Exatecan derivatives and antibody-drug conjugates thereof

Abstract

Disclosed herein, in part, is compound (I), an exatecan derivative having a novel chemical linker containing a cathepsin B cleavable site and attached to a targeting antibody.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 185,736, filed May 7, 2021, U.S. Provisional Patent Application No. 63 / 248,705, filed September 27, 2021, and U.S. Provisional Patent Application No. 63 / 321,187, filed March 18, 2022, the contents of which are hereby incorporated by reference in their entireties. [Background technology]

[0002] Antibody-drug conjugates (ADCs) provide a mechanism to selectively deliver small molecule therapeutics to antigen-positive cancer cells and attenuate the systemic toxicity of cytotoxic drugs to antigen-negative normal cells. The three components of an ADC - the antibody, the cytotoxic payload, and the linker that connects them - are critical in designing an effective therapy. Despite active development, challenges remain, for example, toxicity due to antibody binding to targets in normal tissues and dispersion of the cytotoxic payload in normal tissues due to instability of the ADC linker. Thus, many ADCs have not been successful in clinical trials due to lack of safety and / or efficacy at tolerated doses.

[0003] Topoisomerase I plays a critical role in DNA replication both under normal and pathological conditions, such as cancer. Since inhibition of topoisomerase I leads to cell death, compounds that bind to and inhibit topoisomerase I would be useful as therapeutic agents.

[0004] Camptothecin is a natural product with cytotoxic activity in various cell lines. Binding of its active lactone ring to topoisomerase I inhibits DNA replication, thus triggering cell apoptosis. However, its limitations in drug development include, for example, poor water solubility and the equilibrium between its active lactone form and the inactive ring-open form.

[0005] Exatecan is a water-soluble camptothecin derivative. As a chemotherapy agent, exatecan mesylate was not approved as a drug after several clinical trials due to lack of efficacy or high toxicity at the doses tested. To enable clinical utility of exatecan, efforts have been made to convert it into a prodrug covalently linked to a carboxymethyldextran polyalcohol polymer via a peptide spacer (a substrate for intracellular cathepsin proteases). However, this prodrug has not been successful in clinical trials.

[0006] Thus, there is a need for compounds that are more suitable for clinical development and more likely to succeed in the treatment of human tumors.Furthermore, preferential delivery of topoisomerase I inhibitors to diseased tissues by antibody-drug conjugates may improve safety and efficacy, providing treatment options to more patients and cancer types. Summary of the Invention [Means for solving the problem]

[0007] The present disclosure relates to compounds that are useful for the treatment of cancer.The present disclosure is directed, in part, to the exatecan derivatives that are useful as payloads in drug conjugates (e.g., antibody-drug conjugates), to the linker-payload constructs that are useful for binding payloads to antibodies, and to the drug conjugates that are based on exatecan.For example, the present disclosure provides compounds that represent therapeutic payloads, linker-payload constructs, or drug conjugates.

[0008] For example, the disclosure provides exatecan derivatives for use as therapeutic payloads. Also provided herein are linker-payload constructs and drug conjugates, each containing the disclosed therapeutic payloads. Further provided herein are pharmaceutical uses of the disclosed compounds, methods for their manufacture and pharmaceutical compositions containing them as active ingredients, both alone or in combination with other agents, as well as for their use as pharmaceuticals and / or in the manufacture of medicaments for the treatment of cancer.

[0009] For example, disclosed herein is a therapeutic payload represented by formula (I) or a pharma- ceutically acceptable salt or stereoisomer thereof. JPEG2024518438000002.jpg4267, where X is selected from the group consisting of O and S; Z is a bond, Y is hydrogen, -C 1-3 Alkyl, -CHO, and -C(O)-C 1-3 is selected from the group consisting of alkyl, and R is R 1 , R 2 , R 3 , R 4 , R 5 and hydrogen; or Y and Z, together with the nitrogen atom to which they are attached, each independently represent R Z R is bonded to the heteroaryl, and R is R 6 and R 1 is -C(O)-C 1-3 Alkyl, -C(O)-OC 1-3 Alkyl, C 1-4 Alkyl, -C 1-3 Alkyl-OC 1-3 Alkyl, -C(O)-C 3-4 Alkynyl, -S(O)2-C 1-3 Alkyl, -C(S)-C 1-3 Alkyl, -C 1-3 Alkyl-SC 1-3Alkyl and -C(O)-O-[(CH2)2-O] 1-10 -C alkyl; 1 is substituted by hydroxy, R 11 and optionally substituted with one or more additional substituents each independently selected from R 11 is, for each occurrence, independently selected from halogen, hydroxy, -C 1-3 Alkyl-OH, -C 1-3 Haloalkyl and -C 3-4 cycloalkyl; R 2 is -C(O)-NR a -C 1-3 Alkyl, -C(O)-C 0-3 Alkyl-C(O)-NR a -C 1-3 Alkyl, -C(O)-C 1-3 Alkyl-NR a -C 1-3 Alkyl, -S(O)2-C 1-3 Alkyl-NR a -C(O)-C 1-3 Alkyl and -C(O)NR a -[(CH2)2-O] 1-10 -C alkyl; 2 is substituted by hydroxy, R 22 and optionally substituted with one or more additional substituents each independently selected from R 22 is, for each occurrence, independently selected from halogen, hydroxy, -C 1-3 Alkyl-OH and -C 1-3 haloalkyl; R 3 is -C(O)-C 0-3 Alkyl-R 30 , -C(O)-C 0-3 Alkyl-OC 1-3 Alkyl-R 30 , -C 0-3 Alkyl-R 30 and -C 1-3 Alkyl-OC 1-3Alkyl-R 30 and, when alkyl is present, halogen and -C 1-3 haloalkyl; R 30 , N, NR 31 and O; R is selected from the group consisting of 5-6 membered heteroaryl and 4-10 membered heterocycles having 1, 2 or 3 heteroatoms, each independently selected from the group consisting of 30 are each independently R 33 and optionally substituted on one or more available carbons with one or more substituents selected from R 31 is, for each occurrence, independently, hydrogen, -C 1-3 Alkyl, -C 1-3 Alkyl-OH, -CH(OH)CHOH, -CHO and -C(O)-C 1-3 is selected from the group consisting of alkyl, R 33 For each occurrence, independently, -C 1-3 Alkyl-OH, halogen, hydroxy, oxo and -C 1-3 haloalkyl; R 4 is -C(O)-NR a -C 3-6 Cycloalkyl, -C(O)-C 0-2 Alkyl-C 3-6 Cycloalkyl, -C(S)-C 0-2 Alkyl-C 3-6 Cycloalkyl, -C(O)-NR a -C 3-6 Cycloalkyl and -C 3-6 Cycloalkenyl-NR a -C 1-3 alkyl; R 4 are each independently R 44 and is substituted by one or more substituents selected from R 44 is, for each occurrence, independently selected from hydroxy, halogen, oxo, -C 1-3Alkyl and -C 1-3 alkyl-OH; R 5 is -S(O)2-C 1-3 Alkyl-NR a R b , -C 1-4 Alkyl-NR a R b , -C(O)-C 1-3 Alkyl-O-NR a R b , -N=S(=O)(C 1-3 Alkyl)C 1-3 Alkyl, -C(O)-CH2-phenyl-CH2NR a R b and -[(CH2)2-NR a ] 1-5 -C 1-3 Alkyl-NR a R b each alkyl is independently selected from the group consisting of R 55 and optionally substituted by one or more substituents selected from R 55 represents, independently for each occurrence, a halogen, -C 1-3 Alkyl and -C 1-3 haloalkyl; R 6 are each independently substituted by hydroxy; 66 -C 1-3 is alkyl, R 66 represents, independently for each occurrence, a halogen and -C 1-3 haloalkyl; R Z is halogen, -C 1-3 Alkyl and -C 1-3 alkyl-OH; and R a and R b is, for each occurrence, independently, hydrogen, -C 1-3 Alkyl-OH and -C 1-3haloalkylOH; When X is O and Y is H, R is not hydrogen or -C(O)CHOH.

[0010] Also disclosed herein is a linker-payload construct represented by formula (IIA) or formula (IIB), or a pharma- ceutically acceptable salt or stereoisomer thereof: JPEG2024518438000003.jpg43126 (A is NH or triazolyl, L 1 is -CBP-NH-CH2- or -CBP-, where CBP is a cathepsin B-cleavable peptide or a cathepsin D-cleavable peptide; RR is L 1 and an alkoxy or amino moiety formed from a hydroxy or -NH2 moiety of a therapeutic payload described herein.

[0011] Further disclosed herein is a linker-payload construct represented by formula (IIIA) or (Formula IIIB), or a pharma- ceutically acceptable salt or stereoisomer thereof: JPEG2024518438000004.jpg3360(IIIA) JPEG2024518438000005.jpg3360(IIIB) (L 1 is a cathepsin B or cathepsin D cleavable peptide, and L 2 is the autoimmune part.

[0012] Further disclosed herein is a drug conjugate represented by formula (IVA) or formula (IVB), or a pharma- ceutically acceptable salt or stereoisomer thereof: JPEG2024518438000006.jpg3467(IVA) JPEG2024518438000007.jpg3467(IVB) (X is O or S, A is NH or triazolyl; Lig is the targeting moiety, L 1 is -CBP-NH-CH2- or -CBP-, where CBP is a cathepsin B or cathepsin D cleavable peptide, and RR is L 1 and an alkoxy or amino moiety formed from the hydroxy or -NH2 moiety of R of any one of the therapeutic payloads described herein.

[0013] A method of treating cancer is contemplated herein, comprising administering an effective amount of the disclosed compound to a patient in need thereof.For example, provided herein is a method of treating cancer in a patient in need thereof, comprising administering an effective amount of the disclosed therapeutic payload, the disclosed linker-payload construct, or the disclosed drug conjugate to the patient.

[0014] The pharmaceutical composition comprising at least one of the disclosed compounds and pharmaceutically acceptable carrier is further described herein.For example, provided herein is the pharmaceutically acceptable composition comprising the disclosed compounds, for example, the disclosed therapeutic payload, the disclosed linker-payload construct, or the disclosed drug conjugate, and pharmaceutically acceptable excipient. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] The features and other details of the present disclosure will be described in more detail.Before further description of the present disclosure, the specific terms employed in the present specification, examples and appended claims will be summarized here.These definitions should be read in the light of the remaining parts of the present disclosure as understood by those skilled in the art.Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.

[0016] definition As used herein, the terms "a" and "an" are intended to mean one or more, unless otherwise specified. For example, the term "agent" includes both a single agent and a combination of two or more agents.

[0017] The term "alkenyl" as used herein refers to an unsaturated straight or branched chain hydrocarbon having at least one carbon-carbon double bond. Exemplary alkenyl groups include, but are not limited to, C 26 Alkenyl and C 3-4 These include, but are not limited to, straight or branched groups having 2 to 6 or 3 to 4 carbon atoms, referred to as alkenyl. Exemplary alkenyl groups include, but are not limited to, vinyl, allyl, butenyl, pentenyl, and the like.

[0018] The term "alkoxy" as used herein refers to a straight or branched alkyl group attached to oxygen (alkyl-O-). Exemplary alkoxy groups include, but are not limited to, C 1-6 Alkoxy, and C 2-6 Examples of alkoxy groups include, but are not limited to, alkoxy groups having 1 to 6 or 2 to 6 carbon atoms, referred to as alkoxy. Exemplary alkoxy groups include, but are not limited to, methoxy, ethoxy, isopropoxy, and the like.

[0019] The term "alkoxyalkyl" as used herein refers to a straight or branched alkyl group bonded to a second straight or branched alkyl group and bonded to an oxygen (alkyl-O-alkyl-). Exemplary alkoxyalkyl groups include those described herein as C 1-6 Alkoxy-C 1-6 and alkoxyalkyl groups, each of which independently contains 1 to 6 carbon atoms, referred to as alkyl. Exemplary alkoxyalkyl groups include, but are not limited to, methoxymethyl, 2-methoxyethyl, 1-methoxyethyl, 2-methoxypropyl, ethoxymethyl, 2-isopropoxyethyl, and the like.

[0020] The term "alkoxycarbonyl" as used herein refers to a straight or branched alkyl group bonded to a carbonyl group and attached to an oxygen (alkyl-OC(O)-). Exemplary alkoxycarbonyl groups include those described herein as C 1-6 Examples of alkoxycarbonyl groups include, but are not limited to, alkoxycarbonyl groups having 1 to 6 carbon atoms, referred to as alkoxycarbonyl. Exemplary alkoxycarbonyl groups include, but are not limited to, methoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl, and the like.

[0021] The term "alkenyloxy" as used herein refers to a straight or branched alkenyl group attached to oxygen (alkenyl-O-). Exemplary alkenyloxy groups include those described herein as C 3-6 Examples of alkenyloxy groups include, but are not limited to, groups having an alkenyl group having 3 to 6 carbon atoms, referred to as alkenyloxy. Exemplary alkenyloxy groups include, but are not limited to, allyloxy, butenyloxy, and the like.

[0022] The term "alkynyloxy" as used herein refers to a straight or branched alkynyl group attached to oxygen (alkynyl-O). Exemplary alkynyloxy groups include those described herein as C 3-6 Examples of alkynyloxy groups include, but are not limited to, groups having an alkynyl group having 3 to 6 carbon atoms, referred to as alkynyloxy. Exemplary alkynyloxy groups include, but are not limited to, propynyloxy, butynyloxy, and the like.

[0023] The term "alkyl" as used herein refers to a saturated straight chain or branched hydrocarbon. Exemplary alkyl groups include those described herein as C 1-6 Alkyl, C 1-4 Alkyl, and C 1-3These include, but are not limited to, straight or branched chain hydrocarbons having 1 to 6, 1 to 4, or 1 to 3 carbon atoms, referred to as alkyl. Exemplary alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, 2-methyl-1-butyl, 3-methyl-2-butyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, and the like.

[0024] The term "alkylcarbonyl" as used herein refers to a straight or branched alkyl group attached to a carbonyl group (alkyl-C(O)-). Exemplary alkylcarbonyl groups include those described herein as C 1-6 Examples of alkylcarbonyl groups include, but are not limited to, alkylcarbonyl groups having 1 to 6 atoms, also known as alkylcarbonyl groups. Exemplary alkylcarbonyl groups include, but are not limited to, acetyl, propanoyl, isopropanoyl, butanoyl, and the like.

[0025] "Alkylene" means a linear or branched saturated aliphatic divalent radical having the number of carbon atoms indicated. "Cycloalkylene" means a divalent radical of a carbocyclic saturated hydrocarbon group having the number of carbon atoms indicated.

[0026] The term "alkynyl" as used herein refers to an unsaturated straight or branched chain hydrocarbon having at least one carbon-carbon triple bond. Exemplary alkynyl groups include, but are not limited to, C 2-6 Alkynyl and C 3-6 These, referred to as alkynyl, include, but are not limited to, straight chain or branched groups having 2 to 6 carbon atoms, or 3 to 6 carbon atoms. Exemplary alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, hexynyl, methylpropynyl, and the like.

[0027] As used herein, the term "carbonyl" refers to a -C(O)- group.

[0028] As used herein, the term "cyano" refers to a -CN group.

[0029] The term "cycloalkoxy" as used herein refers to a cycloalkyl group attached to an oxygen (cycloalkyl-O-). Exemplary cycloalkoxy groups include those described herein as C 3-6 Examples of cycloalkoxy groups include, but are not limited to, cycloalkoxy groups having 3 to 6 carbon atoms, referred to as cycloalkoxy groups. Exemplary cycloalkoxy groups include, but are not limited to, cyclopropoxy, cyclobutoxy, cyclohexyloxy, and the like.

[0030] The term "cycloalkyl" or "carbocyclic group" as used herein refers to a saturated or partially unsaturated hydrocarbon group having, for example, 3 to 6, or 4 to 6 carbon atoms, respectively, as used herein. 3-6 Cycloalkyl or C 4-6 Exemplary cycloalkyl groups include, but are not limited to, cyclohexyl, cyclopentyl, cyclopentenyl, cyclobutyl, or cyclopropyl.

[0031] The term “halo” or “halogen” as used herein refers to F, Cl, Br, or I.

[0032] The term "heteroaryl" or "heteroaromatic group" as used herein refers to a monocyclic aromatic 5-6 membered ring system containing one or more heteroatoms, e.g., 1-3 heteroatoms such as nitrogen, oxygen, sulfur, etc. The heteroaryl ring may be linked to adjacent groups through a carbon or nitrogen, if possible. Examples of heteroaryl rings include, but are not limited to, furan, thiophene, pyrrole, thiazole, oxazole, isothiazole, isoxazole, imidazole, pyrazole, triazole, pyridine, or pyrimidine, etc.

[0033] The term "heterocyclyl" or "heterocyclic group" is art-recognized and refers to a saturated or partially unsaturated, monocyclic or bicyclic ring structure, e.g., 4-10 membered, or 4-6 membered, saturated ring structure, including bridged, fused, or spiro rings, whose ring structures contain one to three heteroatoms, such as nitrogen, oxygen, and sulfur. Where possible, heterocyclyl rings may be linked to adjacent radicals through carbon or nitrogen. Examples of heterocyclyl groups include, but are not limited to, pyrrolidine, piperidine, morpholine, thiomorpholine, piperazine, oxetane, azetidine, tetrahydrofuran, or dihydrofuran, and the like.

[0034] The term "heterocyclyloxy" as used herein refers to a heterocyclyl group attached to an oxygen (heterocyclyl-O-).

[0035] The term "heteroaryloxy" as used herein refers to a heteroaryl group attached to an oxygen (heteroaryl-O-).

[0036] As used herein, the terms "hydroxy" and "hydroxyl" refer to an --OH group.

[0037] The term "oxo" as used herein refers to the group ═O.

[0038] "Pharmaceutically or pharmacologically acceptable" includes molecular entities and compositions that do not produce adverse, allergic or other untoward reactions when administered to animals or humans. For human administration, preparations should meet sterility, pyrogenicity, and general safety and purity standards as required by FDA Office of Biologics standards.

[0039] The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" as used herein refers to any solvent, dispersion medium, coating, isotonicity agent, absorption retardant, etc., that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. The composition may also contain other active compounds that provide complementary, additional, or enhanced therapeutic functions.

[0040] As used herein, the term "pharmaceutical composition" refers to a composition comprising at least one compound disclosed herein formulated together with one or more pharma- ceutically acceptable carriers.

[0041] "Individual", "patient", or "subject" are used interchangeably and include any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, or primates, and most preferably humans. Compounds of the present disclosure can be administered to mammals, such as humans, but also to other mammals, such as animals requiring veterinary treatment, for example, livestock animals (e.g., dogs, cats, etc.), farm animals (e.g., cows, sheep, pigs, horses, etc.) and laboratory animals (e.g., rats, mice, guinea pigs, etc.). "Modulation" includes antagonism (e.g., inhibition), agonism, partial antagonism and / or partial agonism.

[0042] "Treating" includes any effect that results in the improvement of a symptom, disease, disorder, or the like, for example, alleviating, reducing, modulating, or eliminating.

[0043] As used herein, the term "therapeutically effective amount" or "effective amount" refers to an amount of a subject compound that elicits a biological or medical response in a tissue, system or animal (e.g., a mammal or human) that is desired by a researcher, veterinarian, medical doctor or other clinician. The compounds of the present disclosure are administered in a therapeutically effective amount to treat a disease. Alternatively, a therapeutically effective amount of a compound is the amount necessary to achieve a desired therapeutic and / or prophylactic effect, such as an amount that causes weight loss.

[0044] As used herein, the term "pharmaceutically acceptable salt(s)" refers to salts of acidic or basic groups that may be present in the compounds used in the compositions. Compounds included in the compositions that are basic can form a wide variety of salts with various inorganic and organic acids. Acids which may be used to prepare pharma- ceutically acceptable acid addition salts of such basic compounds are those which form non-toxic acid addition salts, i.e., salts containing pharma- ceutically acceptable anions, including malate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate, tannate, pantothenate, tartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). Compounds contained in the present composition that are acidic in nature can form base salts with various pharmacologically acceptable cations. Examples of such salts include alkali metal salts or alkaline earth metal salts, particularly calcium salts, magnesium salts, sodium salts, lithium salts, zinc salts, potassium salts, and iron salts. Compounds contained in the present composition that contain a basic or acidic moiety can also form pharma-ceutically acceptable salts with various amino acids. Compounds of the present disclosure may contain both acidic and basic groups, for example, one amino group and one carboxylic acid group. In such cases, the compound may exist as an acid addition salt, zwitterion, or base salt.

[0045] As will be appreciated by those of skill in the art, "H" is the symbol for hydrogen, "N" is the symbol for nitrogen, "S" is the symbol for sulfur, and "O" is the symbol for oxygen. "Me" is an abbreviation for methyl. It will be understood that this disclosure is to be interpreted in accordance with the laws and principles of chemical bonding.

[0046] The compounds of the present disclosure may contain one or more asymmetric centers and therefore exist as stereoisomers. As used herein, the term "stereoisomer" consists of all enantiomers or diastereomers. These compounds may be designated by the symbols "(+)", "(-)", "R" or "S" depending on the arrangement of the substituents around the stereoisomeric carbon atom, but those of skill in the art will recognize that the structure may implicitly exhibit chiral centers. The present disclosure encompasses various stereoisomers of these compounds and mixtures thereof. Mixtures of enantiomers or diastereomers may be designated "(±)" in the nomenclature, but those of skill in the art will recognize that the structure may implicitly exhibit chiral centers.

[0047] The compounds of the present disclosure may contain one or more double bonds and therefore exist as geometric isomers resulting from the arrangement of substituents around a carbon-carbon double bond. JPEG2024518438000008.jpg27 depicts bonds that may be single, double, or triple bonds, as described herein. Substituents around a carbon-carbon double bond are designated to be in the "Z" or "E" configuration, with the terms "Z" and "E" being used in accordance with IUPAC standards. Unless otherwise specified, structures depicting double bonds encompass both the "E" and "Z" isomers. Substituents around a carbon-carbon double bond may be designated as "cis" or "trans", with "cis" referring to substituents on the same side of the double bond and "trans" referring to substituents on opposite sides of the double bond.

[0048] The compounds of the present disclosure may contain carbocyclic or heterocyclic rings and therefore exist as geometric isomers due to the arrangement of the substituents around the ring. The arrangement of the substituents around the carbocyclic or heterocyclic ring is designated as being in the "Z" or "E" configuration, and the terms "Z" and "E" are used according to IUPAC standards. Unless otherwise specified, structures depicting carbocyclic or heterocyclic rings encompass both the "Z" and "E" isomers. The substituents around the carbocyclic or heterocyclic ring may also be referred to as "cis" or "trans", with the term "cis" referring to the substituents on the same side of the plane of the ring and the term "trans" referring to the substituents on opposite sides of the plane of the ring. Mixtures of compounds in which the substituents are arranged on both the same and opposite sides of the plane of the ring are referred to as "cis / trans".

[0049] Individual enantiomers and diastereomers of the compounds of the present disclosure can be prepared synthetically from commercially available starting materials containing asymmetric or stereoisomeric centers, or by preparation of a racemic mixture followed by separation methods well known to those skilled in the art. These separation methods include (1) attaching the mixture of enantiomers to a chiral auxiliary, recrystallizing or chromatographically separating the resulting mixture of diastereomers, and liberating the optically pure products from the auxiliary, (2) forming salts with an optically active resolving agent, (3) directly separating the mixture of optical enantiomers on a chiral liquid chromatography column, or (4) kinetically separating the mixture using stereoselective chemical or enzymatic reagents. Racemic mixtures can also be separated into their constituent enantiomers by well known methods such as chiral phase liquid chromatography or crystallizing the compounds in chiral solvents. Stereoselective synthesis is a chemical or enzymatic reaction in which a single reactant forms an unequal mixture of stereoisomers, either in the production of a new stereoisomer or in the conversion of an existing stereoisomer, and is well known in the art. Stereoselective synthesis encompasses both enantio- and diastereoselective conversions and may involve the use of chiral auxiliaries. See, for example, Carreira and Kvaerno, Classics in Stereoselective Synthesis, Wiley-VCH: Weinheim, 2009.

[0050] The compounds disclosed herein can exist in solvated and unsolvated forms with pharma- ceutically acceptable solvents such as water, ethanol, etc., and it is intended that the present disclosure encompasses both solvated and unsolvated forms.In one embodiment, the compound is amorphous.In one embodiment, the compound is a single polymorph.In another embodiment, the compound is a mixture of polymorphs.In another embodiment, the compound is a crystalline form.

[0051] The present disclosure also encompasses isotopically labeled compounds of the present disclosure that are identical to those described herein, except that one or more atoms are replaced with an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature. Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, e.g., 2 H, 3 H, 13 C. 14 C. 15 N, 18 O. 17 O. 31 P, 32 P, 35 S, 18 F and 36 For example, the compounds of the present disclosure may have one or more H atoms replaced with deuterium.

[0052] Certain isotopically labeled disclosed compounds (e.g., 3 H and 14 C) are useful in compound and / or substrate tissue distribution assays. Tritium isotopes (i.e., 3 H) and carbon-14 isotopes (i.e., 14 C) is particularly preferred because it is easy to prepare and detect. 2 Substitution with heavy isotopes such as H) may confer certain therapeutic advantages resulting from greater metabolic stability (e.g., increased half-life in vivo or reduced dosage requirements) and therefore may be preferred in some circumstances. Isotopically labeled compounds of the present disclosure can generally be prepared following procedures similar to those disclosed in the Examples herein by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0053] The term "prodrug" refers to a compound that is converted in vivo to obtain the disclosed compound or a pharma- ceutically acceptable salt, hydrate, or solvate thereof. This conversion can occur at various sites (intestinal lumen, during intestinal transit, in blood, liver, etc.) and by various mechanisms (esterases, amidases, phosphatases, oxidative metabolism, reductive metabolism, etc.). Prodrugs are well known in the art (see, e.g., Rautio, Kumpulainen, et al, Nature Reviews Drug Discovery 2008, 7, 255). For example, when a compound of the present disclosure or a pharma- ceutically acceptable salt, hydrate, or solvate thereof contains a carboxylic acid functional group, a prodrug can be prepared by replacing a hydrogen atom of the acid group with a carboxylic acid functional group such as (C 1-8 ) alkyl, (C 2-12 ) alkylcarbonyloxymethyl, 1-(alkylcarbonyloxy)ethyl having 4 to 9 carbon atoms, 1-methyl-1-(alkylcarbonyloxy)ethyl having 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having 3 to 6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having 4 to 7 carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having 5 to 8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having 3 to 9 carbon atoms, 1-(N(alkoxycarbonyl)amino)ethyl having 4 to 10 carbon atoms, 3-phthalidyl, 4-crotonolactonyl, γ-butyrolactone-4-yl, di-N,N-(C 1-2 ) Alkylamino(C 2-3 ) alkyl (β-dimethylaminoethyl, etc.), carbamoyl-(C 1-2 ) alkyl, N,N-di(C 1-2 ) alkylcarbamoyl-(C 1-2 ) alkyl and piperidino-, pyrrolidino- or morpholino (C 2-3 ) alkyl or the like.

[0054] Similarly, when the disclosed compounds contain an alcohol functional group, the prodrug may be a prodrug that converts a hydrogen atom of the alcohol group to a hydrogen atom of, for example, (C 1-6) alkylcarbonyloxymethyl, 1-((C 1-6 ) alkylcarbonyloxy) ethyl, 1-methyl 1-((C 1-6 ) alkylcarbonyloxy) ethyl (C 1-6 ) alkoxycarbonyloxymethyl, N(C 1-6 ) alkoxycarbonylaminomethyl, succinoyl, (C 1-6 α-amino (C 1-4 ) alkylcarbonyl, arylalkylcarbonyl and α-aminoalkylcarbonyl, or α-aminoalkylcarbonyl-α-aminoalkylcarbonyl, where each α-aminoalkylcarbonyl group is independently selected from the group consisting of naturally occurring L-amino acids, P(O)(OH), -P(O)(O(C 1-6 ) alkyl) 2 or glycosyl (a group resulting from removal of the hydroxyl group of the hemiacetal form of a carbohydrate).

[0055] When the compounds of the present disclosure incorporate an amine function, prodrugs can be formed, for example, by the generation of amides or carbamates, N-alkylcarbonyloxyalkyl derivatives, (oxodioxolenyl)methyl derivatives, N-Mannich bases, imines or enamines. Secondary amines can also be metabolically cleaved to generate biologically active primary amines, or tertiary amines can be metabolically cleaved to generate biologically active primary or secondary amines. See, for example, Simplicio, et al., Molecules 2008, 13, 519.

[0056] Procedures for preparing the compounds described herein are provided below in the examples and can be supplemented or replaced by procedures known to those skilled in the art. The raw materials used in the examples can be purchased or prepared by methods described in the chemical literature, or adaptations thereof, using methods known to those skilled in the art. The order in which the steps are carried out will vary depending on the groups introduced, the reagents used, and will be apparent to those skilled in the art. Any of the disclosed compounds, or intermediates described herein, can be further derivatized using one or more standard synthetic methods known to those skilled in the art.

[0057] Salts of the compounds disclosed herein can be prepared by reacting the compounds disclosed herein with an appropriate acid or base in a suitable solvent or mixture of solvents (e.g., an ether such as diethyl ether, or an alcohol such as ethanol, or an aqueous solvent, etc.) using conventional procedures. Salts of the compounds disclosed herein can be exchanged for other salts by treatment using conventional ion exchange chromatography procedures.

[0058] compound Disclosed herein, for example, is a therapeutic payload represented by formula (I) or a pharma- ceutically acceptable salt or stereoisomer thereof. JPEG2024518438000009.jpg4267, where X is selected from the group consisting of O and S; Z is a bond, Y is hydrogen, -C 1-3 Alkyl, -CHO, and -C(O)-C 1-3 is selected from the group consisting of alkyl, and R is R 1 , R 2 , R 3 , R 4 , R 5 and hydrogen; or Y and Z, together with the nitrogen atom to which they are attached, each independently represent R ZR is bonded to the heteroaryl, and R is R 6 and R 1 is -C(O)-C 1-3 Alkyl, -C(O)-OC 1-3 Alkyl, C 14 Alkyl, -C 1-3 Alkyl-OC 1-3 Alkyl, -C(O)-C 3-4 Alkynyl, -S(O)2-C 1-3 Alkyl, -C(S)-C 1-3 Alkyl, -C 1-3 Alkyl-SC 1-3 Alkyl, and -C(O)-O-[(CH2)2-O] 1-10 -C alkyl; 1 is substituted by hydroxy, R 11 and optionally substituted with one or more additional substituents each independently selected from R 11 is independently, for each occurrence, halogen, hydroxy, -C 1-3 Alkyl-OH, -C 1-3 Haloalkyl, and -C 3-4 cycloalkyl; R 2 is -C(O)-NR a -C 1-3 Alkyl, -C(O)-C 0-3 Alkyl-C(O)-NR a -C 1-3 Alkyl, -C(O)-C 1-3 Alkyl-NR a -C 1-3 Alkyl, -S(O)2-C 1-3 Alkyl-NR a -C(O)-C 1-3 Alkyl, and C(O)NR a -[(CH2)2-O] 1-10 -C alkyl; 2 is substituted by hydroxy, R 22and optionally substituted with one or more additional substituents each independently selected from R 22 is independently, for each occurrence, halogen, hydroxy, -C 1-3 Alkyl-OH and -C 1-3 haloalkyl; R 3 -C(O)-C 0-3 Alkyl-R 30 , -C(O)-C 0-3 Alkyl-OC 1-3 Alkyl-R 30 , -C 0-3 Alkyl-R 30 , and -C 1-3 Alkyl-OC 1-3 Alkyl-R 30 and, when alkyl is present, halogen and -C 1-3 haloalkyl; R 30 , N, NR 31 R is selected from the group consisting of 5-6 membered heteroaryl and 4-10 membered heterocycles having 1, 2 or 3 heteroatoms, each independently selected from the group consisting of 30 are each independently R 33 and optionally substituted on one or more available carbons with one or more substituents selected from R 31 is, for each occurrence, independently, hydrogen, -C 1-3 Alkyl, -C 1-3 Alkyl-OH, -CH(OH)CHOH, -CHO, and -C(O)-C 1-3 is selected from the group consisting of alkyl, R 33 For each occurrence, independently, -C 1-3 Alkyl-OH, halogen, hydroxy, oxo and -C 1-3 haloalkyl; R 4 is -C(O)-NR a -C 3-6Cycloalkyl, -C(O)-C 0-2 Alkyl-C 3-6 Cycloalkyl, -C(S)-C 0-2 Alkyl-C 3-6 Cycloalkyl, -C(O)-NR a -C 3-6 Cycloalkyl and -C 3-6 Cycloalkenyl-NR a -C 1-3 alkyl; R 4 are each independently R 44 and is substituted by one or more substituents selected from R 44 is, for each occurrence, independently selected from hydroxy, halogen, oxo, -C 1-3 Alkyl and -C 1-3 alkyl-OH; R 5 is -S(O)2-C 1-3 Alkyl-NR a R b , -C 1-4 Alkyl-NR a R b , -C(O)-C 1-3 Alkyl-O-NR a R b , -N=S(=O)(C 1-3 Alkyl)C 1-3 Alkyl, -C(O)-CH2-phenyl-CH2NR a R b , and -[(CH2)2-NR a ] 1-5 -C 1-3 Alkyl-NR a R b each alkyl is independently selected from the group consisting of R 55 and optionally substituted by one or more substituents selected from R 55 represents, independently for each occurrence, a halogen, -C 1-3 Alkyl and -C 1-3 haloalkyl; R 6 are each independently substituted by hydroxy;66 -C 1-3 is alkyl, R 66 represents, independently for each occurrence, a halogen and C 1-3 haloalkyl; R Z is halogen, -C 1-3 Alkyl and -C 1-3 alkyl-OH; and R a and R b is, for each occurrence, independently, hydrogen, -C 1-3 Alkyl-OH and -C 1-3 haloalkylOH; When X is O and Y is H, R is not hydrogen or -C(O)CHOH.

[0059] In some embodiments, X is O. In other embodiments, Z is a bond. In certain embodiments, Y is selected from, for example, hydrogen, -CH3, -CHO, and -COCH3.

[0060] In some embodiments, R is R 1 For example, in some embodiments, R is -C(O)-C1 alkyl, -C(O)-C2 alkyl, -C(O)-O-C2 alkyl, -C(O)-O-C3 alkyl, -C2 alkyl, -C3 alkyl, -C2 alkyl-O-C2 alkyl, -C(S)-C1 alkyl, -S(O)2-C1 alkyl, -S(O)2-C2 alkyl, -S(O)2-C3 alkyl, -C(O)-C3 alkynyl, -C2 alkyl-S-C2 alkyl, and -C(O)-O-[(CH2)2-O] 1-5 -C alkyl, R is substituted by hydroxy, R 11 In certain embodiments, R 11is selected from the group consisting of fluoro, hydroxy, -CH2-OH, -CF3, and cyclopropyl.

[0061] For example, in some embodiments, -N(Y)-ZR can be selected from the group consisting of: JPEG2024518438000010.jpg189170

[0062] In another embodiment, R is R 2 In further embodiments, Y is hydrogen. In some embodiments, for example, R is selected from the group consisting of -C(O)-NH-C2 alkyl, -C(O)-NH-C3 alkyl, -C(O)-C(O)-NH-C2 alkyl, -C(O)-C(O)-NH-C3 alkyl, -C(O)-C1 alkyl-C(O)-NH-C2 alkyl, -C(O)-C2 alkyl-C(O)-NH-C2 alkyl, -C(O)-C2 alkyl-C(O)-NH-C3 alkyl, -S(O)2-C2 alkyl-NH-C(O)-C1 alkyl, -S(O)2-C2 alkyl-NH-C(O)-C2 alkyl, and -C(O)NH-[(CH2)2-O] 1-2 -C alkyl; 2 is replaced by hydroxy, R 22 In a further embodiment, R 22 is selected from the group consisting of fluoro, hydroxy, -CH2-OH and -CF3.

[0063] For example, in some embodiments, -N(Y)-ZR is selected from the group consisting of: JPEG2024518438000011.jpg137170

[0064] In other embodiments, R is R 3In certain embodiments, Y is hydrogen. In further embodiments, R is, for example, -C(O)-triazolyl, -C(O)-C1 alkyl-triazolyl, -C(O)-C2 alkyl-triazolyl, -C(O)-C3 alkyl-triazolyl, -C1 alkyl-triazolyl, -C2 alkyl-triazolyl, -C3 alkyl-triazolyl, -C(O)-O-C1 alkyl-triazolyl, -C(O)-O-C2 alkyl-triazolyl, -C(O)-C1 alkyl-O-C2 alkyl alkyl-triazolyl, -C(O)-C2 alkyl-O-C1 alkyl-triazolyl, -C(O)-C2 alkyl-O-C2 alkyl-triazolyl, -C2 alkyl-O-C1 alkyl-triazolyl, and -C2 alkyl-O-C2 alkyl-triazolyl, wherein alkyl in each occurrence is optionally substituted with 1, 2, or 3 substituents each independently selected from the group consisting of fluoro and -CF3; Triazolyl, if present, is hydrogen, -C 1-3 Alkyl and -C 1-2 substituted on an available nitrogen by a substituent selected from the group consisting of alkyl-OH, and Triazolyl includes chloro, fluoro and -C 1-2 -alkyl-OH.

[0065] For example, in some embodiments, R is selected from the group consisting of: JPEG2024518438000012.jpg213170JPEG2024518438000013.jpg135170

[0066] In other embodiments, R is selected from the group consisting of -C(O)-furanyl, -Calkyl-furanyl, -C(O)-oxazolyl, and -C(O)-pyrazolyl; R is selected from the group consisting of hydroxy and -C 1-2 For example, in one embodiment, R is selected from the group consisting of: JPEG2024518438000014.jpg26170

[0067] In a further embodiment, R is selected from the group consisting of: JPEG2024518438000015.jpg80163

[0068] In a further embodiment, R is R 4 In some embodiments, Y is hydrogen. In other embodiments, R is selected from the group consisting of, for example, -C(O)-C3 cycloalkyl, -C(S)-C3 cycloalkyl, -C(O)-C4 cycloalkyl, -C(O)-C5 cycloalkyl, -C(O)-C6 cycloalkyl, -C(O)-NH-C3 cycloalkyl, -C(O)-NH-C4 cycloalkyl, -C4 cycloalkenyl-NH-C2 alkyl, -C4 cycloalkenyl-NH-C3 alkyl, -C5 cycloalkenyl-NH-C2 alkyl, and -C5 cycloalkenyl-NH-C2 alkyl, wherein cycloalkyl or cycloalkenyl is selected from the group consisting of hydroxy, oxo, -C 1-3 Alkyl and -C 1-2 substituted by one or more substituents each independently selected from the group consisting of alkyl-OH; The alkyl is substituted with 1, 2, or 3 substituents each independently selected from the group consisting of hydroxy and -CH2OH.

[0069] For example, in some embodiments, R is selected from the group consisting of: JPEG2024518438000016.jpg129162

[0070] In some embodiments, R is R 5In other embodiments, Y is selected from the group consisting of hydrogen, -CH3, and -C(O)CH3. In certain embodiments, R is selected from the group consisting of -S(O)2-C2 alkyl-NH2, -S(O)2-C3 alkyl-NH2, -C2 alkyl-NH2, -C3 alkyl-NH2, -C(O)-C1 alkyl-O-NH2, -C(O)-CH2-phenyl-CH2NH2, and -(CH2)2-NH-C2 alkyl-NH2, where the alkyl is optionally substituted with one or two -CH3 groups.

[0071] For example, in some embodiments, -ZN(Y)-R is selected from the group consisting of: JPEG2024518438000017.jpg41170

[0072] In other embodiments, Y and Z together with the nitrogen atom to which they are attached form a triazolyl substituted at a position substitutable by R. In certain embodiments, R is C1 alkyl-OH or C2 alkyl-OH, and R is optionally substituted by -CF3. In further embodiments, -ZN(Y)-R is selected from the group consisting of: JPEG2024518438000018.jpg42170

[0073] In yet other embodiments, X is S. In certain embodiments, Y is hydrogen. In certain embodiments, R is, for example, hydrogen, JPEG2024518438000019.jpg18151.

[0074] In some embodiments, the disclosed therapeutic payloads can be selected from, for example, any one of the compounds set forth in Table 1, or a pharma- ceutically acceptable salt or stereoisomer thereof. [Table 1] JPEG2024518438000021.jpg211152JPEG2024518438000022.jpg196152JPEG2024518438000023.jpg206144JPEG2024518438000024.jpg225157JPEG2024518438000025.jpg227156JPEG2024518438000026.jpg224162JPEG2024518438000027.jpg207155JPEG2024518438000028.jpg200157JPEG2024518438000029.jpg229152JPEG2024518438000030.jpg217150JPEG2024518438000031.jpg196152JPEG2024518438000032.jpg218152JPEG2024518438000033.jpg227156JPEG2024518438000034.jpg210152JPEG2024518438000035.jpg206156JPEG2024518438000036.jpg229164JPEG2024518438000037.jpg200155JPEG2024518438000038.jpg205154JPEG2024518438000039.jpg207157JPEG2024518438000040.jpg208147JPEG2024518438000041.jpg208141JPEG2024518438000042.jpg210140JPEG2024518438000043.jpg222147JPEG2024518438000044.jpg227152JPEG2024518438000045.jpg197141JPEG2024518438000046.jpg216149JPEG2024518438000047.jpg204141JPEG2024518438000048.jpg208143JPEG2024518438000049.jpg205140JPEG2024518438000050.jpg205142JPEG2024518438000051.jpg198146JPEG2024518438000052.jpg216154JPEG2024518438000053.jpg219151JPEG2024518438000054.jpg238164.

[0075] In some embodiments, the disclosed therapeutic payloads can be formed, for example, by contacting a cell or tissue with a drug conjugate of formula (IA) or a pharma- ceutically acceptable salt or stereoisomer thereof at 37° C. and a pH of about 5 to about 7.7. JPEG2024518438000055.jpg3672 (X is O or S, A is NH or triazolyl; Lig is the targeting moiety, L 1 is a linker moiety, and RR is L 1 and an alkoxy or amino moiety formed from the hydroxy or -NH2 moiety of R of any of the therapeutic payloads disclosed herein.

[0076] Disclosed herein is a method of delivering a therapeutically effective amount of a therapeutic payload moiety to a patient in need thereof, comprising administering to the patient a drug conjugate represented by Formula (IA) or a pharma- ceutically acceptable salt or stereoisomer thereof. JPEG2024518438000056.jpg3672 (X is O or S, A is NH or triazolyl; Lig is the targeting moiety, L 1 is a linker moiety, and RR is L 1 and an alkoxy or amino moiety formed from the hydroxy or -NH2 moiety of R of any of the therapeutic payloads disclosed herein.

[0077] Also contemplated herein are drug conjugates represented by: JPEG2024518438000057.jpg3341 (n is 1 to about 10, for example, about 6.5 to 8.5.)

[0078] In some embodiments, Lig is a monoclonal antibody. For example, in some embodiments, Lig is an antibody selected from the group consisting of an anti-TROP2 antibody, an anti-EGRF antibody, an anti-HER2 antibody, an anti-B7-H3 antibody, an anti-CD30 antibody, an anti-CD33 antibody, and an anti-CD70 antibody. In one embodiment, Lig is, for example, an anti-TROP2 antibody.

[0079] In another embodiment, L 1 but, -Succinimidyl-(CH2)2-O-(CH2)2-C(O)-CBP-NH-CH2-, -Succinimidyl-(CH2)2-O-(CH2)2-C(O)-CBP-, -Succinimidyl-(CH2)5-C(O)-CBP-NH-CH2- or -Succinimidyl-(CH2)5-C(O)-CBP-, CBP is a cathepsin B cleavable site or a cathepsin D cleavable site.

[0080] In further embodiments, the CBP is a cathepsin B-cleavable peptide or a cathepsin D-cleavable peptide. In one embodiment, the CBP is -Gly-Gly-Phe-Gly- or -Val-Cit-.

[0081] In some embodiments, L 1 is, for example, selected from the group consisting of: JPEG2024518438000058.jpg157170

[0082] Further disclosed herein is a method of delivering a therapeutically effective amount of a therapeutic payload moiety to a patient in need thereof, comprising administering to the patient a drug conjugate represented by Formula (IB) or a pharma- ceutically acceptable salt or stereoisomer thereof. JPEG2024518438000059.jpg3460 (X is O or S, Lig is the targeting moiety, L 1 is the linking moiety, and L 2 is the autoimmune part.

[0083] In some embodiments, Lig is a monoclonal antibody. For example, in some embodiments, Lig is an antibody selected from the group consisting of an anti-TROP2 antibody, an anti-EGRF antibody, an anti-HER2 antibody, an anti-B7-H3 antibody, an anti-CD30 antibody, an anti-CD33 antibody, and an anti-CD70 antibody. In one embodiment, Lig is an anti-TROP2 antibody.

[0084] In another embodiment, L 1 teeth, -Succinimidyl-(CH2)2-O-(CH2)2-C(O)-CBP- or -succinimidyl-(CH2)5-C(O)-CBP-, CBP is a cathepsin B cleavable site or a cathepsin D cleavable site.

[0085] In some embodiments, the CBP is, for example, a cathepsin B-cleavable peptide or a cathepsin D-cleavable peptide. In one embodiment, the CBP is -Gly-Gly-Phe-Gly- or -Val-Cit-.

[0086] In a further embodiment, L 1 is, for example, selected from the group consisting of: JPEG2024518438000060.jpg115164

[0087] In a further embodiment, L 2 is, for example, selected from the group consisting of: JPEG2024518438000061.jpg108167

[0088] Disclosed herein are, for example, linker-payload constructs of formula (IIA) or formula (IIB), or a pharma- ceutically acceptable salt or stereoisomer thereof. JPEG2024518438000062.jpg42120 (A is NH or triazolyl, L 1 is -CBP-NH-CH2- or -CBP-, where CBP is a cathepsin B-cleavable peptide or a cathepsin D-cleavable peptide; RR is L 1 and a hydroxy or alkoxy or amino moiety formed from the -NH2 moiety of R of any therapeutic payload disclosed herein.

[0089] In some embodiments, L 1 is selected from the group consisting of: JPEG2024518438000063.jpg54141

[0090] In some embodiments, the linker-payload construct is selected from the group consisting of: JPEG2024518438000064.jpg36105JPEG2024518438000065.jpg36105JPEG2024518438000066.jpg36110JPEG2024518438000067.jpg361 10JPEG2024518438000068.jpg4785JPEG2024518438000069.jpg4785JPEG2024518438000070.jpg4675JPEG2024518438000071.jpg4675

[0091] In some embodiments, the disclosed linker-payload constructs can be selected, for example, from any one of the compounds disclosed in Table 2, or a pharma- ceutically acceptable salt or stereoisomer thereof. [Table 2] JPEG2024518438000073.jpg235170JPEG2024518438000074.jpg239170JPEG2024518438000075.jpg224170JPEG2024518438000076.jpg89170

[0092] Further disclosed herein is a linker-payload construct of Formula (IIIA) or Formula (IIIB) or a pharma- ceutically acceptable salt or stereoisomer thereof. JPEG2024518438000077.jpg42124(L 1 is a cathepsin B or cathepsin D cleavable peptide, and L 2 is the autoimmune part.

[0093] In some embodiments, L 1 is selected from the group consisting of: JPEG2024518438000078.jpg29132

[0094] In other embodiments, the linker-payload construct is selected from the group consisting of: JPEG2024518438000079.jpg37110JPEG2024518438000080.jpg37111JPEG2024518438000081.jpg4676JPEG2024518438000082.jpg4677

[0095] In some embodiments, L 2 is selected from the group consisting of: JPEG2024518438000083.jpg109160

[0096] Also disclosed herein is, for example, a drug conjugate represented by formula (IVA) or formula (IVB), or a pharma- ceutically acceptable salt or stereoisomer thereof. JPEG2024518438000084.jpg41136 (X is O or S, A is NH or triazolyl; Lig is the targeting moiety, L 1 is -CBP-NH-CH2- or -CBP-, where CBP is a cathepsin B or cathepsin D cleavable peptide, and RR is L 1 and a hydroxy or -NH2 moiety of R of any therapeutic payload disclosed herein.

[0097] In some embodiments, Lig is a monoclonal antibody. For example, Lig is an antibody selected from the group consisting of an anti-TROP2 antibody, an anti-EGRF antibody, an anti-HER2 antibody, an anti-B7-H3 antibody, an anti-CD30 antibody, an anti-CD33 antibody, and an anti-CD70 antibody. In one embodiment, for example, Lig is an anti-TROP2 antibody.

[0098] In other embodiments, the CBP is, for example, -Gly-Gly-Phe-Gly- or -Val-Cit-.

[0099] In a further embodiment, L 1 is selected from the group consisting of: JPEG2024518438000085.jpg73162

[0100] In further embodiments, the drug conjugate is, for example, selected from the group consisting of: JPEG2024518438000086.jpg36111JPEG2024518438000087.jpg36111JPEG2024518438000088.jpg37117JPEG2024518438000089.jpg371 17JPEG2024518438000090.jpg4691JPEG2024518438000091.jpg4690JPEG2024518438000092.jpg4682JPEG2024518438000093.jpg4782

[0101] Also disclosed herein is a drug conjugate represented by Formula (VA) or Formula (VB), or a pharma- ceutically acceptable salt or stereoisomer thereof. JPEG2024518438000094.jpg41137 (X is O or S, Lig is the targeting moiety, L 1 is a cathepsin B or cathepsin D cleavable peptide, and L 2 is the autoimmune part.

[0102] In some embodiments, Lig is a monoclonal antibody. For example, in some embodiments, Lig is an antibody selected from the group consisting of an anti-TROP2 antibody, an anti-EGRF antibody, an anti-HER2 antibody, an anti-B7-H3 antibody, an anti-CD30 antibody, an anti-CD33 antibody, and an anti-CD70 antibody. In one embodiment, Lig is an anti-TROP2 antibody.

[0103] In another embodiment, L 1 is selected from the group consisting of: JPEG2024518438000095.jpg29138

[0104] In further embodiments, the drug conjugate is, for example, selected from the group consisting of: JPEG2024518438000096.jpg37118JPEG2024518438000097.jpg39113JPEG2024518438000098.jpg4783JPEG2024518438000099.jpg4780

[0105] In a further embodiment, L 2 is selected from the group consisting of: JPEG2024518438000100.jpg108169

[0106] Also disclosed herein is a drug conjugate selected from the group consisting of: JPEG2024518438000101.jpg7391JPEG2024518438000102.jpg7391JPEG2024518438000103.jpg9185JPEG2024518438000104.jpg7291JPEG2024518438000105.jpg8192Lig is the targeting part.

[0107] In some embodiments, Lig is a monoclonal antibody. For example, in some embodiments, Lig is an antibody selected from the group consisting of an anti-TROP2 antibody, an anti-EGRF antibody, an anti-HER2 antibody, an anti-B7-H3 antibody, an anti-CD30 antibody, an anti-CD33 antibody, and an anti-CD70 antibody. In one embodiment, Lig is, for example, an anti-TROP2 antibody.

[0108] The intended targets and corresponding antibodies of this disclosure are shown in Table 3. [Table 3] JPEG2024518438000107.jpg223170JPEG2024518438000108.jpg104160

[0109] method Disclosed herein are, for example, methods of treating cancer in a patient in need thereof comprising administering to the patient an effective amount of a therapeutic payload disclosed herein, wherein the cancer is selected from the group consisting of lung cancer, renal cancer, urothelial cancer, colorectal cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, breast cancer, melanoma, liver cancer, bladder cancer, gastric cancer, and esophageal cancer.

[0110] Also disclosed herein is a method of treating cancer in a patient in need thereof comprising administering to the patient an effective amount of a linker-payload construct disclosed herein, wherein the cancer is selected from the group consisting of lung cancer, renal cancer, urothelial cancer, colorectal cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, breast cancer, melanoma, liver cancer, bladder cancer, gastric cancer, and esophageal cancer.

[0111] Further disclosed herein is a method of treating cancer in a patient in need thereof, comprising administering to the patient an effective amount of a drug conjugate comprising any of the payloads disclosed herein, wherein the cancer is selected from the group consisting of lung cancer, renal cancer, urothelial cancer, colorectal cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, breast cancer, melanoma, liver cancer, bladder cancer, gastric cancer, and esophageal cancer.

[0112] In certain embodiments, the patient is a human.

[0113] In certain embodiments, administration of the disclosed compounds can include subcutaneous administration. In certain embodiments, administration of the disclosed compounds can include intravenous administration. In certain embodiments, administration of the disclosed compounds can include oral administration.

[0114] The provided methods of treatment can include administering the disclosed compounds once, twice, or three times daily, about every other day (e.g., every third day), twice weekly (e.g., every third, fourth, fifth, sixth days, or, for example, with about 2 to about 3 days between doses), once weekly, three times weekly, every other week, twice monthly, monthly, bimonthly, or even less frequently.

[0115] In particular, in certain embodiments, the present disclosure provides a method of treating one or more of the above medical indications, comprising administering to a subject in need thereof a therapeutically effective amount of a compound described herein.

[0116] In certain embodiments, the compound utilized by one or more of the methods disclosed herein is one of the later compounds, sub-later compounds, or specific compounds described herein.

[0117] The compounds of the present disclosure can be administered to patients (animals and humans) in need of such treatment in a dosage that provides optimal pharmaceutical efficacy. It will be understood that the dosage required for use in any particular application will vary from patient to patient, depending not only on the specific compound or composition selected, but also on the route of administration, the nature of the condition being treated, the age and condition of the patient, any concurrent medications or special diets the patient is currently following, and other factors that one skilled in the art would recognize, and that the appropriate dosage is ultimately left to the discretion of the attending physician. To treat the clinical conditions and diseases noted herein, the compounds of the present disclosure can be administered orally, subcutaneously, topically, parenterally, by inhalation spray or rectally in dosage unit formulations that contain conventional non-toxic pharmacologic acceptable carriers, adjuvants and vehicles. Parenteral administration includes subcutaneous injection, intravenous injection, intramuscular injection, or infusion techniques.

[0118] Treatment can be continued for a desired period of time or for a short period of time. Suitable treatment periods can be, for example, at least about 1 week, at least about 2 weeks, at least about 1 month, at least about 6 months, at least about 1 year, or indefinitely. Treatment periods can be terminated when desired results are achieved.

[0119] Pharmaceutical Compositions and Kits Another aspect of the present disclosure provides pharmaceutical compositions comprising compounds as disclosed herein formulated with a pharmaceutically acceptable carrier. In particular, the present disclosure provides pharmaceutical compositions comprising compounds as disclosed herein formulated with one or more pharmaceutically acceptable carriers. These formulations include those suitable for oral, rectal, topical, buccal, parenteral (e.g., subcutaneous, intramuscular, intradermal, or intravenous), vaginal, or aerosol administration, although the most suitable administration form in any given case depends on the extent and severity of the condition being treated and the nature of the particular compound being used. For example, the disclosed compositions can be formulated as a unit dose and / or can be formulated for oral or subcutaneous administration.

[0120] For example, disclosed herein is a pharmaceutical composition comprising a therapeutic payload disclosed herein and a pharma- ceutically acceptable excipient. Also disclosed herein is a pharmaceutical composition comprising a linker-payload construct disclosed herein and a pharma- ceutically acceptable excipient. Further disclosed herein is a pharmaceutical composition comprising a drug conjugate disclosed herein and a pharma- ceutically acceptable excipient.

[0121] Exemplary pharmaceutical compositions of the present disclosure can be used in the form of pharmaceutical preparations, for example, in solid, semi-solid or liquid form, which contain one or more disclosed compounds as active ingredients in admixture with organic or inorganic carriers or excipients suitable for external, enteral or parenteral use. The active ingredients can be combined with conventional non-toxic pharmaceutically acceptable carriers for, for example, tablets, pellets, capsules, suppositories, solutions, emulsions, suspensions, and any other form suitable for use. The active subject compound is included in the pharmaceutical composition in an amount sufficient to produce the desired effect on the disease process or condition.

[0122] To prepare solid compositions such as tablets, the primary active ingredient can be mixed with a pharmaceutical carrier, such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate or gums, and other pharmaceutical diluents, such as water, to form a solid preformulation composition containing a homogenous mixture of the disclosed compound or a non-toxic pharma-ceutically acceptable salt thereof. When these preformulation compositions are referred to as homogenous, it is meant that the active ingredient is uniformly dispersed throughout the composition, so that the composition can be readily subdivided into equally effective unit dosage forms, such as tablets, pills, and capsules.

[0123] In solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules, etc.), the subject compositions are mixed with one or more pharma- ceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and / or silicic acid, (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia, (3) humectants, such as glycerol, (4) disintegrants, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, sodium carbonate, (5) solution retardants such as paraffin, (6) absorption accelerators, such as quaternary ammonium compounds, (7) wetting agents, such as acetyl alcohol and glycerol monostearate, (8) adsorbents, such as kaolin and bentonite clay, (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof, and (10) coloring agents. In the case of capsules, tablets, and pills, the compositions can also include buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar or high molecular weight polyethylene glycols or the like.

[0124] Tablets can be produced by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be produced with binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surface active agents or dispersants. Molded tablets can be produced by molding a mixture of the subject composition moistened with an inert liquid diluent in a suitable machine. Tablets, and other solid dosage forms such as dragees, capsules, pills and granules, can be optionally scored or produced with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulation art.

[0125] Compositions for inhalation or pneumoperitoneum include pharmaceutically acceptable solutions and suspensions in aqueous or organic solvents, or mixtures thereof, and powders.Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.In addition to the subject composition, liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan, cyclodextrin and mixtures thereof.

[0126] Suspending agents may contain, in addition to the subject compositions, suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0127] Formulations for rectal or vaginal administration can be presented as suppositories, which can be prepared by mixing the subject composition with one or more suitable non-irritating excipients or carriers consisting of, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, which are solid at room temperature but liquid at body temperature and therefore will melt in the body cavity and release the active agent.

[0128] Dosage forms for transdermal administration of the subject compositions include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active component may be mixed under sterile conditions with a pharma- ceutically acceptable carrier, and any required preservatives, buffers, or propellants.

[0129] The ointments, pastes, creams and gels may contain, in addition to the subject composition, excipients such as animal or vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc and zinc oxide, or mixtures thereof.

[0130] Powders and sprays can contain, in addition to the subject composition, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0131] The compositions and compounds of the present disclosure can alternatively be administered by aerosol. This is accomplished by producing an aqueous aerosol, liposomal formulation or solid particles containing the compound. A non-aqueous (e.g., fluorocarbon propellant) suspension can be used. Sonic nebulizers can be used to minimize exposure of the drug to shear that may result in degradation of the compounds contained in the subject composition. Typically, aqueous aerosols are produced by formulating an aqueous solution or suspension of the subject composition with conventional pharma- ceutically acceptable carriers and stabilizers. Carriers and stabilizers vary according to the requirements of the particular subject composition, but typically include non-ionic surfactants (Tweens, Pluronics, or polyethylene glycols), non-toxic proteins such as serum albumin, sorbitan esters, oleic acid, lecithin, amino acids such as glycine, buffers, salts, sugars or sugar alcohols. Aerosols are generally produced from isotonic solutions.

[0132] Pharmaceutical compositions of the present disclosure suitable for parenteral administration consist of the compositions of the present application in combination with one or more pharma- ceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions immediately prior to use; these compositions may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0133] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present disclosure include water, ethanol, polyol (glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate and cyclodextrin.Proper fluidity can be maintained, for example, by using coating materials such as lecithin, by maintaining the required particle size in the case of dispersion, and by using surfactants.

[0134] In another aspect, the present disclosure provides an enteral pharmaceutical formulation comprising the disclosed compound and an enteric material, and a pharma- ceutically acceptable carrier or excipient.Enteric material refers to a polymer that is substantially insoluble in the acidic environment of the stomach, and dissolves primarily in intestinal fluids at a certain pH.The small intestine is the part of the digestive tract (intestine) between the stomach and the large intestine, and includes the duodenum, jejunum, and ileum.The pH of the duodenum is about 5.5, the pH of the jejunum is about 6.5, and the pH of the distal ileum is about 7.5. Thus, the enteric material will not dissolve until, for example, the pH is about 5.0, until the pH is about 5.2, until the pH is about 5.4, until the pH is about 5.6, until the pH is about 5.8, until the pH is about 6.0, until the pH is about 6.2, until the pH is about 6.4, until the pH is about 6.6, until the pH is about 6.8, until the pH is about 7.0, until the pH is about 7.2, until the pH is about 7.4, until the pH is about 7.6, until the pH is about 7.8, until the pH is about 8.0, until the pH is about 8.2, until the pH is about 8.4, until the pH is about 8.6, until the pH is about 8.8, until the pH is about 9.0, until the pH is about 9.2, until the pH is about 9.4, until the pH is about 9.6, until the pH is about 9.8, until the pH is about 10.0.Exemplary enteric materials include cellulose acetate phthalate (CAP), hydroxypropyl methylcellulose phthalate (HPMCP), polyvinyl acetate phthalate (PVAP), hydroxypropyl methylcellulose acetate succinate (HPMCAS), cellulose acetate trimellitate, hydroxypropyl methylcellulose succinate, cellulose acetate succinate, cellulose acetate hexahydrophthalate, cellulose propionate phthalate, cellulose acetate maleate, cellulose acetate butyrate, cellulose acetate propionate, copolymers of methyl methacrylic acid and methyl methacrylate, copolymers of methyl acrylate, methyl methacrylate and methacrylic acid, copolymers of methyl vinyl ether and maleic anhydride (Gantrez). ES series), natural resins such as ethyl methacrylate-methyl methacrylate-chlorotrimethylammonium ethyl acrylate copolymers, zein, shellac, copalcohol, and some commercially available enteric dispersions (e.g., Eudragit L30D55, Eudragit FS30D, Eudragit L100, Eudragit S100, Kollicoat EMM30D, Estacryl 30D, Coateric, Aquateric). The solubility of each of the above materials is known or can be readily determined in vivo. While the above is a list of possible materials, one of ordinary skill in the art having the benefit of this disclosure will recognize that it is not comprehensive and that there are other enteric materials that will meet the objectives of the present invention.

[0135] Advantageously, the disclosure also provides kits for use by consumers in need of, for example, treatment of cancer. Such kits include suitable dosage forms, such as those described herein, and instructions describing how to use such dosage forms to mediate, reduce or prevent inflammation. The instructions would instruct the consumer or medical personnel to administer the dosage forms according to modes of administration known to those skilled in the art. Such kits can advantageously be packaged and sold in single or multiple kit units. An example of such a kit is a so-called blister pack. Blister packs are well known in the packaging industry and are widely used for packaging pharmaceutical unit dosage forms (tablets, capsules, etc.). Blister packs generally consist of a sheet of relatively stiff material covered with a foil, preferably of a transparent plastic material. During the packaging process, recesses are formed in the plastic foil. The recesses have the size and shape of the tablets or capsules to be packed. The tablets or capsules are then placed in the recesses and the sheet of relatively stiff material is sealed to the plastic foil with the face of the foil opposite to the direction in which the recesses were formed. As a result, the tablets or capsules are enclosed in the recesses between the plastic foil and the sheet. Preferably, the strength of the sheet is such that the tablets or capsules can be removed from the blister pack by manually applying pressure to the recesses, forming openings in the sheet at the locations of the recesses, and the tablets or capsules can then be removed through said openings.

[0136] It may be desirable to provide a memory aid on the kit, for example in the form of a number next to the tablet or capsule, which number corresponds to the day in the regimen on which the tablet or capsule so designated should be taken. Another example of such a memory aid is a calendar printed on a card, for example: "Week 1, Mon, Tues...etc., Week 2, Mon, Tues...etc." Other variations on memory aids will be readily apparent. A "daily dose" may be one tablet or capsule, or it may be several tablets or capsules to be taken on that day. Also, the daily dose of a first compound may be one tablet or capsule, while the daily dose of a second compound may be several tablets or capsules, or vice versa. The memory aid should reflect this.

[0137] Also contemplated herein are methods and compositions that include a second active agent or methods of administering a second active agent. Contemplated herein are the disclosed compounds in combination with at least one other agent previously shown to treat cancer. EXAMPLES

[0138] The compounds described herein can be prepared in many ways based on the teachings contained herein and the synthetic procedures known in the art. In the description of the synthetic methods described below, it is understood that all reaction conditions proposed, including the selection of solvents, reaction atmosphere, reaction temperature, experimental time, and work-up procedures, can be selected to be standard conditions for the reaction, unless otherwise indicated. It is understood by those skilled in the art of organic synthesis that the functionality present on various parts of the molecule should be compatible with the proposed reagents and reactions. Substituents that are not compatible with the reaction conditions will be clear to those skilled in the art, and therefore alternative methods are shown. The starting materials of the examples are commercially available or can be easily prepared from known materials by standard methods. At least some of the compounds identified herein as "intermediates" are intended as compounds of the present disclosure.

[0139] All reactions were carried out under argon atmosphere in heat gun dried glassware using standard septa techniques unless otherwise noted. All commercially available starting building blocks were purchased from commercial vendors. Reactions were monitored by HPLC-MS analysis using a Shimadzu UFLC-MS-2020 system with ESI and / or thin layer chromatography (TLC) using silica gel 60 F254 plates (Merck) and visualized with UV at 254 nm. Purification was performed using an automated flash chromatography system (ECOM) with prepacked columns containing C18 or modified C18 silica gel (Interchim, PT-15C18AQ, 15 μm Puriflash 200, 5g, 12g, or 25g). Preparative HPLC was performed using an ECOM HPLC system with a modified C18 semi-preparative column (YMC-Actus, Triart Prep C18, 250x20 mm, S-10 μm, 12 nm). HPLC-MS analysis was performed using a Shimadzu UFLC-MS-2020 system with ESI. Column Acquity UPLC BEH C18 1.7 μm, 2.1 x 50 mm. Solvent A: H2O 0.1 % HCOOH; Solvent B: MeCN + 0.1 % HCOOH. Total flow rate 0.6 ml / min. Total method time 10 min. UV-Vis spectra were recorded in the range 200-800 nm using a Shimadzu SPD-M2OA Prominence diode array detector. NMR spectra were recorded on a 400 MHz Bruker AVANCE III spectrometer (1H at 400 MHz) and / or a Bruker AVANCE 500 (1H at 500.0 MHz) using ≥99% deuterated solvents. Chemical shifts (ppm, δ scale) are the solvent signals of the 1H spectra. Intermediates and final products were lyophilized from water or mixtures of dioxane or acetonitrile in water using a Gregory Instruments Freeze Dryer (Model L4-110). Abbreviation JPEG2024518438000109.jpg183170

[0140] Example 1: Synthesis of Compound 2 Step 1 JPEG2024518438000110.jpg41170 Intermediate 1 A mixture of ethanolamine (23 mg, 0.4142 mmol) and dimethoxysquarate (3 equiv., 177 mg, 1.242 mmol) was suspended in 10 mL of 1 M borate buffer (pH = 9) and the mixture was stirred at 55 °C for 16 h. 2 mL of DMF was added and the solvent was evaporated under reduced pressure to a final volume of about 3 mL. The crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 and a gradient of ACN in water (0 → 50% ACN in water). After lyophilization from water, the desired product was recovered as a white powder (46 mg, 65%). C7H 10 NO4 MS calculated: 172.06, Found: 172.25, [M+H] +

[0141] Compound 2 Exatecan mesylate (20 mg, 0.0377 mmol) and previously synthesized intermediate 1 (1.5 equiv., 9.7 mg, 0.0564) were suspended in 5 mL of 1 M borate buffer (pH=9), and the mixture was stirred at 55° C. for 16 h. 2 mL of DMF was added and the solvent was evaporated under reduced pressure to a final volume of about 3 mL. The crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 and a gradient of ACN in water (0→50% ACN in water). After lyophilization from water, the desired product was recovered as a white powder (12 mg, 57%). C 30 H 28 MS calculated for FN4O7: 575.19, found: 575.45, [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 8.45 (s, 1H), 7.83 (d, J = 10.9 Hz, 1H), 7.78 (s, 1H), 7.32 (s, 1H), 5.79 (s, 1H), 5.42 (s, 2H), 5.29 (d, J= 7.5 Hz, 2H), 3.68 - 3.43 (m, 4H), 3.23 (d, J = 7.7 Hz, 2H), 2.42 (d, J= 1.9 Hz, 3H), 2.33 (td, J = 5.7, 4.8, 2.9 Hz, 1H), 1.96 - 1.78 (m, 2H), 1.76 (s, 1H), 1.26 - 1.15 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0142] Example 2: Synthesis of Compound 1001 JPEG2024518438000111.jpg146168Intermediate 1. Ethanolamine (100 mg, 1.637 mmol) and dimethoxysquarate (1.2 equiv., 1.964 mmol, 279 mg) were dissolved in 10 mL of 1 M borate buffer (pH 9). The reaction mixture was stirred at room temperature for 16 h. The solvent was evaporated under reduced pressure and the resulting solid was redissolved in DMF and loaded directly onto the column. The product was purified by reversed-phase flash chromatography using a column containing 40 g of C18 and a gradient of ACN in water (0 -> 20% ACN in water). The desired product was recovered as a white solid after lyophilization from water (205 mg, 73%). C7H 10 MS calculated for NO4: 172.06, Found: 172.17, [M + H] + .

[0143] Intermediate 2. Intermediate 1 (10 mg, 0.058 mmol) and the starting peptide FmocGGFG-OAc (1 equiv, 0.058 mmol, 37 mg) were dissolved in 2 mL of anhydrous DMF under argon atmosphere and 100 μL of HCl (2M in Et2O) was added. The reaction mixture was stirred at room temperature for 1 h and loaded directly onto the column. The product was purified by reversed-phase flash HPLC using a semi-preparative column containing 25 g of diol-modified C18 and a gradient of ACN in water (0→80% ACN in water). The desired product was recovered as a white solid after lyophilization from water (25 mg, 58%). C 38 H 40 N6NaO 10 MS calculated: 763.27, found: 763.80, [M + Na] + .

[0144] Intermediate 3. Intermediate 2 (25 mg, 0.0338 mmol) and exatecan mesylate (1.5 equiv, 0.508 mmol, 27 mg) were suspended in 4 mL of 1 M borate buffer (pH 9) and the reaction mixture was stirred at 55 °C for 4 h. DMF (2 mL) was added and the solvent was evaporated under reduced pressure to a final volume of about 2 mL. The product was purified by reversed-phase flash HPLC using a semi-preparative column containing a diol-modified C18 with a gradient of ACN in water (0 -> 100% ACN in water). The desired product was recovered as a white solid after lyophilization from water-dioxane (11 mg, 28%). C 61 H 59 FN9O 13 MS calculated: 1144.42, Found: 1144.01, [M + H] + .

[0145] Intermediate 4. Intermediate 3 (11 mg, 0.0096 mmol) was dissolved in 1 mL of DMF and morpholine (20 μL) was added. The reaction mixture was stirred at room temperature for 30 min. The mixture was filtered through a 0.2 μm syringe filter and loaded directly onto the column. The product was purified by reversed-phase flash HPLC using a semi-preparative column containing a diol-modified C18 with a gradient of ACN in water (0→100% ACN in water). The desired product was recovered as a yellowish solid after lyophilization from water-dioxane (7.5 mg, 88%). C 46 H 50 FN9O 11 MS calculated: 923.36, Found: 923.75, [M + H] + .

[0146] Compound 1001 Intermediate 4 (7.5 mg, 0.0081 mmol) was dissolved in 1 mL of DMF. 2,5-Dioxopyrrolidin-1-yl 3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)propanoate (2 eq., 0.0163 mmol, 5 mg) and DIPEA (20 μL) were added. The reaction mixture was stirred at room temperature for 30 min. The mixture was filtered through a 0.2 μm syringe filter and loaded directly onto the column. The product was purified by reversed-phase flash HPLC using a semi-preparative column containing a diol-modified C18 with a gradient of ACN in water (ACN 0→80% in water). The desired product was recovered as a white solid after lyophilization from water (7 mg, 77%). C 55 H 58 FN 10 O 15 MS calculated: 1117.41, Found: 1117.44, [M + H] + . 1H NMR (500 MHz, DMSO-d6) δ 8.80 (s, 1H), 8.62 (d, J = 10.9 Hz, 1H), 8.52 (s, 1H), 8.27 (s, 1H), 8.15 (d, J = 33.5 Hz, 1H), 8.02 (d, J= 16.0 Hz, 1H), 7.82 (d, J = 11.5 Hz, 1H), 7.67 (s, 1H), 7.36 - 7.20 (m, 4H), 7.19 - 7.12 (m, 2H), 6.68 (s, 1H), 6.05 (t, J = 20.7 Hz, 1H), 5.86 (s, 1H), 5.75 (s, 1H), 5.30 (d, J = 23.9 Hz, 1H), 5.18 (m, 2H), 4.85 (d, J = 11.1 Hz, 2H), 4.66 - 4.54 (m, 1H), 4.54 (s, 2H), 4.44 (m, 1H), 3.85 - 3.79 (m, 1H), 3.70 - 3.65 (m, 3H), 3.56 - 3.47 (m, 2H), 3.47 - 3.42 (m, 1H), 3.41 - 3.36 (m, 1H), 3.17 (s, 1H), 3.07 - 2.97 (m, 2H), 2.85 - 2.69 (m, 4H), 2.67 - 2.52 (m, 2H), 2.44 - 2.36 (m, 3H), 2.33 - 2.23 (m, 1H), 1.97 (m, 2H), 1.82 (d, J = 7.9 Hz, 2H), 1.23 (s, 2H), 0.87 - 0.81 (m, 3H).

[0147] Example 3: Synthesis of Compound 12 JPEG2024518438000112.jpg78158Intermediate 1 Exatecan mesylate (39 mg, 0.0737 mmol), malonic acid (5 equiv, 0.3687 mmol, 38 mg) and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM, 5 equiv, 0.3687 mmol, 102 mg) were dissolved in a 5:1 mixture of DMF and water (6 mL). Triethylamine (50 equiv, 3.6873 mmol, 514 μL) was added and the reaction mixture was stirred at room temperature for 3 h. The solvent was evaporated under reduced pressure and the crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in 1% TFA (0 → 40% ACN in 1% TFA). The desired product was recovered as a white powder after lyophilization from water (34 mg, 88%). 27 H 23 MS calculated for FN3O7: 520.15, Found: 520.49 [MH] - .

[0148] Intermediate 2: Intermediate 1 (24 mg, 0.0461 mmol), 2-((tert-butyldimethylsilyl)oxy)ethan-1-amine (5 equiv., 0.2303 mmol, 48 μL) and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM, 5 equiv., 0.2303 mmol, 64 mg) were dissolved in a 5:1 mixture of DMF and water (6 mL). Triethylamine (50 equiv., 2.303 mmol, 321 μL) was added and the reaction mixture was stirred at room temperature for 3 h. The solvent was evaporated under reduced pressure and the crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in water (0 to 70% ACN in water). The desired product was recovered as a yellowish foam after lyophilization from water-DMF (7 mg, 22%). 35 H 44 MS calculated for FN4O7S: 679.30, Found: 679.00, [M+H] + .

[0149] Compound 12. The previously synthesized intermediate 2 (7 mg, 0.0103 mmol) was suspended in 1% TFA (2 mL) and the mixture was stirred at room temperature for 1 h. The crude reaction mixture was loaded directly onto a column and purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in 1% TFA (0 to 40% ACN in 1% TFA). The desired product was recovered as a yellowish powder after lyophilization from water (3 mg, 53%). 29 H 30 MS calculated for FN4O7: 565.21, found: 565.70, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.62 (d, J= 8.6 Hz, 1H), 8.05 (t, J = 5.6 Hz, 1H), 7.80 (d, J = 10.9 Hz, 1H), 7.31 (s, 1H), 5.58 - 5.54 (m, 1H), 5.43 (s, 2H), 5.26 (d, J = 5.2 Hz, 2H), 3.38 (t, J = 6.3 Hz, 2H), 3.18 (s, 2H), 3.15 - 3.04 (m, 2H), 2.58 - 2.52 (m, 2H), 2.46 (m, 3H), 2.25 - 2.16 (m, 1H), 2.11 (s, 1H), 1.87 (m, 2H), 1.76 (s, 2H), 0.88 (t, J = 7.4 Hz, 3H).

[0150] Example 4: Synthesis of Compound 1005 JPEG2024518438000113.jpg126170Intermediate 1 FmocGGFG-N3 (23 mg, 0.0350 mmol) was dissolved in 2 mL of dioxane. Pd / C (10% w / w, 5 mg) was suspended in the mixture and H2 was bubbled through the suspension using a balloon while stirring at room temperature for 2 h. The suspension was taken up with a syringe, filtered through a 0.2 μm syringe filter and directly placed in a flask containing a previously prepared solution of malonic acid (5 equiv., 0.1750 mmol, 18 mg), DMTMM (5 equiv., 0,1750 mmol, 48 mg) and DIPEA (100 μL) in ACN (2 mL) and water (0.5 mL). The reaction mixture was stirred at room temperature for 2 h. The solvent was evaporated under reduced pressure and the resulting solid was redissolved in DMF and loaded directly onto the column. The product was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in water (0 to 40% ACN in water). The desired product was recovered as a white solid after lyophilization from water (15 mg, 60%). 36 H 41 NO 10 MS calculated value: 715.27, Measured value: 715.55 [M - H] - .

[0151] Intermediate 2 Intermediate 1 (14 mg, 0.0195 mmol) was dissolved in a mixture of DMF (2 mL) and water (0.5 mL). Exatecan mesylate (1.5 equiv, 0.0293 mmol, 16 mg), DMTMM (3 equiv, 0.0587 mmol, 16 mg) and DIPEA (20 μL) were added to the solution and the reaction mixture was stirred at room temperature for 4 h. The solvent was evaporated under reduced pressure and the resulting solid was redissolved in DMF and loaded directly onto the column. The product was purified by reversed-phase flash HPLC using a semi-preparative column containing a diol-modified C18 with a gradient of ACN in water (0→100% ACN in water). The desired product was recovered as a yellow solid after lyophilization from water. (15 mg, 66%). C 60 H 61 FN9O 13 MS calculated: 1134.44, Found: 1134.40 [M + H]+ .

[0152] Intermediate 3 Intermediate 2 (15 mg, 0.0132 mmol) was dissolved in 1 mL of DMF and morpholine (20 μL) was added. The reaction mixture was stirred at room temperature for 30 min. The mixture was filtered through a 0.2 μm syringe filter and loaded directly onto the column. The product was purified by reversed-phase flash HPLC using a semi-preparative column containing a diol-modified C18 with a gradient of ACN in water (0→100% ACN in water). The desired product was recovered as a yellow solid after lyophilization from water-dioxane (10 mg, 83%). C 45 H 51 FN9O 11 MS calculated: 912.37, Found: 912.91 [M + H] + .

[0153] Compound 1005 Intermediate 3 (10 mg, 0.0110 mmol) was dissolved in 1 mL of DMF. 2,5-Dioxopyrrolidin-1-yl 3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)propanoate (2 eq., 0.0219 mmol, 7 mg) and DIPEA (20 μL) were added. The reaction mixture was stirred at room temperature for 30 min. The mixture was filtered through a 0.2 μm syringe filter and loaded directly onto the column. The product was purified by reversed-phase flash HPLC using a semi-preparative column containing a diol-modified C18 with a gradient of ACN in water (0→80% ACN in water). The desired product was recovered as a yellow solid after lyophilization from water-dioxane (11 mg, 90%). C 54 H 60 FN 10 O 15 MS calculated: 1107.42, Found: 1107.50 [M + H] + . 1H NMR (500 MHz, DMSO-d6) δ 8.67 (m, 1H), 8.61 (m,1H), 8.52 (m, 1H), 8.38 (s, 1H), 8.24 (s, 1H), 8.12 (t, J = 5.6 Hz, 1H), 7.81 (d, J= 11.0 Hz, 1H), 7.67 (s, 1H), 7.28 (m, 4H), 7.25 - 7.24 (m, 2H), 7.212 (s, 1H), 7.15 (m, 1H), 6.68 (s, 1H), 6.09 - 6.00 (m, 1H), 5.58 - 5.47 (m, 1H), 5.19 - 5.08 (m, 2H), 4.88 (d, J = 11.8 Hz, 1H), 4.65 (d, J = 11.7 Hz, 1H), 4.50 (dd, J = 11.7, 6.6 Hz, 1H), 4.47 (m, 2H), 3.78 - 3.63 (m, 3H), 3.63 - 3.52 (m, 4H), 3.19 (m, 3H), 3.11 - 3.01 (m, 4H), 2.63 (p, J = 1.9 Hz, 5H), 2.43 - 2.36 (m, 6H), 2.24 - 2.14 (m, 1H), 1.75 (m, 2H), 1.23 (s, 2H), 0.85 (dd, J = 7.9, 6.3 Hz, 3H).

[0154] Example 5: Synthesis of Compound 16 JPEG2024518438000114.jpg89159Intermediate 1 Exatecan mesylate (20 mg, 0.0376 mmol), succinic acid (5 equiv, 0.1881 mmol, 22 mg) and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM, 5 equiv, 0.1881 mmol, 53 mg) were dissolved in a 5:1 mixture of DMF and water (4 mL). Triethylamine (200 μL) was added and the reaction mixture was stirred at room temperature for 3 h. The solvent was evaporated under reduced pressure and the crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in 1% TFA (0 → 40% ACN in 1% TFA). The desired product was recovered as a brown powder after lyophilization from water-dioxane (18 mg, 89%). 28 H 25 MS calculation for FN3O7: 534.17, Measured value: 534.80 [MH] - .

[0155] Intermediate 2 Previously synthesized intermediate 1 (15 mg, 0.0424 mmol), 2-((tert-butyldimethylsilyl)oxy)ethan-1-amine (3 equiv., 0.1272 mmol, 22 mg) and azabenzotriazole tetramethyluronium hexafluorophosphate (HATU, 3 equiv., 0.1272 mmol, 48 mg) were dissolved in DMF (1 mL). Diisopropylethylamine (50 μL) was added and the reaction mixture was stirred at room temperature for 1 h. The solvent was evaporated under reduced pressure and the crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in water (0→80% ACN in water). The desired product was recovered as a yellow foam after lyophilization from water-dioxane (22 mg, 74%). C 36 H 46 MS calculated for FN4O7Si: 693.31, Found: 693.55, [M+H] + .

[0156] Compound 16. Intermediate 2 (22 mg, 0.0317 mmol) synthesized above was suspended in 1% TFA (2 mL) and the mixture was stirred at room temperature for 1 h. The crude reaction mixture was loaded directly onto a column and purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in 1% TFA (0 to 40% ACN in 1% TFA). The desired product was recovered as a white powder after lyophilization from water (11 mg, 60%). 30 H 32 MS calculated for FN4O7: 579.23, Found: 579.25, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.46 (d, J = 8.7 Hz, 1H), 7.82 (m, 1H), 7.79 (d, J = 11.0 Hz, 1H), 7.31 (s, 1H), 5.56 (t, J = 4.8 Hz, 1H), 5.42 (s, 2H), 5.30 - 5.10 (m, 2H), 3.47 (m, 2H), 3.35 (t, J = 6.2 Hz, 2H), 3.18 (m, 2H), 3.07 (td, J = 6.1, 4.4 Hz, 2H), 2.56 - 2.52 (m, 2H), 2.42 - 2.32 (m, 5H), 2.12 (q, J = 5.3 Hz, 2H), 1.95 - 1.79 (m, 2H), 0.88 (t, J = 7.3 Hz, 3H).

[0157] Example 6: Synthesis of Compound 18 JPEG2024518438000115.jpg40161Intermediate 1 Tris(hydroxymethyl)aminomethane (25 mg, 0.206 mmol) and dimethoxysquarate (3 equiv., 0.619 mmol, 88 mg,) were suspended in 10 mL of 1 M borate buffer (pH = 9) and the mixture was stirred at 55 °C for 16 h. 2 mL of DMF was added and the solvent was evaporated under reduced pressure to a final volume of about 3 mL. The crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in water (0 → 50% ACN in water). The desired product was recovered as a white powder after lyophilization from water (22 m, 48%). C9H 14 NO6 MS calculated value: 232.08, Found value: 232.19, [M+H] + .

[0158] Compound 18. Exatecan mesylate (20 mg, 0.0377 mmol) and previously synthesized intermediate 1 (1.5 equiv., 0.0564 mmol, 13 mg) were suspended in 5 mL of 1 M borate buffer (pH = 9) and the mixture was stirred at 55 °C for 16 h. 2 mL of DMF was added and the solvent was evaporated under reduced pressure to a final volume of about 3 mL. The crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in water (ACN 0 -> 50% in water). The desired product was recovered as a white powder after lyophilization from water (9 mg, 38%). C 32 H 32 MS calculated for FN4O9: 635.22, found: 635.63, [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 8.31 (d, J = 8.9 Hz, 1H), 7.86 (d, J= 10.9 Hz, 1H), 7.32 (s, 1H), 7.14 (s, 1H), 6.54 (s, 1H), 5.86 (dd, J = 8.8, 4.3 Hz, 1H), 5.42 (s, 1H), 5.39 (d, J = 19.0 Hz, 2H), 5.15 (d, J= 19.0 Hz, 2H), 4.70 (t, J = 5.6 Hz, 3H), 3.61 (d, J = 5.7 Hz, 6H), 3.27 (m, 1H), 2.44 (m, 3H), 2.41 (m, 1H), 2.26 (m, 1H), 1.85 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0159] Example 7: Synthesis of Compound 22 Step 1: JPEG2024518438000116.jpg78151Intermediate 1 2-[[tert-Butyl(dimethyl)silyl]oxy]ethanol (1.14 mmol, 200 mg) was dissolved in anhydrous dichloromethane (4 mL) under an argon atmosphere. The reaction mixture was cooled at 0°C and diisopropylethylamine (1.1 equiv, 1.25 mmol, 218 μL) was added followed by triphosgene (1.2 equiv, 0.45 mmol, 135 mg). The reaction mixture was stirred at 0°C for 2 h and additional diisopropylethylamine (1.2 equiv, 1.25 mmol, 218 μL) was added followed by 2-mercaptopyridine (1.1 equiv, 1.25 mmol, 155 mg). After stirring at 0°C for an additional 2 h, the reaction mixture was diluted with 20 mL of dichloromethane and saturated NH4Cl (10 mL) was added. The organic phase was washed with saturated NH4Cl (2 x 100 mL) and brine (100 mL). Silica gel flash chromatography using a gradient of EtOAc in cyclohexane (0 -> 50% EtOAc in cyclohexane) afforded the desired product (100 mg, 28%). 14 H 24NO3SSi MS calculated: 314.12, Found: 314.10, [M+H] + .

[0160] Compound 22 Exatecan mesylate (0.0376 mmol, 20 mg) and triethylamine (2 equiv., 0.0752 mmol, 11 μL) were dissolved in 2 mL of anhydrous DMF under argon. Intermediate 1 (1.5 equiv., 0.0752 mmol, 18 mg) was added and the reaction mixture was stirred at room temperature for 48 h. The solvent was evaporated under reduced pressure and the crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in water (0→40% ACN in water). The fractions containing the desired reaction product (Intermediate 2) were evaporated and the solid was resuspended in 1% TFA and stirred at room temperature for 1 h. The desired product was repurified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 and a gradient of ACN in 1% TFA (0 to 40% ACN in 1% TFA) and recovered as a white powder after lyophilization from water (16 mg, 81% over two steps, calculated with exatecan). 27 H 27 MS calculated for FN3O7: 524.18, Found: 524.20, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 7.97 (d, J= 9.0 Hz, 1H), 7.77 (d, J = 11.0 Hz, 1H), 7.32 (s, 1H), 5.43 (s, 2H), 5.32 - 5.16 (m, 2H), 4.11 (m, 2H), 3.33 (s, 1H), 3.25 (d, J = 6.0 Hz, 1H), 3.11 (d, J = 18.7 Hz, 1H), 2.68 (m, 1H), 2.55 (m, 2H), 2.39 - 2.29 (m, 4H), 2.24 - 2.12 (m, 2H), 1.87 (m, 2H), 0.88 (t, J = 7.3 Hz, 3H).

[0161] Example 8: Synthesis of compound 1007 JPEG2024518438000117.jpg144170Intermediate 1 Compound 22 (28 mg, 0.0535 mmol) and FmocGGFG-OAc (2 equiv., 0.107 mmol, 67 mg) were dissolved in 1 mL of anhydrous DMF. HCl (100 μL, 2M in Et2O) was added and the reaction mixture was stirred at room temperature for 2 h. The mixture was loaded directly onto the column. The product was purified by reversed-phase flash chromatography using a column containing 25 g of C18 and a gradient of ACN in water (0→70% ACN in water). The desired product was recovered as a white solid after lyophilization from water (23 mg, 39%). C 58 H 58 FN8O 13 MS calculated: 1093.41, Found: 1093.63, [M + H] + .

[0162] Intermediate 2 Intermediate 1 (23 mg, 0.0211 mmol) was dissolved in 1 mL of anhydrous DMF and morpholine (100 μL) was added. The reaction mixture was stirred at room temperature for 1 h and then loaded directly onto the column. The product was purified by reversed-phase flash HPLC using a semi-preparative column containing 25 g of diol-modified C18, using a gradient of ACN in water (0→100% ACN in water). The desired product was recovered as a white solid after lyophilization from water (16 mg, 85%). C 43 H 48 FN8O 11 MS calculated: 871.34, Found: 871.44, [M + H] + .

[0163] Compound 1007 Intermediate 2 (16 mg, 0.0179 mmol) was dissolved in 1 mL of DMF. 2,5-Dioxopyrrolidin-1-yl 3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)propanoate (2 eq., 0.0358 mmol, 11 mg) and DIPEA (20 μL) were added. The reaction mixture was stirred at room temperature for 30 min. The mixture was filtered through a 0.2 μm syringe filter and loaded directly onto the column. The product was purified by reversed-phase flash HPLC using a semi-preparative column containing a diol-modified C18 with a gradient of ACN in water (0→100% ACN in water). The desired product was recovered as a white solid after lyophilization from water (7 mg, 37%). C 52 H 57 FN9O 15 MS calculated: 1066.40, Found: 1065.98, [M + H] + . 1H NMR (500 MHz, DMSO-d6) δ 8.79 (s, 1H), 8.65 (d, J = 6.9 Hz, 1H), 8.58 - 8.49 (m, 1H), 8.45 (m, 1H), 8.37 (s, 1H), 8.31 - 8.22 (m, 1H), 8.15 - 8.08 (m, 2H), 8.07 - 7.96 (m, 1H), 7.91 (s, 1H), 7.79 (t, J = 11.5 Hz, 2H), 7.34 - 7.28 (m, 1H), 7.25 - 7.17 (m, 3H), 7.01 (d, J = 10.9 Hz, 2H), 6.67 (s, 1H), 6.61 - 6.46 (m, 1H), 5.42 (s, 1H), 5.39 - 5.28 (m, 1H), 5.26 - 5.23 (m, 2H), 5.15 (d, J= 7.3 Hz, 1H), 4.57 (m, 2H), 4.49 (d, J = 11.1 Hz, 2H), 4.18 (d, J= 4.8 Hz, 2H), 3.77 - 3.63 (m, 3H), 3.61 (m, 2H), 3.58 - 3.42 (m, 3H), 3.25 - 3.17 (m, 1H), 3.04 (s, 2H), 2.77 (dd, J = 8.5, 5.5 Hz, 1H), 2.47 - 2.29 (m, 4H), 2.20 - 2.11 (m, 1H), 2.00 (m, 1H), 1.86 (m, 2H), 1.23 (s, 2H), 0.97 - 0.82 (m, 3H).

[0164] Example 9: Synthesis of Compound 42 JPEG2024518438000118.jpg77163Intermediate 1 2-((tert-Butyldimethylsilyl)oxy)ethan-1-amine (21 mg, 0.121 mmol), triphosgene (0.95 equiv., 0.0383 mmol, 11 mg) and diisopropylethylamine (5 equiv., 0.605 mmol, 105 μL) were dissolved in dichloromethane (2 mL) under argon atmosphere. The reaction mixture was stirred at room temperature for 1 h. Complete reaction conversion to give intermediate 1 was confirmed by LCMS analysis. The reaction product was used in the next step without further purification.

[0165] Compound 42 Exatecan mesylate (0.0602 mmol, 32 mg) and diisopropylethylamine (2 equiv., 0.120 mmol, 21 μL) were dissolved in 1 mL of anhydrous DMF and the solution was cooled at 0 °C. The previously prepared solution of isocyanate intermediate 1 in dichloromethane (2 mL, 0.1150 mmol) was added at 0 °C and the reaction mixture was allowed to reach room temperature and stirred for 1 h. The solvent was evaporated under reduced pressure and the crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in water (0 → 60% ACN in water). The fractions containing the desired reaction product (intermediate 2) were evaporated and the solid was resuspended in 1% TFA and stirred at room temperature for 1 h. The desired product was repurified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in 1% TFA (0 to 40% ACN in 1% TFA) and recovered as a white powder after lyophilization from water (15 mg, 48% over three steps, calculated from exatecan). 27 H 28 MS calculated for FN4O6: 523.20, Found: 523.25, [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 7.76 (dd, J = 11.0, 2.0 Hz, 1H), 7.31 (s, 1H), 6.82 (d, J = 8.9 Hz, 1H), 6.61 (m, 1H), 5.43 (d, J = 2.6 Hz, 2H), 5.40 - 5.30 (m, 2H), 5.22 (s, 1H), 5.17 (s, 1H), 3.48 - 3.40 (m, 2H), 3.21 - 3.01 (m, 3H), 2.38 (d, J = 1.9 Hz, 3H), 2.23 - 2.06 (m, 2H), 1.96 - 1.80 (m, 2H), 1.76 (s, 2H), 0.90 (t, J = 7.3 Hz, 3H).

[0166] Example 10: Synthesis of Compound 1008 JPEG2024518438000119.jpg166168Intermediate 1 Compound 48 (17 mg, 0.0325 mmol) and FmocGGFG-OAc (3 equiv., 0.0976 mmol, 61 mg) were dissolved in 1 mL of anhydrous DMF. HCl (100 μL, 2M in Et2O) was added and the reaction mixture was stirred at room temperature for 2 h. The product was purified by reversed-phase flash chromatography using a column containing 25 g of C18 and a gradient of ACN in water (0→80% ACN in water). The desired product was recovered as a white solid after lyophilization from water (15 mg, 44%). C 58 H 59 FN9O 12 MS calculated: 1092.43, Found: 1093.03, [M + H] + .

[0167] Intermediate 2 Intermediate 1 (15 mg, 0.0135 mmol) was dissolved in 1 mL of anhydrous DMF and morpholine (100 μL) was added. The reaction mixture was stirred at room temperature for 1 h and then loaded directly onto the column. The product was purified by reversed-phase flash HPLC using a semi-preparative column containing 25 g of diol-modified C18, using a gradient of ACN in water (0→100% ACN in water). The desired product was recovered as a white solid after lyophilization from water (10 mg, 87%). C 43 H 49 FN9O 10 MS calculated: 870.36, Found: 870.88, [M + H] + .

[0168] Compound 1008. Intermediate 2 (10 mg, 0.0118 mmol) was dissolved in 1 mL of DMF. 2,5-dioxopyrrolidin-1-yl 3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)propanoate (2 eq., 0.0237 mmol, 7 mg) and DIPEA (20 μL) were added. The reaction mixture was stirred at room temperature for 30 min. The mixture was filtered through a 0.2 μm syringe filter and loaded directly onto the column. The product was purified by reversed-phase flash HPLC using a semi-preparative column containing a diol-modified C18 with a gradient of ACN in water (0→100% ACN in water). The desired product was recovered as a white solid after lyophilization from water (4 mg, 32%). C 52 H 58 FN 10 O 14 MS calculated: 1065.41, Found: 1065.79, [M + H] + .

[0169] Example 11: Synthesis of Compound 48 JPEG2024518438000120.jpg67163Intermediate 1 Benzyl (1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl)carbamate (100 mg, 0.392 mmol), tert-butyldimethylsilyl chloride (1.5 equiv, 1.764 mmol, 266 mg) and imidazole (1.5 equiv, 1.764 mmol, 120 mg) were dissolved in 5 mL of anhydrous DMF. The reaction mixture was stirred overnight at room temperature. The solvent was evaporated and purified by flash chromatography on silica (0→50% EtOAc in hexanes) to give intermediate 1 (188 mg, 80%). C 30 H 60 MS calculated for NO5Si3: 598.38, Found: 598.25, [M+H] + .

[0170] Intermediate 2: The previously prepared intermediate 1 (188 mg, 0.314 mmol) was dissolved in 5 mL of anhydrous dioxane. The solvent was evaporated and purified by flash chromatography on silica (0→50% EtOAc in hexanes) to give the desired intermediate 2 (101 mg, 69%). 22 H 54 MS calculated for NO3Si3: 464.34, Found: 463.98, [M+H] + .

[0171] Compound 48. The previously prepared intermediate 2 (56 mg, 0.121 mmol), triphosgene (1 equiv, 11.4 mg), and diisopropylethylamine (100 μL) were dissolved in 2 mL of anhydrous dichloromethane. The reaction mixture was stirred at room temperature for 1 h and then added to a premixed solution of exatecan (30 mg, 0.0564 mmol) and diisopropylethylamine (30 μL) in anhydrous DMF. The reaction mixture was stirred at room temperature for 24 h. The solvent was evaporated and the crude solid was redissolved in 2 mL of DMF. Water (2 mL) and TFA (1 mL) were added and the mixture was stirred at room temperature for 1 h. The solvent was evaporated under reduced pressure and coevaporated with water three times. The crude solid was redissolved in DMF (2 mL) and purified by reverse-phase flash chromatography on a semi-preparative column (diol-modified C18, 0→70% ACN / 1% TFA in water) to give the product (15 mg, 70%) as a white powder after lyophilization. 29 H 32 MS calculated for FN4O8: 383.22, found: 383.54, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 7.69 (d, J= 10.9 Hz, 1H), 7.30 (s, 1H), 7.13 (d, J = 8.9 Hz, 1H), 5.87 (s, 1H), 5.43 (d, J = 1.6 Hz, 2H), 5.40 - 5.33 (m, 1H), 5.29 (d, J = 19.5 Hz, 1H), 5.12 (d, J = 19.2 Hz, 1H), 3.55 (s, 6H), 3.16 (dt, J = 10.6, 4.3 Hz, 2H), 2.35 (d, J = 1.9 Hz, 3H), 2.26 - 2.15 (m, 1H), 2.14 - 2.03 (m, 1H), 1.86 (hept, J = 7.1 Hz, 2H), 0.88 (t, J = 7.3 Hz, 3H).

[0172] Example 12: Synthesis of Compound 52 JPEG2024518438000121.jpg62170Intermediate 1 Methyl propiolate (1.0 equiv., 55 mg, 0.65 mmol), (2-azidoethoxy)(tert-butyl)dimethylsilane (1.15 equiv., 150 mg, 0.74 mmol) and tris[(1-benzyltriazol-4-yl)methyl]amine (0.15 equiv., 50 mg, 0.094 mmol) were dissolved in DMF (3 ml) and diluted with 1M aqueous CuSO. 4. 5H2O (0.1 equiv., 0.06 mmol, 60 μL) and 2M aqueous sodium ascorbate (0.2 equiv., 0.12 mmol, 60 μL) were added and the resulting mixture was stirred at room temperature for 2 h. DMF was evaporated, the residue was taken up in EtOAc and the organic phase was washed with water, 0.2M aqueous HCl, saturated aqueous NH4Cl and brine and dried over Na2SO4. Purification by flash chromatography (silica, 0%-30% EtOAc / cyclohexane) afforded the triazole intermediate 1 (158 mg, 85%) as a white solid.

[0173] Intermediate 2 To a solution of triazole intermediate 1 (1.0 equiv., 158 mg, 0.55 mmol) in MeOH (2 mL) was added 2M aqueous NaOH (1.0 equiv., 0.55 mL) and the resulting mixture was stirred at room temperature overnight. The solvent was evaporated and the residue was re-evaporated twice with toluene, suspended in EtOAc and filtered. The solid was washed with Et2O and dried in vacuum to give triazole intermediate 2 (135 mg, 84%) as a white solid.

[0174] Intermediate 3. Exatecan mesylate (15 mg, 0.028 mmol), triazole intermediate 2 (2.1 equiv., 0.06 mmol, 17 mg), N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.0 equiv., 0.056 mmol, 11 mg) and 1-hydroxybenzotriazole (2.0 equiv., 0.056 mmol, 8 mg) were dissolved in DMF (1 mL) and diisopropylethylamine (5.0 equiv., 0.14 mmol, 18 mg, 25 μL) was added under argon atmosphere and the mixture was stirred at room temperature for 5 h. LC-MS showed complete consumption of starting material. Purification of the mixture by reversed-phase flash chromatography (25 g, diol-modified C18, ACN 0%–75% in water) afforded triazole intermediate 3 (14 mg, 73%) as a white powder after lyophilization.

[0175] Compound 52. Triazole intermediate 3 (14 mg, 0.02 mmol) was dissolved in ACN / 0.1% aqueous TFA mixture (1:1, 2 ml), then 2 drops of TFA were added and the resulting mixture was stirred at room temperature for 2 h. LC-MS showed complete consumption of intermediate 1 and the solvent was evaporated under reduced pressure. The residue was purified by reversed-phase flash chromatography using a semi-separative column (diol-modified C18, ACN 0→50% in water) to give compound 52 (10 mg, 85%) as a white powder after lyophilization. 29 H 28 MS calculated for FN6O6: 575.20, found: 575.25, [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ: 8.63 (s, 1H), 7.77 (d, J = 10.9 Hz, 1H), 7.29 (s, 1H), 5.73 (dd, J = 7.7, 5.3 Hz, 1H), 5.36 (s, 2H), 5.14 (d, J = 3.7 Hz, 2H), 4.48 (t, J = 5.3 Hz, 2H), 3.81 (t, J = 5.4 Hz, 2H), 3.33 (s, 2H), 3.32 - 3.21 (m, 1H), 3.19 - 3.07 (m, 1H), 2.38 (d, J = 1.9 Hz, 3H), 2.34 - 2.20 (m, 2H), 1.93 - 1.76 (m, 2H), 0.85 (t, J = 7.3 Hz, 3H).

[0176] Example 13: Synthesis of compound 1010 JPEG2024518438000122.jpg183159Intermediate 1 A mixture of compound 52 (1.0 equiv., 30 mg, 0.052 mmol) and FmocGGFG-OAc (2.0 equiv., 0.104 mmol, 66 mg) was dissolved in anhydrous DMF (1.5 mL) followed by the addition of 2M HCl / Et2O (150 μL). The reaction mixture was stirred at room temperature for 4 h during which FmocGGFG-OAc was added in portions (approximately 0.5 equiv. each) to the reaction mixture. The reaction mixture was then purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0% to 75% ACN / H2O). The product-containing fractions were lyophilized and the residue (product + coeluting impurities) was used directly in the next step. C 60 H 59 FN 11 O 12 MS calculated: 1144.43, Found: 1144.40, [M+H] + .

[0177] Intermediate 2 Morpholine (140 μL) was added to a solution of intermediate 1 (obtained in the previous step) in anhydrous DMF (2 ml) and the reaction mixture was stirred at room temperature for 1 h. LC-MS showed complete consumption of the starting material. The mixture was directly purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0% to 60% ACN / H2O) to give the product as a white solid after lyophilization (20 mg, 42%, 2 steps). 45 H 49 FN 11 O 10 MS calculated: 922.36, Found: 922.35, [M+H] + .

[0178] Compound 1010 To a solution of intermediate 2 (1.0 equiv., 20 mg, 0.022 mmol) in anhydrous DMF (1 ml) was added Mal-PEG-NHS ester (1.0 equiv., 0.022 mmol, 6.7 mg) and DIPEA (2.4 equiv., 0.053 mmol, 7 mg, 9 μL). The reaction mixture was stirred at room temperature for 40 min as LC-MS showed complete consumption of starting material. Purification by reversed-phase flash HPLC using a semi-separating column (diol-modified C18, 0% to 60% ACN / H2O) afforded the desired product as a white solid after lyophilization (7.5 mg, 31%). C 54 H 58 FN 12 O 14 MS calculated: 1117.42, Found: 1117.35, [M + H] + . 1H NMR (500 MHz, DMSO-d6) δ: 9.28 (t, J= 9.8 Hz, 1H), 8.65 (d, J = 2.1 Hz, 1H), 8.56 (t, J = 6.7 Hz, 1H), 8.32 (t, J = 5.9 Hz, 1H), 8.15 - 8.08 (m, 2H), 8.00 (t, J = 5.8 Hz, 1H), 7.80 (d, J = 10.8 Hz, 1H), 7.31 (d, J = 3.4 Hz, 1H), 7.28 - 7.21 (m, 5H), 7.20 - 7.15 (m, 1H), 7.00 (s, 1H), 6.51 (s, 1H), 5.78 - 5.71 (m, 1H), 5.41 - 5.33 (m, 2H), 5.25 - 5.11 (m, 2H), 4.63 - 4.60 (m, 2H), 4.58 (d, J = 6.7 Hz, 2H), 4.53 - 4.47 (m, 1H), 3.86 - 3.83 (m, 2H), 3.74 (td, J = 17.3, 5.8 Hz, 2H), 3.67 (d, J = 5.7 Hz, 2H), 3.64 - 3.50 (m, 5H), 3.46 (t, J = 5.7 Hz, 2H), 3.18 - 3.11 (m, 1H), 3.05 (dd, J= 14.0, 4.6 Hz, 1H), 2.84 - 2.75 (m, 1H), 2.40 (s, 3H), 2.33 (t, J = 6.5 Hz, 2H), 2.30 - 2.23 (m, 3H), 1.93 - 1.78 (m, 2H), 0.90 - 0.83 (m, 3H).

[0179] Example 14: Synthesis of Compound 58 JPEG2024518438000123.jpg431544:1 DMF / water mixture (4 mL) was added with exatecan mesylate (20 mg, 0.038 mmol), trans-3-hydroxycyclobutane-1-carboxylic acid (1.25 equiv., 6 mg, 0.048 mmol), DMTMM (2.0 equiv., 21 mg, 0.076 mmol) and diisopropylethylamine (20 μL). The resulting solution was stirred at room temperature for 1 h, when LC-MS showed complete consumption of starting material. The mixture was directly purified by reversed-phase HPLC chromatography using a semi-separated column (diol-modified C18, 0 to 100% ACN / H2O). After lyophilization, the desired product was obtained as a white powder (13 mg, 64%). C 29 H 29 MS calculated for FN3O6: 534.20, Found: 534.21, [M+H] + . 1 H NMR (401 MHz, DMSO-d6) δ 8.38 (d, J= 8.7 Hz, 1H), 7.76 (d, J = 11.0 Hz, 1H), 7.29 (s, 1H), 6.52 (s, 1H), 5.56 (m, 1H), 5.42 (s, 2H), 5.11 (d, J = 6.5 Hz, 2H), 5.07 (d, J= 6.2 Hz, 1H), 4.37 (m, 1H), 3.15 (m, 2H), 2.91 (m, 1H), 2.50 - 2.40 (m, 4H), 2.40 (s, 3H), 2.17 - 2.08 (m, 1H), 2.05 - 1.99 (m, 1H), 1.86 (p, J = 7.0 Hz, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0180] Example 15: Synthesis of Compound 66 JPEG2024518438000124.jpg65156Intermediate 1 To a solution of 3-butynoic acid (1.0 equiv., 17 mg, 0.2 mmol), (2-azidoethoxy)(tert-butyl)dimethylsilane (1.25 equiv., 50 mg, 0.25 mmol) and tris[(1-benzyltriazol-4-yl)methyl]amine (0.15 equiv., 16 mg, 0.03 mmol) in DMF (1 ml) was added triethylamine (1.0 equiv., 0.2 mmol, 20 mg, 28 μL), 1M aqueous CuSO 4.5 HO (0.1 equiv, 0.02 mmol, 20 μL) and 2 M aqueous sodium ascorbate (0.2 equiv, 0.04 mmol, 20 μL) were added and the resulting mixture was stirred at room temperature overnight. The mixture was purified by reverse-phase flash chromatography (25 g, diol-modified C18, 0% to 75% ACN / HO) to give the triazole intermediate 1 (47 mg, 61%).

[0181] Triazole intermediate 2. Exatecan mesylate (10 mg, 0.019 mmol), triazole intermediate 1 (2.0 equiv, 0.038 mmol, 15 mg), N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.0 equiv, 0.038 mmol, 8 mg) and 1-hydroxybenzotriazole (2.0 equiv, 0.038 mmol, 5.5 mg) were dissolved in DMF (0.5 mL) and diisopropylethylamine (5.0 equiv, 0.095 mmol, 12 mg, 17 μL) was added under argon atmosphere and the mixture was stirred at room temperature for 5 h. LC-MS showed complete consumption of starting material. Purification of the mixture by reverse-phase flash chromatography (12 g, diol-modified C18, 0% to 75% ACN / H2O) afforded the triazole intermediate 2 (8 mg, 60%) as a white powder after lyophilization.

[0182] Compound 66. Triazole intermediate 2 (8 mg, 0.011 mmol) was dissolved in ACN / 0.1% aq. TFA mixture (1:1, 1 ml), then 2 drops of TFA were added and the resulting mixture was stirred at room temperature for 2 h. LC-MS showed complete consumption of intermediate 1 and the solvent was evaporated under reduced pressure. The residue was purified by reversed-phase flash chromatography using a semi-separative column (diol-modified C18, 0% to 50% ACN / H2O) to give compound 76 (5 mg, 77%) as a white powder after lyophilization. C 30 H 30 MS calculated for FN6O6: 589.22, found: 589.30, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ: 7.99 (s, 1H), 7.81 (d, J = 11.0 Hz, 1H), 7.31 (s, 1H), 5.56 (q, J = 4.6 Hz, 1H), 5.43 (s, 2H), 5.26 (d, J = 18.9 Hz, 1H), 5.16 (d, J = 18.9 Hz, 1H), 4.41 - 4.34 (m, 2H), 3.80 - 3.73 (m, 2H), 3.61 (s, 2H), 3.34 (s, 2H), 3.24 - 3.13 (m, 2H), 2.41 (s, 3H), 2.24 - 2.07 (m, 2H), 1.95 - 1.78 (m, 2H), 0.88 (t, J = 7.3 Hz, 3H).

[0183] Example 16: Synthesis of Compound 79 JPEG2024518438000125.jpg421593-Hydroxybicyclo[1.1.1]pentane-1-carboxylic acid (5.8 mg, 0.0451 mmol) and exatecan mesylate (0.8 equiv., 20 mg, 0.0376 mmol) were dissolved in 5 mL of 1:4 H2O / DMF mixed solvent containing 38 μL of 1M NaOH solution (0.8 equiv., NaOH). The mixture was stirred at room temperature for 1 h. The solvent was evaporated under reduced pressure to a final volume of about 2 mL. The crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in water (0 → 50% ACN in water). A second purification was performed using a semi-preparative column loaded with diol-modified C18 with a gradient of ACN in water (0 → 80% ACN in water). The desired product was recovered as a white powder after lyophilization from water (14 mg, 68%). 30 H 29 MS calculated for FN3O6: 546.20, Found: 546.22, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.42 (d, J = 8.9 Hz, 1H), 7.72 (d, J= 10.9 Hz, 1H), 7.29 (s, 1H), 6.50 (s, 1H), 6.35 (s, 1H), 5.53 (td, J = 8.9, 4.6 Hz, 1H), 5.42 (s, 2H), 5.19 - 5.08 (m, 1H), 4.96 (d, J = 18.8 Hz, 1H), 3.98 (s, 1H), 3.15 - 3.03 (m, 1H), 2.36 (d, J = 1.8 Hz, 3H), 2.22 - 2.14 (m, 1H), 2.14 - 2.04 (m, 7H), 1.94 - 1.79 (m, J = 7.1 Hz, 2H), 0.88 (t, J = 7.3 Hz, 3H).

[0184] Example 17: Synthesis of Compound 83 JPEG2024518438000126.jpg411644:1To a DMF / water mixture (4 mL) was added exatecan mesylate (20 mg, 0.0376 mmol), 4-hydroxybut-2-inoic acid (2 equiv., 0.0753 mmol, 8 mg), DMTMM (1.5 equiv., 0.0564 mmol, 16 mg) and diisopropylethylamine (20 μL). The resulting solution was stirred at room temperature for 1 h, at which time LC-MS showed complete consumption of the starting material. The mixture was directly purified by reversed-phase HPLC chromatography using a semi-separated column (diol-modified C18, 0→100% ACN / H2O). After lyophilization, the desired product was obtained as a white powder (15 mg, 76 %). C 28 H 25 MS calculated for FN3O6: 518.17, Found: 518.24, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 9.33 (d, J = 8.7 Hz, 1H), 7.78 (d, J = 10.9 Hz, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.58 (dt, J = 8.8, 5.4 Hz, 1H), 5.48 (t, J= 6.0 Hz, 1H), 5.42 (s, 2H), 5.18 (q, J = 18.9 Hz, 2H), 4.23 (d, J= 6.0 Hz, 1H), 3.98 (s, 1H), 3.28 - 3.07 (m, 2H), 2.38 (d, J = 1.8 Hz, 3H), 2.18 (q, J = 6.2 Hz, 2H), 1.97 - 1.77 (m, 2H), 0.88 (t, J = 7.3 Hz, 3H).

[0185] Example 18: Synthesis of Compound 100 JPEG2024518438000127.jpg24167JPEG2024518438000128.jpg45165Intermediate 1 A solution of d-ribose (1.00 g, 6.66 mmol) in pyridine was cooled to 0 °C and tert-butyl(chloro)diphenylsilane (1.2 equiv., 2.20 g, 2.08 mL) was added dropwise. The resulting solution was stirred at 0 °C for 2 h and then at room temperature overnight. Purification by flash chromatography (silica, 0 → 30% EtOAc / cyclohexane) gave intermediate 2 (1.73 g, 67%).

[0186] Intermediate 2 To a solution of intermediate 1 (1.73 g, 4.45 mmol) in DCM (40 mL) was added triethylamine (6.0 equiv, 26.7 mmol, 2.7 g, 3.72 mL) and the resulting solution was cooled to 0 °C. Then, a solution of benzoyl chloride (4.0 equiv, 17.8 mmol, 2.5 g, 2.07 mL) in DCM (10 mL) was added dropwise over 15 min and the reaction mixture was allowed to warm to room temperature and stirred overnight. The resulting mixture was washed with 1 M aqueous hydrochloric acid, saturated aqueous NaHCO3, and brine and the organic phase was dried over Na2SO4. The solvent was removed using a rotary evaporator and the crude product was dissolved in THF (100 ml). To this solution was added TBAF (2.2 equiv, 9.6 mmol, 3.0 g) in one portion and the resulting solution was stirred at room temperature for 2 h. The solvent was evaporated and the residue was partitioned between EtOAc and saturated aqueous NH4Cl. The organic phase was washed with brine and dried over Na2SO4. The crude product was purified by reverse-phase flash chromatography (40 g, diol-modified C18, 0→75% ACN / H2O) to give intermediate 2 (1.27 g, 62%, 2 steps).

[0187] Intermediate 3 To a solution of intermediate 2 (1.27 g, 2.75 mmol) in 30% water / CAN (30 ml) were successively added TEMPO (0.1 equiv., 0.275 mmol, 43 mg) and bis(acetoxy)iodobenzene (2.0 equiv., 5.5 mmol, 1.78 g) and the resulting mixture was stirred at room temperature overnight. The solvent was evaporated and the residue was coevaporated three times with water. It was then taken up in EtOAc and the organic phase was washed with water, brine and dried over Na2SO4. The solvent was removed to give intermediate 3 (1.27 g, 97%).

[0188] Intermediate 4. Exatecan mesylate (30 mg, 0.056 mmol), intermediate 3 (3.0 equiv, 0.168 mmol, 80 mg), N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.5 equiv, 0.14 mmol, 27 mg) and 1-hydroxybenzotriazole (2.5 equiv, 0.14 mmol, 19 mg) were dissolved in DMF (2 mL) and diisopropylethylamine (6.0 equiv, 0.34 mmol, 44 mg, 59 μL) was added under argon atmosphere and the mixture was stirred at room temperature for 1 h. LC-MS showed complete consumption of starting material. The mixture was purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→100% ACN / H2O) to give intermediate 4 (33 mg, 66%) as an off-white powder after lyophilization.

[0189] Compound 100 To a solution of intermediate 4 (33 mg, 0.037 mmol) in methanol (2 ml) was added dry K2CO3 (2.0 equiv., 0.074 mmol, 10 mg) and the resulting mixture was stirred at room temperature for 30 min. LC-MS showed complete consumption of the starting material. The mixture was acidified with 0.1% TFA / water and the solvent was evaporated under reduced pressure. The residue was purified by reversed-phase flash chromatography using a semi-separating column (diol-modified C18, 0→75% ACN / 0.1% TFA in water) to give the product (14 mg, 65%, mixture of stereoisomers) as a white powder after lyophilization. C 29 H 29MS calculated value of FN3O9: 582.19, measured value: 582.20, [M+H] + . 1 1H NMR (400 MHz, DMSO-d6, major isomer) δ: 7.96 (d, J = 9.8 Hz, 1H), 7.79 (d, J = 10.9 Hz, 1H), 7.30 (s, 1H), 6.58 (d, J= 4.6 Hz, 1H), 6.52 (s, 1H), 5.66 - 5.52 (m, 1H), 5.42 (s, 2H), 5.25 (d, J= 6.8 Hz, 1H), 5.23 - 5.18 (m, 1H), 5.09 (d, J = 4.6 Hz, 1H), 5.06 (dd, J= 4.5, 1.7 Hz, 1H), 4.27 - 4.20 (m, 1H), 4.15 (d, J = 5.7 Hz, 1H), 3.69 (td, J = 4.6, 1.7 Hz, 1H), 3.17 (t, J = 6.5 Hz, 2H), 2.38 (s, 3H), 2.20 - 2.11 (m, 2H), 1.95 - 1.77 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0190] Example 19: Synthesis of Compound 103 JPEG2024518438000129.jpg40158Step 1: Compound 103 Exatecan mesylate (10 mg, 0.019 mmol), 5-(hydroxymethyl)furan-2-carboxylic acid (3 eq, 8 mg, 0.057 mmol), N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.5 eq, 10 mg, 0.048 mmol) and 1-hydroxybenzotriazole (2.5 eq, 7 mg, 0.048 mmol) were dissolved in DMF (1 mL), and diisopropylethylamine (5 eq, 17 μL, 0.095 mmol) was added under argon atmosphere and the mixture was stirred at room temperature for 40 min. LC-MS showed complete consumption of starting material. Purification by reversed-phase flash chromatography (25 g, diol-modified C18, 0→70% ACN / H2O) followed by further purification on a semi-preparative column (diol-modified C18, 0→70% ACN / H2O) afforded the product (7 mg, 66%) as a yellowish powder after lyophilization. 30 H 27 MS calculated for FN3O7: 560.18, Found: 560.15, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ: 8.90 (d, J = 8.6 Hz, 1H), 7.79 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 7.17 (d, J= 3.4 Hz, 1H), 6.50 (s, 1H), 6.45 (d, J = 3.4 Hz, 1H), 5.77 - 5.67 (m, 1H), 5.40 - 5.33 (m, 3H), 5.18 (d, J = 18.9 Hz, 1H), 5.10 (d, J = 19.0 Hz, 1H), 4.44 (d, J = 5.7 Hz, 2H), 3.29 - 3.09 (m, 1H), 2.40 (d, J= 1.9 Hz, 3H), 2.24 (q, J = 6.2 Hz, 2H), .94 - 1.75 (m, 2H), 0.86 (t, J= 7.3 Hz, 3H).

[0191] Example 20: Synthesis of Compound 1012 JPEG2024518438000130.jpg198150Intermediate 1. To a solution of FmocGGFGG-OAc (1.0 equiv., 50 mg, 0.079 mmol) and 5-(hydroxymethyl)furan-2-carboxylic acid (1.2 equiv., 14 mg, 0.095 mmol) in anhydrous DMF (0.5 ml) was added 2M HCl / Et2O (70 μL) and the resulting mixture was stirred at room temperature for 2 h. The volatiles were removed and the residue was purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→60% ACN / H2O) to give intermediate 1 as a white solid after lyophilization (23 mg, 41%). C 37 H 36 FN5O 10 MS calculated value: 710.25, Measured value: 710.25, [MH] - .

[0192] Intermediate 2 To a mixture of intermediate 1 (1.05 equiv., 23 mg, 0.032 mmol), exatecan mesylate (1.0 equiv., 16.3 mg, 0.031 mmol) and DMTMM (1.05 equiv., 8.9 mg, 0.032 mmol), DMF / water (5:1, 1.2 ml) and diisopropylethylamine (2.1 equiv., 12 μL, 0.065 mmol) were added and the resulting mixture was stirred at room temperature for 40 min, since LC-MS analysis showed complete consumption of starting material. The reaction mixture was purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→75% ACN / H2O) to give intermediate 2 as a white solid after lyophilization (27 mg, 77 %). C 61 H 58 FN8O 13 MS calculated: 1129.41, Found: 1129.45, [M+H] + .

[0193] Intermediate 3 To a solution of intermediate 2 (1.0 equiv., 27 mg, 0.024 mmol) in anhydrous DMF (1 ml) was added morpholine (50 μL) and the reaction mixture was stirred at room temperature for 1.5 h. LC-MS showed complete consumption of the starting material. The mixture was directly purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→60% ACN / H2O) to give the product as a white solid after lyophilization (15 mg, 69 %). 46 H 48 FN8O 11 MS calculated: 907.34, Found: 907.35, [M+H] + .

[0194] Compound 1012. To a solution of intermediate 3 (1.0 equiv., 15 mg, 0.017 mmol) in anhydrous DMF (1 ml) was added Mal-PEG-NHS ester (1.0 equiv., 0.017 mmol, 5.2 mg) and DIPEA (1.05 equiv., 0.0173 mmol, 3.05 μL). The reaction mixture was stirred at room temperature for 1.5 h as LC-MS showed complete consumption of starting material. Purification by reversed-phase flash HPLC using a semi-separating column (diol-modified C18, 0→60% ACN / H2O) afforded the desired product as a white solid after lyophilization (11 mg, 60%). C 55 H 57 FN9O 15 MS calculated: 1102.40, Found: 1102.45, [M+H] + . 1H NMR (500 MHz, DMSO-d6) δ: 9.06 - 8.93 (m, 1H), 8.59 (t, J = 6.4 Hz, 1H), 8.31 (t, J = 5.9 Hz, 1H), 8.16 - 8.05 (m, 2H), 8.04 - 7.94 (m, 1H), 7.80 (d, J = 10.8 Hz, 1H), 7.30 (s, 1H), 7.27 - 7.11 (m, 7H), 6.99 (s, 1H), 6.59 (d, J = 3.4 Hz, 1H), 6.51 (s, 1H), 5.78 - 5.63 (m, 1H), 5.38 (s, 2H), 5.28 - 4.97 (m, 2H), 4.59 (d, J= 7.0 Hz, 2H), 4.53 - 4.37 (m, 3H), 3.84 - 3.63 (m, 5H), 3.63 - 3.48 (m, 5H), 3.45 (t, J = 5.8 Hz, 2H), 3.29 - 3.22 (m, 1H), 3.20 - 3.08 (m, 1H), 3.04 (dd, J = 13.9, 4.6 Hz, 1H), 2.79 (dd, J = 13.9, 9.6 Hz, 1H), 2.40 (s, 3H), 2.32 (t, J = 6.6 Hz, 2H), 2.27 - 2.19 (m, 2H), 1.96 - 1.77 (m, 2H), 0.86 (t, J = 7.5 Hz, 3H).

[0195] Example 21: Synthesis of Compound 105 JPEG2024518438000131.jpg34157 Compound 105 To a suspension of exatecan mesylate (30 mg, 0.056 mmol) in DMF (1 mL) under argon atmosphere, diisopropylethylamine (3.5 eq, 0.196 mmol, 34 μL) and 2-bromoethanol (2 eq, 0.112 mmol, 14 mg, 8 μL) were added and the mixture was heated to 80 °C for 2 days. LC-MS showed complete consumption of starting material. Purification by reversed-phase flash chromatography on a semi-preparative column (diol-modified C18, 0 → 50% ACN / 1% TFA in water) afforded the product (16 mg, 48%) as a white powder after lyophilization. C 26 H 27 MS calculated for FN3O5: 480.19, Found: 480.25, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ: 9.03 (s br, 1H), 8.80 (s br, 1H), 7.88 (d, J = 10.8 Hz, 1H), 7.34 (s, 1H), 6.57 (s, 1H), 5.57 - 5.36 (m, 4H), 5.30 (s, 1H), 5.16 - 5.02 (m, 1H), 3.69 (t, J = 5.5 Hz, 2H), 3.31 - 3.06 (m, 3H), 2.84 - 2.71 (m, 1H), 2.41 (s, 3H), 2.26 - 2.10 (m, 1H), 1.99 - 1.76 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0196] Example 22: Synthesis of compound 1013 JPEG2024518438000132.jpg162169Intermediate 1 A mixture of compound 105 (1.0 equiv, 20 mg, 0.034 mmol) and FmocGGFG-OAc (2.0 equiv, 0.068 mmol, 43 mg) was dissolved in anhydrous DMF (1 mL), followed by the addition of 2M HCl / Et2O (100 μL). The reaction mixture was stirred at room temperature for 4 h, during which time FmocGGFG-OAc was added in portions (approximately 0.5 equiv each) to the reaction mixture. The reaction mixture was then purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0 → 75% ACN / H2O). The product-containing fractions were lyophilized, and the residue (product + coeluting impurities) was used directly in the next step. C 57 H 58 FN8O 11 MS calculated: 1049.42, Found: 1049.40, [M+H] + .

[0197] Intermediate 2 Morpholine (80 μL) was added to a solution of intermediate 1 (obtained in the previous step) in anhydrous DMF (2 ml) and the reaction mixture was stirred at room temperature for 1.5 h. LC-MS showed complete consumption of the starting material. The mixture was directly purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→60% ACN / H2O) to give the product as a white solid after lyophilization (12 mg, 43%). 42 H 48 MS calculated for FN8O9: 827.35, found: 827.30, [M+H] + .

[0198] Compound 1013 To a solution of intermediate 2 (12 mg, 0.015 mmol) in anhydrous DMF (1 ml) was added Mal-PEG-NHS ester (1.0 equiv., 0.015 mmol, 4.3 mg) and DIPEA (2.2 equiv., 0.032 mmol, 4.2 mg, 5.7 μL). The reaction mixture was stirred at room temperature for 40 min as LC-MS showed complete consumption of starting material. Purification by reversed-phase flash HPLC using a semi-preparative column (diol-modified C18, 0→60% ACN / H2O) afforded the desired product as a white solid after lyophilization (6.3 mg, 41%). C 51 H 57 FN9O 13 MS calculated: 1022.41, Found: 1022.40, [M + H] + . 1 H NMR (500 MHz, DMSO-d6) δ: 8.65 (t, J= 6.7 Hz, 1H), 8.33 - 8.26 (m, 1H), 8.15 - 8.07 (m, 2H), 8.03 - 7.96 (m, 1H), 7.76 - 7.70 (m, 1H), 7.60 (s, 1H), 7.26 - 7.18 (m, 5H), 7.18 - 7.12 (m, 1H), 6.99 (s, 2H), 5.49 - 5.24 (m, 3H), 4.81 (d, J = 11.8 Hz, 1H), 4.71 - 4.55 (m, 3H), 4.48 (ddd, J = 9.6, 8.1, 4.5 Hz, 1H), 4.28 (t, J = 4.2 Hz, 1H), 3.81 - 3.67 (m, 3H), 3.65 (d, J = 5.7 Hz, 2H), 3.62 - 3.47 (m, 6H), 3.47 - 3.41 (m, 2H), 3.22 - 3.12 (m, 1H), 3.06 - 2.90 (m, 2H), 2.87 - 2.71 (m, 2H), 2.38 - 2.29 (m, 5H), 2.26 - 1.98 (m, 4H), 1.26 - 1.16 (m, 1H), 0.90 - 0.82 (m, 3H).

[0199] Example 23: Synthesis of Compound 106 JPEG2024518438000133.jpg38149Compound 106 Compound 107 Trifluoroacetic acid (20 mg, 0.0364 mmol), silyl-protected glycolic acid (1 equiv., 0.0364 mmol, 11.5 mg), and DMTMM (1 equiv., 0.0364 mmol, 10 mg) were dissolved in 2 mL of a 1:4 water / DMF mixed solvent. DIPEA (20 μL) was added and the reaction mixture was stirred at room temperature for 1 h, and LCMS analysis showed complete conversion. The solvent was evaporated under vacuum and the crude reaction mixture was redissolved in 1 mL of DCM and 3 mL of TFA. After stirring at room temperature for 16 h, the solvent was evaporated and the crude reaction product was redissolved in 1 mL of DMF and directly purified by reversed-phase semi-separated flash chromatography (diol-modified C18, 0→80% ACN / water). The desired product was obtained as a 1:1 mixture of two inseparable isomers, both forms in equilibrium (10 mg, 54%, orange solid). 26 H 25 MS calculated for FN3O5S: 510.15, Found: 510.44, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.51 (dd, J = 15.9, 9.1 Hz, 1H), 7.83 - 7.74 (m, 2H), 7.63 - 7.55 (m, 1H), 7.39 - 7.24 (m, 1H), 6.69 (s, 1H), 5.92 (m, 1H), 5.64 (d, J = 6.7 Hz, 1H), 5.56 - 5.44 (m, 2H), 5.30 (t, J = 19.0 Hz, 1H), 4.17 - 3.99 (m, 1H), 3.26 (m, 1H), 3.19 - 3.08 (m, 1H), 2.60 - 2.52 (m, 1H), 2.38 (d, J= 1.9 Hz, 3H), 2.21 (d, J = 7.2 Hz, 2H), 1.94 - 1.83 (m, 1H), 0.91 - 0.81 (m, 3H).

[0200] Example 24: Synthesis of Compound 107 JPEG2024518438000134.jpg98156Intermediate 1 Exatecan mesylate (52 mg, 0.0978 mmol) was dissolved in 3 mL of anhydrous pyridine and heated to 80 °C. tert-Butyldimethylsilyl trifluoromethanesulfonate (10 equiv., 0.978 mmol, 259 mg) was added and the reaction mixture was stirred at 80 °C for 3 h, as LCMS analysis showed complete conversion to the product. The reaction mixture was cooled to room temperature and 9-fluorenylmethyloxycarbonyl chloride (2 equiv., 0.196 mmol, 51 mg) was added. The reaction mixture was stirred at room temperature for 2 h. The residue was purified by reversed-phase flash chromatography (diol-modified C18, 0→100% ACN) to give the desired intermediate 1 (45 mg, 60%) as a pale yellow solid. C 45 H 47 MS calculated for FN3O6Si: 772.32, Found: 772.30, [M+H] + .

[0201] Intermediate 2: Intermediate 1 (0.0583 mmol, 45 mg) prepared above and Lawesson's reagent (5 equiv., 0.146 mmol, 59 mg) were dissolved in toluene (5 mL) and the reaction mixture was stirred at 100 °C for 3 h. The solvent was evaporated and the residue was directly purified by reversed-phase flash chromatography (diol-modified C18, 0→100% ACN / water) to give the desired product Intermediate 2 (41 mg, 89%) as a yellow solid. 45 H 47 MS calculated for FN3O5SSi: 788.30, Found: 788.33, [M+H] + .

[0202] Intermediate 3: Intermediate 2 (41 mg, 0.0520 mmol) prepared above was dissolved in 2 mL of anhydrous DMF and morpholine (100 μL) was added. The reaction mixture was stirred at room temperature for 1 h. The residue was directly purified by reversed-phase flash chromatography (diol-modified C18, 0→100% CAN in water) to give Intermediate 3 (23 mg, 79%) as a bright yellow solid. 30 H 37MS calculated for FN3O3SSi: 566.23, Found: 566.45, [M+H] + .

[0203] Compound 107 TFA The previously prepared intermediate 3 (23 mg, 0.0407 mmol) was dissolved in 2 mL of anhydrous dichloromethane and 2 mL of trifluoroacetic acid was added. The reaction mixture was stirred at room temperature for 12 h. The solvent was evaporated under reduced pressure and the crude reaction product was redissolved in 2 mL of DMF and directly purified by reversed-phase flash chromatography (diol-modified C18, 0→100% ACN / water). A second reversed-phase HPLC purification (semi-separate HPLC, diol-modified C18, 0→100% ACN in water) afforded compound 107 trifluoroacetate as a 1:1 mixture of two inseparable isomers, with both forms in equilibrium (11 mg, 60%, light yellow solid). C 24 H 23 MS calculated for FN3O3S: 452.14, Found: 452.15, [M+H] + . 1 H NMR (500 MHz, DMSO-d6) δ 8.55 (d, J = 5.8 Hz, 3H), 7.97 - 7.91 (m, 1H), 7.84 (s, 1H), 7.32 (d, J = 37.9 Hz, 1H), 6.70 (d, J = 26.1 Hz, 1H), 6.07 - 5.91 (m, 2H), 5.71 (d, J = 20.1 Hz, 1H), 5.56 (dd, J = 16.6, 3.7 Hz, 1H), 3.32 (m, 1H), 3.14 (m, 1H), 2.44 (s, 3H), 1.91 (h, J = 6.9 Hz, 2H), 1.26 (q, J = 7.1 Hz, 2H), 0.88 (t, J = 7.3 Hz, 3H).

[0204] Example 25: Synthesis of Compound 108 JPEG2024518438000135.jpg46170 Diisopropylethylamine (2.5 equiv, 0.07 mmol, 9 mg, 13 μL) and propargyl bromide (2.5 equiv, 0.07 mmol, 8.5 mg, 9 μL of an 80% solution in toluene) were added to a suspension of exatecan mesylate (1.0 equiv, 15 mg, 0.028 mmol) in DMF (0.2 ml) and the resulting solution was stirred for 48 h. Then, 2-azidoethanol (5.0 equiv, 0.14 mmol, 12 mg, 11 μL), tris(benzyltriazolylmethyl)amine (1.5 equiv, 0.042 mmol, 22 mg), CuSO 4. 5H2O (1.0 equiv., 0.028 mmol, 140 μL 2M aqueous solution) and sodium ascorbate (2.0 equiv., 0.056 mmol, 56 μL 1M aqueous solution) were added sequentially to the reaction mixture and the solution was stirred overnight. The crude reaction mixture was loaded directly onto a column and purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0% to 50% ACN / H2O) to give compound 108 (12 mg, 77%) as a white powder after lyophilization. 29 H 30 MS calculated for FN6O5: 561.23, Found: 561.30, [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ: 7.97 (s, 1H), 7.72 (d, J = 11.0 Hz, 1H), 7.29 (s, 1H), 5.43 (s, 2H), 5.28 (d, J = 19.0 Hz, 1H), 5.19 (d, J = 19.0 Hz, 1H), 4.39 (t, J = 5.5 Hz, 2H), 4.25 (t, J = 4.1 Hz, 1H), 3.97 (q, J= 13.9 Hz, 2H), 3.80 - 3.75 (m, 2H), 3.33 (s, 2H), 3.24 (ddd, J = 15.8, 10.4, 4.3 Hz, 1H), 3.01 (dt, J = 16.8, 4.8 Hz, 1H), 2.39 - 2.27 (m, 6H), 2.09 - 1.98 (m, 1H), 1.95 - 1.78 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0205] Example 26: Synthesis of compound 1015 JPEG2024518438000136.jpg101170Intermediate 1 Compound 108 (1.0 equiv., 30 mg, 0.0536 mmol) and FmocGGFG-OAc (2.0 equiv., 0.107 mmol, 67 mg) were dissolved in anhydrous DMF (1.5 mL), followed by the addition of 2M HCl / Et2O (150 μL). The reaction mixture was stirred at room temperature for 4 h, during which time FmocGGFG-OAc was added in portions (approximately 0.5 equiv. each) to the reaction mixture. The reaction mixture was then purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→75% ACN / H2O). The product-containing fractions were lyophilized, and the residue (product + coeluting impurities) was used directly in the next step. C 60 H 61 FN 11 O 11 MS calculated: 1130.45, Found: 1130.40, [M+H] + .

[0206] Intermediate 2 Morpholine (100 μL) was added to a solution of intermediate 1 (obtained in the previous step) in anhydrous DMF (2 ml) and the reaction mixture was stirred at room temperature for 1 h. LC-MS showed complete consumption of the starting material. The mixture was directly purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→50% ACN / H2O) to give the product as a white solid after lyophilization (14 mg, 29% over two steps). 45 H 51 FN 11 MS calculated for O9: 908.39, found: 908.55, [M+H] + .

[0207] Compound 1015. To a solution of intermediate 2 (1 eq., 14 mg, 0.0154 mmol) in anhydrous DMF (1 ml) was added Mal-PEG-NHS ester (1 eq., 0.0154 mmol, 5 mg) and DIPEA (2.5 eq., 0.039 mmol, 5 mg, 6.6 μL). The reaction mixture was stirred at room temperature for 30 min as LC-MS showed complete consumption of starting material. Purification by reversed-phase flash HPLC using a semi-separative column (diol-modified C18, 0→100% CAN in water) afforded the desired product as a white solid (7 mg, 41%) after lyophilization from water-acetonitrile. C 54 H 60 FN 12 O 13 MS calculated: 1103.44, Found: 1103.61, [M + H] + . 1H NMR (500 MHz, DMSO-d6) δ 8.54 (m, 1H), 8.30 (m, 1H), 8.12 (m, 2H), 8.01 (m, 1H), 7.97 (s, 1H), 7.73 (m, 1H), 7.65 (s, 1H), 7.29 (s, 1H), 7.25 - 7.17 (m, 4H), 7.17 - 7.12 (m, 1H), 7.01 (d, J = 5.9 Hz, 1H), 6.99 (s, 2H), 6.51 (s, 1H), 5.43 (s, 1H), 5.34 - 5.06 (m, 2H), 4.55 (m, 2H), 4.51 (m, 1H), 4.49 - 4.43 (m, 1H), 4.25 (d, J = 17.1 Hz, 1H), 4.10 - 3.88 (m, 2H), 3.80 (q, J = 4.9 Hz, 2H), 3.76 - 3.63 (m, 5H), 3.45 (t, J = 5.9 Hz, 2H), 3.22 (d, J = 10.9 Hz, 1H), 3.04 - 2.98 (m, 2H), 2.87 (t, J= 6.0 Hz, 1H), 2.82 - 2.73 (m, 2H), 2.59 (s, 1H), 2.36 (m, 4H), 2.32 (m, 2H), 2.06 - 1.97 (m, 2H), 1.86 (m 1H), 1.29 - 1.21 (m, 2H), 0.86 (m, 3H).

[0208] Example 27: Synthesis of Compound 109 JPEG2024518438000137.jpg46165Intermediate 1 Exatecan mesylate (20 mg, 0.0376 mmol) was dissolved in 2 mL of DMF containing diisopropylethylamine (5 equiv., 0.188 mmol, 33 μL). The reaction mixture was stirred at room temperature overnight. The crude reaction mixture was loaded directly onto a column and purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in water (0% to 60% ACN in water). The desired product was recovered as a white powder after lyophilization from water (16 mg, 76%). C 27 H25 MS calculated for FN3O4: 474.51, found: 474.66, [M+H] + .

[0209] Compound 109. The previously prepared intermediate 1 (16 mg, 0.0286 mmol) and CpRu(COD)cl (0.1 equiv., 1.3 mg, 0.0029 mmol) were suspended in 10 mL of anhydrous dichloromethane. 2-Azidoethanol (3.2 equiv., 0.115 mmol, 10 mg) was added and the mixture was refluxed at 50° C. for 16 h. The solvent was evaporated and the crude product was redissolved in DMF (2 mL). The catalyst was filtered through a 2 μm syringe filter and the crude reaction mixture was purified by reversed-phase HPLC using a semi-separated column containing a diol-modified C18 with a gradient of ACN in water (0% to 80% ACN in water). The desired product was recovered as a white powder after lyophilization from water (8 mg, 50%). C 29 H 30 MS calculated for FN6O5: 561.23, Found: 561.75, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 7.74 (d, J = 11.0 Hz, 1H), 7.69 - 7.62 (m, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.43 (d, J = 3.5 Hz, 2H), 5.41 - 5.28 (m, 1H), 4.44 (m, 2H), 4.30 (t, J = 4.6 Hz, 1H), 4.16 (d, J = 14.3 Hz, 1H), 4.02 (d, J= 14.3 Hz, 1H), 3.76 (t, J = 5.5 Hz, 2H), 3.27 - 3.19 (m, 1H), 3.02 (m, 1H), 2.68 (m, 2H), 2.37 (m, 3H), 2.25 (m, 2H), 2.18 - 1.99 (m, 1H), 1.86 (m, 2H), 0.86 (dt, J = 9.2, 7.3 Hz, 3H).

[0210] Example 28: Synthesis of Compound 110 JPEG2024518438000138.jpg72160Intermediate 1 Exatecan mesylate (30 mg, 0.066 mmol) and diisopropylethylamine (2.5 equiv., 0.164 mmol, 29 μL) were dissolved in 2 mL of DMF. 3-(benzyloxy)propane-1-sulfonyl chloride (1.2 equiv., 0.788 mmol, 196 mg) was added and the reaction mixture was stirred at room temperature for 3 h. The solvent was evaporated under reduced pressure and the crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in 1% TFA (ACN 0→60% in 1% TFA). The desired product was recovered as a white powder after lyophilization from water (16 mg, 37%). C 34 H 35 MS calculated for FN3O7S: 648.22, Found: 647.99 [M+H] + .

[0211] Compound 110. The previously synthesized intermediate 1 (16 mg, 0.0247 mmol) was dissolved in 3 mL of dioxane. Pd / C (10% w / w, 5 mg) was suspended in the mixture and H2 was bubbled through the suspension using a balloon while stirring at room temperature for 2 h. The dioxane was evaporated under reduced pressure and the crude product was redissolved in 2 mL of DMF and purified by reversed-phase flash HPLC using a semi-preparative column containing a diol-modified C18 and a gradient of ACN in water (0 -> 80% ACN in water). The desired product was recovered as a yellow solid after lyophilization from water-dioxane (6 mg, 44%). C 27 H 29 MS calculated for FN3O7S: 558.17, Found: 558.66 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 7.78 (dd, J = 11.0, 6.5 Hz, 1H), 7.31 (s, 1H), 5.46 - 5.38 (m, 3H), 5.08 (t, J = 5.2 Hz, 1H), 3.55 (td, J = 6.1, 2.3 Hz, 2H), 3.32 - 3.28 (m, 2H), 3.16 (dt, J = 16.6, 5.9 Hz, 1H), 2.68 (p, J = 1.8 Hz, 1H), 2.40 - 2.35 (m, 4H), 2.33 (m, 2H), 2.26 (q, J = 6.2 Hz, 2H), 1.89 (m, 4H), 1.76 (s, 1H), 0.88 (t, J = 7.3 Hz, 3H).

[0212] Example 29: Synthesis of Compound 111 JPEG2024518438000139.jpg39164 Compound 111 Methyl 2-(hydroxymethyl)cyclopropane-1-carboxylate (25 mg, 0.175 mmol) was dissolved in 1 mL of methanol and 870 μL of 1M NaOH (1 eq.) was added. The mixture was stirred at room temperature for 5 h, the solvent was evaporated and the crude product was lyophilized from water. To the resulting solid was added exatecan mesylate (46 mg, 0.5 eq., 0,874 mmol), DMTMM (48 mg, 1 eq., 0.175 mmol) and 10 mL of a 4:1 DMF / water mixture. The mixture was stirred at room temperature for 30 min. The solvent was evaporated under reduced pressure to a final volume of about 2 mL. The crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in water (ACN 0→60% in water). A second purification was performed using a semi-preparative column loaded with diol-modified C18 using a gradient of ACN in water (0-80% ACN in water). Two isomers were separated during the semi-preparative purification. The products were recovered separately as white powders after lyophilization from water / dioxane (42 mg total, 91% calculated from exatecan). 29 H 29MS calculated value of FN3O6: 534.20, measured value: 534.10, [M+H] + .

[0213] Isomer A: 1 H NMR (400 MHz, DMSO-d6) δ 8.67 (d, J = 8.8 Hz, 1H), 7.78 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.56 (q, J = 6.6 Hz, 1H), 5.42 (s, 2H), 5.16 (d, J = 2.9 Hz, 2H), 4.63 (t, J = 5.5 Hz, 1H), 3.48 - 3.38 (m, 1H), 3.31 - 3.25 (m, 2H), 3.21 - 3.08 (m, 1H), 2.30 (m, 2H), 2.24 - 2.07 (m, 2H), 1.96 - 1.77 (m, 2H), 1.58 (d, J = 4.4 Hz, 1H), 1.49 (m, 1H), 0.99 (dt, J = 8.4, 4.3 Hz, 1H), 0.88 (t, J = 7.3 Hz, 3H), 0.71 (m, 1H).

[0214] Isomer B (containing 7% of isomer A, by NMR integration): 1 H NMR (400 MHz, DMSO-d6) δ 8.72 (d, J = 8.9 Hz, 1H), 7.77 (d, J = 10.9 Hz, 1H), 7.32 (s, 1H), 6.53 (s, 1H), 5.56 (m, 2H), 5.44 (s, 2H), 5.31 - 5.05 (m, 2H), 4.52 (dd, J = 6.1, 5.0 Hz, 1H), 3.45 (m 1H), 3.26 (m, 1H), 3.20 - 3.08 (m, 1H), 2.38 (m, 2H), 2.24 - 2.00 (m, 2H), 1.87 (m, 2H), 1.76 (s, 1H), 1.60 - 1.48 (m, 1H), 1.01 (m, 1H), 0.88 (t, J = 7.3 Hz, 3H), 0.75 (m, 1H).

[0215] Example 30: Synthesis of Compound 1016 JPEG2024518438000140.jpg144170Intermediate 1 2-((benzyloxy)methyl)cyclopropane-1-carboxylic acid (27.6 mg, 0.1338 mmol) was dissolved in 3 mL of anhydrous dioxane. Pd / C (10%) was added and hydrogen was bubbled through for 5 h at room temperature with stirring. The solution was filtered through a 0.2 μm syringe filter and the flask was washed with acetonitrile. The filtrate was evaporated, redissolved in dioxane and lyophilized overnight. The crude filtrate was redissolved in 2 mL of anhydrous DMF and FmocGGFG-OAc (1 equiv, 0.1338 mmol, 90 mg) was added followed by 200 μL of a 2 M solution of hydrochloric acid in ethyl ether. The reaction mixture was stirred at room temperature for 1 h and loaded directly onto a column for purification. Purification was performed by reversed-phase flash chromatography (25 g, diol-modified C18, 0→75% ACN / H2O). The fractions containing the product were lyophilized from water (35 mg, 51%). 36 H 40 MS calculated for N5O9: 686.28, found: 686.66, [M+H] + .

[0216] Intermediate 2 To a solution of the previously prepared intermediate 1 (35 mg, 0.0505 mmol) in DMF (2 mL) was added exatecan mesylate (1 equiv, 0.0505 mmol, 27 mg), DMTMM (1.2 equiv, 0.0607 mmol, 17 mg), DIPEA (10 μL) and water (200 μL). The reaction mixture was stirred at room temperature for 30 min and loaded directly onto the column for purification. Purification was performed by reversed-phase flash chromatography (25 g, diol-modified C18, 0→60% ACN / H2O). The fractions containing the product were lyophilized from water (28 mg, 50%). C 60 H 60 FN8O 12 MS calculated: 1103.43, Found: 1103.88, [M+H] + .

[0217] Intermediate 3 The intermediate 2 (28 mg, 0.0254 mmol) prepared above was dissolved in DMF (2 mL) and morpholine (100 μL) was added. The reaction mixture was stirred at room temperature for 1 h. Purification was carried out by reversed-phase HPLC chromatography (semi-separated, diol-modified C18, 0→100% ACN / H2O). The fractions containing the product were lyophilized from water (11 mg, 51%). C 45 H 50 FN8O 10 MS calculated: 881.36, Found: 881.12, [M+H] +

[0218] Compound 1016 The previously prepared intermediate 3 (11 mg, 0.0130 mmol) was dissolved in 2 mL of anhydrous DMF. 2,5-Dioxopyrrolidin-1-yl 3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)propanoate (1.1 equiv., 0.0143 mmol, 4 mg) and DIPEA (1.1 equiv., 0.0143 mmol, 2.5 μL) were added and the reaction mixture was stirred at room temperature for 1 h. Purification was performed by reversed-phase HPLC chromatography (semi-separated, diol-modified C18, 0→100% ACN / H2O). The fractions containing the product were lyophilized from water (7 mg, 50%). C 54 H 59 FN9O 14 MS calculated: 1076.42, Found: 1076.56, [M+H] + . 1H NMR (500 MHz, DMSO-d6) δ 8.80 - 8.70 (m, 2H), 8.53 (m, 1H), 8.45 (m, 1H), 8.27 (m, 1H), 8.10 (m, 2H), 7.99 (m, 2H), 7.93 (d, J = 6.1 Hz, 1H), 7.89 (d, J = 8.4 Hz, 1H), 7.80 (m, 2H), 7.34 - 7.20 (m, 2H), 7.18 - 7.08 (m, 2H), 6.67 (s, 2H), 6.51 (s, 1H), 5.62 (d, J = 10.0 Hz, 1H), 5.55 (s, 2H), 5.43 (t, J = 5.7 Hz, 2H), 5.19 - 5.07 (m, 3H), 5.06 - 5.02 (m, 1H), 4.74 (d, J = 6.0 Hz, 1H), 4.61 - 4.44 (m, 3H), 3.72 (m, 2H), 3.57 (m, 2H), 3.16 (d, J = 8.7 Hz, 1H), 3.08 - 3.00 (m, 1H), 2.79 (m, 2H), 2.39 (m, 3H), 2.34 (d, J = 1.8 Hz, 3H), 2.24 - 2.07 (m, 2H), 1.96 - 1.77 (m, 2H), 1.55 (m, 1H), 1.51 - 1.44 (m, 1H), 0.99 (m, 1H), 0.88 (t, J= 7.3 Hz, 3H), 0.83 - 0.71 (m, 1H).

[0219] Example 31: Synthesis of Compound 1017 JPEG2024518438000141.jpg168159Intermediate 1 (1S,2S)-2-(hydroxymethyl)cyclopropane-1-carboxylic acid (1.0 equiv., 5.7 mg, 0.049 mmol), Fmoc-GE(OBn)VCit-NH-CH2-OAc (1.5 equiv., 62 mg, 0.074 mmol) were dissolved in anhydrous DMF (0.7 ml), 2M HCl / Et2O (100 μl) was added, and the resulting mixture was stirred at room temperature for 1 h. The reaction mixture was purified by reversed-phase flash chromatography using a semi-separating column (diol-modified C18, 0→75% ACN / 0.1% HCl). The fractions containing the product (coeluted with impurity) were lyophilized to give 31 mg of impure intermediate 1. C 46 H 56 FN7O 12 MS calculated value: 898.40, Measured value: 898.40, [MH] - .

[0220] Intermediate 2 To a mixture of intermediate 1 (1.0 equiv., 31 mg, 0.034 mmol), exatecan mesylate (0.9 equiv., 17 mg, 0.031 mmol) and DMTMM (1.0 equiv., 10 mg, 0.034 mmol) was added DMF / water (5:1, 1.2 ml) and diisopropylethylamine (2.0 equiv., 12 μl, 0.068 mmol), and the resulting mixture was stirred at room temperature for 1 h, since LC-MS analysis showed complete consumption of the starting material. The reaction mixture was purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→75% ACN / H2O) to give intermediate 2 as a white solid after lyophilization (25 mg, 39% (2 steps)). C 70 H 78 FN 10 O 15 MS calculated: 1317.56, Found: 1317.55, [M+H] + .

[0221] Intermediate 3 To a solution of intermediate 2 (1.0 equiv., 25 mg, 0.019 mmol) in a mixture of dioxane (1.0 ml) and DMF (0.5 ml), 10% Pd / C (5 mg) was added and the reaction mixture was hydrogenated (balloon) at room temperature for 2 h. LC-MS showed complete consumption of starting material. The mixture was filtered through a pad of Celite and the filtrate was concentrated on a rotary evaporator to remove dioxane. Morpholine (40 μl) was added to the resulting solution and the reaction mixture was stirred at room temperature for 1 h. Purification by reversed-phase flash chromatography (25 g, diol-modified C18, 0→60% ACN / H2O) afforded the product intermediate 3 as a white solid after lyophilization (8 mg, 42%). C 48 H 62 FN 10 O 13 MS calculated value: 1005.43, Found value: 1005.40, [M+H] + .

[0222] Compound 1017. To a solution of intermediate 3 (1.0 equiv., 8 mg, 0.008 mmol) in anhydrous DMF (0.5 ml) was added Mal-PEG-NHS ester (1.0 equiv., 0.008 mmol, 2.5 mg) and DIPEA (1.05 equiv., 0.008 mmol, 1.5 μL). The reaction mixture was stirred at room temperature for 1.5 h as LC-MS showed complete consumption of the starting material. Purification by reversed-phase flash chromatography using a semi-separative column (diol-modified C18, 0→70% ACN / 0.1% TFA) afforded the desired product as a pale yellow solid after lyophilization (5 mg, 52%). C 57 H 71 FN 11 O 17 MS calculated value: 1200.50, Found value: 1200.50, [M+H] + . 1H NMR (500 MHz, DMSO-d6) δ: 12.09 (s br, 1H), 8.71 (d, J = 8.7 Hz, 1H), 8.58 - 8.49 (m, 1H), 8.05 (t, J = 5.2 Hz, 1H), 8.01 - 7.92 (m, 2H), 7.83 - 7.70 (m, 2H), 7.31 (s, 1H), 7.00 (s, 2H), 6.58 (s br, 1H), 5.92 (s, 1H), 5.62 - 5.49 (m, 1H), 5.50 - 5.33 (m, 2H), 5.28 - 5.12 (m, 2H), 4.60 - 4.40 (m, 2H), 4.39 - 4.25 (m, 1H), 4.21 - 4.02 (m, 2H), 3.77 - 3.62 (m, 3H), 3.31 - 3.19 (m, 2H), 3.20 - 3.09 (m, 1H), 3.04 - 2.82 (m, 2H), 2.40 (s, 3H), 2.35 - 2.26 (m, 2H), 2.27 - 2.10 (m, 4H), 1.99 - 1.79 (m, 4H), 1.76 - 1.64 (m, 1H), 1.63 - 1.41 (m, 4H), 1.40 - 1.23 (m, 3H), 1.05 - 0.93 (m, 1H), 0.92 - 0.84 (m, 3H), 0.84 - 0.75 (m, 6H), 0.75 - 0.69 (m, 2H).

[0223] Example 32: Synthesis of Compound 115 JPEG2024518438000142.jpg47170Intermediate 1 3-Amino-1,2-propanediol (25 mg, 0.274 mmol) and dimethoxysquarate (3 equiv., 0.823 mmol, 117 mg) were suspended in 10 mL of 1 M borate buffer (pH = 9) and the mixture was stirred at 55 °C for 16 h. 2 mL of DMF was added and the solvent was evaporated under reduced pressure to a final volume of about 3 mL. The crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in water (0 → 50% ACN in water). The desired product was recovered as a white powder after lyophilization from water (33 mg, 57%). C8H 12 MS calculated for NO5: 202.07, Found: 202.18, [M+H] + .

[0224] Compound 115. Exatecan mesylate (20 mg, 0.0377 mmol) and previously synthesized intermediate 1 (1.5 equiv., 0.0564 mmol, 12 mg) were suspended in 5 mL of 1 M borate buffer (pH = 9) and the mixture was stirred at 55 °C for 16 h. 2 mL of DMF was added and the solvent was evaporated under reduced pressure to a final volume of about 3 mL. The crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in water (0 -> 50% ACN in water). The desired product was recovered as a white powder after lyophilization from water (15 mg, 66%). C 31 H 30 MS calculated for FN4O8: 605.20, Found: 605.22, [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 8.04 (s, 1H), 7.85 (d, J = 10.9 Hz, 1H), 7.32 (s, 1H), 6.53 (s, 1H), 5.79 (s, 1H), 5.42 (s, 2H), 5.39 (s, 2H), 3.74 (s, 2H), 3.55 (s, 1H), 3.52 - 3.31 (m, 6H), 3.32 - 3.20 (m, 1H), 2.43 (d, J= 1.9 Hz, 3H), 2.34 - 2.26 (m, 1H), 1.96 - 1.77 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0225] Example 33: Synthesis of Compound 117 JPEG2024518438000143.jpg50167Intermediate 1 Exatecan succinamide (1 equiv., 16 mg, 0.030 mmol), (2,2-dimethyl-1,3-dioxolan-4-yl)methanamine (3 equiv., 12 mg, 0.090 mmol) and diisopropylethylamine (5 equiv., 0.150 mmol, 26 μL) were dissolved in DMF / water (5:1, 1 mL), DMTMM (1.3 equiv., 13 mg, 0.039 mmol) was added under argon atmosphere and the reaction mixture was stirred at room temperature for 0.5 h. LC-MS showed complete consumption of starting material. The mixture was purified by reversed-phase flash chromatography (diol-modified C18, 0→75% ACN / H2O) to give intermediate 1 (16 mg, 82%) as an off-white powder after lyophilization.

[0226] Compound 117. To a solution of intermediate 1 (16 mg, 0.025 mmol) in dioxane / water (1:1, 4 ml), two drops of TFA were added and the resulting mixture was stirred at 40° C. for 5 h. LC-MS showed complete consumption of intermediate 1, and the solvent was evaporated under reduced pressure. The residue was purified by reversed-phase flash chromatography (diol-modified C18, 0→50% ACN / 1% TFA in water) to give the product (11 mg, 72%) as a white powder after lyophilization. 31 H 34MS calculated value of FN4O8: 609.24, measured value: 609.25, [M+H] + . 1 1H NMR (400 MHz, DMSO-d6) δ: 8.47 (d, J = 8.7 Hz, 1H), 7.86 - 7.81 (m br, 1H), 7.79 (d, J = 11.0 Hz, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.56 (dt, J = 9.0, 4.8 Hz, 1H), 5.42 (s, 2H), 5.22 (d, J = 19.1 Hz, 1H), 5.15 (d, J = 19.1 Hz, 1H), 4.68 (dd, J= 5.0, 2.1 Hz, 1H), 4.47 (t, J = 5.7 Hz, 1H), 3.48 - 3.40 (m, 1H), 3.29 - 3.22 (m, 2H), 3.20 - 3.09 (m, 3H), 2.99 - 2.87 (m, 1H), 2.46 - 2.33 (m, 6H), 2.21 - 2.05 (m, 2H), 1.95 - 1.78 (m, 2H), 0.88 (t, J = 7.3 Hz, 3H).

[0227] Example 34: Synthesis of Compound 118 JPEG2024518438000144.jpg51169Intermediate 1 To a solution of (2,2-dimethyl-1,3-dioxolan-4-yl)methanamine (2 equiv., 15 mg, 0.114 mmol) and diisopropylethylamine (5 equiv., 0.285 mmol, 50 μL) in dichloromethane (3 mL), triphosgene (0.6 equiv., 11 mg, 0.035 mmol) was added and the reaction mixture was stirred at room temperature for 2 h, A solution of (2,2-dimethyl-1,3-dioxolan-4-yl)methanamine (2 equiv., 15 mg, 0.114 mmol) and diisopropylethylamine (5 equiv., 0.285 mmol, 50 μL) in dichloromethane (3 mL) was added and the reaction mixture was stirred at rt for 2 h, A solution of (2,2-dimethyl-1,3-dioxolan-4-yl)methanamine (2 equiv., 15 mg, 0.114 mmol) and diisopropylethylamine (1 equiv., 0.057 mmol, 30 mg) in anhydrous DMF (1 mL) was added and the resulting mixture was stirred at rt for 2 h. The reaction was quenched by the addition of methanol (1 mL) and the solvent was evaporated under reduced pressure. The residue was purified by reverse-phase flash chromatography (diol-modified C18, 0→60% ACN / H2O) to give intermediate 1 (29 mg, 86%) as an off-white powder after lyophilization, which was used in the next step.

[0228] Compound 118. To a solution of intermediate 1 (29 mg, 0.049 mmol) in dioxane / water (1:1, 4 ml), 2 drops of TFA were added and the resulting mixture was stirred at 40° C. overnight. LC-MS showed complete consumption of intermediate 1, and the solvent was evaporated under reduced pressure. The residue was purified by reversed-phase flash chromatography (diol-modified C18, 0→50% ACN / 1% TFA in water) to give the product compound 187 (15 mg, 55%) as a white powder after lyophilization. 28 H 30 MS calculated for FN4O7: 553.21, found: 553.20, [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ: 7.76 (d, J = 10.9 Hz, 1H), 7.30 (d, J = 1.6 Hz, 1H), 6.78 (dd, J = 8.9, 6.6 Hz, 1H), 6.52 (d, J = 1.1 Hz, 1H), 5.42 (s, 2H), 5.39 - 5.28 (m, 1H), 5.22 (d, J = 19.3, 1H), 4.81 (dd, J = 9.3, 4.9 Hz, 1H), 4.57 (q, Jj= 5.9, 1H), 3.56 - 3.43 (m, 1H), 3.38 - 3.23 (m, 4H), 3.16 (m br, 2H), 3.04 - 2.91 (m, 1H), 2.38 (s, 3H), 2.24 - 2.06 (m, 2H), 1.94 - 1.79 (m, 2H), 0.88 (t, J= 7.3 Hz, 3H).

[0229] Example 35: Synthesis of Compound 122 JPEG2024518438000145.jpg43157 Compound 122 Intermediate 1 of compound 12 (20 mg, 0.0373 mmol), intermediate 2 of compound 48 (2 eq, 0.0747 mmol, 35 mg), DMTMM (2 eq, 0.0747 mmol, 21 mg) and diisopropylamine (20 μL) were dissolved in 2 mL of 4:1 DMF-water mixed solvent and the reaction mixture was stirred at room temperature for 1 h. The solvent was evaporated under reduced pressure and the crude product was redissolved in 2 mL of DCM. TFA (1 mL) and water (1 mL) were added and the mixture was stirred at room temperature for 1 h. The solvent was evaporated and the crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 and a gradient of ACN in 1% TFA (ACN 0 → 40% in 1% TFA). The desired product was recovered as a white powder after lyophilization from water-acetonitrile (34 mg, 88%). 32 H 36 MS calculated for FN4O9: 639.25, Found: 639.29 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 8.50 (d, J = 8.7 Hz, 1H), 7.99 (s, 1H), 7.81 (d, J = 11.0, 1H), 7.32 (d, J = 9.0 Hz, 1H), 7.21 (d, J= 11.7 Hz, 1H), 6.54 (s, 1H), 5.57 (m, 1H), 5.43 (d, J = 1.6 Hz, 2H), 5.21 (m, 2H), 4.20 - 4.08 (m, 1H), 3.55 (m, 1H), 3.49 (s, 6H), 3.21 - 3.16 (m, 2H), 2.77 - 2.65 (m, 1H), 2.43 - 2.34 (m, 3H), 2.21 - 2.06 (m, 2H), 1.94 - 1.79 (m, 2H), 0.88 (td, J = 7.4, 3.8 Hz, 3H).

[0230] Example 36: Synthesis of Compound 129 JPEG2024518438000146.jpg42162Intermediate 1 Exatecan mesylate (25 mg, 0.0466 mmol), dimethoxysquarate (3 equiv., 0.140 mmol, 20 mg) and sodium methoxide (5 mg) were suspended in 2 mL of anhydrous MeOH and the mixture was stirred at room temperature for 16 h. Methanol was removed under reduced pressure and the crude reaction product was redissolved in 2 mL of DMF. The crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in 1% TFA (0→40% ACN in 1% TFA). The desired product was recovered as a white powder after lyophilization from water (16 mg, 63%). C 29 H 25 MS calculated for FN3O7: 546.17, Found: 545.98, [M+H] + .

[0231] Compound 129. Intermediate 1 prepared above (16 mg, 0.0293 mmol) was suspended in 10 mL of 1M borate buffer (pH = 9) and the mixture was stirred at 55 °C for 16 h. 2 mL of DMF was added and the solvent was evaporated under reduced pressure to a final volume of about 3 mL. The crude reaction mixture was purified by reversed-phase flash chromatography using a column containing 25 g of diol-modified C18 with a gradient of ACN in 1% TFA (ACN 0 -> 40% in 1% TFA). The desired product was recovered as a white powder after lyophilization from water (7 mg, 45%). C 28 H 23 MS calculated for FN3O7: 532.15, found: 532.20, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 9.03 (d, J = 8.6 Hz, 1H), 7.81 (d, J = 10.9 Hz, 1H), 7.31 (s, 1H), 5.58 (q, J = 6.4 Hz, 1H), 5.41 (s, 2H), 5.24 (q, J = 18.9 Hz, 2H), 3.99 (bs, 2H), 3.27 (dt, J = 16.8, 6.4 Hz, 1H), 3.14 (dt, J= 17.0, 5.8 Hz, 1H), 2.40 (d, J = 1.9 Hz, 3H), 2.33 (q, J = 6.1 Hz, 2H), 1.95 - 1.81 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0232] Example 37: Synthesis of Compound 130 JPEG2024518438000147.jpg70159Intermediate 1 Exatecan mesylate (1 equiv., 40 mg, 0.075 mmol), 2-(((tert-butoxycarbonyl)amino)oxy)acetic acid (1.2 equiv., 18 mg, 0.090 mmol) and DMTMM (1.2 equiv., 25 mg, 0.090 mmol) were added to DMF / water (5:1, 2 ml) and diisopropylethylamine (2.2 equiv., 0.165 mmol, 29 μL) and the resulting mixture was stirred at room temperature for 0.5 h. LC-MS indicated complete consumption of starting material. Purification of the mixture by reversed-phase flash chromatography (diol-modified C18, 0→100 % ACN / H2O) afforded intermediate 1 (43 mg, 94 %) as a white powder after lyophilization.

[0233] Compound 130. Intermediate 1 (43 mg, 0.070 mmol) was dissolved in 4M hydrochloric acid / dioxane (2 ml) and the mixture was stirred at room temperature for 1.5 h. The resulting suspension was filtered and the solid was washed with dioxane and Et2O to give the hydrochloride salt of compound 130 (35 mg, 90%) as a yellow powder. 26 H 26 MS calculated for FN4O6: 509.18, Found: 509.20, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ: 10.98 (s br, 3H), 8.89 (d, J = 8.5 Hz, 1H), 7.81 (d, J = 11.0 Hz, 1H), 7.32 (s, 1H), 5.62 (dt, J = 8.5, 4.2 Hz, 1H), 5.43 (s, 2H), 5.37 - 5.24 (m, 2H), 4.63 - 4.52 (m, 2H), 3.20 (dd, J = 7.9, 4.7 Hz, 2H), 2.41 (d, J = 1.9 Hz, 3H), 2.32 - 2.21 (m, 1H), 2.21 - 2.09 (m, 1H), 1.96 - 1.77 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0234] Example 38: Synthesis of compound 1018 JPEG2024518438000148.jpg132170Intermediate 1 To a mixture of compound 130 (1.0 equiv., 30 mg, 0.055 mmol), FmocGGFGGG-OH (1.25 equiv., 47 mg, 0.069 mmol) and DMTMM (1.25 equiv., 19 mg, 0.069 mmol) was added DMF / water (5:1, 2.4 ml) and diisopropylethylamine (2.25 equiv., 22 μL, 0.124 mmol), and the resulting mixture was stirred at room temperature for 1 h, since LC-MS analysis showed complete consumption of the starting material. The reaction mixture was purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→75% ACN / H2O) to give intermediate 1 as a white solid after lyophilization (48 mg, 75%). C 60 H 60 FN 10 O 14 MS calculated: 1163.43, Found: 1163.40, [M+H] + .

[0235] Intermediate 2 To a solution of intermediate 1 (1.0 equiv., 48 mg, 0.041 mmol) in anhydrous DMF (1.5 ml) was added morpholine (100 μL) and the reaction mixture was stirred at room temperature for 1 h. LC-MS showed complete consumption of the starting material. The mixture was directly purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→60% ACN / H2O) to give the product as a white solid after lyophilization (26 mg, 67 %). 45 H 50 FN 10 O 12 MS calculated: 941.36, Found: 941.40, [M+H] + .

[0236] Compound 1018. To a solution of intermediate 2 (1.0 equiv., 26 mg, 0.027 mmol) in anhydrous DMF (1 ml) was added Mal-PEG-NHS ester (1.0 equiv., 0.027 mmol, 8.4 mg) and DIPEA (1.1 equiv., 0.030 mmol, 5.2 μL). The reaction mixture was stirred at room temperature for 40 min as LC-MS showed complete consumption of starting material. DMF was removed and the residue was concentrated from a mixture of 0.1% aq. TFA and ACN. Purification by reversed-phase flash HPLC using a semi-separated column (diol-modified C18, 0→60% ACN / H2O) afforded the desired product as a white solid after lyophilization (17 mg, 55%). C 54 H 59 FN 11 O 16 MS calculated: 1136.41, Found: 1136.45, [M+H] + . 1 H NMR (500 MHz, DMSO-d6) δ: 11.45 (s, 1H), 8.96 - 8.68 (m, 1H), 8.26 (t, J = 5.8 Hz, 1H), 8.22 - 8.04 (m, 3H), 8.04 - 7.93 (m, 2H), 7.87 - 7.75 (m, 1H), 7.31 (s, 1H), 7.23 (d, J = 6.9 Hz, 4H), 7.19 - 7.12 (m, 1H), 6.99 (s, 1H), 6.59 - 6.44 (m, 1H), 5.64 - 5.54 (m, 1H), 5.42 (s, 2H), 5.35 - 5.10 (m, 2H), 4.50 (td, J = 9.8, 9.1, 4.6 Hz, 1H), 4.45 - 4.28 (m, 2H), 3.79 - 3.39 (m, 17H), 3.25 - 3.10 (m, 2H), 3.09 - 2.99 (m, 1H), 2.88 - 2.72 (m, 1H), 2.39 (s, 3H), 2.32 (t, J = 6.5 Hz, 2H), 2.27 - 2.02 (m, 2H), 1.96 - 1.78 (m, 2H), 0.86 (t, J = 7.3 Hz, 3H).

[0237] Example 39: Synthesis of compound 1019 JPEG2024518438000149.jpg121167Intermediate 1 To a mixture of compound 130 (1.0 equiv., 35 mg, 0.064 mmol), FmocGGFGGP-OH (1.2 equiv., 55 mg, 0.077 mmol) and DMTMM (1.2 equiv., 22 mg, 0.077 mmol) was added DMF / water (5:1, 2.4 ml) and diisopropylethylamine (2.2 equiv., 25 μL, 0.141 mmol), and the resulting mixture was stirred at room temperature for 1 h, since LC-MS analysis showed complete consumption of the starting material. The reaction mixture was purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→75% ACN / H2O) to give intermediate 1 as a white solid after lyophilization (65 mg, 84 %). C 63 H 64 FN 10 O 14 MS calculated: 1203.46, Found: 1203.50, [M+H] + .

[0238] Intermediate 2 To a solution of intermediate 1 (1.0 equiv., 65 mg, 0.054 mmol) in anhydrous DMF (1.7 ml) was added morpholine (130 μL) and the reaction mixture was stirred at room temperature for 1 h. LC-MS showed complete consumption of the starting material. The mixture was directly purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→60% ACN / H2O) to give the product as a white solid after lyophilization (34 mg, 64%). 48 H 54 FN 10 O 12 MS calculated: 981.39, Found: 981.40, [M+H] + .

[0239] Compound 1019 To a solution of intermediate 2 (1.0 equiv., 34 mg, 0.035 mmol) in anhydrous DMF (1 ml) was added Mal-PEG-NHS ester (1.0 equiv., 0.035 mmol, 10.8 mg) and DIPEA (1.05 equiv., 0.037 mmol, 6.4 μL). The reaction mixture was stirred at room temperature for 1 h as LC-MS showed complete consumption of starting material. DMF was removed and the residue was concentrated from a mixture of 0.1% TFA, CAN and water. Purification by reversed-phase flash HPLC using a semi-separated column (diol-modified C18, 0→60% ACN / H2O) afforded the desired product as a white solid after lyophilization (18 mg, 44%). C 57 H 63 FN 11 O 16 MS calculated: 1176.44, Found: 1176.45, [M+H] + . 1H NMR (500 MHz, DMSO-d6) δ: 11.50 (s, 1H), 8.85 (d, J = 8.7 Hz, 1H), 8.31 - 8.25 (m, 1H), 8.16 - 8.02 (m, 3H), 7.96 (t, J = 5.7 Hz, 1H), 7.88 - 7.77 (m, 2H), 7.74 (t, J = 5.2 Hz, 1H), 7.35 - 7.28 (m, 2H), 7.28 - 7.18 (m, 5H), 7.18 - 7.11 (m, 1H), 6.99 (s, 1H), 6.57 - 6.46 (m, 1H), 5.70 - 5.58 (m, 1H), 5.43 (s, 2H), 5.35 - 5.11 (m, 3H), 4.54 - 4.46 (m, 1H), 4.32 (s, 2H), 4.08 - 4.03 (m, 1H), 3.90 - 3.39 (m, 14H), 3.24 - 3.10 (m, 2H), 3.03 (dd, J = 13.9, 4.4 Hz, 1H), 2.77 (dd, J = 13.9, 9.8 Hz, 1H), 2.40 (s, 3H), 2.32 (t, J = 6.5 Hz, 2H), 2.26 - 2.11 (m, 2H), 1.97 - 1.77 (m, 3H), 1.69 - 1.51 (m, 1H), 0.86 (t, J= 7.4 Hz, 3H).

[0240] Example 40: Synthesis of Compound 136 JPEG2024518438000150.jpg40170Intermediate 1 To a suspension of exatecan mesylate (1.0 equiv, 30 mg, 0.056 mmol) in DMF (1.2 mL) under argon atmosphere was added diisopropylethylamine (4.5 equiv, 0.25 mmol, 45 μL) and (2-bromoethoxy)-tert-butyldimethylsilane (3.3 equiv, 0.19 mmol, 45 mg, 40 μL) and the mixture was heated to 80 °C for 3 days. Purification by reversed-phase flash chromatography (25 g, diol-modified C18, 0→60% ACN / H2O) afforded the product intermediate 1 (12 mg, 36%) as an off-white powder after lyophilization.

[0241] Compound 136 To a solution of intermediate 1 (1.0 equiv., 12 mg, 0.02 mmol) in DCM (1.5 mL), acetic anhydride (2.2 equiv., 0.042, 4.3 mg, 4 μL) and diisopropylethylamine (2.2 equiv., 0.042 mmol, 8 μL) were added, and the resulting mixture was stirred at room temperature for 40 h. The volatiles were then removed on a rotary evaporator, and the residue was dissolved in a mixture of water and acetonitrile (1:1, 2 mL), followed by the addition of two drops of TFA. The resulting mixture was stirred at room temperature for 2.5 h, and LC-MS showed complete consumption of the starting material. Purification by reversed-phase flash chromatography using a semi-separated column (diol-modified C18, 0→70% ACN / H2O) afforded the product (9 mg, 86%) as a white powder after lyophilization. C 28 H 29 MS calculated for FN3O6: 522.20, Found: 522.20, [M+H] + . 1 H NMR (400 MHz, DMSO-d6, mixture of isomers) δ: 7.84 - 7.71 (m, 1H), 7.34 - 7.28 (m, 1H), 6.55 - 6.48 (m, 1H), 5.64 - 5.44 (m, 1H), 5.41 (s, 2H), 5.20 - 5.01 (m, 1H), 4.99 - 4.81 (m, 2H), 3.69 - 3.37 (m, 3H), 3.19 - 2.92 (m, 1H), 2.46 - 2.30 (m, 4H), 2.31 - 2.11 (m, 3H), 1.97 - 1.77 (m, 2H), 0.97 - 0.78 (m, 3H).

[0242] Example 41: Synthesis of Compound 1022 JPEG2024518438000151.jpg111158Intermediate 1 (S,S)-3-Fluoropyrrolidine-2-carboxylic acid (62.5 mg, 0.47 mmol) was dissolved in 1,4-dioxane (1 mL) and water (3 mL) and cooled to 0 °C. K2CO3 (162 mg, 1.18 mmol) was added, followed by Fmoc-Cl (115 mg, 0.45 mmol). The mixture was stirred at RT overnight and water (10 mL) was added. The mixture was acidified to pH 2-3 with aqueous HCl (1M) and extracted with DCM (2 × 10 mL). The combined organic layers were dried with Na2SO4 and concentrated to dryness to give the product as a white solid (122 mg, 76% yield). C 20 H 19 MS calculated for FNO4: 356.12, Found: 356.22, [M+H] + .

[0243] Intermediate 2 Exatecan mesylate (30 mg, 0.056 mmol), previously prepared intermediate 1 (5 equiv, 100 mg) and DIPEA (200 μL) were dissolved in a 5:1 mixture of DMF and water (3 mL). DMTMM (5 equiv, 78 mg) was added and the reaction mixture was stirred at room temperature for 30 min and directly loaded onto the column for purification. Purification was performed by reversed-phase flash chromatography (25 g, diol-modified C18, 0→70% ACN / H2O). The fractions containing the product were lyophilized from water (35 mg, 81%). C 44 H 39 F2N4O8 + MS calculated: 789.27, Found: 789.43, [M+H] + .

[0244] Intermediate 3 The intermediate 2 prepared above (35 mg, 0.044 mmol) was dissolved in DMF (2 mL) and morpholine (100 μL) was added. The reaction mixture was stirred at room temperature for 1 h. Purification was carried out by reversed-phase HPLC chromatography (25 g, diol-modified C18, 0→50% ACN / H2O). The fractions containing the product were lyophilized from water (19 mg, 76%). C 29 H 29 F2N4O6+ MS calculated: 567.20, Found: 567.57, [M+H] + .

[0245] Intermediate 4: Intermediate 3 (19 mg, 0.035 mmol), Fmoc GGFGG-COOH (2 equiv, 47 mg) and DIPEA (150 μL) were dissolved in a 5:1 mixture of DMF and water (3 mL). DMTMM (2 equiv, 19.4 mg) was added and the reaction mixture was stirred at room temperature for 30 min and directly loaded onto the column for purification. Purification was achieved by reversed-phase flash chromatography (semi-separated, diol-modified C18, 0→50% ACN / H2O). The fractions containing the product were lyophilized from water (32 mg, 80%). C 61 H 60 F2N9O 12 + MS calculated: 1149.43, Found: 1149.25, [M+H] + .

[0246] Intermediate 5: Intermediate 4 (70 mg, 0.061 mmol) prepared above was dissolved in DMF (2 mL) and morpholine (100 μL) was added. The reaction mixture was stirred at room temperature for 1 h. Purification was carried out by reversed-phase HPLC chromatography (semi-separated, diol-modified C18, 0→50% ACN / H2O). The fractions containing the product were lyophilized from water (21 mg, 38%). C 46 H 50 F2N9O 10 + MS calculated value: 926.36, Found value: 926.78, [M+H] + .

[0247] Compound 1022 The previously prepared intermediate 5 (21 mg, 0.023 mmol) was dissolved in 1.5 mL of anhydrous DMF. 2,5-Dioxopyrrolidin-1-yl 3-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)propanoate (1.1 equiv., 9.6 mg) and DIPEA (10 μL) were added and the reaction mixture was stirred at room temperature for 1 h. Purification was achieved by reversed-phase HPLC chromatography (semi-separated, diol-modified C18, ACN 0→50% in water). The fractions containing the product were lyophilized from water (10 mg, 39%). C 54 H 59 FN9O 14 MS calculated: 1122.42, Found: 1122.45, [M+H] + . 1H NMR (500 MHz, DMSO-d6) δ 8.50 (d, J = 8.5 Hz, 1H), 8.29 (t, J = 5.9 Hz, 1H), 8.13 - 8.07 (m, 2H), 7.97 (t, J = 5.7 Hz, 1H), 7.84 - 7.76 (m, 2H), 7.34 (s, 1H), 7.24 (m, 5H), 7.17 (m, 1H), 7.00 (s, 2H), 6.52 (d, J = 4.1 Hz, 1H), 5.55 (dt, J = 8.5, 4.2 Hz, 1H), 5.51 - 5.39 (m, 2H), 5.38 - 5.32 (m, 1H), 5.26 - 5.23 (m, 2H), 4.57 - 4.45 (m, 2H), 4.07 (m, 1H), 3.86 (m, 1H), 3.75 (m, 3H), 3.67 (d, J = 5.6 Hz, 2H), 3.61 (d, J = 5.7 Hz, 1H), 3.55 (m, 4H), 3.46 (t, J = 5.8 Hz, 2H), 3.21 - 3.14 (m, 1H), 3.04 (m, 2H), 2.79 (m, 1H), 2.45 - 2.40 (m, 4H), 2.33 (t, J = 6.6 Hz, 3H), 2.21 (m, 1H), 2.15 (d, J = 5.0 Hz, 1H), 2.11 - 2.04 (m, 1H), 1.87 (m, 2H), 0.89 (t, J = 7.4 Hz, 3H).

[0248] Example 42: Synthesis of Compound 140 JPEG2024518438000152.jpg39159 Compound 140 To a solution of compound 105 trifluoroacetate (1.0 equiv., 14 mg, 0.024 mmol) in TFA formic acid (0.45 ml) was added 37% aqueous formaldehyde (0.12 ml) and the resulting mixture was stirred at 50° C. for 6 h. Then water was added and the mixture was concentrated on a rotary evaporator. The residue was purified by reversed-phase flash chromatography using a semi-separating column (diol-modified C18, 0→60% ACN / 0.1% TFA) to give, after lyophilization, the trifluoroacetate of compound 141 (4.5 mg, 31%) as a white powder. C 27 H 29 MS calculated for FN3O5: 494.21, Found: 494.25, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ: 7.97 - 7.73 (m, 1H), 7.34 (s, 1H), 6.74 - 6.33 (m, 1H), 5.58 - 5.26 (m, 4H), 5.17 - 4.88 (m, 1H), 3.87 - 3.50 (m, 7H), 3.30 - 2.94 (m, 2H), 2.90 - 2.59 (m, 1H), 2.39 (s, 3H), 2.30 - 2.16 (m, 1H), 1.96 - 1.78 (m, 2H), 0.89 (t, J= 7.3 Hz, 3H).

[0249] Example 43: Synthesis of Compound 147 JPEG2024518438000153.jpg39168 Compound 147 To a suspension of exatecan mesylate (1.0 equiv., 40 mg, 0.075 mmol) in DMF (2 mL) under argon atmosphere, diisopropylethylamine (3.5 equiv., 0.26 mmol, 45 μL) and 2-(2-bromoethoxy)ethanol (2.0 equiv., 26 mg) were added and the mixture was heated to 80 °C for 2 days. Purification of the residue by reversed-phase flash chromatography using a semi-preparative column (diol-modified C18, 0→60% ACN / 0.1% TFA) afforded the trifluoroacetate salt of compound 147 (8 mg, 17%) as a yellowish powder after lyophilization. C 28 H 31 MS calculated for FN3O6: 524.22, Found: 524.25, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ: 9.07 (s br, 1H), 8.87 (s br, 1H), 7.88 (d, J = 10.7 Hz, 1H), 7.35 (s, 1H), 6.56 (s, 1H), 5.52 (d, J = 19.1 Hz, 1H), 5.45 (s, 2H), 5.41 (d, J = 19.1 Hz, 1H), 5.09 (s br, 1H), 4.71 (s br, 1H), 3.80 - 3.64 (m, 2H), 3.63 - 3.56 (m, 2H), 3.56 - 3.51 (m, 2H), 3.27 - 3.10 (m, 2H), 2.83 - 2.71 (m, 1H), 2.41 (s, 3H), 2.24 - 2.13 (m, 1H), 1.96 - 1.79 (m, 2H), 0.87 (t, J= 7.3 Hz, 4H).

[0250] Example 44: Synthesis of Compound 148 JPEG2024518438000154.jpg87164Intermediate 1 Glycolic acid (100 mg, 1.316 mmol) was coevaporated with anhydrous pyridine three times and dissolved in 2 mL of anhydrous pyridine under an argon atmosphere. tert-Butyldiphenylsilyl trifluoromethanesulfonate (2 eq., 2.632 mmol, 723 mg, 556 μL) was added and the reaction mixture was stirred at room temperature for 12 h. The reaction mixture was cooled at 0° C. and water (5 mL) was added. The residue was purified by reversed-phase flash chromatography (diol-modified C18, 0→60% ACN / 0.1% TFA) to give intermediate 1 (339 mg, 82%) as a colorless liquid. C 18 H 21 MS calculation for O3Si: 313.12, Measured value: 313.20, [MH] - .

[0251] Intermediate 2 Exatecan mesylate (100 mg, 0.188 mmol), previously prepared intermediate 1 (2 equiv., 0.376 mmol, 118 mg), DMTMM (1 equiv., 0.226 mmol, 62 mg), diisopropylethylamine (100 μL) and water (500 μL) were mixed in 4 mL of DMF and the reaction mixture was stirred at room temperature for 1 h. The residue was directly purified by reversed-phase flash chromatography (diol-modified C18, 0→100% ACN / water) to give intermediate 2 (97 mg, 70%) as a yellow solid. C 42 H 43 MS calculated for FN3O6Si: 732.29, Found: 732.33, [M+H] + .

[0252] Intermediate 3: Intermediate 2 (97 mg, 0.132 mmol) prepared above was coevaporated three times with anhydrous pyridine and dissolved in 5 mL of anhydrous pyridine under an argon atmosphere. tert-Butyldimethyl(chloro)silane (10 equiv., 1.32 mmol, 199 mg) was added and the reaction mixture was stirred at 80 °C for 48 h. The residue was directly purified by reversed-phase flash chromatography (diol-modified C18, 0 → 100% ACN / water) to give Intermediate 3 (53 mg, 47%) as a yellow solid. C48 H 57 MS calculated for FN3O6Si2: 846.38, Found: 846.12, [M+H] + .

[0253] Intermediate 4 Intermediate 3 prepared above (25 mg, 0.0296 mmol) was dissolved in 5 mL of anhydrous toluene and Lawesson's reagent (4 equiv., 0.0592, 24 mg) was added. The reaction mixture was stirred at 100° C. for 4 h. The reaction mixture was allowed to warm to room temperature and the toluene was evaporated under reduced pressure. The crude reaction product was redissolved in 2 mL of DMF and the residue was directly purified by reversed-phase flash chromatography (diol-modified C18, 0→100% ACN / water) to give Intermediate 4 (18 mg, 76%) as a yellow solid. C 48 H 57 MS calculated for FN3O4S2Si2: 878.33, Found: 878.50, [M+H] + .

[0254] Compound 148. Intermediate 4 prepared above (18 mg, 0.0205 mmol) was dissolved in 2 mL of anhydrous dichloromethane and 1 mL of trifluoroacetic acid was added. The reaction mixture was stirred at room temperature for 2 h. The solvent was evaporated under reduced pressure and the crude reaction product was redissolved in 2 mL of DMF and directly purified by reversed-phase flash chromatography (diol-modified C18, 0→100% ACN / water). A second reversed-phase HPLC purification (semi-separate HPLC, diol-modified C18, 0→100% ACN in water) gave a 1:1 mixture of two inseparable isomers, with both forms in equilibrium (7 mg, 65%, yellow solid). C 26 H 25 MS calculated for FN3O4S2: 526.13, found: 526.15, [M+H] + . 1H NMR (500 MHz, DMSO-d6, mixture of two isomers), δ 10.51 (m, 1H), 7.85 (d, J = 10.8 Hz, 1H), 7.81 (d, J = 2.9 Hz, 1H), 6.69 (d, J = 9.1 Hz, 1H), 6.50 (q, J = 9.4, 8.2 Hz, 1H), 6.05 (m, 1H), 5.91 (m, 1H), 5.51 (m, 1H), 5.47 - 5.31 (m, 2H), 4.60 - 4.44 (m, 2H), 3.16 (t, J = 13.2 Hz, 1H), 2.51 (q, J = 1.9 Hz, 2H), 2.40 (s, 3H), 2.28 (m, 1H), 1.96 - 1.83 (m, J = 7.1 Hz, 2H), 0.96 - 0.84 (m, 3H).

[0255] Example 45: Synthesis of Compound 159 JPEG2024518438000155.jpg35162 Compound 159 To a mixture of exatecan mesylate (1.0 equiv., 20 mg, 0.038 mmol), (3-hydroxyoxetan-3-yl)carboxylic acid (1.25 equiv., 6 mg, 0.048 mmol) and HATU (2.0 equiv., 29 mg, 0.076 mmol), dry DMF (1.5 mL) and diisopropylethylamine (4.0 equiv., 0.152 mmol, 27 μL) were added, and the resulting solution was stirred at room temperature under argon atmosphere for 1 h. The mixture was directly purified by reversed-phase flash chromatography using a semi-separated column (diol-modified C18, 0→70% ACN in water) to give the product (17 mg, 84%) as a white powder after lyophilization. C 28 H 27 MS calculated for FN3O7: 536.18, Found: 536.20, [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ: 8.52 (d, J = 9.1 Hz, 1H), 7.68 (d, J = 10.8 Hz, 1H), 7.27 (s, 1H), 6.94 (s, 1H), 6.51 (s, 1H), 5.60 (q, J = 7.6 Hz, 1H), 5.47 - 5.33 (m, 2H), 5.04 (d, J= 18.8 Hz, 1H), 4.99 (d, J = 6.5 Hz, 1H), 4.94 - 4.83 (m, 2H), 4.58 (d, J= 6.5 Hz, 1H), 4.52 (d, J = 6.3 Hz, 1H), 3.27 - 3.16 (m, 1H), 3.16 - 3.02 (m, 1H), 2.34 (s, 3H), 2.23 - 2.13 (m, 2H), 1.93 - 1.78 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0256] Example 46: Synthesis of Compound 163 JPEG2024518438000156.jpg37170Intermediate 1 To a suspension of exatecan mesylate (1.0 equiv, 30 mg, 0.056 mmol) in DMF (1.2 mL) under argon atmosphere was added diisopropylethylamine (4.5 equiv, 0.25 mmol, 45 μL) and (2-bromoethoxy)-tert-butyldimethylsilane (3.3 equiv, 0.19 mmol, 45 mg, 40 μL) and the mixture was heated to 80° C. for 3 days. Purification by reversed-phase flash chromatography (25 g, diol-modified C18, 0→60% ACN in water) afforded the product intermediate 1 (13 mg, 39%) as an off-white powder after lyophilization.

[0257] Compound 163 Acetic formic anhydride (2.2 equiv., 0.048, 4.2 μL) (Huffman, CWJ Org. Chem. 1958, 23 (5), 727-729.) and diisopropylethylamine (2.5 equiv., 0.055 mmol, 10 μL) were added to a solution of intermediate 1 (1.0 equiv., 13 mg, 0.022 mmol) in DCM (1.5 ml) and the resulting mixture was stirred at room temperature for 2 h. The volatiles were then removed on a rotary evaporator and the residue was purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→80% ACN in water). The fractions containing the TBS-protected product were combined, acidified with TFA (50 μL) and evaporated to dryness (2x). Purification of the residue by reversed-phase flash chromatography using a semi-preparative column (diol-modified C18, 0→70% ACN in water) afforded the product (6 mg, 54%) as a white powder after lyophilization. 27 H 27 MS calculated for FN3O6: 508.19, Found: 508.20, [M+H] + . 1 H NMR (400 MHz, DMSO-d6, mixture of isomers) δ: 8.28 (s, 0.55H), 8.17 (s, 0.45H), 7.78 (dd, J = 13.3, 10.8 Hz, 1H), 7.30 (s, 1H), 6.51 (s, 1H), 5.59 - 5.47 (m, 1H), 5.41 (s, 3H), 5.25 (d, J = 18.8 Hz, 0.45H), 5.17 - 5.00 (m, 1H), 4.97 - 4.87 (m, 1H), 4.77 (t, J = 5.2 Hz, 0.55H), 3.68 - 3.38 (m, 2H), 3.28 - 2.96 (m, 2H), 2.46 - 2.17 (m, 4H), 1.98 - 1.77 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H).

[0258] Example 47: Synthesis of Compound 164 JPEG2024518438000157.jpg67170Intermediate 1 Exatecan mesylate (30 mg, 0.056 mmol), N-(Fmoc)-2-aminoacetaldehyde (24 mg, 0.085 mmol) and DIPEA (40 μL) were dissolved in dry DMF (2 mL). The mixture was stirred at 60 °C for 1 h. NaBH3CN (30 mg, 0.47 mmol) was then added and the reaction mixture was stirred at 60 °C for 2 h. The mixture was directly purified by reversed-phase flash chromatography (25 g, diol-modified C18, ACN 0 → 70% in water). The fractions containing the product were lyophilized from water (28 mg, 72%). C 41 H 38 FN4O6 + MS calculated value: 701.27, Found value: 701.43, [M+H] + .

[0259] Intermediate 2 The previously prepared intermediate 1 (28 mg, 0.04 mmol) was dissolved in DMF (2 mL) and morpholine (100 μL) was added. The reaction mixture was stirred at room temperature for 1 h. Purification was achieved by reversed-phase HPLC chromatography (25 g, diol-modified C18, 0→50% ACN in water (TFA)). The fractions containing the product were lyophilized from water (14 mg, 76%). 26 H 28 FN4O4 + MS calculated: 479.20, Found: 479.32, [M+H] + .

[0260] Intermediate 3 Intermediate 2 (17 mg, 0.036 mmol) prepared above and N-(Fmoc)-2-aminoacetaldehyde (5 mg, 0.018 mmol) were dissolved in dry DMF (2 mL). The mixture was stirred at 60 °C for 1 h. NaBH3CN (10 mg, 0.16 mmol) was then added and the reaction mixture was stirred at 60 °C for 2 h. The mixture was directly purified by reversed-phase flash chromatography (25 g, diol-modified C18, 0→70% ACN in water). The fractions containing the product were lyophilized from water (10 mg, 37%). C 43 H43 FN5O6 + MS calculated: 744.31, Found: 744.45, [M+H] + .

[0261] Compound 164: The intermediate 3 prepared above (10 mg, 0.013 mmol) was dissolved in DMF (2 mL) and morpholine (100 μL) was added. The reaction mixture was stirred at room temperature for 1 h. Purification was achieved by reversed-phase HPLC chromatography (semi-separated, diol-modified C18, 0→50% ACN in water (TFA)). The fractions containing the product were lyophilized from water (3 mg, 44%). 28 H 33 FN5O4 + MS calculated: 522.24, Found: 522.30, [M+H] + . 1 H NMR (500 MHz, DMSO-d6) δ 7.81 (d, J = 10.8 Hz, 1H), 7.35 (s, 1H), 6.57 (s, 2H), 5.45 (s, 2H), 4.46 (s, 1H), 3.90 (m, 1H), 3.77 (m 1H), 3.18 - 3.14 (m, 2H), 3.12 (d, J= 10.2 Hz, 4H), 3.04 - 2.96 (m, 4H), 2.41 (s, 3H), 2.08 (m, 1H), 1.88 (m, 2H), 0.88 (t, J = 7.3Hz, 3H).

[0262] Example 48: Synthesis of Compound 166 JPEG2024518438000158.jpg45153Exatecan mesylate (30 mg, 0.056 mmol), 5-hydroxy-1H-pyrazole-3-carboxylic acid (11 mg, 0.084 mmol), EDC (22 mg, 0.115 mmol), HOBt (18 mg, 0.117 mmol) and diisopropylamine (30 μL) were added to DMF (2 mL). The resulting solution was stirred at room temperature under argon for 16 h. The mixture was directly purified by reversed-phase HPLC chromatography using a semi-separated column (diol-modified C18, ACN 0→100% in water). After lyophilization from water, the desired product was obtained as a yellow powder (7 mg, 23%). C 28 H 25 FN5O6: MS calculated: 546.17, Found: 546.25, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 10.15 (s, 1H), 8.79 (d, J = 8.7 Hz, 1H), 7.80 (d, J = 10.8 Hz, 1H),7.31 (s, 1H), 6.68 (s, 1H), 6.02 (s, 1H), 5.72 (q, J = 6.6 Hz, 1H), 5.39 (s, 2H), 5.17 (s, 2H), 3.27 (m, 1H), 3.19 - 3.10 (m, 1H), 2.41 (d, J= 1.9 Hz, 3H), 2.25 (m, 2H), 1.94 - 1.81 (m, 2H), 1.77 (s, 1H), 0.87 (t, J= 7.3 Hz, 3H).

[0263] Example 49: Synthesis of Compound 167 JPEG2024518438000159.jpg421594:1To a DMF / water mixture (4 mL) was added exatecan mesylate (20 mg, 0.038 mmol), 2-(hydroxymethyl)oxazole-4-carboxylic acid (1.25 equiv., 7 mg, 0.048 mmol), DMTMM (2.0 equiv., 21 mg, 0.076 mmol) and diisopropylethylamine (20 μL). The resulting solution was stirred at room temperature for 1 h, when LC-MS showed complete consumption of starting material. The mixture was directly purified by reversed-phase HPLC chromatography using a semi-separated column (diol-modified C18, ACN 0→100% in water). After lyophilization from water, the desired product was obtained as a white powder (17 mg, 80%). C 29 H 26 MS calculated for FN4O7: 561.18, found: 561.20, [M+H] + .1H NMR (400 MHz, DMSO-d6) δ 9.01 (d, J = 8.8 Hz, 1H), 8.71 (s, 1H), 7.76 (d, J = 10.9 Hz, 1H), 7.30 (s, 1H), 6.50 (s, 1H), 5.78 (t, J = 6.2 Hz, 1H), 5.73 - 5.64 (m, 1H), 5.37 (s, 2H), 5.10 (s, 2H), 4.54 (d, J = 6.2 Hz, 2H), 3.98 (s, 1H), 3.30 - 3.23 (m, 1H), 3.17 - 3.09 (m, 1H), 2.38 (d, J = 1.9 Hz, 3H), 2.33 - 2.19 (m, 1H), 1.95 - 1.80 (m, 2H), 0.86 (t, J = 7.3 Hz, 3H).

[0264] Example 50: Synthesis of Compound 168 JPEG2024518438000160.jpg471654:1To a DMF / water mixture (4 mL) was added exatecan mesylate (20 mg, 0.038 mmol), 5-(hydroxymethyl)-1H-pyrazole-3-carboxylic acid (1.25 equiv., 7 mg, 0.048 mmol), DMTMM (2.0 equiv., 21 mg, 0.076 mmol) and diisopropylamine (20 μL). The resulting solution was stirred at room temperature under argon for 1 h, when LC-MS showed complete consumption of starting material. The mixture was directly purified by reversed-phase HPLC chromatography using a semi-separated column (diol-modified C18, ACN 0→100% in water). After lyophilization from water, the desired product was obtained as a white powder (14 mg, 66%). C 29 H 27 MS calculated for FN5O6: 560.19, found: 560.30, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 13.25 - 13.20 (m, 1H), 8.85 (d, J = 8.9 Hz, 1H), 7.76 (d, J= 10.9 Hz, 1H), 7.29 (s, 1H), 6.62 (d, J = 1.9 Hz, 1H), 6.51 (d, J= 3.4 Hz, 1H), 5.69 (q, J = 7.6 Hz, 1H), 5.38 (d, J = 12.2 Hz, 2H), 5.28 - 5.01 (m, 3H), 4.54 (d, J = 5.6 Hz, 2H), 3.29 - 3.01 (m, 1H), 2.43 - 2.36 (m, 4H), 2.29 - 2.22 (m, 2H), 1.95 - 1.74 (m, 2H), 0.86 (t, J= 7.3 Hz, 3H).

[0265] Example 51: Synthesis of Compound 175 JPEG2024518438000161.jpg89170Intermediate 1 To a solution of compound 11 (1.0 equiv, 52 mg, 0.097 mmol) and imidazole (2.5 equiv, 0.243 mmol, 17 mg) in DMF (2 mL) was added tert-butyl(chloro)diphenylsilane (2.5 equiv, 0.243 mmol, 68 mg, 63 μL) under argon atmosphere and the mixture was stirred at room temperature for 4 h. Purification by reversed-phase flash chromatography (25 g, diol-modified C18, ACN 0→80% in water) afforded the product intermediate 1 (61 mg, 81%) as an off-white powder after lyophilization.

[0266] Intermediate 2. To a solution of intermediate 1 (1.0 equiv., 61 mg, 0.078 mmol) in dry pyridine (3 mL) under argon atmosphere, tert-butyldimethylsilyl trifluoromethanesulfonate (14.0 equiv., 1.09 mmol, 287 mg, 250 μL) was added and the mixture was heated to 80 °C for 5 h. It was then concentrated on a rotary evaporator and the residue was purified by reversed-phase flash chromatography (25 g, diol-modified C18, ACN in water 0→100%) to give the product intermediate 2 (56 mg, 81%) as a white powder after lyophilization.

[0267] Compound 175. To a mixture of intermediate 2 (1.0 equiv., 56 mg, 0.063 mmol) and Lawesson's reagent (5.0 equiv., 0.316 mmol, 128 mg), anhydrous toluene (8 ml) was added and the resulting mixture was heated to 100 °C for 6 h. The reaction progress was monitored by LC-MS. Further Lawesson's reagent was added (2.0 equiv., 0.126 mmol, 52 mg) and the mixture was heated to 100 °C for another 3 h. The volatiles were then removed on a rotary evaporator and the residue was purified by reversed-phase flash chromatography (25 g, diol-modified C18, ACN 0→100% in water). The fractions containing intermediate 3 were combined and concentrated on a rotary evaporator. The residue was dissolved in dioxane (2 ml), followed by the addition of water (1 ml) and trifluoroacetic acid (1 ml). The resulting mixture was stirred at room temperature for 40 h and the reaction progress was monitored by LC-MS. The solvent was then removed by rotary evaporation and the residue was purified by reversed-phase flash chromatography using a semi-preparative column (diol-modified C18, 0→70% ACN in water) to give the product (12 mg, 34%) as a yellow powder after lyophilization. 29 H 29 MS calculated for FN3O4S2: 566.16, Found: 566.20, [M+H] + . 1H NMR (500 MHz, DMSO-d6) δ: 10.62 (dd, J = 15.6, 8.6 Hz, 1H), 7.85 (d, J = 10.8 Hz, 1H), 7.80 (d, J = 2.8 Hz, 1H), 6.68 (s, 1H), 6.53 - 6.41 (m, 1H), 5.91 (dd, J = 16.6, 3.3 Hz, 1H), 5.55 - 5.45 (m, 1H), 5.46 - 5.30 (m, 2H), 3.71 (dd, J = 11.2, 4.6 Hz, 1H), 3.50 (dd, J = 11.5, 5.3 Hz, 1H), 3.31 - 3.10 (m, 2H), 2.40 (s, 3H), 2.36 - 2.17 (m, 1H), 2.17 - 2.06 (m, 1H), 2.04 - 1.82 (m, 3H), 1.54 - 1.34 (m, 1H), 1.10 - 0.93 (m, 1H), 0.86 (t, J = 7.3 Hz, 3H).

[0268] Example 52: Synthesis of Compound 176 JPEG2024518438000162.jpg88170Intermediate 1 Glycolic acid (100 mg, 1.316 mmol) was coevaporated with anhydrous pyridine three times and dissolved in 2 mL of anhydrous pyridine under an argon atmosphere. tert-Butyldiphenylsilyl trifluoromethanesulfonate (2 eq., 2.632 mmol, 723 mg, 556 μL) was added and the reaction mixture was stirred at room temperature for 12 h. The reaction mixture was cooled at 0° C. and water (5 mL) was added. The residue was purified by reversed-phase flash chromatography (diol-modified C18, 0→60% ACN / 0.1% TFA) to give intermediate 1 (339 mg, 82%) as a colorless liquid. C 18 H 21 MS calculation for O3Si: 313.12, Measured value: 313.20, [MH] - .

[0269] Intermediate 2 Exatecan mesylate (100 mg, 0.188 mmol), previously prepared intermediate 1 (2 equiv., 0.376 mmol, 118 mg), DMTMM (1.2 equiv., 0.226 mmol, 62 mg), diisopropylethylamine (100 μL) and water (500 μL) were mixed in 4 mL of DMF and the reaction mixture was stirred at room temperature for 1 h. The residue was directly purified by reversed-phase flash chromatography (diol-modified C18, 0→100% ACN / water) to give intermediate 2 (97 mg, 70%) as a yellow solid. C 42 H 43 MS calculated for FN3O6Si: 732.29, Found: 732.33, [M+H] + .

[0270] Intermediate 3: Intermediate 2 (97 mg, 0.132 mmol) prepared above was coevaporated three times with anhydrous pyridine and dissolved in 5 mL of anhydrous pyridine under an argon atmosphere. tert-Butyldimethyl(chloro)silane (10 equiv., 1.32 mmol, 199 mg) was added and the reaction mixture was stirred at 80 °C for 48 h. The residue was directly purified by reversed-phase flash chromatography (diol-modified C18, 0 → 100% ACN / water) to give Intermediate 3 (53 mg, 47%) as a yellow solid. C 48 H 57 MS calculated for FN3O6Si2: 846.38, Found: 846.12, [M+H] + .

[0271] Intermediate 4: Intermediate 3 (25 mg, 0.0296 mmol) prepared above was dissolved in 5 mL of anhydrous toluene and Lawesson's reagent (1 equiv., 0.0296, 6 mg) was added. The reaction mixture was stirred at 100° C. for 4 h. The reaction mixture was allowed to warm to room temperature and the toluene was evaporated under reduced pressure. The crude reaction product was redissolved in 2 mL of DMF and the residue was directly purified by reversed-phase flash chromatography (diol-modified C18, 0→100% ACN / water) to give Intermediate 4 (22 mg, 86%) as a yellow solid. C 48 H 57MS calculated for FN3O5SSi2: 862.35, Found: 862.40, [M+H] + .

[0272] Compound 176. Intermediate 4 prepared above (22 mg, 0.0255 mmol) was dissolved in 1 mL of anhydrous dichloromethane and 2 mL of trifluoroacetic acid was added. The reaction mixture was stirred at room temperature for 16 h. The solvent was evaporated under reduced pressure and the crude reaction product was redissolved in 1 mL of DMF and directly purified by reversed-phase flash chromatography (diol-modified C18, 0→80% ACN / water). A second reversed-phase HPLC purification (semi-separate HPLC, diol-modified C18, 0→100% ACN in water) afforded compound 177 as a 1:1 mixture of two inseparable isomers, with both forms in equilibrium (11 mg, 84%, orange solid). 26 H 25 MS calculated for FN3O5S: 510.15, Found: 510.15, [M+H] + . 1 H NMR (401 MHz, DMSO-d6) δ 10.40 (d, J = 8.8 Hz, 1H), 7.81 (d, J = 10.9 Hz, 1H), 7.32 (s, 1H), 6.53 (s, 1H), 6.44 - 6.38 (m, 1H), 5.98 (t, J = 5.8 Hz, 1H), 5.42 (s, 2H), 5.12 (d, J = 3.4 Hz, 2H), 4.40 (d, J = 5.8 Hz, 2H), 3.23 - 3.13 (m, 2H), 2.41 (d, J = 1.9 Hz, 3H), 2.30 - 2.19 (m, 1H), 1.86 (dq, J = 14.2, 7.1 Hz, 2H), 1.24 (s, 1H), 0.87 (t, J = 7.3 Hz, 3H).

[0273] Example 53: Synthesis of Compound 188 JPEG2024518438000163.jpg46153To a 4:1 DMF / water mixture (4 mL), exatecan mesylate (20 mg, 0.0376 mmol), 3-(1,3-dioxolan-2-yl)propanoic acid (2 equiv., 0.0753 mmol, 11 mg), DMTMM (1.5 equiv., 0.0564 mmol, 16 mg) and diisopropylethylamine (20 μL) were added. The resulting solution was stirred at room temperature for 1 h, when LC-MS showed complete consumption of starting material. The mixture was directly purified by reversed-phase HPLC chromatography using a semi-separated column (diol-modified C18, ACN 0→100% in water). After lyophilization, the desired product was obtained as a white powder (19 mg, 89%). C 30 H 31 MS calculated for FN3O7: 564.21, Found: 564.34, [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.46 (d, J = 8.7 Hz, 1H), 7.78 (d, J = 10.9 Hz, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.56 (dt, J = 9.3, 5.0 Hz, 1H), 5.43 (s, 2H), 5.31 - 5.11 (m, 2H), 4.83 (t, J = 4.5 Hz, 1H), 3.89 - 3.80 (m, 2H), 3.79 - 3.69 (m, 2H), 3.24 - 3.10 (m, 2H), 2.40 (d, J = 1.9 Hz, 3H), 2.26 (t, J= 7.6 Hz, 2H), 2.14 (q, J = 7.3, 6.6 Hz, 2H), 1.98 - 1.77 (m, 4H), 0.88 (t, J = 7.3 Hz, 3H).

[0274] Example 54: Synthesis of Compound 1002 Step 1: JPEG2024518438000164.jpg32170 Intermediate 1 is dissolved in anhydrous DMF in a magnetically stirred flask and copper(II) acetate and lead tetraacetate are added. The flask is heated in a 60 °C oil bath for 20 min. The oil bath is removed and the reaction mixture is cooled to room temperature. The mixture is purified on a C18 RP column to give 260 Intermediate 2. Step 2: JPEG2024518438000165.jpg30170

[0275] Intermediate 2 and benzyl 3-hydroxypropionate are suspended in a cold solution of 20% TFA in dichloromethane and stirred at room temperature for 60 min. The solvent is evaporated and the residue is purified on a C18 RP to give intermediate 3. Step 3: JPEG2024518438000166.jpg30170

[0276] To a stirred solution of intermediate 3 in DMF is added morpholine. After 1.5 h, the reaction mixture is loaded onto a C18 RP column and eluted to give intermediate 4. Step 4: JPEG2024518438000167.jpg26170

[0277] To a solution of intermediate 4 in 5:95 deionized water:methanol (35 mL) is added 10% palladium on carbon (0.09 g). The mixture is hydrogenated at 30 PSI H2 for 80 minutes, filtered, and evaporated in vacuo to give intermediate 5. Step 5: JPEG2024518438000168.jpg32170

[0278] To a stirred solution of intermediate 5 in anhydrous DMF is added DIPEA and Mal-PEG1-NHS ester. The mixture is stirred at room temperature for 30 min and applied to a C18 RP column for elution to give intermediate 6. Step 6: JPEG2024518438000169.jpg33170

[0279] Exatecan mesylate DMTMM, triethylamine and intermediate 6 in 20% DMF / water solution are stirred for 30 minutes at 37° C. The reaction mixture is cooled to room temperature and purified by silica gel column chromatography to give compound 1002. Step 7: JPEG2024518438000170.jpg33170

[0280] To a 5 mg / ml solution of anti-huTrop2 antibody, TCEP in 50 mM EPPS (pH 7.4, containing 5 mM EDTA) is added. After stirring for 90 min at 37 °C, the mixture is cooled to ~25 °C and the conjugation reaction is initiated by adding 12 equivalents of compound 1002 dissolved in a ~40 °C solution of 50 mM EPPS (pH 7.4, containing 20% ​​DMSO). The reaction mixture is stirred at room temperature for 2 h, then mixed with 10 mM acetic acid, 10% sucrose, 0.01% Tween-20 pH 5.0 and concentrated. The resulting mixture is purified by size exclusion column chromatography to give compound 2000.

[0281] Example 55: Synthesis of Compound 1003 Step 1: JPEG2024518438000171.jpg29170 A suspension of intermediate 2 from Example 54 and 2,2-difluoro-3-((2-hydroxyethyl)amino)-3-oxopropanoic acid in 20% TFA / DCM is stirred at room temperature for 60 min. The solvent is evaporated and the residue is purified by reverse phase column chromatography to give intermediate 9. Step 2: JPEG2024518438000172.jpg38170

[0282] Exatecan mesylate, DMTMM, triethylamine and intermediate 9 in 20% DMF / water solution are stirred for 30 minutes at 35° C. The reaction mixture is purified by silica gel column chromatography to give intermediate 10. Step 3: JPEG2024518438000173.jpg42170

[0283] To a stirred solution of intermediate 10 in DMF is added morpholine. After 1.5 h, the reaction mixture is loaded onto a C18 RP column and eluted to give intermediate 11. Step 4: JPEG2024518438000174.jpg32165

[0284] To a stirred solution of intermediate 11 in anhydrous DMF is added DIPEA and Mal-PEG1-NHS ester. The mixture is stirred at RT for 30 min and purified by reverse phase column chromatography to give compound 1003. Step 5: JPEG2024518438000175.jpg35170

[0285] To a 5 mg / ml solution of anti-huTrop2 antibody, TCEP in 50 mM EPPS (pH 7.4, containing 5 mM EDTA) is added. After stirring for 90 min at 37 °C, the mixture is cooled to ~25 °C and the conjugation reaction is initiated by adding 12 equivalents of compound 1003 dissolved in a ~40 °C solution of 50 mM EPPS (pH 7.4, containing 20% ​​DMSO). The reaction mixture is stirred at room temperature for 2 h, then mixed with 10 mM acetic acid, 10% sucrose, 0.01% Tween-20 pH 5.0 and concentrated. The resulting mixture is purified by size exclusion column chromatography to give compound 2001.

[0286] Example 56: Synthesis of Compound 1004 Step 1: JPEG2024518438000176.jpg26170 A suspension of intermediate 2 from example 54 and benzyl 2,2-difluoro-3-hydroxypropanoate is stirred at room temperature for 60 min. The solvent is evaporated and the residue is purified by reverse phase column chromatography to give intermediate 14. Step 2: JPEG2024518438000177.jpg28170

[0287] To a solution of intermediate 14 in 5:95 deionized water:methanol (35 mL) is added 10% palladium on carbon (0.09 g). The mixture is hydrogenated at 30 PSI H2 for 80 minutes, filtered, and evaporated in vacuo to give intermediate 15. Step 3: JPEG2024518438000178.jpg26170

[0288] Exatecan mesylate, DMTMM, triethylamine and intermediate 15 in 20% DMF / water solution are stirred for 30 minutes at 35° C. The reaction mixture is purified by silica gel column chromatography to give intermediate 16. Step 4: JPEG2024518438000179.jpg28162

[0289] To a stirred solution of intermediate 16 in DMF is added morpholine. After 1.5 h, the reaction mixture is loaded onto a C18 RP column and eluted to give intermediate 17. Step 5: JPEG2024518438000180.jpg27167

[0290] To a stirred solution of intermediate 17 in anhydrous DMF is added DIPEA and Mal-PEG1-NHS ester. The mixture is stirred at RT for 30 min and purified by reverse phase column chromatography to give compound 1004. Step 6: JPEG2024518438000181.jpg27167

[0291] To a 5 mg / ml solution of anti-huTrop2 antibody, TCEP in 50 mM EPPS (pH 7.4, containing 5 mM EDTA) is added. After stirring for 90 min at 37°C, the mixture is cooled to ~25°C and the conjugation reaction is initiated by adding 12 equivalents of compound 1004 dissolved in a ~40°C solution of 50 mM EPPS (pH 7.4, containing 20% ​​DMSO). The reaction mixture is stirred at room temperature for 2 h, then mixed with 10 mM acetic acid, 10% sucrose, 0.01% Tween-20 pH 5.0 and concentrated. The resulting mixture is purified by size exclusion column chromatography to give compound 2002.

[0292] Example 57: Synthesis of Compound 1020 Step 1: JPEG2024518438000182.jpg33170 Exatecan mesylate, DMTMM, triethylamine and intermediate 25 in 20% DMF / water solution are stirred at 35° C. for 30 minutes. The reaction mixture is purified by silica gel column chromatography to give compound 1020.

[0293] Step 2: JPEG2024518438000183.jpg34170 To a 5 mg / ml solution of anti-huTrop2 antibody, TCEP in 50 mM EPPS (pH 7.4, containing 5 mM EDTA) is added. After stirring for 90 min at 37 °C, the mixture is cooled to ~25 °C and the conjugation reaction is initiated by adding 12 equivalents of compound 1020 dissolved in a ~40 °C solution of 50 mM EPPS (pH 7.4, containing 20% ​​DMSO). The reaction mixture is stirred at room temperature for 2 h, then mixed with 10 mM acetic acid, 10% sucrose, 0.01% Tween-20 pH 5.0 and concentrated. The resulting mixture is purified by size exclusion column chromatography to give compound 2003.

[0294] Example 58: Synthesis of Compound 1021 Step 1: JPEG2024518438000184.jpg29170 Exatecan mesylate, DMTMM, triethylamine and intermediate 27 in 20% DMF / water solution are stirred at 35° C. for 30 minutes. The reaction mixture is purified by silica gel column chromatography to give compound 1021.

[0295] Step 2: JPEG2024518438000185.jpg35170To a 5 mg / ml solution of anti-huTrop2 antibody, TCEP in 50 mM EPPS (pH 7.4, containing 5 mM EDTA) is added. After stirring for 90 min at 37 °C, the mixture is cooled to ~25 °C and the conjugation reaction is initiated by adding 12 equivalents of compound 1020 dissolved in a ~40 °C solution of 50 mM EPPS (pH 7.4, containing 20% ​​DMSO). The reaction mixture is stirred at room temperature for 2 h, then mixed with 10 mM acetic acid, 10% sucrose, 0.01% Tween-20 pH 5.0 and concentrated. The resulting mixture is purified by size exclusion column chromatography to give compound 2004.

[0296] Example 59: Synthesis of Compound 1006 Step 1: Intermediate 12 and chloroethanol are suspended in 20% TFA in dichloromethane (12 mL) and stirred at room temperature for 60 min. The solvent is evaporated and the residue is purified by reverse phase column chromatography to give intermediate 45.

[0297] Step 2: Powdered potassium thiol acetate is added to a solution of intermediate 45 and tetrabutylammonium iodide in DMF stirred at 0 °C. The reaction mixture is gradually warmed to room temperature, rt, and stirred overnight. The reaction mixture is quenched with water and the residue is purified by reverse phase column chromatography to give intermediate 46.

[0298] Step 3: JPEG2024518438000188.jpg50170To a solution of intermediate 46 in acetonitrile at 10°C, a solution of NCS in 4:1 CH3CN / 2N HCl is added dropwise. The reaction mixture is stirred at 10°C for 2 hours and dried over molecular sieves at 10°C overnight. The solvent is evaporated and the residue is diluted with dry acetonitrile and evaporated again. After three cycles of dilution and evaporation, intermediate 47 is dissolved in dry THF, cooled in an ice bath and a solution of exatecan and triethanolamine in 5 ml of dry ethyl acetate is added. The reaction mixture is stirred at ice bath temperature for 2 hours and then at room temperature overnight. The solvent is evaporated and the residue is purified by reverse phase column chromatography to give intermediate 48.

[0299] Step 4: To a stirred solution of intermediate 16 in DMF is added morpholine. After 1.5 h, the reaction mixture is loaded onto a C18 RP column and eluted to give intermediate 17.

[0300] Step 5: To a stirred solution of intermediate 49 in anhydrous DMF is added DIPEA and Mal-PEG1-NHS ester. The mixture is stirred at RT for 30 min and purified by reverse phase column chromatography to give compound 1006.

[0301] Step 6: JPEG2024518438000191.jpg26168To a 5 mg / ml solution of anti-huTrop2 antibody, TCEP in 50 mM EPPS (pH 7.4, containing 5 mM EDTA) is added. After stirring for 90 min at 37 °C, the mixture is cooled to ~25 °C and the conjugation reaction is initiated by adding 12 equivalents of compound 1006 dissolved in a ~40 °C solution of 50 mM EPPS (pH 7.4, containing 20% ​​DMSO). The reaction mixture is stirred at room temperature for 2 h, then mixed with 10 mM acetic acid, 10% sucrose, 0.01% Tween-20 pH 5.0 and concentrated. The resulting mixture is purified by size exclusion column chromatography to give compound 2005.

[0302] Example 60: Synthesis of Compound 1009 Step 1: JPEG2024518438000192.jpg26161 Intermediate 2 from the propargyl alcohol of Example 54 (192.6 mg, 3.44 mmol) is suspended in a cold solution of 20% TFA in dichloromethane and stirred at room temperature for 60 min. The solvent is evaporated and the residue is purified by reverse phase column chromatography to give intermediate 66.

[0303] Step 2: JPEG2024518438000193.jpg31148 To a solution of exatecan in methanol was added imidazole-1-sulfonyl azide, HCl salt, K2CO3 and copper sulfate pentahydrate dissolved in water. The reaction mixture was stirred at 37° C. for 18 hours and the residue was purified by column chromatography to give intermediate 67.

[0304] Step 3: JPEG2024518438000194.jpg41170 To a suspension of azide 67 in DMSO is added intermediate compound 66. Bis(triphenylphosphine)copper(I) acetate and BTTAA are added and the reaction mixture is stirred at room temperature for 2 h and at 40° C. for an additional 5 h. The solvent is evaporated and the residue is purified by reverse phase column chromatography to give intermediate 68.

[0305] Step 4: JPEG2024518438000195.jpg39168 To a stirred solution of intermediate 68 in DMF is added morpholine. After 1.5 h, the reaction mixture is loaded onto a C18 RP column and eluted to give intermediate 69.

[0306] Step 5: JPEG2024518438000196.jpg35168 To a stirred solution of intermediate 69 in anhydrous DMF is added DIPEA and Mal-PEG1-NHS ester. The mixture is stirred at RT for 30 min and purified by reverse phase column chromatography to give compound 1009.

[0307] Example 61: Synthesis of Compound 1011 Step 1: JPEG2024518438000197.jpg35168 Chloro(pentamethylcyclopentadienyl)(cyclooctadiene)ruthenium(II), azide 67 and intermediate 66 are dissolved in acetonitrile. The reaction mixture is stirred at ambient temperature for 2 h and then at 40 °C for an additional 4 h. The solvent is evaporated and the residue is purified by reverse phase column chromatography to give intermediate 71.

[0308] Step 2: To a stirred solution of intermediate 68 in DMF is added morpholine. After 1.5 h, the reaction mixture is loaded onto a C18 RP column and eluted to give intermediate 72.

[0309] Step 3: JPEG2024518438000199.jpg26170 To a stirred solution of intermediate 69 in anhydrous DMF is added DIPEA and Mal-PEG1-NHS ester. The mixture is stirred at RT for 30 min and purified by reverse phase column chromatography to give compound 1011.

[0310] Example 62: Synthesis of Compound 1023 JPEG2024518438000200.jpg25170 Intermediate 2 To a solution of compound 1 (350 mg, mmol) in DMF (3 mL) were added Cu(OAc)2, AcOH and Pb(OAc)4 in that order. The reaction mixture was stirred at 60° C. for 40 min and then cooled at room temperature. The crude reaction mixture was directly loaded onto a C18 column (40 g C18) and eluted with a gradient of ACN in water (0→60% ACN in water). Fractions containing the desired product were combined and the solvent was partially evaporated under reduced pressure to a final volume of about 10 mL and lyophilized. The desired product was obtained as a white powder (285 mg, HPLC purity 80%). C 33 H 35 N5NaO8: 652.24, Measured value: 652.42 [M+Na] + . JPEG2024518438000201.jpg25170

[0311] Intermediate 3 Intermediate 2 (150 mg, 0.238 mmol) was dissolved in DMF (2 mL). Azidoethanol (2 eq., 0.457 mmol, 40 mg) was added followed by 100 μL of a 2M solution of HCl in dioxane. The reaction mixture was stirred at room temperature for 1 h. The crude reaction mixture was directly loaded onto a C18 column (40 g C18) and eluted with a gradient of ACN in water (0→60% ACN in water). Fractions containing the desired product were combined and the solvent was partially evaporated under reduced pressure to a final volume of approximately 10 mL and lyophilized. The desired product was obtained as a white powder (94 mg, HPLC purity 65%). C 33 H 36 MS calculated for N8NaO7: 679.26, Found: 679.28 [M+Na] + . JPEG2024518438000202.jpg168170

[0312] Intermediate 4 To a solution of intermediate 3 (1.0 equiv, 26 mg, 0.040 mmol) in dioxane (1.5 ml) was added Pd / C (10% w / w, 5 mg) and the resulting suspension was hydrogenated and the hydrogenation reaction mixture was hydrogenated (balloon) at room temperature for 1.5 h. LC-MS analysis confirmed complete consumption of starting material. The suspension was filtered to give the desired product as a solution in dioxane. C 33 H 39 MS calculated for N6O7: 631.29, found: 631.30, [M+H] + .

[0313] Intermediate 6 To a mixture of exatecan mesylate (1.0 equiv, 20 mg, 0.037 mmol) and succinic anhydride (1.1 equiv, 4.1 mg, 0.041 mmol), DMF (0.5 mL) and diisopropylethylamine (2.2 equiv, 14 μL, 0.082 mmol) were added and the mixture was stirred at room temperature for 30 min. Then, a solution of intermediate 4 in dioxane (obtained in the previous step) was added to the reaction mixture, followed by DMTMM (1.0 equiv, 11 mg, 0.037 mmol), diisopropylethylamine (1.0 equiv, 7 μL, 0.037 mmol) and water (0.25 mL) and the resulting solution was stirred at room temperature for 1 h. The mixture was concentrated on a rotary evaporator and the residue was purified by reverse-phase flash chromatography (25 g, diol-modified C18, 0→60% ACN / 0.1% TFA in water) to give the desired product (15 mg, 35%) as a pale yellow solid after lyophilization. 61 H 63 FN9O 13 MS calculated: 1148.45, Found: 1148.45, [M+H] + .

[0314] Compound 1023 To a solution of intermediate 6 (1.0 equiv., 15 mg, 0.013 mmol) in DMF (1 mL) was added morpholine (35 μL) and the reaction mixture was stirred at room temperature for 1.5 h. Then, DMF and excess morpholine were evaporated on a rotary evaporator. The residue was redissolved in DMF (0.5 mL) followed by addition of Mal-PEG-NHS ester (1.1 equiv., 4.4 mg, 0.014 mmol) and diisopropylethylamine (1.1 equiv., 2.5 μL, 0.014 mmol) and the resulting solution was stirred at room temperature for 1 h. Purification by reversed-phase flash chromatography using a semi-separated column (diol-modified C18, 0→ 70% ACN / 0.1% TFA in water) afforded the desired product (7 mg, 48%) as a pale yellow solid after lyophilization. C 52 H 62 FN 10 O 15 MS calculated: 1121.44, Found: 1121.45, [M+H] + .

[0315] Example 63: Synthesis of Compound 71 JPEG2024518438000203.jpg39170 Intermediate 1 Bromoacetic acid (0.715 g, 10 mmol) was dissolved in 5 mL of water, sodium azide (0.696 g, 5 mmol) was added, and the solution was stirred at room temperature overnight. The solution was acidified with hydrochloric acid to pH=1, and the desired product was extracted with diethyl ether. The solvent was dried over Na2SO4, filtered, and evaporated to give the reaction product, which was used in the next step without further purification.

[0316] Intermediate 2 Exatecan mesylate (100 mg, 0.188 mmol), 2-azidoacetic acid (1.1 equiv., 0.207 mmol, 21 mg), DMTMM (1.3 equiv., 0.244 mmol, 68 mg) and DIPEA (50 μL) were dissolved in 5 mL of 4:1 DMF / water mixed solvent. The reaction mixture was stirred at room temperature for 1 h. The mixture was directly purified by reversed-phase flash chromatography (diol-modified C18, 25 g, 0→100% ACN in water). After lyophilization, the desired product was obtained as a white powder (79 mg, 74 %). C 26 H 24 MS calculated for FN6O5: 519.18, Found: 519.41, [M+H] + .

[0317] Compound 71. The previously prepared intermediate 2 (10 mg, 0.0193 mmol), propargyl alcohol (1.2 equiv., 0.0232 mmol, 1.35 μL), and catalyst CpRu(COD)Cl (10 %, 0.7 mg) were suspended in anhydrous DCM (2 mL) under argon atmosphere. The mixture was stirred at 40° C. for 16 h. The crude reaction product was directly purified by reversed-phase HPLC chromatography (semi-isolated diol-modified C18, 0→100% ACN in water). After lyophilization from water-ACN, the desired product was obtained as a yellowish powder (9 mg, 90 %). C 29 H 28 MS calculated for FN6O6: 575.21, found: 575.14, [M+H] + .

[0318] Example 64: Synthesis of Compound 72 JPEG2024518438000204.jpg41134 Compound 72 Intermediate 2 from Example 63 previously prepared (10 mg, 0.0193 mmol), propargyl alcohol (1.2 equiv., 0.0232 mmol, 1.35 μL), sodium ascorbate (0.2 equiv., 0.00386 mmol, 2M in water, 1.93 μL), copper sulfate pentahydrate (0.1 equiv., 0.00193 mmol, 1M in water, 1.93 μL) and TBTA (0.15 equiv., 0.0029 mmol, 1.5 mg) were dissolved in 2 mL of a 4:1 mixture of DMF / water. The reaction mixture was stirred at room temperature for 2 h. The crude reaction product was directly purified by reversed-phase HPLC chromatography (semi-isolated diol-modified C18, 0→100% ACN in water). After lyophilization from water-ACN, the desired product was obtained as a yellowish powder (10 mg, 95%). 29 H 28 MS calculated for FN6O6: 575.21, Found: 575.55, [M+H] + .

[0319] Example 65: Synthesis of Compound 73 JPEG2024518438000205.jpg49134 Compound 73 Previously prepared intermediate 2 from Example 63 (10 mg, 0.0193 mmol), 3-butyn-1-ol (1.2 equiv., 0.0232 mmol, 1.50 μL), and catalyst CpRu(COD)Cl (10%, 0.7 mg) were suspended in anhydrous DCM (2 mL) under argon atmosphere. The mixture was stirred at 40° C. for 16 h. The crude reaction product was directly purified by reversed-phase HPLC chromatography (semi-isolated diol-modified C18, 0→100% ACN in water). After lyophilization from water-ACN, the desired product was obtained as a yellowish powder (6 mg, 54%). C 30 H 30 MS calculated for FN6O6: 589.22, found: 589.23, [M+H] + .

[0320] Example 66: Synthesis of Compound 74 JPEG2024518438000206.jpg43162 Compound 74 Intermediate 2 from Example 63 previously prepared (10 mg, 0.0193 mmol), 3-butyn-1-ol (1.2 eq, 0.0232 mmol, 1.50 μL), sodium ascorbate (0.2 eq, 0.00386 mmol, 2 M in water, 1.93 μL), copper sulfate pentahydrate (0.1 eq, 0.00193 mmol, 1 M in water, 1.93 μL) and TBTA (0.15 eq, 0.0029 mmol, 1.5 mg) were dissolved in 2 mL of a 4:1 mixture of DMF / water. The reaction mixture was stirred at room temperature for 2 h. The crude reaction product was directly purified by reversed-phase HPLC chromatography (semi-isolated diol-modified C18, 0 → 100% ACN in water). After lyophilization from water-ACN, the desired product was obtained as a white powder (7 mg, 62%). 30 H 30 MS calculated for FN6O6: 589.22, found: 589.05, [M+H] + .

[0321] Example 67: Synthesis of Compound 173 JPEG2024518438000207.jpg461624:1 DMF / water mixture (4 mL) was added with exatecan mesylate (20 mg, 0.038 mmol), trans-3-hydroxymethylcyclobutane-1-carboxylic acid, lithium salt (1.25 equiv., 6.6 mg, 0.048 mmol), DMTMM (2.0 equiv., 21 mg, 0.076 mmol) and diisopropylethylamine (10 μL). The resulting solution was stirred at room temperature for 1 h, when LC-MS showed complete consumption of starting material. The mixture was directly purified by reversed-phase HPLC chromatography using a semi-separated column (diol-modified C18, 0→100% ACN / 1% TFA). After lyophilization, the desired product was obtained as a white powder (16 mg, 77%). C 30 H 31 MS calculated for FN3O6: 548.22, found: 548.31, [M+H] + .

[0322] Example 68 Kinetic Solubility of Compounds Based on Turbidity Turbidity-Based Aqueous Solubility (Kinematic Solubility) Procedure In vitro kinetic solubility of compounds in PBS pH 7.4 buffer at 25 °C was measured by diluting the compounds from 100% dimethyl sulfoxide (DMSO) into PBS buffer and measuring the absorbance at 490 nm, 590 nm, and 650 nm. Stock concentrations in 100% DMSO were provided at 1-6 mM. Solubility was measured by diluting test compounds from 100% DMSO into PBS pH 7.4 buffer in clear flat-bottom polystyrene assay plates as 10-point 2-fold serial dilutions starting with a 100-fold dilution of the DMSO stock solution into PBS buffer. The total assay volume was 200 microliters. The solutions were mixed by shaking the plate and incubated for 30 min at 25 °C before measuring the absorbance at 490, 590, and 650 nm. 1% (v / v) DMSO in PBS buffer was used as a blank. For each test compound, the blank-corrected sum of absorbance at 490 nm, 590 nm, and 650 nm was measured twice and averaged to determine the mean optical density (OD) at each concentration. The turbidity threshold optical density value was determined as the mean absorbance at 490 nm, 590 nm, and 650 nm of 1% (v / v) DMSO in PBS pH 7.4 buffer plus two standard deviations. The maximum soluble concentration (micromolar) corresponded to the highest concentration at which the mean optical density was below the optical density value set for the turbidity threshold (Table 4). Amiodarone and propranolol were used as low- and high-solubility controls, respectively. [Table 4] JPEG2024518438000209.jpg50153

[0323] Example 69: Evaluation of compound polarity based on retention time in reversed-phase liquid chromatography Reverse phase liquid chromatography retention times were measured by two independent experiments. In the first experiment, HPLC-MS analysis was performed on a Shimadzu UFLC-MS-2020 system equipped with ESI. Column Acquity UPLC BEH C18 1.7 μm, 2.1 x 50 mm. Solvent A: 0.1% formic acid in water; Solvent B: 0.1% formic acid in acetonitrile. Gradient: 0% B 0.8 min, 0% B to 100% B 4.2 min, 100% B 3 min. Total flow rate 0.6 ml / min. Total method time 10 min. UV-Vis spectra were recorded in the range 200-800 nm using a Shimadzu SPD-M2OA Prominence diode array detector. In the second experiment, lyophilized compounds were dissolved in dry DMSO at 2 mM and aliquots were frozen and stored at -80 °C. Compounds dissolved in DMSO were analyzed using RP-HPLC (UV / VIS, MS ELSD) Agilent 1100 platform with 1200 DAD and SofTA ELSD detectors, Agilent 6150 MS system. Shimadzu 3.0mm x 30mm XR ODS 2.2μm column was used at 50 °C and 1.5 mL / min. Solvent A: 0.1% formic acid (water), Solvent B: 0.08% formic acid (methanol), gradient: 5%-100% B in 3.0 min, 100% solvent B in 0.3 min. Retention times of compounds in reversed phase liquid chromatography using both are shown in Table 5. [Table 5] JPEG2024518438000211.jpg133147nc - Data not collected *-HPLC, Solvent A: water + 0.1% formic acid, Solvent B: acetonitrile + 0.1% formic acid. Gradient: 0% B 0.8 min, 0% B to 100% B 4.2 min, 100% B 3 min **-HPLC, Solvent A: water + 0.1% formic acid, Solvent B: methanol + 0.1% formic acid. Gradient: 5% to 100% solvent B in 3.0 min, 100% solvent B in 0.3 min. RT - RP HPLC retention time (min) The a,b conformer was not isolated. The A,B stereoisomers were either isolated or synthesized from stereochemically pure building blocks. C, D Separated and purified conformers

[0324] Example 70 In vitro cytotoxicity test Human tumor cell lines, SK-BR-3, NCI-H292, HT-29, MCF-7, NCI-N87 and FaDu were obtained from ATCC. NCI-H292, HT-29, MCF-7 and NCI-N87 cells were cultured in RPMI-1640 medium (Gibco, Life Technologies) supplemented with 10% v / v heat-inactivated FBS (Corning), and FaDu cells were maintained in DMEM medium (Gibco, Life Technologies) supplemented with 10% v / v heat-inactivated FBS (Corning). SK-BR-3 cells were maintained in McCoys 5A medium (Gibco, Life Technologies) supplemented with 10% v / w heat-inactivated FBS (Corning) at 37°C in a humidified incubator containing 5% CO2.

[0325] Compound preparation Lyophilized compounds were dissolved in 100% dry DMSO and aliquots were frozen and stored at -80 °C. Concentration and purity of compound stock solutions in DMSO were measured by RP- HPLC Agilent 1100 platform with 1200 DAD detector and SofTA ELSD detector, Agilent 6150 MS system (UV / VIS, ELSD purity, MS compound identity confirmation). Shimadzu 3.0mm x 30mm XR ODS 2.2μm column was used at 50 °C and 1.5 mL / min. Solvent A: 0.1% formic acid in water, Solvent B: 0.08% formic acid in methanol - gradient: 5% - 100% B in 3.0 min, 100% solvent B in 0.3 min. Compound concentrations in 100% DMSO measured by ELS were 1-6 mM. Compound purity measured by UV-VIS and ELS is shown in Table 5.

[0326] Cytotoxicity assay Cells were plated at 2.0 × 103 cells per well in 100 μL of medium in 96-well white flat-bottom plates (Corning). After 24 h of incubation, test compounds were added at 2- or 3-fold dilutions in a 10-point serial dilution. After an additional 6 days of incubation at 37 °C and 5% CO2, cell viability was assessed using the CellTiter-Glo Luminescent Cell Viability Assay (Promega). Luminescence was measured using a GloMax instrument (Promega). Luminescence values ​​were plotted against log concentration of test compound, and IC50 values ​​were calculated as best fit values ​​using a 4-parameter dose-response curve fit by GraphPad Prism 9, with R-squared values ​​ranging from 0.97-0.999. For a subset of payloads, each treatment was independently replicated 2–8 times, and IC50 values ​​were averaged. Table 6 shows the mean and standard deviation (stdev) of IC50 values ​​for treatments replicated 2–8 times. Standard deviation (stdev) is shown as (N / A) for treatments performed in duplicate. [Table 6] JPEG2024518438000213.jpg219170JPEG2024518438000214.jpg93170

[0327] Incorporation by Reference All publications and patents mentioned herein are hereby incorporated by reference in their entirety for all purposes to the same extent as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In the case of conflict, the present application, including definitions herein, will control.

[0328] Equivalent While specific embodiments of the subject invention have been described, the above specification is illustrative and not restrictive. Many variations of the disclosure will become apparent to those skilled in the art upon review of this specification. The full scope of the disclosure, along with such variations, along with its full scope of equivalents, should be determined by reference to the claims and the specification. Unless otherwise indicated, all numerical values ​​expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are understood to be modified in all instances by the term "about." Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and appended claims are approximations that may vary depending upon the desired properties sought to be obtained by the present disclosure.

Claims

1. A compound represented by formula (I) or a pharma- ceutically acceptable salt or stereoisomer thereof. (X is O, Z is a bond; Y is hydrogen, -C １－３ Alkyl, -CHO, and -C(O)-C １－３ is selected from the group consisting of alkyl, and R is R ２ , R ３ and R ４ or Y and Z, together with the nitrogen atom to which they are attached, each independently represent R Ｚ R is bonded to the heteroaryl, forming a 5-6 membered heteroaryl optionally substituted with 1, 2 or 3 substituents selected from R is R ６ or -N(Y)-Z-R is is selected from the group consisting of R ２ is -C(O)-NR ａ -C １－３ Alkyl, —C(O)—C ０－３ Alkyl-C(O)-NR ａ -C １－３ Alkyl, -S(O) ２ -C １－３ Alkyl-NR ａ -C(O)-C １－３ Alkyl, and -C(O)NR ａ - [(CH ２ ) ２ -O] １－１０ -C ２ alkyl; R ２ is substituted by hydroxy, R ２２ and R ２２ is independently, for each occurrence, halogen, hydroxy, -C １－３ Alkyl-OH and -C １－３ haloalkyl; R ３ is -C(O)-C ０－３ Alkyl-R ３０ , -C(O)-C ０－３ Alkyl-O-C １－３ Alkyl-R ３０ , -C ０－３ Alkyl-R ３０ , and -C １－３ Alkyl-O-C １－３ Alkyl-R ３０ and when alkyl is present, it is selected from the group consisting of halogen and -C １－３ haloalkyl; R ３０ , N, NR ３１ and O; R is a 5-6 membered heteroaryl having 1, 2 or 3 heteroatoms independently selected from the group consisting of ３０ Each independently represents R ３３ and optionally substituted on one or more available carbons with one or more substituents selected from R ３１ is, for each occurrence, independently, hydrogen, -C １－３ Alkyl, -C １－３ Alkyl-OH, -CH(OH)CH ２ OH, -CHO, and -C(O)-C １－３ is selected from the group consisting of alkyl, R ３３ represents, independently for each occurrence, -C １－３ Alkyl-OH, halogen, hydroxy, oxo and -C １－３ haloalkyl; or R ３ teeth, is selected from the group consisting of R ４ teeth, is selected from the group consisting of R ６ are substituted by hydroxy, each independently R ６６ -C １－３ is alkyl, R ６６ represents, for each occurrence, independently, a halogen and -C １－３ haloalkyl; R Ｚ is halogen, -C １－３ Alkyl and -C １－３ alkyl-OH; and R ａ is hydrogen, -C １－３ Alkyl-OH and -C １－３ haloalkyl-OH.

2. The compound according to claim 1, wherein Z is a bond.

3. Y is hydrogen, -CH ３ , —CHO and —COCH ３ 2. The compound of claim 1 selected from the group consisting of:

4. R is R ２ The compound according to claim 1,

5. The compound of claim 1, wherein Y is hydrogen.

6. R is -C(O)-NH-C ２ Alkyl, -C(O)-NH-C ３ Alkyl, -C(O)-C(O)-NH-C ２ Alkyl, -C(O)-C(O)-NH-C ３ Alkyl, —C(O)—C １ Alkyl-C(O)-NH-C ２ Alkyl, —C(O)—C ２ Alkyl-C(O)-NH-C ２ Alkyl, —C(O)—C ２ Alkyl-C(O)-NH-C ３ Alkyl, -S(O) ２ -C ２ Alkyl-NH-C(O)-C １ Alkyl, -S(O) ２ -C ２ Alkyl-NH-C(O)-C ２ Alkyl and -C(O)NH-[(CH ２ ) ２ -O] １－２ -C ２ alkyl; R ２ is substituted by hydroxy, R ２２ 2. The compound of claim 1, optionally substituted with one or more additional substituents each independently selected from:

7. R ２２ Fluoro, hydroxy, -CH ２ -OH and -CF ３ 2. The compound of claim 1 selected from the group consisting of:

8. 2. The compound of claim 1, wherein -N(Y)-Z-R is selected from the group consisting of:

9. R is R ３ The compound according to claim 1,

10. The compound of claim 1, wherein Y is hydrogen.

11. R is -C(O)-triazolyl, -C(O)-C １ Alkyl-triazolyl, -C(O)-C ２ Alkyl-triazolyl, -C(O)-C ３ Alkyl-triazolyl, -C １ Alkyl-triazolyl, -C ２ Alkyl-triazolyl, -C ３ Alkyl-triazolyl, —C(O)—O—C １ Alkyl-triazolyl, —C(O)—O—C ２ Alkyl-triazolyl, -C(O)-C １ Alkyl-O-C ２ Alkyl-triazolyl, -C(O)-C ２ Alkyl-O-C １ Alkyl-triazolyl, -C(O)-C ２ Alkyl-O-C ２ Alkyl-triazolyl, -C ２ Alkyl-O-C １ Alkyl-triazolyl, and -C ２ Alkyl-O-C ２ alkyl-triazolyl, where alkyl at each occurrence is independently selected from the group consisting of fluoro and -CF ３ and optionally substituted with 1, 2 or 3 substituents selected from the group consisting of: Triazolyl, when present, is hydrogen, —C １－３ Alkyl and -C １－２ substituted on an available nitrogen by a substituent selected from the group consisting of alkyl-OH; Triazolyl includes chloro, fluoro and -C １－２ 2. The compound of claim 1, optionally substituted on an available carbon with a substituent selected from the group consisting of: -alkyl-OH.

12. 2. The compound of claim 1, wherein R is selected from the group consisting of:

13. R is -C(O)-furanyl, -C １ R is selected from the group consisting of hydroxy and -C(O)-alkyl-furanyl, -C(O)-oxazolyl and -C(O)-pyrazolyl. １－２ The compound of claim 1 , substituted with a substituent selected from the group consisting of alkyl-OH.

14. 2. The compound of claim 1, wherein R is selected from the group consisting of:

15. 2. The compound of claim 1, wherein Y and Z together with the nitrogen atom to which they are attached form a triazolyl substituted with R at any substitutable position.

16. R is C １ Alkyl-OH or -C ２ alkyl-OH, R is -CF ３ 2. The compound of claim 1, which is optionally substituted by:

17. 2. The compound of claim 1, wherein -N(Y)-Z-R is selected from the group consisting of:

18. 2. The compound of claim 1, wherein the compound is selected from the group consisting of:

19. A compound represented by formula (IIA) or a pharma- ceutically acceptable salt or stereoisomer thereof. A is NH or triazolyl; L １ teeth, is selected from the group consisting of RR is L １ and R of claim 1 is hydroxy or -NH ２ and an alkoxy or amino moiety formed from a moiety.

20. 20. The compound of claim 19, wherein the compound is selected from the group consisting of:

21. A compound selected from the group consisting of:

22. A pharmaceutical composition for the treatment of cancer, for administering to a patient in need thereof an effective amount of a compound according to any one of claims 1 to 21, The pharmaceutical composition, wherein the cancer is selected from the group consisting of lung cancer, renal cancer, urothelial cancer, colorectal cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, breast cancer, melanoma, liver cancer, bladder cancer, gastric cancer and esophageal cancer.

23. A pharmaceutical composition comprising a compound according to any one of claims 1 to 21 and a pharma- ceutically acceptable excipient.

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