), 4.05 - 3.91 (m, 2H), 3.91 - 3.68 (m, 2H), 3.17 (s, 3H), 1.92 - 1.72 (m, 2H), 1.39 (s, 9H), 1.36 - 1.22 (m, 2H). [000397]Step 7: Synthesis of tert-butyl N-[(3,20-difluoro-13-oxa-5,7,18,25- tetrazatetracyclo[17.3.1.12,6.18,12]pentacosa-1(22),2,4,6(25),8,10,12(24),19(23),20-nonaen-10- yl)methyl-methyl-oxo-λ⁶-sulfanylidene]carbamate (Q78-7)[000398]To a solution of Q78-6 (4.6 g, 6.18 mmol, 1 eq) in DMF (80 mL) was added phenylsulfanylsodium (1.8 g, 13.59 mmol, 2.2 eq) at 20°C under N2. The mixture was stirred at 20°C for 16 hours. The reaction mixture was poured into water (50 mL). The aqueous phase was extracted with dichloromethane (2x100 mL). The combined organic phase was washed with brine (2x100 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=10:1 to 0:1) to give Q78-7 (2.85 g, 4.77 mmol, 77.1% yield, 93.6% purity) as a light-yellow solid.1H NMR (400 MHz, DMSO-d6) δ 9.87 (s, 1H), 8.59 (d, J = 4.2 Hz, 1H), 8.08 (s, 1H), 7.64 (br d, J = 7.8 Hz, 1H), 7.30 - 7.22 (m, 1H), 7.16 (dd, J = 8.4, 11.4 Hz, 1H), 6.89 (s, 1H), 6.84 - 6.74 (m, 1H), 5.96 (br s, 1H), 4.77 (s, 2H), 4.13 (br t, J = 4.8 Hz, 2H), 3.25 - 3.10 (m, 5H), 1.80 (br dd, J = 7.8, 16.2 Hz, 2H), 1.71 - 1.54 (m, 2H), 1.39 (s, 9H). [000399]Step 8: Synthesis of (3,20-difluoro-13-oxa-5,7,18,25- tetrazatetracyclo[17.3.1.12,6.18,12]pentacosa-1(22),2,4,6(25),8,10,12(24),19(23),20-nonaen-10- yl)methyl-imino-methyl-oxo-λ⁶-sulfane (Q78-8)[000400]Q78-7 (2.85 g, 5.09 mmol, 1 eq) was dissolved in TFA (3 mL) and DCM (30 mL) at 20°C under N2. The mixture was stirred at 20°C for 16 hours. The reaction mixture was poured into NaHCO3 (50 mL) and ethyl acetate (50 mL). The mixture was stirred for 0.5 hours, then the mixture was filtered,P368919WO / 59362-725.601 and the filter cake was dried in vacuo. The crude product was washed with EtOAc (30 mL) to give Q78-8 (2.1 g, 4.30 mmol, 84.3% yield, 94% purity) as a light-yellow solid.1H NMR (400 MHz, DMSO-d6) δ 9.81 (s, 1H), 8.59 (d, J = 4.4 Hz, 1H), 8.03 (s, 1H), 7.66 (br d, J = 7.6 Hz, 1H), 7.30 - 7.23 (m, 1H), 7.23 - 7.11 (m, 1H), 6.86 (s, 1H), 6.84 - 6.78 (m, 1H), 5.97 (br t, J = 5.1 Hz, 1H), 4.37 - 4.24 (m, 2H), 4.14 (br t, J = 4.8 Hz, 2H), 3.62 (s, 1H), 3.29 - 3.14 (m, 2H), 2.85 (s, 3H), 1.87 - 1.72 (m, 2H), 1.72 - 1.55 (m, 2H). [000401]Step 9: Chiral separation of Q78-4[000402]Compound Q78-8 was separated by prep-SFC (column: DAICEL CHIRALPAK AD (250mm*30mm, 10um);mobile phase: [ACN / EtOH(0.1%NH3H2O)]; B%: 60%-60%,45min) to give B28 (803.38 mg, 38.2% yield) as a light yellow solid and B29 (762.33 mg, 36.3% yield) as a light-yellow solid. Note: The R / S configuration of these 2 compounds were not confirmed. [000403]Intermediate Q78: 1H NMR (400 MHz, DMSO-d6) δ 9.82 (s, 1H), 8.59 (d, J = 4.4 Hz, 1H), 8.03 (t, J = 1.8 Hz, 1H), 7.66 (dd, J = 1.8, 8.8 Hz, 1H), 7.32 - 7.19 (m, 1H), 7.19 - 7.06 (m, 1H), 6.84 (br d, J = 18.4 Hz, 2H), 6.02 - 5.92 (m, 1H), 4.39 - 4.20 (m, 2H), 4.20 - 4.08 (m, 2H), 3.61 (s, 1H), 3.26 - 3.11 (m, 2H), 2.84 (s, 3H), 1.86 - 1.68 (m, 2H), 1.67 - 1.52 (m, 2H). The desired enantiomer (optical rotation 3.54˚ ± 0.00˚, 20C, 589 nm) was obtained with 100% ee at Rt 2.75-3.70 min. LC-MS: Rt = 2.33 min; MS (ESIpos): m / z = 460 [M+H]+. Single (+) isomer, absolute stereochemistry unknown. [000404]Intermediate Q78*: 1H NMR (400 MHz, DMSO-d6) δ 9.81 (s, 1H), 8.59 (d, J = 4.2 Hz, 1H), 8.03 (s, 1H), 7.65 (br d, J = 7.4 Hz, 1H), 7.30 - 7.21 (m, 1H), 7.17 (dd, J = 8.4, 11.6 Hz, 1H), 6.88 - 6.83 (m, 1H), 6.81 (s, 1H), 5.97 (br t, J = 5.4 Hz, 1H), 4.37 - 4.21 (m, 2H), 4.21 - 4.06 (m, 2H), 3.61 (s, 1H), 3.28 - 3.14 (m, 2H), 2.84 (s, 3H), 1.95 - 1.73 (m, 2H), 1.66 - 1.52 (m, 2H). The desired enantiomer (optical rotation -4.33˚ ± 0.28˚, 20C, 589 nm) was obtained with 99.62% ee at Rt 3.70-4.80 min. LC-MS: R = 2.33 min + t ; MS (ESIpos): m / z = 460 [M+H] . Single (-) isomer, absolute stereochemistry unknown. Intermediate Q79: [000405]N-{18-[(2,5-dioxopyrrolidin-1-yl)oxy]-14,18-dioxo-4,7,10-trioxa-13-azaoctadecanan-1-oyl}- L alpha aspartyl L prolyl N [(R*) {[320 difluoro 13 oxa 571825tetraazatetracyclo[17.3.1.12,6.18,12]pentacosa-1(23),2(25),3,5,8(24),9,11,19,21-nonaen-10- yl]methyl}(methyl)oxo-lambda6-sulfanylidene]-L-valinamide[000406]Intermediate Q79 (was synthesized using Intermediate Q78 following the same general procedure as described previously to synthesize Intermediate Q42. Intermediate Q79 was obtained (14.7 mg, 90% purity, 62% yield). LC-MS (Method 1): Rt= 4.20 min; MS (ESIpos): m / z = 1185 [M+H]+. Intermediate Q80: [000407]Trifluoroacetic acid.N-[(benzyloxy)carbonyl]-N-methyl-L-valyl-N-[(3R,4S,5S)-3-methoxy-1- {(2S)-2-[(1R,2R)-1-methoxy-2-methyl-3-oxo-3-{[(1S,2R)-1-phenyl-1-(L-valyloxy)propan-2- yl]amino}propyl]pyrrolidin-1-yl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide (1 / 1)[000408]Step 1: To a solution of Monomethyl auristatin E (N-methyl-L-valyl-N-[(3R,4S,5S)-1-{(2S)-2- [(1R,2R)-3-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]amino}-1-methoxy-2-methyl-3- oxopropyl]pyrrolidin-1-yl}-3-methoxy-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide, 200 mg, 279 µmol) in DMF (28 ml) were added 1-{[(benzyloxy)carbonyl]oxy}pyrrolidine-2,5-dione (83.3 mg, 334 µmol) and DIEA (150 µl, 840 µmol). The mixture was stirred at rt for 20 h and then concentrated under reduced pressure. The residue was purified by preparative HPLC and lyophilized to afford N- [(benzyloxy)carbonyl]-N-methyl-L-valyl-N-[(3R,4S,5S)-1-{(2S)-2-[(1R,2R)-3-{[(1S,2R)-1-hydroxy-1-P368919WO / 59362-725.601 phenylpropan-2-yl]amino}-1-methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl}-3-methoxy-5-methyl-1- oxoheptan-4-yl]-N-methyl-L-valinamide (224 mg, 100 % purity, 94 % yield). LC-MS (Method 2): Rt = 2.29 min; MS (ESIpos): m / z = 853 [M+H]+. [000409]Step 2: To a solution of N-[(benzyloxy)carbonyl]-N-methyl-L-valyl-N-[(3R,4S,5S)-1-{(2S)-2- [(1R,2R)-3-{[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]amino}-1-methoxy-2-methyl-3- oxopropyl]pyrrolidin-1-yl}-3-methoxy-5-methyl-1-oxoheptan-4-yl]-N-methyl-L-valinamide (222 mg, 100 % purity, 260 µmol) in DCM (40 ml) were added tert-butyl (4S)-4-methyl-2,5-dioxo-1,3- oxazolidine-3-carboxylate (190 mg, 781 µmol) and DMAP (63.6 mg, 521 µmol). The mixture was refluxed for 16 h and then concentrated under reduced pressure. The residue was purified by preparative HPLC and lyophilized to afford N-[(benzyloxy)carbonyl]-N-methyl-L-valyl-N-[(3R,4S,5S)-1-{(2S)-2- [(1R,2R)-3-{[(1S,2R)-1-{[N-(tert-butoxycarbonyl)-L-valyl]oxy}-1-phenylpropan-2-yl]amino}-1- methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl}-3-methoxy-5-methyl-1-oxoheptan-4-yl]-N-methyl-L- valinamide (267 mg, 100 % purity, 98 % yield). LC-MS (Method 3): Rt= 6.35 min; MS (ESIpos): m / z = 1052 [M+H]+. [000410]Step 3: To a solution of N-[(benzyloxy)carbonyl]-N-methyl-L-valyl-N-[(3R,4S,5S)-1-{(2S)-2- [(1R,2R)-3-{[(1S,2R)-1-{[N-(tert-butoxycarbonyl)-L-valyl]oxy}-1-phenylpropan-2-yl]amino}-1- methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl}-3-methoxy-5-methyl-1-oxoheptan-4-yl]-N-methyl-L- valinamide (323 mg, 100 % purity, 307 µmol) in DCM (50 ml), was added TFA (5.0 ml). The mixture was stirred at rt for 2 h and then concentrated under reduced pressure. The residue was dissolved in ACN / H2O and lyophilized to afford Intermediate Q80 (339 mg, 100 % purity, quant.) as an amorphous residue. LC-MS (Method 2): R = 1.80 min; MS (ESIpos + t ): m / z = 952 [M+H] . Intermediate Q81: [000411](1S,2R)-2-({(2R,3R)-3-methoxy-3-[(2S)-1-{(3R,4S,5S)-3-methoxy-5-methyl-4-[methyl(N- methyl-L-valyl-L-valyl)amino]heptanoyl}pyrrolidin-2-yl]-2-methylpropanoyl}amino)-1-phenylpropyl N- (3-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}propanoyl)-L-alpha-aspartyl-L-prolyl-L-valinate[000412]Step 1: To a solution of trifluoroacetic acid.N-[(benzyloxy)carbonyl]-N-methyl-L-valyl-N- [(3R,4S,5S)-3-methoxy-1-{(2S)-2-[(1R,2R)-1-methoxy-2-methyl-3-oxo-3-{[(1S,2R)-1-phenyl-1-(L- valyloxy)propan-2-yl]amino}propyl]pyrrolidin-1-yl}-5-methyl-1-oxoheptan-4-yl]-N-methyl-L- valinamide (1 / 1) (339 mg 100 % purity 318 µmol) (Intermediate Q80) in DMF (20 ml) were addedP368919WO / 59362-725.601 (2S)-1-[(19S)-19-(2-tert-butoxy-2-oxoethyl)-2,2-dimethyl-4,17,20-trioxo-3,8,11,14-tetraoxa-5,18- diazaicosan-20-yl]pyrrolidine-2-carboxylic acid (206 mg, 350 µmol) (Intermediate Q39), HATU (193 mg, 508 µmol) and DIEA (170 µl, 950 µmol). The mixture was stirred at rt for 1h30 and then concentrated under reduced pressure. The residue was purified by preparative HPLC and lyophilized to afford tert-butyl (19S)-19-[(2S)-2-({(2S)-1-[(1S,2R)-2-{[(2R,3R)-3-{(2S)-1-[(5S,8S,11S,12R)-11-[(2S)- butan-2-yl]-12-methoxy-4,10-dimethyl-3,6,9,14-tetraoxo-1-phenyl-5,8-di(propan-2-yl)-2-oxa-4,7,10- triazatetradecan-14-yl]pyrrolidin-2-yl}-3-methoxy-2-methylpropanoyl]amino}-1-phenylpropoxy]-3- methyl-1-oxobutan-2-yl}carbamoyl)pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa- 5,18-diazahenicosan-21-oate (447 mg, 100 % purity, 92 % yield) as an amorphous residue. LC-MS (Method 3): Rt= 6.26 min; MS (ESIpos): m / z = 1523 [M+H]+. [000413]Step 2: To a solution of tert-butyl (19S)-19-[(2S)-2-({(2S)-1-[(1S,2R)-2-{[(2R,3R)-3-{(2S)-1- [(5S,8S,11S,12R)-11-[(2S)-butan-2-yl]-12-methoxy-4,10-dimethyl-3,6,9,14-tetraoxo-1-phenyl-5,8- di(propan-2-yl)-2-oxa-4,7,10-triazatetradecan-14-yl]pyrrolidin-2-yl}-3-methoxy-2- methylpropanoyl]amino}-1-phenylpropoxy]-3-methyl-1-oxobutan-2-yl}carbamoyl)pyrrolidine-1- carbonyl]-2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa-5,18-diazahenicosan-21-oate (52.9 mg, 34.7 µmol) in TFA (10 ml). The mixture was stirred at rt for 72 h and then concentrated under reduced pressure. The residue was dissolved in ACN / H2O and lyophilized to afford Intermediate Q81 (10.5 mg, 77 % purity, 19% yield). LC-MS (Method 1): Rt= 2.73 min; MS (ESIpos): m / z = 1233 [M+H]+. Intermediate Q82: [000414](1S,2R)-2-({(2R,3R)-3-methoxy-3-[(2S)-1-{(3R,4S,5S)-3-methoxy-5-methyl-4-[methyl(N- methyl-L-valyl-L-valyl)amino]heptanoyl}pyrrolidin-2-yl]-2-methylpropanoyl}amino)-1-phenylpropyl N- [1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2,15-dioxo-6,9,12-trioxa-3-azapentadecan-15-yl]-L-alpha- aspartyl-L-prolyl-L-valinate[000415]Intermediate Q81 (10.4 mg, 77 % purity, 6.52 µmol), 1-{2-[(2,5-dioxopyrrolidin-1-yl)oxy]-2- oxoethyl}-1H-pyrrole-2,5-dione (3.29 mg, 13.0 µmol) and DIEA (3.4 µl, 20 µmol) were dissolved in DMF (2.0 ml) and the reaction was stirred at RT for 1h. DMF was evaporated in vacuo and the residue was purified by preparative HPLC, then lyophilized to give Intermediate Q82 (4.8 mg, 100% purity, 54% yield). LC-MS (Method 1): R + t = 3.25 min; MS (ESIpos): m / z = 1369 [M+H] .Intermediate Q83: [000416]Trifluoroacetic acid.N1-(2-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}ethyl)-N4-[4-({2-[(2S)-2- cyano-4,4-difluoropyrrolidin-1-yl]-2-oxoethyl}carbamoyl)quinolin-8-yl]butanediamide (1 / 1)[000417]Step 1: To a solution of 4-{[4-({2-[(2S)-2-cyano-4,4-difluoropyrrolidin-1-yl]-2- oxoethyl}carbamoyl)quinolin-8-yl]amino}-4-oxobutanoic acid (40.0 mg, 100 % purity, 87.1 µmol) in DMF (2.0 ml), were added tert-butyl (2-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}ethyl)carbamate (50.9 mg, 174 µmol), HATU (43.0 mg, 113 µmol) and DIEA (61 µl, 350 µmol). The reaction was stirred for 30minutes at RT and then concentrated in vacuo. The residue was separated by prep. HPLC and lyophilized to give tert-butyl (16-{[4-({2-[(2S)-2-cyano-4,4-difluoropyrrolidin-1-yl]-2- oxoethyl}carbamoyl)quinolin-8-yl]amino}-13,16-dioxo-3,6,9-trioxa-12-azahexadecan-1-yl)carbamate (53.7 mg, 100 % purity, 84% yield). LC-MS (Method 2): Rt = 1.44 min; MS (ESIpos): m / z = 734 [M+H]+. [000418]Step 2: tert-butyl (16-{[4-({2-[(2S)-2-cyano-4,4-difluoropyrrolidin-1-yl]-2- oxoethyl}carbamoyl)quinolin-8-yl]amino}-13,16-dioxo-3,6,9-trioxa-12-azahexadecan-1-yl)carbamate (53.4 mg, 100 % purity, 72.8 µmol) was dissolved in DCM (5.0 ml), then TFA (1.0 ml) was added. The reaction was stirred for 1h at RT. The reaction was concentrated at RT on the oil pump. The resulting residue was dissolved in ACN / water and freeze-dried to give Intermediate Q83 (52.9 mg, 100% purity, 97% yield). LC-MS (Method 2): Rt= 0.81 min; MS (ESIpos): m / z = 634 [M+H]+. Intermediate Q84: [000419]N1-[4-({2-[(2S)-2-cyano-4,4-difluoropyrrolidin-1-yl]-2-oxoethyl}carbamoyl)quinolin-8-yl]- N4-{17-[(2,5-dioxopyrrolidin-1-yl)oxy]-13,17-dioxo-3,6,9-trioxa-12-azaheptadecan-1-yl}butanediamideP368919WO / 59362-725.601 [000420] Intermediate Q83 (68.9 mg, 211 µmol), 1,1'-[(1,5-dioxopentane-1,5- diyl)bis(oxy)]di(pyrrolidine-2,5-dione) (68.9 mg, 211 µmol) and DIEA (37 µl, 210 µmol) were dissolved in DMF (2.0 ml) and the reaction was stirred at RT for 1h. DMF was evaporated in vacuo and the residue was purified by preparative HPLC, then lyophilized to give Intermediate Q84 (40.3 mg, 90% purity, 61% yield). LC-MS (Method 2): R = 1.15 min; M + t S (ESIpos): m / z = 845 [M+H] . Intermediate Q85: (3S,19S,37S)-3,37-bis{[(2S)-2-{[(2S)-1-{[(4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H- pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl]oxy}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidin-1- yl]carbonyl}-19-({21-[(2,5-dioxopyrrolidin-1-yl)oxy]-17,21-dioxo-4,7,10,13-tetraoxa-16-azahenicosan- 1-oyl}amino)-5,18,22,35-tetraoxo-8,11,14,26,29,32-hexaoxa-4,17,23,36-tetraazanonatriacontane-1,39- dioic acid[000421] Intermediate 86 (30.0mg, 13.3umol) was dissolved in DMF (3.0ml) and then disuccinimidyl glutarate (13.03mg, 39.9umol) and DIEA (9.27ul, 53.2umol) were added. The reaction was stirred for 2h at RT. The residue was purified by prep HPLC and lyophilised to give Intermediate 85 (21.3 mg, 100% purity, 68% yield). LC-MS (Method 1): Rt= 4.31 min; MS (ESIpos): m / z = 1176 [M+2H]2+. Intermediate Q86: (3S,19S,37S)-19-[(15-amino-4,7,10,13-tetraoxapentadecan-1-oyl)amino]-3,37-bis{[(2S)-2-{[(2S)-1- {[(4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-P368919WO / 59362-725.601 yl]oxy}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidin-1-yl]carbonyl}-5,18,22,35-tetraoxo- 8,11,14,26,29,32-hexaoxa-4,17,23,36-tetraazanonatriacontane-1,39-dioic acid trifluoroacetate (1:1)[000422] Intermediate 87 (77.0 mg, 34.4 umol) was dissolved in DCM (6.0 ml), then TFA (1.5ml) was added and the solution wa stirred at RT for 1h. The reaction was concentrated in vacuo, dissolved in ACN / H2O and lyophilised to give Intermediate 86 (75.2 mg, 97% purity, 94% yield) as an amorphous residue. LC-MS (Method 1): Rt= 3.73 min; MS (ESIpos): m / z = 1070 [M+2H]2+. Intermediate Q87: (3S,19S,37S)-3,37-bis{[(2S)-2-{[(2S)-1-{[(4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro-1H- pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl]oxy}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidin-1- yl]carbonyl}-19-[(2,2-dimethyl-4,20-dioxo-3,8,11,14,17-pentaoxa-5-azaicosan-20-yl)amino]-5,18,22,35- tetraoxo-8,11,14,26,29,32-hexaoxa-4,17,23,36-tetraazanonatriacontane-1,39-dioic acid[000423] Intermediate Q88 (105.0mg, 0.051mmol) was dissolved in DMF (4.0ml), then tert-butyl {15- [(2,5-dioxopyrrolidin-1-yl)oxy]-15-oxo-3,6,9,12-tetraoxapentadec-1-yl}carbamate (28.1mg, 0.061mmol) and DIEA (26.4ul, 0.152mmol) were added. The reaction was stirred for 2h at RT, which was then purified by prep HPLC and lyophilized to give Intermediate Q87 (77.30 mg, 100% purity, 68% yield). LC-MS (Method 1): Rt= 4.63 min; MS (ESIpos): m / z = 1120 [M+2H]2+. Intermediate Q88: (3S,19S,37S)-19-amino-3,37-bis{[(2S)-2-{[(2S)-1-{[(4S)-4,11-diethyl-3,14-dioxo-3,4,12,14-tetrahydro- 1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl]oxy}-3-methyl-1-oxobutan-2- yl]carbamoyl}pyrrolidin-1-yl]carbonyl}-5,18,22,35-tetraoxo-8,11,14,26,29,32-hexaoxa-4,17,23,36-P368919WO / 59362-725.601 tetraazanonatriacontane-1,39-dioic acid trifluoroacetate (1:1)[000424] Intermediate Q89 (102.0mg, 51.2 umol) was dissolved in DCM (8.0ml), TFA (2.0ml) was added and the reaction was stirred at RT for 1h. The reaction wasn concentrated in vacuo, dissolved in ACN / H2O and lyophilised to give Intermediate Q88 (105.2 mg, 97% purity, 99% yield) as an amorphous residue. LC-MS (Method 1): Rt= 3.78 min; MS (ESIpos): m / z = 947 [M+2H]2+. Intermediate Q89: (3S,19S,37S)-19-[(tert-butoxycarbonyl)amino]-3,37-bis{[(2S)-2-{[(2S)-1-{[(4S)-4,11-diethyl-3,14- dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl]oxy}-3-methyl-1- oxobutan-2-yl]carbamoyl}pyrrolidin-1-yl]carbonyl}-5,18,22,35-tetraoxo-8,11,14,26,29,32-hexaoxa- 4,17,23,36-tetraazanonatriacontane-1,39-dioic acid[000425] Intermediate Q90 (43.9mg, 0.10mmol) was dissolved in DMF (10.0ml), then Intermediate 11 (200.0mg, 0.199mmol) and DIEA (104.0ul, 597.0 umol) were added and the reaction was stirred for 1h at RT. The reaction was concentrated in vacuo and the residue was purified twice by prep HPLC and then lyophilized to give Intermediate Q89 (102.4 mg, 100% purity, 52% yield) as an amorphous residue. LC-MS (Method 1): Rt= 4.56 min; MS (ESIpos): m / z = 996 [M+2H]2+. Intermediate Q90: bis(2,5-dioxopyrrolidin-1-yl) N-(tert-butoxycarbonyl)-L-glutamate[000426] To a solution of Boc-L-glutamic acid (250.0mg, 1.011mmol) and N-hydroxysuccinimide (290.9mg, 2.53mmol) in DMF (2.5ml) was added 1-(3-dimethylaminopropyl)-3- ethylcarbodiimidhydrochlorid (484.6mg, 2.53mmol) and the reaction was stirred overnight at RT. The reaction was concentrated in vacuo, water was added and the solution was extracted with ethyl acetateP368919WO / 59362-725.601 (3x). The combined organic phases were washed with 10% citric acid (2x), brine, sat. NaHCO3 (3x) and brine. The organic phase was dried over MgSO4 and then concentrated in vacuo and under high vacuum to give Intermediate Q90 (420.7 mg, 100% purity, 94% yield) as an amorphous residue. LC-MS (Method 2): R = 1.36 min; MS (ESIneg): m / z = 343 [ - t M(- NHS ester fragment)-H] . Intermediate Q91: N-[1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2,15-dioxo-6,9,12-trioxa-3-azapentadecan-15-yl]-L-alpha- aspartyl-L-prolyl-N-[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2-yl]-2,2- dimethylpropyl}(glycoloyl)amino]-1-(methylamino)-1-oxobutan-2-yl]-L-valinamide[000427]Intermediate Q92 (10.0mg, 0.009mmol) was dissolved in DMF (1.0ml). AMAS (3.27mg, 12.98umol) and DIEA (3.02ul, 0.017mmol) were added and the reaction was stirred for 2.5h at RT. The reaction was reduced in vacuo. The residues was purified by HPLC and lyophilised to give Intermediate Q91 (10.0 mg, 100% purity, 98% yield) as a white foam. LC-MS (Method 1): Rt= 4.46 min; MS (ESIpos): m / z = 1178 [M+H]+. Intermediate Q92: N-(3-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}propanoyl)-L-alpha-aspartyl-L-prolyl-N-[(2S)-4-[{(1R)-1-[1- benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2-yl]-2,2-dimethylpropyl}(glycoloyl)amino]-1-(methylamino)- 1-oxobutan-2-yl]-L-valinamide trifluoroacetate (1:1)[000428]Intermediate Q93 (143.4mg, 0.120mmol) was dissolved in 2,2,2-trifluoroethanol (4.0ml). ZnCl2 (97.9mg, 0.719mmol) was added and the reaction was stirred for 2h at 50C.3 further equivalents of ZnCl2 were added and and the reaction was stirred for 1h at 50C. EDTA (210.0mg, 0.719mmol) and H20 + 0.1% TFA (2ml) were added. The residue was purified by prep HPLC and then lyophilised to give Intermediate Q92 (125.6mg, 100% purity, 91% yield). LC-MS (Method 2): Rt = 1.54 min; MS (ESIpos): m / z = 1041 [M+H]+. Intermediate Q93: tert-butyl (19S)-19-{[(2S)-2-{[(2S)-1-{[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2- yl]-2,2-dimethylpropyl}(glycoloyl)amino]-1-(methylamino)-1-oxobutan-2-yl]amino}-3-methyl-1- oxobutan-2-yl]carbamoyl}pyrrolidin-1-yl]carbonyl}-2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa-5,18- diazahenicosan-21-oateP368919WO / 59362-725.601 [000429]To a solution of Intermediate Q94 (94.4 mg, 0.128mmol) and Intermediate Q39 (82.8 mg, 0.14 mmol) in DMF (3.0 ml) were added HATU (77.6 mg, 0.204 mmol) and DIEA (66.7 ul, 0.383 mmol). The reaction was stirred at RT for 1.5h. The residue was purified by prep HPLC and then lyophilised to give Intermediate Q93 (143.7 mg, 100% purity, 94% yield). LC-MS (Method 2): Rt= 2.41 min; MS (ESIpos): m / z = 1197 [M+H]+. Intermediate Q94: N-[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2-yl]-2,2- dimethylpropyl}(glycoloyl)amino]-1-(methylamino)-1-oxobutan-2-yl]-L-valinamide trifluoroacetate (1:1)[000430]Intermediate Q95 (99.2 mg, 0.137 mmol) was dissolved in 2,2,2-trifluoroethanol (4.0 ml). ZnCl2 (111.8 mg, 0.82 mmol) was added and the reaction was stirred for 2h at 50C. EDTA (239.6 mg, 0.82 mmol) and H20 + 0.1% TFA (2ml) were added and the residue was purified by prep HPLC and then lyophilised to give Intermediate Q94 (94.6 mg, 100% purity, 94% yield). LC-MS (Method 2): Rt = 1.53 min; MS (ESIpos): m / z = 626 [M+H]+. Intermediate Q95: tert-butyl N-[(1S)-1-[[(1S)-3-[[(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)pyrrol-2-yl]-2,2-dimethyl- propyl]-(2-hydroxyacetyl)amino]-1-(methylcarbamoyl)propyl]carbamoyl]-2-methyl-propyl]carbamateP368919WO / 59362-725.601[000431](2S)-2-amino-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2-yl]-2,2- dimethylpropyl}(glycoloyl)amino]-N-methylbutanamide trifluoroacetate (1:1) (100.0 mg, 0.155 mmol), the synthesis of which is described in WO2015096982, was dissolved in DMF (2.5 ml). N-(tert- butoxycarbonyl)valine (40.5 mg, 0.187 mmol), HATU (88.7 mg, 0.233 mmol) and DIEA (81.2 ul, 0.466 mmol) were added and the reaction was stirred at RT for 3.5h. The residues was purified by prep HPLC and then lyophilised to give Intermediate Q95 (99.4 mg, 100% purity, 88% yield). LC-MS (Method 2): Rt= 2.43 min; MS (ESIpos): m / z = 724 [M-H]- . Intermediate Q96: N-{18-[(2,5-dioxopyrrolidin-1-yl)oxy]-14,18-dioxo-4,7,10-trioxa-13-azaoctadecan-1-oyl}-L-alpha- aspartyl-L-prolyl-N-[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2-yl]-2,2- dimethylpropyl}(glycoloyl)amino]-1-(methylamino)-1-oxobutan-2-yl]-L-valinamide[000432]Intermediate Q92 (10.0 mg, 0.009 mmol) was dissolved in DMF (1.0 ml). Di(N- succinimidyl)glutarate (8.47 mg, 0.026 mmol) and DIEA (6.03 ul, 0.035 mmol) were added. The reaction was stirred for 2.5h at RT. The reaction was purified by prep HPLC and then lyophilised to give Intermediate Q96 (11.0 mg, 100% purity, quantitative yield) as a white foam. LC-MS (Method 1): Rt = 4.50 min; MS (ESIpos): m / z = 1252 [M+H]+.Intermediate Q97: (3S,19S,37S)-3,37-bis[(2S)-2-{[(2S)-1-{[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2- yl]-2,2-dimethylpropyl}(hydroxyacetyl)amino]-1-(methylamino)-1-oxobutan-2-yl]amino}-3-methyl-1- oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-19-({5-[(2,5-dioxopyrrolidin-1-yl)oxy]-5- oxopentanoyl}amino)-5,18,22,35-tetraoxo-8,11,14,26,29,32-hexaoxa-4,17,23,36- tetraazanonatriacontane-1,39-dioic acid[000433]Intermediate Q98 (10.0 mg, 4.33 µmol) was dissolved in DMF (2.0 ml). Di(N- succinimidyl)glutarate (4.24 mg, 13.0 µmol) and DIEA (3.0 µl, 17 µmol) were added. The reaction was stirred for 1.5h at RT. The reaction was purified by prep HPLC and then lyophilised to give Intermediate Q97 (8.8 mg, 96% purity, 81% yield). LC-MS (Method 1): Rt= 5.48 min; MS (ESIpos): m / z = 1202 [M+2H]2+. Intermediate Q98: (3S,19S,37S)-19-amino-3,37-bis[(2S)-2-{[(2S)-1-{[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)- 1H-pyrrol-2-yl]-2,2-dimethylpropyl}(hydroxyacetyl)amino]-1-(methylamino)-1-oxobutan-2-yl]amino}- 3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-5,18,22,35-tetraoxo-8,11,14,26,29,32- hexaoxa-4,17,23,36-tetraazanonatriacontane-1,39-dioic acid.trifluoroacetic acid (1 / 1)P368919WO / 59362-725.601bis(2,5-dioxopyrrolidin-1-yl) N-(tert-butoxycarbonyl)-L-glutamate (15.3 mg, 34.6 µmol) in DMF (4.0 ml), were added Intermediate Q92 (80.0 mg, 69.2 µmol)) and N,N-diisopropylethylamine (36 µl, 210 µmol). The mixture was stirred at rt for 1h and then concentrated in vacuo. The residue was purified by preparative HPLC and lyophilized to yield (3S,19S,37S)-3,37-bis[(2S)-2-{[(2S)-1-{[(2S)-4-[{(1R)-1-[1-benzyl-4- (2,5-difluorophenyl)-1H-pyrrol-2-yl]-2,2-dimethylpropyl}(hydroxyacetyl)amino]-1-(methylamino)-1- oxobutan-2-yl]amino}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-19-[(tert- butoxycarbonyl)amino]-5,18,22,35-tetraoxo-8,11,14,26,29,32-hexaoxa-4,17,23,36- tetraazanonatriacontane-1,39-dioic acid (71.1 mg, 100% purity, 90% yield). LC-MS (Method 1): Rt = 5.65 min; MS (ESIpos): m / z = 2293 [M+H]+ Step 2: To a solution of (3S,19S,37S)-3,37-bis[(2S)-2-{[(2S)-1-{[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5- difluorophenyl)-1H-pyrrol-2-yl]-2,2-dimethylpropyl}(hydroxyacetyl)amino]-1-(methylamino)-1- oxobutan-2-yl]amino}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-19-[(tert- butoxycarbonyl)amino]-5,18,22,35-tetraoxo-8,11,14,26,29,32-hexaoxa-4,17,23,36- tetraazanonatriacontane-1,39-dioic acid (70.9 mg, 100 % purity, 30.9 µmol) in trifluoroethanol (4.0 ml), was added zinc chloride (33.7 mg, 247 µmol). The mixture was stirred at 50°C for 2 h. The resulting mixture was diluted in water + 0.1% TFA (2 ml) and EDTA (72.3 mg, 247 µmol;) was then added. The resulting mixture was concentrated, purified over preparative HPLC and lyophilized to afford Intermediate Q98 (371 mg, 100 % purity, 82% yield). LC-MS (Method 1): Rt = 4.75 min; MS (ESIpos): m / z = 2193 [M+H]+P368919WO / 59362-725.601 Intermediate Q99: (3S,19S,37S)-19-[(15-amino-4,7,10,13-tetraoxapentadecanan-1-oyl)amino]-3,37-bis[(2S)-2-{[(2S)-1- {[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2-yl]-2,2- dimethylpropyl}(hydroxyacetyl)amino]-1-(methylamino)-1-oxobutan-2-yl]amino}-3-methyl-1-oxobutan- 2-yl]carbamoyl}pyrrolidine-1-carbonyl]-5,18,22,35-tetraoxo-8,11,14,26,29,32-hexaoxa-4,17,23,36- tetraazanonatriacontane-1,39-dioic acid.trifluoroacetic acid (1 / 1)Step 1: To a solution of Intermediate Q98 (30 mg, 13 µmol) in DMF (3.0 ml), were added tert-butyl {15- [(2,5-dioxopyrrolidin-1-yl)oxy]-15-oxo-3,6,9,12-tetraoxapentadecan-1-yl}carbamate (7.22 mg, 15.6 µmol) and N,N-diisopropylethylamine (6.8 µl, 39 µmol;). The mixture was stirred overnight at rt and then concentrated in vacuo. The residue was purified by preparative HPLC and lyophilized to yield (3S,19S,37S)-3,37-bis[(2S)-2-{[(2S)-1-{[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)-1H-pyrrol-2- yl]-2,2-dimethylpropyl}(hydroxyacetyl)amino]-1-(methylamino)-1-oxobutan-2-yl]amino}-3-methyl-1- oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-19-[(2,2-dimethyl-4,20-dioxo-3,8,11,14,17-pentaoxa- 5-azaicosan-20-yl)amino]-5,18,22,35-tetraoxo-8,11,14,26,29,32-hexaoxa-4,17,23,36- tetraazanonatriacontane-1,39-dioic acid (28.8 mg, 98% purity, 86% yield). LC-MS (Method 1): Rt= 5.69 min; MS (ESIpos): m / z = 2540 [M+H]+ Step 2: To a solution of (3S,19S,37S)-3,37-bis[(2S)-2-{[(2S)-1-{[(2S)-4-[{(1R)-1-[1-benzyl-4-(2,5- difluorophenyl)-1H-pyrrol-2-yl]-2,2-dimethylpropyl}(hydroxyacetyl)amino]-1-(methylamino)-1- oxobutan-2-yl]amino}-3-methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-19-[(2,2-dimethyl- 4,20-dioxo-3,8,11,14,17-pentaoxa-5-azaicosan-20-yl)amino]-5,18,22,35-tetraoxo-8,11,14,26,29,32- hexaoxa-4,17,23,36-tetraazanonatriacontane-1,39-dioic acid (28.8 mg, 96 % purity, 10.9 µmol) in Trifluoroethanol (3.0 ml), was added zinc chloride (11.9 mg, 87.3 µmol). The mixture was stirred at 50°C for 2 h. The resulting mixture was diluted in water + 0.1% TFA (2 ml) and EDTA (25.5 mg, 87.3 µmol) was then added. The resulting mixture was concentrated, purified over preparative HPLC and lyophilizedP368919WO / 59362-725.601 to afford Intermediate Q99 (24.1 mg, 100 % purity, 86% yield). LC-MS (Method 1): Rt = 4.68 min; MS (ESIpos): m / z = 2440 [M+H]+ [000434] Intermediate Q100: (3S)-4-[(2S)-2-[[(1S)-1-[[(1S)-3-[[(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)pyrrol-2-yl]-2,2-dimethyl- propyl]-(2-hydroxyacetyl)amino]-1-(methylcarbamoyl)propyl]carbamoyl]-2-methyl- propyl]carbamoyl]pyrrolidin-1-yl]-3-[3-[2-[2-[2-[[(4S)-5-[2-[2-[2-[3-[[(1S)-2-[(2S)-2-[[(1S)-1-[[(1S)-3- [[(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)pyrrol-2-yl]-2,2-dimethyl-propyl]-(2-hydroxyacetyl)amino]-1- (methylcarbamoyl)propyl]carbamoyl]-2-methyl-propyl]carbamoyl]pyrrolidin-1-yl]-1-(carboxymethyl)-2- oxo-ethyl]amino]-3-oxo-propoxy]ethoxy]ethoxy]ethylamino]-4-[3-[2-[2-[2-[2-[[5-(2,5-dioxopyrrolidin- 1-yl)oxy-5-oxo-pentanoyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]propanoylamino]-5-oxo- pentanoyl]amino]ethoxy]ethoxy]ethoxy]propanoylamino]-4-oxo-butanoic acid[000435]Intermediate Q99 (12.0 mg, 4.70 µmol) was dissolved in DMF (2.0 ml). Di(N- succinimidyl)glutarate (4.60 mg, 13.1 µmol) and DIEA (3.27 µl, 18.8 µmol) were added. The reaction was stirred for 1.5h at RT. The reaction was purified by prep HPLC and then lyophilised to give Intermediate Q100 (10.3 mg, 96% purity, 80% yield). LC-MS (Method 1): Rt= 5.41 min; MS (ESIpos): m / z = 1326 [M+2H]2+. Intermediate Q101: (3S)-4-[(2S)-2-[[(1S)-1-[[(1S)-3-[[(1R)-1-[1-benzyl-4-(2,5-difluorophenyl)pyrrol-2-yl]-2,2-dimethyl- propyl]-(2-hydroxyacetyl)amino]-1-(methylcarbamoyl)propyl]carbamoyl]-2-methyl- propyl]carbamoyl]pyrrolidin-1-yl]-3-[3-[2-[2-[2-[[(4S)-5-[2-[2-[2-[3-[[(1S)-2-[(2S)-2-[[(1S)-1-[[(1S)-3- [[(1R)-1-[1-benzyl-4-(25-difluorophenyl)pyrrol-2-yl]-22-dimethyl-propyl]-(2-hydroxyacetyl)amino]-1-P368919WO / 59362-725.601 (methylcarbamoyl)propyl]carbamoyl]-2-methyl-propyl]carbamoyl]pyrrolidin-1-yl]-1-(carboxymethyl)-2- oxo-ethyl]amino]-3-oxo-propoxy]ethoxy]ethoxy]ethylamino]-4-[3-[2-[2-[2-[2-[[2-(2,5-dioxopyrrol-1- yl)acetyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]propanoylamino]-5-oxo- pentanoyl]amino]ethoxy]ethoxy]ethoxy]propanoylamino]-4-oxo-butanoic acid[000436]Intermediate Q99 (12.0 mg, 4.70 µmol) was dissolved in DMF (2.0 ml). AMAS (2.37 mg, 9.4 µmol) and DIEA (2.45 µl, 14.1 µmol) were added. The reaction was stirred for 1h at RT. The reaction was purified by prep HPLC and then lyophilised to give Intermediate Q101 (10.7 mg, 96% purity, 84% yield). LC-MS (Method 1): Rt= 5.40 min; MS (ESIpos): m / z = 1289 [M+2H]2+. Intermediate Q102: N-{18-[(2,5-dioxopyrrolidin-1-yl)oxy]-14,18-dioxo-4,7,10-trioxa-13-azaoctadecanan-1-oyl}-L-alpha- aspartyl-L-prolyl-N-{[(4S)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H- pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl]methyl}-L-valinamide[000437]Intermediate Q103 (10.0 mg, 9.94 µmol) was dissolved in DMF (2.0 ml). Di(N- succinimidyl)glutarate (9.73 mg, 29.8 µmol) and DIEA (6.9 µl, 40 µmol) were added. The reaction was stirred for 1.5h at RT. The reaction was purified by prep HPLC and then lyophilised to give IntermediateP368919WO / 59362-725.601 Q102 (3.8 mg, 100% purity, 35% yield). LC-MS (Method 2): Rt = 1.22 min; MS (ESIneg): m / z = 1004[M(- NHS ester fragment)-H]- . Intermediate Q103: N-(3-{2-[2-(2-aminoethoxy)ethoxy]ethoxy}propanoyl)-L-alpha-aspartyl-L-prolyl-N-{[(4S)-4-ethyl-4- hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl]methyl}- L-valinamide.trifluoroacetic acid (1 / 1)Step1: To a solution of Intermediate Q104 (31.2 mg, 52.8 µmol) and Intermediate Q39 (34.3 mg, 58.1 µmol) in DMF (3.0 ml) were added HATU (32.1 mg, 84.5 µmol) and DIEA (28 µl, 160 µmol). The reaction was stirred at RT for 1.5h and then concentrated under reduced pressure. The residue was purified by prep HPLC and then lyophilised to give tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-({[(4S)-4-ethyl-4-hydroxy-3,14- dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl]methyl}amino)-3- methyl-1-oxobutan-2-yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa- 5,18-diazahenicosan-21-oate (40.7 mg, 100% purity, 73% yield). LC-MS (Method 2): Rt = 1.80 min; MS (ESIpos): m / z = 1049 [M+H]+. Step2: Tert-butyl (19S)-19-[(2S)-2-{[(2S)-1-({[(4S)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro- 1H-pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl]methyl}amino)-3-methyl-1-oxobutan-2- yl]carbamoyl}pyrrolidine-1-carbonyl]-2,2-dimethyl-4,17-dioxo-3,8,11,14-tetraoxa-5,18-diazahenicosan- 21-oate (40.5 mg, 38.6 µmol) was dissolved in DCM (4.0 mL), then TFA (1 ml) was added. The reaction was stirred overnight at rt and then concentrated in vacuo. The residue was dissolved in ACN / water andP368919WO / 59362-725.601 freeze-dried to give Intermediate Q103 (37.1 mg, 100% purity, 95% yield) as an amorphous residue. LC- MS(Method 6): R = 0.98 min; MS (ESIpos): m / z + t = 892 [M+H] . Intermediate Q104: Trifluoroacetic acid.N-{[(4S)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H- pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl]methyl}-L-valinamide (1 / 1)Step1: To a solution of (4S)-11-(aminomethyl)-4-ethyl-4-hydroxy-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinoline-3,14(4H,12H)-dione (CAS Nr = 154380-16-2, synthesis described in WO2022198232)(50.0 mg, 132 µmol) in DMF (3.0 ml), were added N-(tert-butoxycarbonyl)-L-valine (31.7 mg, 146 µmol), HATU (65.5 mg, 172 µmol) and DIEA (69 µl, 400 µmol). The reaction was stirred at RT for 1 h and then concentrated under reduced pressure. The residue was purified by prep HPLC to give tert-butyl [(2S)-1- ({[(4S)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolizino[1,2- b]quinolin-11-yl]methyl}amino)-3-methyl-1-oxobutan-2-yl]carbamate (31.9 mg, 100% purity, 42% yield). LC-MS (Method 2): Rt= 1.61 min; MS (ESIpos): m / z = 577 [M+H]+. Step2: Tert-butyl [(2S)-1-({[(4S)-4-ethyl-4-hydroxy-3,14-dioxo-3,4,12,14-tetrahydro-1H- pyrano[3',4':6,7]indolizino[1,2-b]quinolin-11-yl]methyl}amino)-3-methyl-1-oxobutan-2-yl]carbamate (31.5 mg, 54.6 µmol) was dissolved in DCM (5.0 mL), then TFA (1 ml) was added. The reaction was stirred at rt for 1 h and then concentrated in vacuo. The residue was dissolved in ACN / water and freeze- dried to give Intermediate Q104 (37.1 mg, 100% purity, 95% yield) as an amorphous residue. LC- MS(Method 6): Rt= 0.91 min; MS (ESIpos): m / z = 477 [M+H]+. Antibody-Binder-Conjugate (ABC) Precursors employed for further payload conjugation Intermediate R1P368919WO / 59362-725.601[000438]To a solution of 15 mg of the isotype antibody TPP-5657 in 1.5 mL PBS buffer (c = 10 mg / mL) presented under an argon atmosphere, 0.086 mg TCEP dissolved in 150 µl PBS-Puffer was added. After stirring for 30 min at RT, 7 Eq (0.59 mg) of Intermediate Q41 dissolved in 150 µL DMSO was added and stirring at RT was continued for another 90 min. Subsequently, the mixture was diluted with PBS Puffer (pH7.2) to a volume of 2.5 mL, applied to a PD-10 column (Sephadex® G-25 GE Healthcare) and eluted with 3.5 mL PBS buffer. The obtained solution was then concentrated by ultracentrifugation and rediluted with PBS buffer (pH 7.2) to a volume of 3.5 mL and again concentrated and re-deluted to a final volume of 1.5ml. The ADC batch obtained was characterized as shown in the table below.Intermediate R2[000439]To a solution of 5 mg of the isotype antibody TPP-5657 in 0.5 mL PBS buffer (c = 10 mg / mL) presented under argon atmosphere, 5 Eq (0.14 mg) of Intermediate Q9 dissolved in 50µL DMSO were added. After stirring at RT for 30 min, 5 Eq (0.14 mg) of Intermediate Q9, dissolved in 50 µL DMSOP368919WO / 59362-725.601 were added and stirring at RT was continued for another 30min. Subsequently, the mixture was diluted with PBS Puffer (pH 7.2) to a volume of 2.5 mL, applied to a PD-10 column (Sephadex® G-25 GE Healthcare) and eluted with 3.5 mL PBS buffer. The obtained solution was then concentrated by ultracentrifugation and rediluted with PBS buffer (pH 7.2) to a volume of 2.5 mL. The ADC batch obtained was characterized as shown in the table below.The following intermediates are prepared and incorporated into ADCs of the present disclosure according to organic synthesis methods known in the art, and adapted from the procedures disclosed herein: Intermediate R3Intermediate R4Intermediate R5Intermediate R6Intermediate R7. General Procedures for the Preparation and Characterization of Antibody-Drug Conjugates (ADCs) B-1 General Process for Expressing AntibodiesP368919WO / 59362-725.601 [000440]DNA-sequences that encode the protein sequences (amino acid sequences) of the used antibodies, for example TPP-5657, TPP-2658, TPP-1015, TPP-6013 and TPP-9476 were inserted into an transient expression vector by someone skilled in the arts as described by Tom et ah, Chapter 12 in Methods Express: Expression Systems edited by Micheal R. Dyson and Yves Durocher, Scion Publishing Ltd, 2007. B-2. General Process for Expressing Antibodies in Mammalian Cells [000441]The antibodies, for example TPP-5657, TPP-2658, TPP-1015, and TPP-9476 were produced in transient mammalian cell cultures as described by Tom et ah, Chapter 12 in Methods Express: Expression Systems edited by Micheal R. Dyson and Yves Durocher, Scion Publishing Ltd, 2007. B-3. General Process for Purifying Antibodies from Cell Supernatants [000442]The antibodies, for example TPP-5657, TPP-2658, TPP-1015, and TPP-9476, were obtained from the cell culture supernatants. The cell supernatants were clarified by centrifugation of cells. The cell supernatant was then purified by affinity chromatography on a MabSelect Sure (GE Healthcare) chromatography column. To this end, the column was equilibrated in DPBS pH 7.4 (Sigma / Aldrich), the cell supernatant was applied and the column was washed with about 10 column volumes of DPBS pH 7.4 + 500 mM sodium chloride. The antibodies were eluted in 50 mM sodium acetate pH 3.5 + 500 mM sodium chloride and then purified further by gel filtration chromatography on a Superdex 200 column (GE Healthcare) in DPBS pH 7.4. [000443]The commercially available antibodies were purified from the commercial products by standard chromatography methods (protein A chromatography, preparative gel filtration chromatography (SEC— size exclusion chromatography)). General Procedures for Antibody-Coupling [000444]The ADCs shown in the structural formulae of the Working examples, which were coupled to the cysteine side chains of the antibodies via maleimide radicals, are, depending on the linker and the coupling procedure, mainly present in the ring-opened or ring-closed forms shown in each case. However, the preparation may comprise a small proportion of the respective other form. The coupling reactions were carried out under argon. [000445]The concentrations of the antibodies or ADCs which are indicated in the experimental procedures may differ without a major impact on the outcome. Antibody or ADC concentrations in the range of 1-20 mg / mL can also be used in the coupling reactions, preferred is a concentration range in the coupling reactions of 5-15 mg / mL. Also the precursor molecules can be added in excesses which may deviate from what is given in the procedures which then may have an impact on the DAR. An excess of 2- 20 equivalents of the payload precursor can be used, preferred is the range 4-15 equivalents. B-4. General procedures for cystein-couplingP368919WO / 59362-725.601 [000446]Typically, under argon, a solution of 0.029 mg of TCEP in 0.05 ml of PBS buffer pH 7.2 was added to 5 mg of the respective antibody dissolved in 0.5 ml of PBS (c=10 mg / ml). (The concentration of the antibody solution can also vary from the standard procedure in a range between 1 mg / ml and 30 mg / ml). The mixture was stirred at RT for 30 min, and then an excess of 2-20 equivalents, preferably 2-10 equivalents, typically 7 equivalents (0.000233 mmol) of the respective intermediate (ADC precursor molecule) dissolved in 0.05 mL DMSO was added. After stirring at RT for a further 90 min, the mixture was diluted with PBS buffer pH 7.2 to a volume of 2.5 ml and then passed through a PD 10 column (Sephadex® G-25, GE Healthcare) equilibrated with PBS buffer pH 7.2 and eluted with PBS buffer pH 7.2. This was followed by concentration by ultracentrifugation and redilution with PBS buffer (pH 7.2) to a volume of about 1-5 mL. B-4a. General procedure for cysteine-coupling and subsequent opening of thiosuccinimide ring [000447]Typically, under argon, a solution of 0.029 mg of TCEP in 0.05 ml of PBS buffer pH 7.2 was added to 5 mg of the respective antibody dissolved in 0.5 ml of PBS (c=10 mg / ml). (The concentration of the antibody solution can also vary from the standard procedure in a range between 1 mg / ml and 30 mg / ml). The mixture was stirred at RT for 30 min, and then an excess of 2-20 equivalents, preferably 2-10 equivalents, typically 7 equivalents (0.000233 mmol) of the respective intermediate (ADC precursor molecule) dissolved in 0.05 mL DMSO was added. After stirring at RT for a further 90 min, the mixture was diluted to a volume of 2.5 ml with PBS buffer which had been adjusted to pH 8 beforehand and then passed through a PD 10 column (Sephadex® G-25, GE Healthcare) equilibrated with PBS buffer pH 8, and eluted with PBS buffer pH 8. The eluate was stirred at RT under argon overnight at pH 8. This was followed by concentration by ultracentrifugation and redilution with PBS buffer (pH 7.2) to a volume of about 1-5 mL. [000448]The following antibodies were used in the exemplified coupling protocols as described above: Examples i: Isotype control mAb (TPP-5657) Examples k: Anti-TWEAKR mAb (TPP-2658) Examples c: Anti-CD123 mAb (TPP-9476) Example p: Avelumab mAb (p-Avelu) [000449]To obtain thiol nucleophiles, an antibody can be reduced by state of the art methodologies as employed also in the examples described below. Alternatively, free cysteine residues can be site- specifically engineered into an antibody. The coupling reactions with the electrophiles described in the section ‘ADC precursor molecules‘ were usually carried out under argon as described in the general procedures in the section ‘ADC examples’. [000450]For biological tests, if required, the concentrations of the final ADC samples were optionally adjusted to the range of 0.5-15 mg / ml by redilution. The respective protein concentrations, stated in the working examples, of the ADC solutions were determined. Furthermore, antibody loading (drug / mAb ratio) was determined using the methods described under B-6.[000451]Depending on the linker, the ADCs shown in the examples may also be present to a lesser or higher degree in the form of the hydrolysed open-chain succinamides attached to the antibodies. [000452]In particular the KSPi-A2DCs attached through the linker substructureto thiol groups of the antibodies, may optionally also be hydrolized after the coupling by rebuffering and stirring at pH 8 for about 20-24 h to obtain ADCs attached via open-chain succinamides. #1 represents the sulphur bridge to the antibody, and #2 the point of attachment to the modified KSP inhibitor. [000453]Other potentially hydrolysis-sensitive thianylsuccinimide bridges to the antibody may contain the following linker substructures, where #1 represents the thioether linkage to the antibody, and #2 the point of attachment to the modified KSP inhibitor:[000454]In the structural formula described in the examples antibody AK (alternatively, “mAb”) linked via cysteine residues can mean: Examples i: isotype control mAb (TPP-5657), partially reduced Examples e: Anti-Her2 mAb (TPP1015), partially reduced Examples k-7007: Anti-TWEAKR mAb (TPP-7007), partially reduced Examples k-2658: Anti-TWEAKR mAb (TPP-2658), partially reduced Examples k-2090: Anti-TWEAKR mAb (TPP-2090), partially reduced whereP368919WO / 59362-725.601 §1 represents the linkage to the succinimide group or to any isomeric hydrolysed open-chain succinamides or the alkylene radical resulting therefrom, and S represents the sulphur atom of a cysteine residue of the partially reduced antibody [000455]In addition, the Cysteine-modified partially reduced antibody may further be acylated at one or more lysine residues as disclosed herein. B-5. General procedures for lysine-coupling [000456]To a solution of 5 mg of the respective antibody typically in 0.5 mL PBS buffer (c = 10 mg / mL) pH 7.2 presented under an Argon atmosphere (the concentration of the antibody solution can also vary from the standard procedure in a range between 1 mg / ml and 30 mg / ml), an excess of 2-10 equivalents, preferably 2-5 equivalents, typically 5 equivalents (0.00165 mmol) of the respective intermediate (ADC precursor molecule) dissolved in 0.5 mL of DMSO was added. After stirring at RT for 30-60 min, the same amount of the ADC precursor molecule, dissolved in 0.05 mL DMSO was added and stirring at RT was continued for another 30-60 min. Subsequently, the mixture was diluted with PBS Puffer (pH 7.2) to a volume of 2.5 mL, applied to a PD-10 column (Sephadex® G-25 GE Healthcare) and eluted with 3.5 mL PBS buffer. The obtained solution was then concentrated by ultracentrifugation to a volume of about 0.300 mL and rediluted with PBS buffer (pH 7.2) to a volume of about 1-5 mL. [000457]The following antibodies were used in the exemplified coupling protocols as described above: Examples i: Isotype control mAb (TPP-5657) Examples k: Anti-TWEAKR mAb (TPP-2658) Examples c: Anti-CD123 mAb (TPP-9476) Example p: Avelumab mAb (p-Avelu) [000458]In the structural formula described in the examples the lysine linked antibody can mean: Example i: Isotype control antibody (TPP-5657) Examples e: Anti-Her2 mAb (TPP1015) Examples k-7007: Anti-TWEAKR mAb (TPP-7007) Examples k-2658: Anti-TWEAKR mAb (TPP-2658) Examples k-2090: Anti-TWEAKR mAb (TPP-2090) Examples p: Avelumab mAb (p-Avelu) where §2 represents the linkage to the carbonyl group and NH represents the side-chain amino group of a lysine residue of the antibody. [000459]The antibodies can also be partially reduced and S-modified (e.g., at one or more cysteine residues) as disclosed herein. B-6. Analytical characterization of ADCs Drug load determination and purity assessment by SEC-UV[000460]The drug-to-antibody ratio (DAR) and purity of each ADC was determined using size exclusion chromatography (SEC). For the analysis, 50 μL of ADC solution was used. SEC analysis was carried out on an Agilent 1200 HPLC system monitored at 260 nm and 280 nm. A Superdex 20010 / 300 GL column (GE Healthcare) was operated at room temperature with an isocratic gradient using phosphate buffered saline (PBS) at a flow rate of 0.5 mL / min. The dimer and aggregate contents were determined from the integration results of UV peaks of monomeric, dimeric, and aggregated species. For calculating the DAR, the areas under the curve of the SEC-peaks at 260 nm (Adrug) and 280 nm (A280) (sum of aggregates, dimers, and monomers) were calculated using the following equation:(Ab = antibody, D = drug, ldrug = drug-related wavelength (260 nm for the KSP toxophore), ε = the absorption coefficients, A = the absorption at the particular wavelengths). [000461]In another method, the DAR is determined using SEC as described above, but carried out on an Agilent 1100 HPLC system using a size exclusion column (SUPERDEX 200 INCREASE 10 / 300 GL, Cytiva). Samples were detected using a UV detector set at 280 nm. Elution was performed with 10 mM sodium phosphate buffer system and 150 mM NaCl, pH 7.4 using a flow rate of 0.7 mL / min. [000462]For the toxophores, experimentally determined absorption coefficients were used. For all antibodies a common set of absorption coefficients was used (see following table). Absorption coefficients for the antibodies and drugs.Determination of the ADC concentrations [000463]The concentration of each ADC (CA2DC) was determined by measuring the absorption at 280 nm. The concentration was calculated using the absorption coefficient of the respective antibody. To take into account the absorbance of the toxophore at 280 nm, the concentration was corrected using the following equation: CA2DC= preliminary concentration / (1 + DARUV* [εdrug 280 nm / εAntibody 280 nm]) wherein, preliminary concentration = the concentration calculated using only the extinction coefficient of the antibody, DARUV = drug load of the respective ADC determined by UV absorption, εdrug 280 nm = extinction coefficient of the drug at 280 nm, and εAntibody 280 nm = extinction coefficient of the antibody at 280 nm).P368919WO / 59362-725.601 Characterization of ADCs by mass spectrometry [000464]For identity check of the antibody and the toxophore species as well as an alternative method for DAR determination, the ADCs have been characterized by mass spectrometric analysis. [000465]Molecular weight analysis was performed using a combination of HPLC and ESI-Q-TOF consisting of e.g. an I-class HPLC (Waters) for sample desalting and separation and an Impact HD mass spectrometer (Bruker Daltonik, Bremen) equipped with instrument control and acquisition software HyStar 3.2, ESI Compass 1.7 and Maximum Entropy Deconvolution Option for MS analysis. [000466]Chromatographic system and conditions: column: Acquity UPLC BEH300C41.7µm, 1.0x50mm; column temperature: 70°C; flow: 200 µl / min; mobile phase solution A: 0.1% formic acid, 94,9% water, 5% ACN; mobile phase solution B: 0.1% formic acid, 9.9% water, 10% ACN and 80% 2- propanol. [000467]Binary gradient profile for complete antibody: 2min 5% B, 2,5min 50% B, 3,5min 50% B, 5min 95%B, 5,1min 5% B, 5,6min 95% B, 5,7min 5% B, 6,2min 95% B, 6,3min 5% B, 7,5min 5% B. [000468]Binary gradient profile for reduced antibody: 2min 5% B, 4min 30% B, 5min 50% B, 7,5min 50%B, 8,5min 95% B, 8,6min 5% B, 9,1min 95% B, 9,3min 5% B, 9,8min 95% B, 9,9min 5% B, 12min 5% B. [000469]For Cys-coupled ADCs determination of the molecular weights of the individual conjugate species have been measured after deglycosylation and reduction. About 160 pmol of the conjugate was diluted with 20mM sodium phosphate pH6.5 in a final volume of 25µl.1µl PNGase F was added and the sample was incubated overnight at 37°C with gentle shaking (Thermomixer).10µl of the deglycosylated sample were denatured with 20µl 5M Gu*HCl (Guanidine hydrochloride in 50mM Tri- ethylammoniumhydrogencarbonat) by boiling the sample for 5 min at 60°C. To reduce the protein, 0.5µl of 255mM DTT dissolved in water was added and the mixture was incubated about 10 min at 60°C. The cooled sample was acidified with 2µl of 10% formic acid / water and then analyzed by mass spectrometry as described above. For DAR determinatio...

Claims

P368919WO / 59362-725.601 CLAIMS 1. An antibody-drug conjugate of Formula (I):or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; P1 is a protease-cleavable linker; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1, 2, 3, 4, or 5.

2. The antibody-drug conjugate of claim 1, or a pharmaceutically acceptable salt thereof, wherein the protease is expressed extracellularly in a tumor microenvironment.

3. The antibody-drug conjugate of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein the protease is neutrophil elastase.

4. The antibody-drug conjugate of claim 1, or a pharmaceutically acceptable salt thereof, wherein P1 is a tripeptide linker having the formula: -(AA1)-(AA2)-(AA3)-; AA1 is Ala, Leu, Ile, or Val; AA2 is Pro; AA3 is Asn, Asp, Gly, or Asp*; and Asp* is an ester prodrug of Asp.

5. The antibody-drug conjugate of claim 4, or a pharmaceutically acceptable salt thereof, wherein each of AA1, AA2, and AA3 is a naturally-occurring amino acid (e.g., an L-amino acid).

6. The antibody-drug conjugate of any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein P1 is a neutrophil elastase-cleavable tripeptide linker having a sequence: (L-Asp)-(L-Pro)-(L-Val)-§, (L-Asn)-(L-Pro)-(L-Val)-§, (Gly)-(L-Pro)-(L-Val)-§, (L-Ala)-(L-Pro)-(L-Val)-§, (L-Nva)-(L-Pro)-(L-Val)-§, (L-His)-(L-Pro)-(L-Val)-§, (L-Asp)-(L-Pro)-(L-Ala)-§, (L-Asn)-(L-Pro)-(L-Ala)-§, (L-Asp)-(L-Pro)-(L-Ile)-§, (L-Asn)-(L-Pro)-(L-Ile)-§, (Gly)-(L-Pro)-(L-Ile)-§, (L-Ala)-(L-Pro)-(L-Ile)-§, (L-Nva)-(L-Pro)-(L-Ile)-§, (L-His)-(L-Pro)-(L-Ile)-§, (L-Asp)-(L-Pro)-(L-Leu)-§,P368919WO / 59362-725.601 (L-Asn)-(L-Pro)-(L-Leu)-§, (Gly)-(L-Pro)-(L-Leu)-§, (L-Ala)-(L-Pro)-(L-Leu)-§, (L-Nva)-(L-Pro)-(L-Leu)-§, or (L-His)-(L-Pro)-(L-Leu)-§; wherein § represents a bond to the therapeutic payload (D1).

7. The antibody-drug conjugate of any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein P1 is a neutrophil elastase-cleavable tripeptide linker having a sequence: -(L-Asp)-(L-Pro)-(L-Val)-§, -(L-Asp)-(L-Pro)-(L-Ala)-§, -(L-Asp*)-(L-Pro)-(L-Val)-§, -(L-Asn)-(L-Pro)-(L-Val)-§, -(L-Asn)-(L-Pro)-(L-Ala)-§, or -(Gly)-(L-Pro)-(L-Val)-§; wherein § represents a bond to the therapeutic payload (D1); and Asp* represents an optionally substituted C1-12alkyl ester of L-Asp.

8. The antibody-drug conjugate of any one of claims 1-3, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II):or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH CONH 3 2 2, -CH2COOH, or -CH2COOR ; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R3 is C 4 1-12 alkyl substituted with 0-6 R ; R4 is hydrogen, halogen, -C(O)OR5, -C(O)N(R5)2, -N(R5)2, -N(R5)3+, -OR5, -SR5, -S(O)R5, -S(O)2R5, -S(O)2N(R5), -S(O)2N(R5)C(O) R5, -S(O)2N(R5)C(O)OR5, -N(R5)S(O)2N(R5)2, -N(R5)S(O)2N(R5)C(O)R5, or -N(R5)S(O)2N(R5)C(O)OR5; R5 is in each instance, independently, hydrogen or C1-6alkyl; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1, 2, 3, 4, or 5.

9. The antibody-drug conjugate of claim 1 or claim 8, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-1), (II-2), (II-3), (II-4), (II-5), or (II-6):or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof comprising cysteine and lysine; B1 and B2 are in each instance, independently, a small molecule target protein binder; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; wherein: L1' denotes L1, bonded via a sulfur atom of a cysteine side-chain of AK; and L1'' denotes L1, bonded via a nitrogen atom of a lysine side-chain of AK; R1 is hydrogen, -CHCONH, - 3 2 2 CH2COOH, or -CH2COOR; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R3 is C 4 1-12 alkyl substituted with 0-6 R; R4 is hydrogen, halogen, -C(O)OR5, -C(O)N(R5), -N(R5), -N(R5)+, -OR5, -SR5, -S(O) 5 2 2 3 R, -S(O)R5, -S(O)N(R5), -S(O)N(R5)C(O) R5, -S(O)N 5 5 5 5 2 2 2 2 (R)C(O)OR, -N(R)S(O)2N(R)2, -N(R5)S(O)N(R5)C( 5 5 5 5 2 O)R, or -N(R)S(O)2N(R)C(O)OR; R5 is in each instance, independently, hydrogen or C1-6 alkyl; m is an integer from 1 to 20;P368919WO / 59362-725.601 m' is an integer from 1 to 10; m'' is an integer from 1 to 10; and n is an integer from 1 to 20.

10. The antibody-drug conjugate of claim 9, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-1):or a pharmaceutically acceptable salt thereof: n is an integer from 1 to 20.

11. The antibody-drug conjugate of claim 9, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-2):or a pharmaceutically acceptable salt thereof, wherein: n is an integer from 1 to 20.

12. The antibody-drug conjugate of claim 9, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-3):or a pharmaceutically acceptable salt thereof.P368919WO / 59362-725.601 13. The antibody-drug conjugate of any one of claims 1-12, or a pharmaceutically acceptable salt thereof, wherein the target protein is a protein that is over-expressed by a tumor cell or in a tumor microenvironment.

14. The antibody-drug conjugate of any one of claims 1-13, or a pharmaceutically acceptable salt thereof, wherein the target protein is expressed on the surface of a tumor cell.

15. The antibody-drug conjugate of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, wherein the target protein of binder B1 and / or B2 is a protein selected from alpha-v beta-3 (“αvß3” or “avß3”) integrin, alpha-v beta-6 (“αvß6” or “avß6”) integrin, carbonic anhydrase IX (“CA9” or “CA IX”), fibroblast activating protein (“FAP”), prostate specific membrane antigen (“PSMA”), heat shock protein 90 (“Hsp 90”), folic acid receptor, glucose transporter 1, somatostatin receptor, aminopeptidase N (APN), low density lipoprotein receptor-related protein 1 (LRP1), bombesin receptor, gonadotropin releasing hormone (GnRH or LHRH) receptor, p32, membrane type 1 matrix metalloprotease (MT1- MMP), Sortilin, or Nectin-4.

16. The antibody-drug conjugate of any one of claims 1-15, or a pharmaceutically acceptable salt thereof, wherein B1 and B2 are each independently: ,, , bsent.

17. The antibody-drug conjugate of any one of claims 1-3, 8, or 13-16, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-B):P368919WO / 59362-725.601(II-B) or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; D1 is a therapeutic payload; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH 3 2CONH2, -CH2COOH, or CH2COOR; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R4 is hydrogen, halogen, -C(O)OR5, -C(O)N(R5) 5 5 + 5 5 2, -N(R)2, -N(R)3, -OR, -SR, -S(O)R5, -S(O) 5 5 5 5 5 5 2R, -S(O)2N(R), -S(O)2N(R)C(O) R, -S(O)2N(R)C(O)OR, -N(R5)S(O)N(R5), -N(R5)S(O)N(R5)C(O)R5, or -N(R5)S(O) 5 5 2 2 2 2N(R)C(O)OR; R5 is in each instance, independently, hydrogen or C1-6 alkyl; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1, 2, 3, 4, or 5.

18. The antibody-drug conjugate of any one of claims 1-11, and 13-17, or a pharmaceutically acceptable salt thereof, wherein n is an integer from 1 to 8.

19. The antibody-drug conjugate of any one of claims 1-11, and 13-17, or a pharmaceutically acceptable salt thereof, wherein L2 comprises the group:wherein: * denotes a bond to a sulfur atom of a cysteine side-chain of AK; and ** denotes a bond to a nitrogen atom of a lysine side-chain of AK; and which may further comprise a polymeric group of the formula (CH2CH2O)1-8CH2CH2X, wherein X is NH, N(CH3), CO, NHCO, N(CH3)CO, CONH, or CON(CH3), and X forms a bond with B2.

20. The antibody-drug conjugate of any one of claims 1-8, or a pharmaceutically acceptable salt thereof, wherein n is 0.

21. The antibody-drug conjugate of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-4):or a pharmaceutically acceptable salt thereof.

22. The antibody-drug conjugate of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-5):P368919WO / 59362-725.601or a pharmaceutically acceptable salt thereof.

23. The antibody-drug conjugate of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-6):or a pharmaceutically acceptable salt thereof; wherein: AK is an antibody or antigen binding fragment thereof comprising cysteine and lysine; L1' is a non-cleavable linker, bonded via a sulfur of the cysteine of AK; L1'' is a non-cleavable linker, bonded via a nitrogen of the lysine of AK; m' is an integer from 1 to 10; and m'' is an integer from 1 to 10.

24. The antibody-drug conjugate of any one of claims 1-23, or a pharmaceutically acceptable salt thereof, wherein each D1 is independently:,or a pharmaceutically acceptable salt thereof, wherein: each R6 and R7 is independently hydrogen, halogen, CN, -C1-6 alkyl, or C1-6 haloalkyl; R8 is hydrogen, halogen, CN, -C1-6 alkyl, C1-6 haloalkyl, C3-6 cycloalkyl, or 5- to 7-membered heterocycloalkyl; R9 is hydrogen, halogen, CN, C1-6alkyl, -C(O)NH2, -C(O)NHC1-6alkyl, -C(O)N(C1-6alkyl)2, -C(O)NHC1-6alkyl-C(O)NHC1-6alkyl, -C(O)NHC1-6alkyl-NHC(O)C1-6alkyl, - NH2, -NHC1-6 alkyl, -N(C1-6 alkyl)2, -NHC(O)C1-6 alkyl, -OH, or -OC1-6 alkyl; wherein each C al 10 1-6 kyl is substituted with 0-5 R ; R10 is in each instance independently selected from the group consisting of hydrogen, halogen, CN, -COOH, -CONH2, -NH2, -NHCH3, -N(CH3)2, -OH, and -OCH3; R11 and R12 are each independently hydrogen, halogen, C1-6alkyl, C1-6haloalkyl, or -OH; or R11 and R12, taken together, form a 5- or 6-membered heterocycle; R13 and R14 are each independently hydrogen, C1-6alkyl, or C1-6alkylamine; or R13 and R14, taken together, form a C6carbocycle substituted with -N(R15)2; each R15 is independently hydrogen, C1-6alkyl, -C(O)C1-6alkyl, -C(O)NHC1-6alkyl or -C(O)OC1-6alkyl ; wherein the C1-6alkyl of R15 is optionally substituted with halogen, hydroxy, phenyl, or heteroaryl; or R15 is a cleavable prodrug group; R16 is hydrogen, halogen, C1-6alkyl, C1-6haloalkyl, -OH, -OCH3, or -OCF3; R17 is hydrogen, halogen, C1-6alkyl, C1-6haloalkyl, -OH, -OCH3, or -OCF3; or R16 and R17, taken together, form a heteroalkylene group of the formula: -O-C2-10alkylene-O-, -NH-C2-10alkylene-O-, or -NH-C2-10alkylene-NH-; wherein the heteroalkylene group is optionally substituted with R21; R18 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, -OH, -OCH3, or -OCF3; or R16 and R18, taken together, form a heteroalkylene group of the formula: -O-C2-10alkylene-O-, -NH-C2-10alkylene-O-, or -NH-C2-10alkylene-NH-; wherein the heteroalkylene group is optionally substituted with R21; R19 is hydrogen, halogen, C1-6alkyl, C1-6haloalkyl, -OH, -OCH3, or -OCF3; R20 is hydrogen or methyl; R21 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, or OH; R22 is hydrogen or C 24 1-6 alkyl, wherein the C1-6 alkyl is unsubstituted or substituted with R ; R23 is hydrogen, C1-6 alkyl, or benzyl, wherein the C1-6 alkyl or benzyl is unsubstituted or substituted with one, two, or three R25 groups; R24 is -OH -O(C1 6 alkyl) -NH2 -NH(C1 6 alkyl) -N(C16 alkyl)2 -SH or -S(C16 alkyl);P368919WO / 59362-725.601 R25 is -OH, -O(C1-6 alkyl), -NH2, -NH(C1-6 alkyl), -N(C1-6 alkyl)2, -NHS(O)2(C1-6 alkyl), C1-6 alkyl, C1-6 aminoalkyl, or OCH2CH2NHC(O)(C1-6 aminoalkyl); each Y1, Y2, Y3, and Y4 is independently -CH, -CF, or N; Y5 is CH2, NH, or O; q is 0, 1, 2, 3, 4, or 5; and r is 0, 1, 2, 3, 4, or 5.

25. The antibody-drug conjugate of any one of claims 1-24, or a pharmaceutically acceptable salt thereof, wherein D1 is a membrane-permeable therapeutic payload.

26. The antibody-drug conjugate of any one of claims 1-25, or a pharmaceutically acceptable salt thereof, wherein D1 is a cytotoxic agent or an immunostimulatory agent.

27. The antibody-drug conjugate of any one of claims 1-26, or a pharmaceutically acceptable salt thereof, wherein D1 is a topoisomerase inhibitor, a kinesin spindle protein inhibitor, a tubulin polymerization inhibitor, a cyclin dependent kinase 9 inhibitor, or an agonist of toll like receptor 7 and / or toll like receptor 8.

28. The antibody-drug conjugate of any one of claims 1-8, and 24-27, or a pharmaceutically acceptable salt thereof, having a structure of Formula (III):or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); each R6 and R7 is independently hydrogen, halogen, CN, -C1-6 alkyl, or C1-6 haloalkyl;R8 is hydrogen, halogen, CN, -C1-6 alkyl, C1-6 haloalkyl, C3-6 cycloalkyl, or 5- to 7-membered heterocycloalkyl; R9 is hydrogen, halogen, CN, C1-6 alkyl, -C(O)NH2, -C(O)NHC1-6 alkyl, -C(O)N(C1-6 alkyl)2, -C(O)NH-C1-6 alkyl-C(O)NH-C1-6 alkyl, -C(O)NHC1-6 alkyl-NHC(O)-C1-6 alkyl, -NH2, - NHC1-6 alkyl, -N(C1-6 alkyl)2, -NHC(O)C1-6 alkyl, -OH, or -OC1-6 alkyl; wherein each C1-6 alkyl is substituted with 0-5 R10; R10 is in each instance independently selected from the group consisting of hydrogen, halogen, CN, -COOH, -CONH2, -NH2, -NHCH3, -N(CH3)2, -OH, and -OCH3; m is an integer from 1 to 20; n is an integer from 0 to 20; p is 1, 2, 3, 4, or 5; q is 0, 1, 2, 3, 4, or 5; and r is 0, 1, 2, 3, 4, or 5.

29. The antibody-drug conjugate of any one of claims 24-28, or a pharmaceutically acceptable salt thereof, wherein each R6 and R7 are hydrogen or halogen; q is 0, 1, or 2; and r is 0, 1, or 2.

30. The antibody-drug conjugate of any one of claims 24-29, or a pharmaceutically acceptable salt thereof, wherein q is 0, and r is 2.

31. The antibody-drug conjugate of any one of claims 24-30, or a pharmaceutically acceptable salt thereof, wherein R8 is -C1-6alkyl.

32. The antibody-drug conjugate of any one of claims 1-8, and 24-27, or a pharmaceutically acceptable salt thereof, having a structure of Formula (III-A):or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance independently a small molecule target protein binder;L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R9 is hydrogen, C1-6 alkyl, -C(O)NH2, -C(O)NHC1-6 alkyl, or -C(O)N(C1-6 alkyl)2; wherein each C1-6 alkyl is optionally substituted with 0-5 halogen; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1 or 2.

33. The antibody-drug conjugate of any one of claims 1-8, and 24-27, or a pharmaceutically acceptable salt thereof, wherein R9 is H, -CH3, -CH2F, -CHF2, -CF3, -C(O)NH2, -C(O)NHCH3, or - C(O)N(CH3)2.

34. The antibody-drug conjugate of any one of claims 1-8, 24-27, and 32-33, or a pharmaceutically acceptable salt thereof, wherein R9 is H, -CHF2, or -C(O)NHCH3.

35. The antibody-drug conjugate of any one of claims 1-8, and 24-27, or a pharmaceutically acceptable salt thereof, having a structure of Formula (IV):or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R11 and R12 are each independently hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, or -OH; or R11 and R12, taken together, form a 5- or 6-membered heterocycle; R13 and R14 are each independently hydrogen, C1-6 alkyl, or C1-6 alkylamine;P368919WO / 59362-725.601 or R13 and R14, taken together, form a C 15 6 carbocycle substituted with -N(R )2; each R15 is independently hydrogen, C1-6 alkyl, -C(O)C1-6 alkyl, -C(O)NHC1-6 alkyl, or -C(O)OC1-6 alkyl; wherein the C1-6 alkyl of R15 is optionally substituted with halogen, hydroxy, phenyl, or heteroaryl; or R15 is a cleavable prodrug group; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1, 2, 3, 4, or 5.

36. The antibody-drug conjugate of claim 35, or a pharmaceutically acceptable salt thereof, wherein R11 is hydrogen, halogen, C1-6 alkyl, or -OH; R12 is hydrogen, halogen, C1-6alkyl, or -OH; or R11 and R12, taken together, form a 5- or 6-membered cyclic ether; R13 is hydrogen, C1-6alkyl, or C1-6alkylamine; R14 is hydrogen, C1-6alkyl, or C1-6alkylamine; or R13 and R14, taken together, form a C6carbocycle substituted with -N(R15)2.

37. The antibody-drug conjugate of any one of claims 1-8, and 35-36, or a pharmaceutically acceptable salt thereof, having a structure of Formula (IV-A), Formula (IV-B), Formula (IV-C), or Formula (IV-D):601or a pharmaceutically acceptable salt thereof wherein:P368919WO / 59362-725.601 AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); each R15 is independently hydrogen, C1-6 alkyl, -C(O)C1-6 alkyl, -C(O)C1-6 haloalkyl, -C(O)-heteroaralkyl, or -C(O)O-aralkyl; or R15 is a cleavable (prodrug) group; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1 or 2.

38. The antibody-drug conjugate of any one of claims 1-8, 24, and 35-37, or a pharmaceutically acceptable salt thereof, wherein each R15 is independently hydrogen, -C1-6alkyl, -C(O)C1-6alkyl, or - C(O)C1-6haloalkyl.

39. The antibody-drug conjugate of any one of claims 1-8, 24, and 35-38, or a pharmaceutically acceptable salt thereof, wherein each R15 is independently hydrogen, -CH3, -CH2CH3, or -C(O)CH3.

40. The antibody-drug conjugate of any one of claims 1-8, and 24-27 or a pharmaceutically acceptable salt thereof, having a structure of Formula (V):or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R16 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, -OH, -OCH3, or -OCF3; R17 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, -OH, -OCH3, or -OCF3; or R16 and R17, taken together, form a heteroalkylene group of the formula:P368919WO / 59362-725.601 -O-C2-10 alkylene-O-, -NH-C2-10 alkylene-O-, or -NH-C2-10 alkylene-NH-; R18 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, -OH, -OCH3, or -OCF3; or R16 and R18, taken together, form a heteroalkylene group of the formula: -O-C2-10 alkylene-O-, -NH-C2-10 alkylene-O-, or -NH-C2-10 alkylene-NH-; R19 is hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl, -OH, -OCH3, or -OCF3; each Y1, Y2, Y3, and Y4 is independently -CH, -CF, or N; Y5 is CH2, NH, or O; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 12, 3, 4, or 5.

41. The antibody-drug conjugate of any one of claims 1-8, and 40, or a pharmaceutically acceptable salt thereof, having a structure of Formula (V-A) or Formula (V-B):(V-B) or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3);R16 and R17 taken together form the group -O-C2-10 alkylene-O- or -NH-C2-10 alkylene-O-; or R16 and R18 taken together form the group -NH-C2-10 alkylene-O-; Y3 is CH or N; Y4 is CH or N; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1 or 2.

42. The antibody-drug conjugate of any one of claims 1-8 or a pharmaceutically acceptable salt thereof, having a structure of Formula (VI):or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R20 is hydrogen or -CH3; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1 or 2.

43. The antibody-drug conjugate of any one of claims 1-8 or a pharmaceutically acceptable salt thereof, having a structure of Formula (VII):P368919WO / 59362-725.601 Formula VII or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; L1 and L2 are in each instance, independently, a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); R22 is hydrogen or C alkyl, wherein the C alkyl is unsubstitut 24 1-6 1-6 ed or substituted with R ; R23 is hydrogen, C1-6 alkyl, or benzyl, wherein the C1-6 alkyl or benzyl is unsubstituted or substituted with one, two, or three R25 groups; R24 is -OH, -O(C1-6alkyl), -NH2, -NH(C1-6alkyl), -N(C1-6alkyl)2, -SH, or -S(C1-6alkyl); R25 is -OH, -O(C1-6alkyl), -NH2, -NH(C1-6alkyl), -N(C1-6alkyl)2, -NHS(O)2(C1-6alkyl), C1-6alkyl, C1-6aminoalkyl, or OCH2CH2NHC(O)(C1-6aminoalkyl); m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1 or 2.

44. The antibody-drug conjugate of any one of claims 1-43, or a pharmaceutically acceptable salt thereof, wherein D1 is:P368919WO / 59362-725.601or a pharmaceutically acceptable salt thereof.

45. The antibody-drug conjugate of any one of claims 1-8, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-D) or Formula (II-E):or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B1 and B2 are in each instance, independently, a small molecule target protein binder; D1 is a therapeutic payload;L2 is a non-cleavable linker; L3 is a protein-coupled group; L4 is a bond or a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); Ra is hydrogen or -CH3; Rb is in each instance, independently, hydrogen or CH3; or two Rb, together with the carbon to which they are attached, form a carbonyl; Rc is hydrogen or -CH3; m is an integer from 1 to 20; n is an integer from 0 to 20; and p is 1 or 2.

46. The antibody-drug conjugate of claim 45, or a pharmaceutically acceptable salt thereof, wherein L3 is:wherein: * denotes a bond to a sulfur atom of a cysteine side-chain of AK; and ** denotes a bond to a nitrogen atom of a lysine side-chain of AK.

47. The antibody-drug conjugate of claim 45 or 46, or a pharmaceutically acceptable salt thereof, wherein L3 is:wherein:* denotes a bond to a sulfur atom of a cysteine side-chain of AK; and ** denotes a bond to a nitrogen atom of a lysine side-chain of AK.

48. The antibody-drug conjugate of any one of claims 45-47, or a pharmaceutically acceptable salt thereof, wherein L4 is:or ; wherein *** denotes a bond to L3.

49. The antibody-drug conjugate of any one of claims 45-47, or a pharmaceutically acceptable salt thereof, wherein L4 is a bond.

50. The antibody-drug conjugate of any one of claims 1-8, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-F) or Formula (II-G):P368919WO / 59362-725.601 or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; B2 is a small molecule target protein binder; D1 is a therapeutic payload; L2 is a non-cleavable linker; L3 is a protein-coupled group; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); Ra is hydrogen or -CH3; Rb is in each instance, independently, hydrogen or CH3; or two Rb, together with the carbon to which they are attached, form a carbonyl; Rc is hydrogen or -CH3; m is an integer from 1 to 20; and n is an integer from 0 to 20.

51. The antibody-drug conjugate of any one of claims 1-8, 17, and 24-50, or a pharmaceutically acceptable salt thereof, wherein p is 1.

52. The antibody-drug conjugate of any one of claims 1-8, 17, and 24-50, or a pharmaceutically acceptable salt thereof, wherein p is 2.

53. The antibody-drug conjugate of any one of claims 1-52, or a pharmaceutically acceptable salt thereof, wherein R2 is -CH3.

54. The antibody-drug conjugate of any one of claims 1-52, or a pharmaceutically acceptable salt thereof, wherein R2 is -CH(CH3)2.

55. The antibody-drug conjugate of any one of claims 1-54, or a pharmaceutically acceptable salt thereof, wherein R1 is hydrogen.

56. The antibody-drug conjugate of any one of claims 1-54, or a pharmaceutically acceptable salt thereof, wherein R1 is -CH2CONH2.

57. The antibody-drug conjugate of any one of claims 1-54, or a pharmaceutically acceptable salt thereof, wherein R1 is -CH2COOH.

58. The antibody-drug conjugate of any one of claims 1-54, or a pharmaceutically acceptable salt thereof, wherein R1 is -CH 3 2COOR .P368919WO / 59362-725.601 59. The antibody-drug conjugate of claim 58, or a pharmaceutically acceptable salt thereof, wherein R3 is C alkyl substit 5 5 + 5 1-12 uted with -N(R)2 or -N(R)3; and each R is independently hydrogen, CH3, or CH2CH3.

60. An antibody-drug conjugate having the following structure: ,,P368919WO / 59362-725.601 ,, or a pharmaceutically acceptable salt thereof, wherein: AK is an antibody or antigen binding fragment thereof; m is an integer from 1 to 20; and n is an integer from 1 to 20.

61. The antibody-drug conjugate of any one of claims 1-60, or a pharmaceutically acceptable salt thereof, wherein AK binds to a target antigen expressed on a tumor cell or in a tumor microenvironment.

62. The antibody-drug conjugate of claim 61, or a pharmaceutically acceptable salt thereof, wherein the target antigen is PD-L1, Her2, TWEAKR, Tenascin-C, αvβ6, or CD123.P368919WO / 59362-725.601 63. The antibody-drug conjugate of any one of claims 1-62, or a pharmaceutically acceptable salt thereof, wherein AK is an antibody selected from: ANP-TPP-5657, Her2-TPP-1015, Tweak-TPP-2658, or CD123-TPP-9476.

64. A pharmaceutical composition comprising the antibody-drug conjugate of any one of claims 1 to 63, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

65. A method of treating a disease or disorder, comprising administering to a patient in need thereof an antibody-drug conjugate of any one of claims 1 to 63.

66. A method of treating a hyperproliferative disorder or an angiogenic disorder, comprising administering to a patient in need thereof an antibody-drug conjugate of any one of claims 1 to 63.

67. A compound of Formula (X):or a pharmaceutically acceptable salt thereof, wherein: PR1 is a protein-reactive group; B1 is a target protein binder; L1a is a non-cleavable linker; P1 is a neutrophil elastase cleavable linker; D1 is a therapeutic payload; and p is 1, 2, 3, 4, or 5.

68. The compound of claim 67, having the structure of Formula (XI):or a pharmaceutically acceptable salt thereof, wherein: PR1 is a protein-reactive group; B1 is a target protein binder; L1a is a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); D1 is a therapeutic payload; and p is 1, 2, 3, 4, or 5.

69. The compound of claim 67 or 68, or a pharmaceutically acceptable salt thereof, having the structure of Formula (XI-A) or Formula (XI-B):or a pharmaceutically acceptable salt thereof, wherein: PR1 is a protein-reactive group; B1 is a target protein binder; L1b is a bond or a non-cleavable linker; R1 is hydrogen, -CH2CONH2, or -CH2COOH; R2 is -CH3, -CH(CH3)2, -CH2CH(CH3)2, or -CH(CH3)(CH2CH3); Ra is hydrogen or -CH3; Rb is in each instance, independently, hydrogen or CH3; or two Rb, together with the carbon to which they are attached, form a carbonyl; Rc is hydrogen or -CH3; and p is 1 or 2.

70. The compound of claim 69, or a pharmaceutically acceptable salt thereof, wherein PR1 is:

71. The compound of claim 67 or 68, or a pharmaceutically acceptable salt thereof, having the structure:P368919WO / 59362-725.601or a pharmaceutically acceptable salt thereof, wherein: Ra is hydrogen or -CH3; R1 is hydrogen, -CH2C(O)NH2, or -CH2C(O)OH; R2 is -CH3 or -CH(CH3)2; and D1 is a therapeutic payload.

72. The compound of claim 71, or a pharmaceutically acceptable salt thereof, wherein D1 is a cytotoxic agent or an immunostimulatory agent.

73. The compound of claim 71, or a pharmaceutically acceptable salt thereof, wherein D1 is a topoisomerase inhibitor, a kinesin spindle protein inhibitor, a tubulin polymerization inhibitor, or a cyclin dependent kinase 9 inhibitor.

74. The compound of claim 71, or a pharmaceutically acceptable salt thereof, wherein D1 is:or a pharmaceutically acceptable salt thereof.

75. A compound of Formula (IX):or a pharmaceutically acceptable salt thereof, wherein: B2 is a small molecule target protein binder; L2a is a non-cleavable linker; and PR2 is a protein-reactive group.

76. The compound of claim 70, or a pharmaceutically acceptable salt thereof, wherein PR2 is:and L2a is an alkyl or heteroalkyl linker, optionally comprising one or more polymeric groups (e.g., each independently selected from -CH2CH2O-, -CH2CH2N(CH3)-, and -CH2CON(CH3)-); alkyl groups; or heteroalkyl groups (e.g., -NH-(CH2)1-6-NH-, -C(O)-(CH2)1-6-C(O)-).

77. The compound of claim 70, or a pharmaceutically acceptable salt thereof, wherein B is a alpha-v beta-3 (“αvß3” or “avß3”) integrin binder, a carbonic anhydrase IX (“CA9” or “CA IX”) binder, a fibroblast activating protein (“FAP”) binder, a prostate specific membrane antigen (“PSMA”) binder, a heat shock protein 90 (“Hsp 90”) binder, a folic acid receptor binder, a glucose transporter 1 binder, a somatostatin receptor binder, an aminopeptidase N (“APN”) binder, a low density lipoprotein receptor- related protein 1 (“LRP1”) binder, a bombesin receptor binder, a gonadotropin releasing hormone (“GnRH” or “LHRH”) receptor binder, a p32 binder, a membrane type 1 matrix metalloprotease (“MT1- MMP”) binder, a Sortilin binder, or Nectin-4 binder.

78. The compound ofany one of claims 70-72, or a pharmaceutically acceptable salt thereof, having the structure:or a pharmaceutically acceptable salt thereof.

79. The antibody drug conjugate of any one of claims 1-55, or a pharmaceutically acceptable salt thereof, wherein AK is an antibody or antigen binding fragment thereof comprising: a) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 2; a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 3; a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 4; a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 6; a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 7; and / or a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 8; b) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 12; a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 13; a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 14; a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 16; a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 17; and / or a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 18;P368919WO / 59362-725.601 c) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 22; a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 23; a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 24; a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 26; a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 27; and / or a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 28; d) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 32; a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 33; a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 34; a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 36; a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 37; and / or a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO:

38. e) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 42; a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 43; a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 44; a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 46; a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 47; and / or a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO:

48. f) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 52; a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 53; a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 54; a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 56; a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 57; and / or a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 58.P368919WO / 59362-725.601 g) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 62; a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 63; a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 64; a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 66; a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 67; and / or a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO:

68. h) a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 72; a heavy chain complementarity determining region 2 (HCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 73; a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence set forth in SEQ ID NO: 74; a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence set forth in SEQ ID NO: 76; a light chain complementarity determining region 2 (LCDR2) comprising the amino acid sequence set forth in SEQ ID NO: 77; and / or a light chain complementarity determining region 3 (LCDR3) comprising the amino acid sequence set forth in SEQ ID NO:

78. i) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 1; and / or an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 5; j) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 1; and / or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 5; k) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 9; and / or an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 10; l) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 9; and / or an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 10; m) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 11; and / or an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 15;P368919WO / 59362-725.601 n) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 11; and / or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 15; o) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 19; and / or an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 20; p) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 19; and / or an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 20; q) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 21; and / or an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 25; r) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 21; and / or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 25; s) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 29; and / or an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 30; t) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 29; and / or an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 30; u) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 31; and / or an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 35; v) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 31; and / or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 35; w) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 39; and / or an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 40;P368919WO / 59362-725.601 x) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 39; and / or an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 40; y) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 41; and / or an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 45; z) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 41; and / or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 45; aa) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 49; and / or an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 50; bb) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 49; and / or an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 50; cc) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 51; and / or an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 55; dd) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 51; and / or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 55; ee) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 59; and / or an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 60; ff) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 59; and / or an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 60; gg) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 61; and / or an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 65;P368919WO / 59362-725.601 hh) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 61; and / or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 65; ii) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 69; and / or an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 70; jj) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 69; and / or an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 70; kk) an immunoglobulin heavy chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 71; and / or an immunoglobulin light chain variable region comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 75; ll) an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 71; and / or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 75; mm) an immunoglobulin heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 79; and / or an immunoglobulin light chain comprising an amino acid sequence at least 85%, 90%, 95%, 97%, 98%, or 99% identical to SEQ ID NO: 80; or nn) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 79; and / or an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO:

80.

80. A pharmaceutical composition comprising the antibody-drug conjugate of any one of claims 1 to 73, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

81. A method of treating a disease or disorder, comprising administering to a patient in need thereof the antibody-drug conjugate of any one of claims 1 to 73.

82. A method of treating a hyperproliferative disorder or an angiogenic disorder, comprising administering to a patient in need thereof the antibody-drug conjugate of any one of claims 1 to 73.

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