US009295731B2

(12) United States Patent (10) Patent No.: US 9.295,731 B2 Nguyen (45) Date of Patent: Mar. 29, 2016

(54) CLEAVABLE DRUG CONJUGATES, (52) U.S. Cl. COMPOSITIONS THEREOF AND METHODS CPC ...... A61K47/48284 (2013.01); A61K 45/06 OF USE (2013.01); A61 K47/4843 (2013.01); A61 K 47/48061 (2013.01); A61 K47/48092 (2013.01); (71) Applicant: Mark Quang Nguyen, San Jose, CA A61K 47/48246 (2013.01) (US) (58) Field of Classification Search None (72) Inventor: Mark Quang Nguyen, San Jose, CA See application file for complete search history.

(73) Assignee: Mark Quang Nguyen, San Jose, CA (56) References Cited (US) U.S. PATENT DOCUMENTS (*) Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35 2003/0022876 A1 1/2003 Ashton et al...... 514, 176 U.S.C. 154(b) by 0 days. * cited by examiner (21) Appl. No.: 13/854,296 Primary Examiner — Alicia L Otton (22) Filed: Apr. 1, 2013 (57) ABSTRACT (65) Prior Publication Data Base-labile crosslinkers, base-labile conjugates comprising US 2014/O294.851A1 Oct. 2, 2014 Such crosslinkers, methods of their synthesis and use are (51) Int. Cl disclosed. A6 IK 47/48 (2006.01) A6 IK 45/06 (2006.01) 10 Claims, No Drawings US 9,295,731 B2 1. 2 CLEAVABLE DRUG CONJUGATES, Mertansine, a derivative of maytansine, is an experimental COMPOSITIONS THEREOF AND METHODS cytotoxic agent. Because of its extreme toxicity, it is not used OF USE as a stand-alone therapeutic agent. To minimize side effects and to maximize efficacy, the compound is conjugated to FIELD tumor-targeting monoclonal antibodies such as lorVotu Zumab and trastuzumab. The disclosure relates to multi-functional crosslinkers Vedotin, an auristatin, is another potent cytotoxic agent. To capable of coupling primary or secondary amine-containing minimize side effects and to maximize efficacy, it is conju therapeutic agents to ligands. The multi-functional crosslink gated to tumor-targeting monoclonal antibody brentuximab. ers are cleavable upon exposure to esterases or to basic con 10 Recombinant interferon alfa-2a (RoferonRA) is used for ditions to release the amine-containing therapeutic agents. the treatment of chronic hepatitis C, hairy cell leukemia, and The disclosure also relates to conjugates comprising the chronic myelogenous leukemia. The recommended dosing crosslinkers, a therapeutic agent, and a ligand. regimen for RoferonRA for the treatment of chronic hepatitis BACKGROUND 15 C is three times per week administered subcutaneously for 12 months. Drugs can often cause severe side effects and/or require PegasyS(R), a pegylated formulation of interferon alfa-2a, multiple dosing regimens. Some of these side effects and requires less frequent dosing. The recommended dosing regi inconvenient dosing regimens can be controlled or reduced men for PegasySR for treating chronic hepatitis C is once by the use of improved formulations and/or improved deliv weekly for 48 weeks by subcutaneous administration. ery methods. Zalbin R, an interferon alfa-2b conjugated to human serum Paclitaxel, for example, is highly effective in treating albumin, is an alternative to unconjugated interferon alfa-2b breast and lung cancers by inhibiting the growth of cancer (IntronRA). Instead of subcutaneously or intramuscularly cells by binding to microtubules. However, because the drug dosing the non-conjugated drug three times a week for up to has a low aqueous solubility, paclitaxel is typically formu 25 24 months, Zalbin R) is administered only once every two lated using the toxic solvent Cremophor REL (polyoxyethy weeks. lated castor oil), which limits the amount of drug that can be Enfluvirtide, an FDA-approved HIV drug, is a fusion administered to a patient. Patients taking the drug often inhibitor having a linear 36-amino acid synthetic peptide. require premedication with Steroids or antihistamines for Due to its short half-life of four hours, treatment requires hypersensitivity reactions caused by the solvent. 30 twice-daily Subcutaneous injections. This inconvenient dos Abraxane R, a new paclitaxel formulation, does not employ ing schedule discourages its widespread use. a toxic solvent. When formulated with human serum albumin, the drug can be suspended in aqueous media. Thus, patients SUMMARY can better tolerate higher paclitaxel doses (up to 260 mg/m vs. 175 g/m) and thereby achieve an improved response rate. 35 Thus, there is a need for improved drug formulations that Doxorubicin is another highly effective cancer drug can reduce or eliminate severe side effects and enhance the belonging to a class of anthracycline compounds. However, tolerability, efficacy, and/or compliance of the drug. More the drug can cause severe cardiotoxicity and death. Patients specifically, many pharmacological compounds having taking a cumulative dose of 300 mg/mhave a 1-2% chance of amine groups can benefit from conjugation to other mol developing cardiomyopathy, with the incidence increasing 40 ecules or moieties provided that the crosslinking group is with increasing dose (400 mg/m,3-5%;450 mg/m, 5 to 8%; cleavable in vivo to release the drug. Thus, there is a need for 500 mg/m, 6-20%). The use of Doxil(R), a pegylated liposo new crosslinkers having a functional group that can form a mal formulation of doxorubicin, results in a reduced inci cleavable bond with amine-containing molecules upon expo dence of cardiotoxicity. Doxil R has a half-life in the blood of Sure to esterases and/or under mild basic conditions (e.g., pH about 50 hours compared to about 10 minutes for doxorubi 45 7.4 to 9) in vivo. cin. The slow-release of doxorubicin may explain the lower Cleavable cross-linkers, intermediates of cleavable cross toxicity of the Doxil R formulation. Myocet(R), a non-pegy linkers, cleavable conjugates comprising the cleavable cross lated liposomal formulation of doxorubicin, also reduces the linkers, and methods of synthesizing the cross-linkers, inter incidence of cardiotoxicity. Furthermore, its use also reduces mediates, and conjugates are disclosed. Therapeutic the incidence of hand-foot syndrome associated with Doxil (R). 50 conjugates provided by the present disclosure comprising the Myocet(R) is approved in Europe and is being evaluated in the cleavable cross-linkers can reduce the side effects, enhance United States. the efficacy, and/or improve the convenience of treatment for Mitoxantrone is used for the treatment of acute non-lym the parent drug. phocytic leukemia. It is also indicated for reducing neuro In a first aspect, compounds of Formula (II) are disclosed: logic disability and/or the frequency of clinical relapses in 55 patients with secondary (chronic) progressive, progressive relapsing, or worsening relapsing-remitting multiple Sclero (II) sis (MS). Although one of the most effective drugs for treating O R1 R2 O MS, mitoxantrone is not recommended for long-term use. Patients who exceed a cumulative dose of 140 mg/m have an 60 AlNx11 X2Nx lsO X O increased risk of acquiring irreversible cardiotoxicity and death. There is no liposomal formulation of this drug. Bleomycin, in combination with other chemotherapeutic or a salt thereof, wherein: agents, is effective in treating Squamous cell carcinoma, non each A' is independently selected from a thiol-reactive Hodgkin’s lymphoma, and testicular carcinoma. However, 65 group, an amine-reactive group, an avidin-binding patients taking a cumulative dose of 400 units (~400 mg) risk group, a photoreactive group, an alkyne-reactive group, developing pulmonary fibrosis. and an azide-reactive group; US 9,295,731 B2 3 4 each X and X is independently selected from a covalent tuted C. heterocycloalkanediyl, Cao alkanecycloal bond, Co alkanediyl. Substituted Co alkanediyl. kanediyl. Substituted Coalkanecycloalkanediyl, Cao Co heteroalkanediyl. Substituted Co. heteroal heteroalkanecycloalkanediyl. Substituted Cao het kanediyl, C-2 cycloalkanediyl. Substituted C. eroalkanecycloalkanediyl, Co arenediyl. Substituted cycloalkanediyl. C. heterocycloalkanediyl. Substi Co-oarenediyl, C-2 heteroarenediyl. Substituted C-20 tuted C. heterocycloalkanediyl, Cao alkanecycloal heteroarenediyl, Co alkanearenediyl. Substituted kanediyl. Substituted Coalkanecycloalkanediyl, Cao Czo alkanearenediyl, Co heteroalkanearenediyl. heteroalkanecycloalkanediyl. Substituted Cao het Substituted Co. heteroalkanearenediyl, and eroalkanecycloalkanediyl, Co arenediyl. Substituted (CH2)—(CH2—CH2—O) (CH), , Coarenediyl. Coheteroarenediyl. Substituted Co 10 wherein: heteroarenediyl, C-2 alkanearenediyl. Substituted each n1 and n3 is independently an integer selected from Czo alkanearenediyl, Co heteroalkanearenediyl. 0 to 5; and Substituted Co. heteroalkanearenediyl, and each n2 is independently an integer selected from 1 to (CH2)—(CH2—CH2—O) -(CH2) , 25; wherein: 15 each X* is independently selected from a covalent bond, each n1 and n3 is independently an integer selected from O— —S——N , —N-, - N=N , —N–C , 0 to 5; and SO— —SO , —SON.— —SS—, —C(O)—, each n2 is independently an integer selected from 1 to C(O)O-, - C(O)N , —C(O)S –C(O)N 25; N=, OP(O)(OH)O OC(O)O OC(O)N , each X* is independently selected from a covalent bond, - NC(O)N , and - NC(S)N: O— —S , —N , —N-, - N=N , —N–C , each R" and R is independently selected from hydrogen, SO— —SO , —SON.— —SS—, —C(O)—, C. alkyl, Substituted C. alkyl, C. heteroalkyl, Sub C(O)O-, - C(O)N , —C(O)S –C(O)N stituted C. heteroalkyl, Co aryl, Substituted Co N=, OP(O)(OH)O OC(O)O OC(O)N , aryl, Coheteroaryl, and substituted Coheteroaryl; - NC(O)N , and - NC(S)N: 25 each D is independently selected from therapeutic agent each R" and R is independently selected from hydrogen, having at least one primary amine group, a therapeutic C. alkyl, Substituted C. alkyl, C. heteroalkyl, Sub agent having at least one secondary amine group, a stituted C. heteroalkyl, Co aryl, Substituted Co therapeutic agent having at least one hydroxy group, and aryl, Coheteroaryl, and Substituted Coheteroaryl; a therapeutic agent having at least one thiol group; and D is selected from a therapeutic agent having at least one 30 nó is an integer selected from 1 to 20. primary amine group, a therapeutic agent having at least In a third aspect, pharmaceutical compositions are dis one secondary amine group, a therapeutic agent having closed comprising a compound of Formula (II), a compound at least one hydroxy group, and a therapeutic agent of Formula (III), or a pharmaceutically acceptable salt of any having at least one thiol group; and of the foregoing, and a pharmaceutically acceptable carrier. n5 is an integer selected from 1 to 20. 35 In a fourth aspect, methods of treating cancer, an autoim In a second aspect, compounds of Formula (III) are dis mune disease, oran infectious disease are disclosed, compris closed: ing administering to a patient in need of Such treatment a therapeutically effective amount of a compound of Formula (II), Formula (III), a pharmaceutical salt of any of the fore (III) 40 going, or a pharmaceutical composition comprising any one O R1 R2 O of the foregoing. In a fifth aspect, methods of synthesizing compounds of Q Y 1nA 11 X n1ls No X O ul DJ, Formula (I), Formula (II), and Formula (III), or a pharmaceu tically acceptable salt of any of the foregoing are disclosed. 45 or a pharmaceutically acceptable salt thereof, wherein: DETAILED DESCRIPTION Q is selected from a ligand having at least one thiol group, a ligand having at least one primary amine group, a Definitions ligandhaving at least one secondary amine group, ligand having at least one biotin-binding group, aligand having 50 A dash (“-”) that is not between two letters or symbols is at least one photoreactive group, a ligand having at least used to indicate a point of attachment for a substituent. For one alkyne group, and a ligand having at least one azide example, —CONH is attached through the carbonatom. group; Compounds provided by the present disclosure are encom each Y is independently selected from a covalent bond and passed by structural formulae disclosed herein and include a hydrogen bond; 55 any specific compounds within these formulae. Compounds may be identified either by their chemical structure and/or each A is independently selected from a covalent bond, an chemical name. When the chemical structure and chemical amide group, an avidin-binding group, a carbamate name conflict, the chemical structure is determinative of the group, a carbonyl group, a disulfide group, an ester identity of the compound. The compounds described herein group, a hydraZone group, an imine group, a Succinim may contain one or more chiral centers and/or double bonds ide group, a Sulfonamide group, a Sulfone group, a Sul 60 and therefore, may exist as stereoisomers, such as double foxide group, a thioether group, a triazole group, and a bond isomers (i.e., geometric isomers), enantiomers or dias urea group: tereomers. Accordingly, the chemical structures depicted each X and X is independently selected from a covalent herein encompass all possible enantiomers and stereoisomers bond, Co alkanediyl. Substituted Co alkanediyl. of the illustrated compounds including the stereoisomerically Co heteroalkanediyl. Substituted Co. heteroal 65 pure form (e.g., geometrically pure, enantiomerically pure or kanediyl, C-2 cycloalkanediyl. Substituted C. diastereomerically pure) and enantiomeric and stereoiso cycloalkanediyl. C. heterocycloalkanediyl. Substi meric mixtures. Enantiomeric and stereoisomeric mixtures US 9,295,731 B2 5 6 can be resolved into their component enantiomers or stereoi The term “alkanecycloalkanediyl refers to a diradical Somers using separation techniques or chiral synthesis tech hydrocarbon group derived by the removal of two hydrogen niques well known to one skilled in the art. The compounds atoms from a single carbonatom or by the removal of a single may also exist in several tautomeric forms including the enol hydrogen atom from two carbon atoms from a parent form, the keto form and mixtures thereof. Accordingly, the alkanecycloalkane group. In certain embodiments an chemical structures depicted herein encompass all possible alkanecycloalkanediyl is selected from 4-methylcyclohex tautomeric forms of the illustrated compounds. The com ane-diyl: pounds described also include isotopically labeled com pounds where one or more atoms have an atomic mass dif ferent from the atomic mass conventionally found in nature. Examples of isotopes that may be incorporated into the com 10 pounds disclosed herein include, but are not limited to, H. H, C, C, N, O, O, etc. Compounds may exist in unsolvated forms as well as Solvated forms, including hydrated forms and as N-oxides. Accordingly, compounds may be hydrated, Solvated, or N-oxides. Certain compounds 15 may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses con The term “alkoxy' refers to an alkyl-O group where templated herein and are intended to be within the scope of alkyl is as defined herein. Examples of alkoxy groups include the present disclosure. Further, it should be understood, when methoxy, ethoxy, n-propoxy, isopropoxy, and n-butoxy. In partial structures of the compounds are illustrated, that brack certain embodiments, an alkoxy group is Cls alkoxy, C. ets indicate the point of attachment of the partial structure to the rest of the molecule. alkoxy, Calkoxy, and in certain embodiments C alkoxy. The term “acyl means an H C(O)—, alkyl-C(O)—, or The term “alkyl refers to a monoradical of a saturated or cycloalkyl-C(O)—group wherein alkyl and cycloalkyl areas unsaturated, branched, or straight-chain acyclic hydrocarbon defined herein. In certain embodiments, an acyl group is Cls group having, for example, from 1 to 20 carbon atoms, from acyl, C-acyl, C-acyl, and in certain embodiments, C 25 1 to 10 carbon atoms, from 1 to 6 carbon atoms, from 1 to 4 acyl. carbonatoms, or from 1 to 3 carbonatoms. Examples of alkyl The term "alkanearene' refers to a hydrocarbon group in groups include methyl, ethyl, n-propyl, iso-propyl. n-butyl, which an alkyl group is bonded to an aromatic group, wherein iso-butyl, n-hexyl, n-decyl, tetradecyl, and the like. The term alkyl and aromatic group are as defined herein. In certain “alkyl is specifically intended to include groups having any embodiments, an alkanearene group is C7-o alkanearene, 30 degree or level of Saturation, i.e., groups having exclusively C. alkanearene, and in certain embodiments Co alka single carbon-carbon bonds, groups having one or more nearenediyl. double carbon-carbon bonds, groups having one or more The term "alkanearenediyl refers to a diradical hydrocar triple carbon-carbon bonds and groups having mixtures of bon group derived by the removal of two hydrogen atoms single, double and triple carbon-carbon bonds. from a single carbon atom or by the removal of a single 35 The term “alkyne-reactive group’ as used herein refers to a hydrogen atom from two carbon atoms from a parent alka functional group capable of reacting with an alkyne in the nearene group. In certain embodiments, an alkanearenediyl presence or absence of a catalyst to form a triazole. Example group is Czo alkanearenediyl, C7-12 alkanearenediyl, and in of a catalyst includes Copper(I). Examples of alkyne-reactive certain embodiments C-o alkanearenediyl. groups include an azide group. In certain embodiments, an The term "alkanediyl refers to a diradical of a saturated or 40 alkyne-reactive group is selected from Formula (A'el), For unsaturated, branched, or straight-chain acyclic hydrocarbon mula (A"e2), and Formula (A'd3): group, having, for example, from 1 to 20 carbon atoms, from 1-10 carbonatoms, from 1-6 carbonatoms, from 1 to 4 carbon atoms, or from 1 to 3 hydrocarbon atoms. Examples of (Ale1) alkanediyl groups include methane-diyl (—CH2—), ethane 45 1.2-diyl (—CHCH ), propane-1,3-diyl and iso-propane 1.2-diyl (e.g., —CH2CH2CH2— and —CH(CH)CH2—), butane-1,4-diyl ( CHCHCHCH ), pentane-1,5-diyl C, ( CHCHCHCHCH ), hexane-1,6-diyl (Ale2) (—CH2CH2CH2CH2CHCH ), heptane-1,7-diyl, octane 1,8-diyl. nonane-1,9-diyl, decane-1,10-diyl, dodecane-1,12 50 diyl, and the like. ~-X and The term “alkanecycloalkane' refers to a hydrocarbon (Ad3) group in which an alkyl group is bonded to a cycloalkane group, wherein alkyland cycloalkane areas defined herein. In certain embodiments, an alkanecycloalkane group is C7-2 55 alkanecycloalkane, C7- alkanecycloalkane, and in certain embodiments C- alkanecycloalkane. In certain embodi N ments an alkanecycloalkane is selected from methylcyclo hexane: 60 The term “amine-reactive group’ as used herein refers to a functional group capable of reacting with a primary amine group, a secondary amine group, a hydrazine group, or a Substituted hydrazine group to form an amide bond, a urea bond, a thiourea bond, a Sulfonamide bond, a carbamate 65 bond, an imine, or a hydrazone. Examples of an amine-reac tive group include an aldehyde group, a ketone group, an NHS-ester group, a Substituted phenylester group, an isocy US 9,295,731 B2 7 8 anate group, an isothiocyanate group, and an alkyl imidate tives interchelate with DNA and RNA, inhibit the enzyme group. In certain embodiments, an amine-reactive group is topoisomerase II, and generate DNA-damaging free oxygen selected from Formula (Ab1), Formula (Ab2). Formula radicals. Examples of anthracycline derivatives include beru (Ab3), and Formula (Ab4): bicin, daunorubicin, doxorubicin, epirubicin, idarubicin, amrubicin, pirarubicin, Zorubicin, mitoxantrone, banox antrone, and Sabarubicin. (Ab1) The term “anthracenedione derivative' refers to a com O pound having the basic structural scaffolds:

L2 10

(Ab2) O R8 R9 O

L2 ls O X O 15

(Ab3) O 2O and is effective in treating metastatic breast cancer, acute 21 lymphoblastic leukaemia, acute myeloid leukemia, non Hodgkin’s lymphoma, and/or multiple Sclerosis. Anthracene N and dione derivatives interchelate with DNA and inhibit the enzyme topoisomerase II. Examples of anthracenedione O (Ab4 25 derivatives include to ledoxantrone, mitoxantrone, nortopix antrone, pixantrone, piroXantrone, and topixantrone. The term “antibiotic” refers to a compound that kills or H slows the growth of bacteria, fungi, and/or other microorgan isms. Examples of antibiotics include amikacin, amphoteri 30 cin B, arbekacin, astromicin, bacitracin, balofloxacin, bed erocin, bekanamycin, besifloxacin, brodimoprim, wherein each R and R is independently selected from ciprofloxacin, clinafloxacin, colistin, daptomycin, dibekacin, hydrogen, Calkyl, substituted C alkyl, Cheteroalkyl, enoxacin, framycetin, garenoxacin, gatifloxacin, gemifloxa Substituted Cheteroalkyl, Co aryl, Substituted Co aryl, cin, gentamicin, gentamicin, grepafloxacin, hamycin, hexeti Coheteroaryl, and substituted Co-o heteroaryl; and L is a 35 dine, hygromycin B, ibacitabine, iclaprim, isepamicin, kana leaving group Such as a halogen, an N-hydroxySuccinimidyl, mycin, lomefloxacin, lucimycin, lymecycline, mepartricin, a Substituted N-hydroxysuccinimidyl, a phenol-yl, a Substi moxifloxacin, natamycin, nemonoxacin, neomycin B, neo tuted phenol-yl, a hydroxybenzotriazolyl, a substituted mycin C, netilmicin, norfloxacin, nystatin, omadacycline, hydroxybenzotriazolyl, an imidazolyl, and a Substituted imi oritavancin, paromomycin, paZufloxacin, perimycin A, peri dazolyl. 40 mycin B, perimycin C, pipemidic acid, polymyxin B, puro The term "amino acid side chain' includes the side chains mycin, radeZolid, retaspimycin, ribostamycin, rimocidin, of naturally occurring standard amino acids, side chains of Sisomicin, sitafloxacin, sparfloxacin, spectinomycin, strepto naturally occurring non-standard amino acids, and side mycin, Sulfacetamide, Sulfadiazine, Sulfadimethoxine, Sul chains of non-naturally occurring amino acid derivatives. In fadimidine, Sulfafurazole, Sulfalene, SulfamaZone, Sulfam certain embodiments, amino acid side chain includes natu- 45 erazine, sulfamethizole, Sulfamethoxazole, rally occurring standard amino acid side chains. Sulfamethoxypyridazine, Sulfametomidine, Sulfametoxydi The term “anthracycline derivative' refers to a compound azine, Sulfametrole, Sulfamoxole, Sulfanilamide, Sulfaperin, having the basic structural scaffold: Sulfaphenazole, Sulfapyridine, Sulfathiazole, Sulfathiourea, Sulfisomidine, teicoplanin, telavancin, tanespimycin, tema 50 floxacin, tetroXoprim, tigecycline, tobramycin, toSufloxacin, t OH O trimethoprim, trimethoprim, trovafloxacin, tyrothricin, uli floxacin, Valnemulin, Vancomycin, Verdamicin, and Zabof ul loxacin. O The term “antifolate” refers to a compound that is an ana 55 log offolic acid that inhibits the enzyme dihydrofolate reduc tase, thymidylate synthase, and/or other enzyme associated O OH O with the metabolism of nucleotides. Antifolates are known to be useful in treating cancer, rheumatoid arthritis, lupus, scle roderma, psoriasis, asthma, sarcoidosis, primary biliary cir O 60 rhosis, polymyositis, and/or inflammatory bowel disease. Examples of antifolates include aminopterin, folitix.orin, N methotrexate, pemetrexed, pralatrexate, raltitrexed, pelitr exol, pyrimethamine, talotrexin, and trimethoprim. O The term “arenediyl refers to an aromatic hydrocarbon 65 diradical derived by the removal of two hydrogenatoms from and is effective in treating leukemias, lymphomas, and breast, a single carbon atom or by the removal of a single hydrogen uterine, ovarian, and/or lung cancers. Anthracycline deriva atom from two carbonatoms of a parent aromatic ring system. US 9,295,731 B2 10 In certain embodiments, an arenediyl group is Co. arene -continued diyl, C-2 arenediyl, Co arenediyl, and in certain embodi ments, Cesarenediyl. Examples of arenediyl groups include O. O (Ac2) benzene-1,2-diyl, benzene-1,3-diyl, benzene-1,4-diyl, naph thalene-1,6-diyl, and the like. The term “aryl refers to a monovalent aromatic hydrocar X > bon radical derived by the removal of one hydrogen atom Hill H from a single carbon atom of a parent aromatic ring system. Aryl benzene: bicyclic ring systems wherein at least one ring 10 is carbocyclic and aromatic, for example, naphthalene, "" O and indane, and tetralin; and tricyclic ring systems wherein at least one ring is carbocyclic and aromatic, for example, fluo (A'c3) rene. Aryl encompasses multiple ring systems having at least one carbocyclic aromatic ring fused to at least one carbocy 15 clic aromatic ring, cycloalkyl ring, or heterocycloalkyl ring. For example, aryl includes a phenyl ring fused to a 5-to 7-membered heterocycloalkyl ring containing one or more " heteroatoms chosen from N, O, and S. For such fused, bicy O clic ring systems wherein only one of the rings is a carbocy clic aromatic ring, the radical carbon atom may be at the carbocyclic aromatic ring or at the heterocycloalkyl ring. The term “azide-reactive group’ as used herein refers to a Examples of aryl groups include, but are not limited to, functional group capable of reacting with an azide in the 25 presence or absence of a catalyst to form a triazole. Example groups derived from aceanthrylene, acenaphthylene, aceph of a catalyst includes Copper(I). Examples of azide-reactive enanthrylene, anthracene, azulene, benzene, chrysene, coro groups include a terminal alkyne group and an internal alkyne nene, fluoranthene, fluorene, hexacene, hexaphene, hexylene, group. In certain embodiments, an azide-reactive group is as-indacene, S-indacene, indane, indene, naphthalene, octa selected from Formula (A' fl), Formula (A'f2), Formula cene, octaphene, octalene, ovalene, penta-2,4-diene, penta 30 (A'f3), Formula (A"fA), and Formula (Af5): cene, pentalene, pentaphene, perylene, phenalene, phenan threne, picene, pleiadene, pyrene, pyranthrene, rubicene, triphenylene, trinaphthalene, and the like. In certain embodi (Alf1) ments, an aryl group can have from 6 to 20 carbon atoms 35 (C-o), from 6 to 12 carbon atoms (C-2), from 6 to 10 carbonatoms (C-o), and in certain embodiments from 6 to 8 i-- carbon atoms (Cs). Aryl, however, does not encompass or overlap in any way with heteroaryl, separately defined herein. 40 The term “aurora kinase inhibitor” refers to a compound (Af2) that inhibits the enzyme aurora kinase, which regulates cel lular mitosis. Aurora kinase inhibitors are being investigated for the treatment of cancer. Examples of aurora kinase inhibi tors include barasertib, hesperadin, and toZasertib. The term “avidin-binding group’ as used herein refers to a 45 biotin, a biotin derivative such as desthiobiotin, iminobiotin, bisnorbiotin, tetranorbiotin, biotin sulfoxide, biotin sulfone, (Af3) and iminobiotin trifluoroacetamide, or a Strep-tag (a syn thetic peptide consisting of an N-terminal or C-terminal eight amino acid sequence: Trp-Ser-His-Pro-Gln-Phe-Glu-Lys) 50 capable of binding to avidin, deglycosylated avidin, Strepta vidin, Strep-Tactin or related proteins. In certain embodi ments, an avidin-binding group is selected from Formula (A'c1), Formula (Ac2), and Formula (A'c3): 55 C

(Alfa) 60

65 F o and US 9,295,731 B2 11 12 -continued embodiments, a cycloalkane group is C-12 cycloalkane, Cas (AlfS) cycloalkane, C. cycloalkane, and in certain embodiments, Cs cycloalkane. The term "cycloalkanediyl refers to a diradical cyclic or %. polycyclic hydrocarbon group. In certain embodiments, a N cycloalkane-diyl group is C-12 cycloalkane-diyl. Cas cycloalkane-diyl. C. cycloalkanediyl, and in certain embodiments, Cse cycloalkanediyl. Examples of cycloal kanediyl groups include cyclohexane-1,4-diyl, cyclohexane 10 1,3-diyl, and cyclohexane-1,2-diyl. The term “cycloalkyl refers to a saturated or unsaturated The term “camptotheca derivative' refers to a compound cyclic or polycyclic hydrocarbon monoradical group. In cer having the basic structural scaffold: tain embodiments, a cycloalkyl group is C. cycloalkyl, Cs cycloalkyl, C. cycloalkyl, and in certain embodiments,

15 Cs cycloalkyl. The term “dipeptidyl peptidase IV inhibitor refers to a compound that inhibits the enzyme dipeptidyl peptidase IV. which is expressed on the Surface of most cell types and is associated with immune regulation, signal transduction and apoptosis. Examples of dipeptidyl peptidase IV inhibitors include tallabostat. The term “dolastatin derivative' refers to a compound that is isolated or derived from the seahare Dolabella auricularia or a structural analog thereof that binds to tubulin B subunit and is effective intreating colon, ovarian, and/or lung cancers. 25 and inhibits microtubule assembly. Dolastatin derivatives are Camptotheca derivatives inhibit the enzyme topoisomerase I potentially effective in treating cancer. Examples of dolasta and interfere with the unwinding of DNA. Examples of camp tin derivatives include auristatin E. monomethyl auristatin E, totheca derivatives include belotecan, atiratecan, exatecan, auristatin F. monomethyl auristatin F. dolastatin 10, monom namitecan, 7-nbutyl-10-amino-camptothecin, and 7-n-butyl ethyl dolastatin 10, dolastatin 12, dolastatin 15, soblidotin, 9-amino-10, 11-methylenedixoy-camptothecin. 30 monomethyl and dolastatin 16. The term “cathepsin Kinhibitor refers to a compound that The term “duocarmycin derivative' refers to a compound inhibits the enzyme cathepsin K, which is associated with that is isolated orderived from Streptomyces sp. or a structural bone resorption. Cathepsin K inhibitors are being investi analog thereof that alkylates DNA at minor groove sites. gated for the treatment of bone cancer metastases. Examples Duocarmycin derivatives are potentially effective in treating 35 cancer. Examples of duocarmycin derivatives include duocar of cathepsin K inhibitors include dutacatib and odanacatib. mycin A, duocarmycin SA, duocarmycin B, and duocarmy The term “colchicine derivative' refers to a compound cin B1. having the basic structural scaffold: The term “ecteinascidin derivative' refers to a compound that is isolated or derived from the sea squirt Ecteinascidia 40 turbinata or a structural analog thereof that produces Super

oxide, resulting in DNA cleavage. Ectienascidin derivatives are potentially effective in treating cancer. Examples of ect einascidin derivatives include trabectedin. The term “enediyne derivative' refers to a compound that 45 is isolated or derived from a bacterial natural product or a structural analog thereof characterized by either nine- and ten-membered rings containing two triple bonds separated by a double bond. Enediyne derivatives bind to and cleave DNA and are potentially effective in treating cancer. Examples of 50 enediyne derivatives include calicheamicinyl, calicheamicin T, esperamicin A1, esperamicin C, esperamicin D, dynemicin A dynemicin H, dynemicin M, dynemicin N, dynemicin O, and is potentially effective in treating cancer. Colchicine dynemicin P. dynemicin Q, dynemicin S, neocarzinostatin derivatives bind to tubulin P subunit and inhibit microtubule chromophore, and uncialamycin. assembly. Examples of colchicine derivatives include deme 55 The term “epothilone derivative refers to a compound that colcine. is isolated or derived from a myxobacterium Sorangium cel The term “crosslinker” as used herein refers to any chemi lulosum or a structural analog thereof that interferes with cal agent that joins two or more molecules through covalent mitotic spindle assembly. Epothilone derivatives are poten bond(s) or through strong hydrogen-bonding(s). tially effective in treating cancer, such as breast cancer. The term “cyclin-dependent kinase inhibitor refers to a 60 Examples of epothilone derivatives include ixabepilone and compound that inhibits the enzyme cyclin-dependent kinase, 21-aminoepothilone B. which regulates cellular mitosis and transcription. Cyclin The term “halichondrin derivative' refers to a compound dependent kinase inhibitors are being investigated for the isolated or derived from the marine sponge Halichondria treatment of cancer. Examples of cyclin-dependent kinase Okadai or a structural analog thereof that interferes with for inhibitors include dinaciclib and seliciclib. 65 mation of microtubules. Halichondrin derivatives are effec The term “cycloalkane' refers to a saturated or partially tive in treating breast cancer. Examples of halichondrin saturated cyclic or polycyclic hydrocarbon group. In certain derivatives include eribulin. US 9,295,731 B2 13 14 The term “heat shock protein 90 (Hsp90) inhibitor” refers The term “kahalalide derivative' refers to a compound that to a compound that binds to and interferes with Hsp90, which is isolated or derived from to the marine mollusk Elysia plays an important role in oncogenesis. Examples of Hsp90 rufescens or a structural analog thereof that is active against inhibitors include geldanamycin and alvespimycin. certain human prostate and breast cancer cell lines. Kahala The term “hemiasterlin derivative' refers to a compound lide derivatives are potentially effective in treating cancer. that is isolated or derived from marine sponges (Cymbastela Examples of kahalalide derivatives include kahalalide F and sp., Hemiasterella minor, Siphonochalina sp., and Auletta elisidepsin. sp.) or a structural analog thereof that depolymerizes micro The term "kinesin-related motor protein Eg5 inhibitor tubules and interferes with cell division. Hemiasterlin deriva refers to a compound that inhibits the motor protein Eg5, 10 which is involved in the regulation of spindle formation dur tives are potentially effective in treating cancer. Examples of ing mitosis. Kinesin-related motor protein Eg5 inhibitors are hemiasterlin derivatives include taltobulin and HTI-286. being investigated for the treatment of cancer. Examples of The term "heteroalkanearenediyl refers to an alkanene kinesin-related motor protein Eg5 inhibitor include litrone arenediyl group in which one or more of the carbonatoms are sib. replaced with a heteroatom (e.g., N, O, S, P, or Si). 15 The term "kinesin spindle protein inhibitor refers to a The term "heteroalkanecycloalkanediyl refers to an compound that inhibits the kinesin spindle protein, which alkanecycloalkanediyl group in which one or more of the involves in the assembly of the bipolar spindle during cell carbonatoms are replaced with a heteroatom (e.g., N, O, S. P. division. Kinesin spindle protein inhibitors are being investi or Si). gated for the treatment of cancer. Examples of kinesin spindle The term "heteroalkanediyl refers to an alkanediyl group protein inhibitor include ispinesib. in which one or more of the carbon atoms is replaced with a The term “ligand’ as used herein refers to an organic com heteroatom (e.g., N, O, S. P. or Si). pound or inorganic Substrate that contains or is functionalized The term "heteroalkyl refers to an alkyl group in which to contain at least one thiol group, at least one primary amine one or more of the carbon atoms are replaced with a heteroa group, at least one secondary amine group, at least one biotin tom (e.g., N, O, S. P. or Si). 25 binding group, at least one photoreactive group, at least one The term "heteroarenediyl refers to an arenediyl group alkyne group, at least one azide group, or a combination of wherein one or more of the carbon atoms are replaced with a any of the foregoing. The function of the ligand, for example, heteroatom (e.g., N, O, S. P. or Si). Examples of heteroare is to prolong the in-vivo half-life of the drug, reduce dosing nediyl groups include furane-diyl and pyridine-diyl. frequency, lower toxicity, enhance the targeted delivery of the The term "heteroaryl” refers to an aryl group wherein one 30 drug, and/or enable parallel in-vitro analysis. The inorganic or more of the ring carbon atoms are replaced with a heteroa Substrate may be, for example, a glass bead or Surface and a tom (e.g., N, O, S, P. or Si). Examples of heteroaryl groups gold bead or surface. The organic compound may be, for include, but are not limited to, monoradicals of acridine, example, a C- hydrocarbon, a natural or modified peptide, arsindole, carbazole, B-carboline, chromane, chromene, cin a natural or modified protein, a natural or modified antibody, noline, furan, imidazole, indazole, indole, indoline, indoliz 35 a natural or modified nucleoside, a natural or modified nucle ine, isobenzofuran, isochromene, isoindole, isoindoline, iso otide, a natural or modified oligonucleotide, a Sugar, a natural quinoline, isothiazole, isoxazole, naphthyridine, oxadiazole, or modified oligosaccharide, an aminoglycoside (antibiotic), oxazole, perimidine, phenanthridine, phenanthroline, phena or a natural or modified polymer or copolymer Such as a Zine, phthalazine, pteridine, purine, pyran, pyrazine, pyra polylactide, a polystyrene Surface, a polystyrene bead, a den Zole, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine, 40 drimer, a polyalkylene oxide, or a polyethylene oxide. A quinazoline, quinoline, quinolizine, quinoxaline, tetrazole, ligand can contain a thiol group, an amine group, a biotin thiadiazole, thiazole, thiophene, triazole, Xanthene, and the binding group, an alkyne group, an azide group, and/or a like. In certain embodiments, a heteroaryl group is Cso photoreactive group, or may be functionalized to contain thiol heteroaryl, C-2 heteroaryl, Css heteroaryl, and in certain group, an amine group, a biotin-binding group, an alkyne embodiments, Cse heteroaryl. In certain embodiments het 45 group, an azide group, and/or a photoreactive group. eroaryl groups are those derived from thiophene, pyrrole, Examples of peptides include glutathione, carnosine, and benzothiophene, benzofuran, indole, pyridine, quinoline, pantetheline. imidazole, oxazole, and pyrazine. Examples of proteins include bovine serum albumin, The term "heterocycloalkanediyl refers to a cycloal human serum albumin, avidin, and strepavidin. kanediyl group in which one or more of the carbonatoms are 50 Examples of antibodies include polyclonal antibodies, replaced with a heteroatom (e.g., N, O, S, P, or Si). monoclonal antibodies, murine monoclonal antibodies, chi The term "heterocycloalkyl refers to a cycloalkyl group in meric monoclonal antibodies, fusion proteins, humanized which one or more of the carbon atoms are replaced with a monoclonal antibodies, and human monoclonal antibodies. heteroatom (e.g., N, O, S. P. or Si). In certain embodiments, an antibody is a humanized mono The term “histone deacetylase inhibitor refers to a com 55 clonal antibody and is selected from Afutuzumab, Alemtu pound that interferes with the enzyme histone deacetylase, Zumab, Bevacizumab, Bivatuzumab, Cantuzumab, Citatu which involves with the coiling of DNA around histones. Zumab, DacetuZumab, Elotuzumab, Etaracizumab, Histone diacetylase inhibitors are being investigated for the FarletuZumab, Gemtuzumab ozogamicin, InotuZumab ozo treatment of lymphoma, breast, and lung cancers. Examples gamicin, LabetuZumab, LintuZumab, MatuZumab.S. Milatu of histone deacetylase inhibitors include entinostat, moceti 60 Zumab, Nimotuzumab, Oportuzumab monatox, Pertuzumab, nostat, panobinostat, and tacedinaline. SibrotuZumab, TacatuZumab tetraxetan, TigatuZumab, Tras The term “immunomodulator” refers to a compound that tuzumab, Tucotuzumab celmoleukin, Veltuzumab, Aseli stimulates and/or suppresses the immune system. Examples Zumab, Apolizumab, Benralizumab, Cedelizumab, Certoli of immunomodulators include apilimod, balamapimod, dil Zumab, Daclizumab, Eculizumab, Efalizumab, mapimod, epetirimod, fingolimod, golotimod, imiquimod, 65 Epratuzumab, Erlizumab, Fontolizumab, Mepolizumab, lenalidomide, loSmapimod, myriocin, pamapimod, residui Natalizumab, Ocrelizumab, Omalizumab, Pascolizumab, mod, gusperimus, pomalidomide, and Sotirimod. Pexelizumab, PRO 140, Reslizumab, Rontalizumab, Roveli US 9,295,731 B2 15 16 Zumab, Ruplizumab, Siplizumab, Talizumab, Teplizumab, Examples of dendrimers include poly(amidoamine) den Tocilizumab, Toralizumab, Vedolizumab, Visilizumab, trimers (PAMAM) and poly(propylenimine) dentrimers TGN1412, Ibalizumab, Tefibazumab, Alacizumab pegol, (PPI). Bevacizumab/Ranibizumab, Etaracizumab, Tadocizumab, The term “microtubule interference compounds’ refers to BapineuZumab, SolaneZumab, Tanezumab, UrtoxaZumab, a class of compounds that disrupt the polymerization or depo Felvizumab, Motavizumab, Palivizumab, Lebrikizumab, and lymerization of microtubules during mitosis. Microtublue RanibiZumab. In certain embodiments, an antibody is a interference compounds are being used and investigated for murine monoclonal antibody and is selected from Abago the treatment of various cancers. Examples of microtubule Vomab, Igovomab, Oregovomab, Afelimomab, Elsilimomab, interference compounds include Vinca alkaloid derivatives, Faralimomab, Gavilimomab, Inolimomab, Maslimomab, 10 cevipabulin, denibulin, and ombrabulin. Nerelimomab, Odulimomab, Telimomab aritox, Vepalimo The term "parent aromatic ring system” refers to an unsat mab, Zolimomabaritox. Altumomab pentetate, Anatumomab urated cyclic or polycyclic ring system having a conjugated at mafenatox, Arcitumomab, Bectumomab, Blinatumomab, electron system. Included within the definition of “parent CC49, Detumomab, Ibritumomab tiuxetan, Minretumomab, aromatic ring system” arefused ring systems in which one or Mitumomab, Naptumomab estafenatox, Nofetumomab mer 15 more of the rings are aromatic and one or more of the rings are pentan, Pemtumomab, Pintumomab, Satumomab pendetide, saturated or unsaturated. Such as, fluorene, indane, indene, Taplitumomab paptox, Tenatumomab, Tositumomab, 3F8, phenalene, etc. Examples of parent aromatic ring systems Besilesomab, Fanolesomab, Lemalesomab, Sulesomab, include, but are not limited to, aceanthrylene, acenaphthyl Biciromab, Imciromab, Capromab pendetide, Edobacomab, ene, acephenanthrylene, anthracene, aZulene, benzene, chry Edrecolomab, and Nacolomab tafenatox. In certain embodi sene, coronene, fluoranthene, fluorene, hexacene, hexaphene, ments, an antibody is a chimeric monoclonal antibody and is hexalene, as-indacene, S-indacene, indane, indene, naphtha selected from Bavituximab, Brentuximab vedotin, Cetux lene, octacene, octaphene, octalene, ovalene, penta-2,4-di imab, Siltuximab, Rituximab, Abciximab, Volociximab, ene, pentacene, pentalene, pentaphene, perylene, phenalene, Basiliximab, Clenoliximab, Galiximab, Gomiliximab, Inflix phenanthrene, picene, pleiadene, pyrene, pyranthrene, imab, Keliximab, Lumiliximab, Priliximab, Teneliximab, 25 rubicene, triphenylene, and trinaphthalene. Vapaliximab, Ecromeximab, and Pagibaximab. In certain The term “parent heteroaromatic ring system” refers to a embodiments, an antibody is a human monoclonal antibody parent aromatic ring system in which one or more carbon and is selected from Adalimumab, Atorolimumab, Fresoli atoms (and any associated hydrogen atoms) are indepen mumab, Golimumab, Lerdelimumab, Metelimumab, Moroli dently replaced with the same or different heteroatom. mumab, Ipilimumab, Tremelimumab, Bertilimumab, Zanoli 30 Examples of heteroatoms to replace the carbon atoms mumab, Briakinumab, Canakinumab, Ustekinumab, include, but are not limited to, N. P. O, S, Si, etc. Specifically Adecatumumab, Belimumab, Cixutumumab, Conatu included within the definition of "parent heteroaromatic ring mumab, Figitumumab, Iratumumab, Lexatumumab, Lucatu systems' are fused ring systems in which one or more of the mumab, Mapatumumab, Necitumumab, Ofatumumab, rings are aromatic and one or more of the rings are saturated Olaratumab, Panitumumab, Pritumumab, Robatumumab, 35 or unsaturated, such as, arsindole, benzodioxan, benzofuran, Votumumab, Zalutumumab, Denosumab, Stamulumab, chromane, chromene, indole, indoline, Xanthene, etc. Efungumab, Exbivirumab, Foravirumab, Libivirumab, Examples of parent heteroaromatic ring systems include, but Rafivirumab, Regavirumab, Sevirumab, Tuvirumab, are not limited to, arsindole, carbazole, B-carboline, chro Nebacumab, Panobacumab, Raxibacumab, Ramucirumab, mane, chromene, cinnoline, furan, imidazole, indazole, Gantenerumab, and Glembatumumab. 40 indole, indoline, indolizine, isobenzofuran, isochromene, Examples of nucleosides include thymidine, cytidine, uri isoindole, isoindoline, isoquinoline, isothiazole, isoxazole, dine, adenosine, and guanosine. naphthyridine, oxadiazole, oxazole, perimidine, phenanthri Examples of oligonucleotides include single-stranded and dine, phenanthroline, phenazine, phthalazine, pteridine, double-stranded oligoribonucleotides, oligoribonucleotide purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyri derivatives, oligodeoxyribonucleotides, and oligodeoxyribo 45 midine, pyrrole, pyrrolizine, quinazoline, quinoline, quino nucleotide derivatives such as phosphorothioates, phospho lizine, quinoxaline, tetrazole, thiadiazole, thiazole, ramidates, and phosphorothioamidates. thiophene, triazole, and Xanthene. Examples of oligonucleotides also include oblimersen and The term “patient includes mammals, such as for imetelstat. example, humans. The terms “patient and “patient” are used Examples of Sugars include glucosamine. 50 interchangeably. Examples of aminoglycosides include streptomycin, neo The term “pharmaceutical composition” refers to at least mycin, framycetin, paromomycin, ribostamycin, kanamycin, one compound and a pharmaceutically acceptable vehicle amikacin, arbekacin, bekanamycin, dibekacin, tobramycin, with which the compound is administered to a patient. spectinomycin, hygromycin B, paromomycin Sulfate, gen The term “pharmaceutically acceptable' refers to tamicin, netilmicin, sisomicin, isepamicin, Verdamicin, and 55 approved or approvable by a regulatory agency of the Federal astromicin. or a state government or listed in the U.S. Pharmacopoeia or Examples of oligosaccharides include polyglucosamine other generally recognized pharmacopoeia for use in animals, (chitosan). including humans. Examples of polymers include polyethylene glycol (PEG), The term “pharmaceutically acceptable salt” refers to a salt monomethyl polyethylene glycol (MPEG), polypropylene 60 of a compound, which possesses the desired pharmacological glycol (PPG), polylactide, N-(2-hydroxypropyl)methacryla activity of the parent compound. Such salts include acid addi mide (HPMA) copolymer, and poly(styrene-co-maleic acid). tion salts, formed with inorganic acids such as hydrochloric In certain embodiments, the polymer is MPEG having a num acid, hydrobromic acid, Sulfuric acid, nitric acid, phosphoric ber average molecular weight from about 200 to about 60,000 acid, and the like; or formed with organic acids such as acetic Daltons, from about 1,000 to about 40,000 Daltons, and in 65 acid, propionic acid, hexanoic acid, cyclopentanepropionic certain embodiments, from about 2,000 to about 12,500 Dal acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, tOnS. Succinic acid, malic acid, maleic acid, fumaric acid, tartaric US 9,295,731 B2 17 18 acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic -continued acid, cinnamic acid, mandelic acid, methanesulfonic acid, (Aid4) ethanesulfonic acid, 1.2-ethane-disulfonic acid, 2-hydroxy O ethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzene Sulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo2.2.2-oct-2- ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropi onic acid, trimethylacetic acid, tertiary butylacetic acid, lau (Ad5) ryl Sulfuric acid, gluconic acid, glutamic acid, NO hydroxynaphthoic acid, salicylic acid, Stearic acid, muconic 10 acid, and the like; and salts formed when an acidic proton present in the parent compound is replaced by a metal ion, e.g., an alkali metalion, an alkaline earthion, oran aluminum ion; or coordinates with an organic base such as ethanola 15 mine, diethanolamine, triethanolamine, N-methylglucamine, (Adé) and the like. In certain embodiments, a pharmaceutically F acceptable salt is the hydrochloride salt. In certain embodi F ments, a pharmaceutically acceptable salt is the Sodium salt. The term “pharmaceutically acceptable salt includes hydrates and other Solvates, as well as salts in crystalline or N F non-crystalline form. F The term “pharmaceutically acceptable vehicle' refers to a (Ad7) pharmaceutically acceptable diluent, a pharmaceutically N O O acceptable adjuvant, a pharmaceutically acceptable excipi 25 ent, a pharmaceutically acceptable carrier, or a combination of any of the foregoing with which a compound can be admin 2 istered to a patient and which does not destroy the pharma and cological activity thereof and which is nontoxic when admin 30 (Ad8) istered in doses sufficient to provide a therapeutically effective amount of the compound. The term “photoreactive group’ as used herein refers to a functional group capable of reacting with a primary, second ary or tertiary amine upon exposure to actinic radiation, Such 35 as ultraviolet light, to form at least one covalent bond. Examples of photoreactive groups include an azide group, a Examples of photoreactive groups are also disclosed in U.S. diaziridine group, a coumarin group, and a psoralen group. In Publication No. 2001/00022761. certain embodiments, a photoreactive group is selected from 40 The term “poly (ADP-ribose) polymerase (PARP) inhibi Formula (A'd1), Formula (A'd2), Formula (A'd3), Formula tor” refers to a compound that inhibits the enzyme PARP, (A'd4), Formula (A'd5), Formula (A'd6). Formula (A"d7), which is associated with DNA repair and programmed cell and Formula (Ad8): death. Examples of a PARP inhibitors include veliparib. The term “purine analog refers to a compound that mim 45 ics the nucleoside adenosine, nucleoside guanosine, nucleo (Ad1) base adenine, or nucleobase guanine and interferes with the DNA replication. Purine analogs are effective in treating cer tain cancers. Examples of purine analogs include deoxyco X formycin, cladribine, clofarabine, fludarabine, thioguanine, 50 and mercaptopurine. O O O The term "pyrimidine analog refers to a compound that mimics the deoxynucleoside thymidine, nucleoside cytidine, N / nucleoside uridine, nucleobase thymine, nucleobase cytosine or nucleobase uracil and interferes with the DNA replication. (Ad2) 55 Pyrimidine analogs are effective in treating certain cancers. Examples of pyrimidine analogs include azacitidine, cytara bine, decitabine, and gemcitabine. The term "pyrrolobenzodiazepine” refers to a compound N.--N that is isolated orderived from Streptomyces sp. or a structural 60 analog thereof that alkylates DNA at minor groove sites. (Ad3) Pyrrolobenzodiazepine is potentially effective in treating cancer. Examples of pyrrolobenzodiazepine include abbey mycin, anthramycin, centanamycin, chicamycin, mazethra mycin, porothramycin A, porothramycin B, Sibanomycin, 65 and Sibiromycin. N The term “streptomyces” refers to a compound that is iso lated or derived from the bacteria Streptomyces or a structural US 9,295,731 B2 19 20 analog thereof that has antibacterial, antifungal, anti-para -continued Sitic, and/or antineoplastic properties. Examples of strepto (Aa3) myces include actinomycin D, 7-aminoactinomycin D, bleo and mycin, mitomycin, and staurosporine. The term “substituted” refers to a group in which one or 5 tsu, more hydrogen atoms are each independently replaced with (A'a4) the same or different substituent(s). In certain embodiments, a substituent is selected from halogen, -S(O),OH, Ns-s-X S —S(O). C. alkyl, -SH, -S-C alkyl, -COOH, 10 —CONH2, N, NO, NH, -NH(C. alkyl). 2 —N(C. alkyl). —CN, =O, C-alkyl, -CF, -OH, Cs aryl, Coheteroalkyl, Css heteroaryl, Calkoxy, C-acyl, and —COR where R is C alkyl. In certain embodiments, a wherein each R and R is independently selected from substituent is chosen from -OH, -NH, and C alkyl. hydrogen, Calkyl, substituted Calkyl, Cheteroalkyl, The term “therapeutic agent” or “drug” refers to a com 15 Substituted C. heteroalkyl, Co aryl, substituted C-aryl, pound known to be or believed to be effective in treating a Coheteroaryl, and substituted Coheteroaryl; and each E disease in a patient. is selected from F, Cl, Br, and I. The term “therapeutically effective amount” refers to the The term “topoisomerase inhibitor” refers to a compound amount of a compound that, when administered to a patient that inhibits the enzyme topoisomerase, which involves in the for treating a disease in a patient, is sufficient to reduce, unwinding and winding of DNA during transcription and minimize, and/or prevent the disease. A “therapeutically replication. Topoisomerase inhibitors are being used and effective amount” can vary depending, for example, on the investigated for the treatment of various cancers. Examples of compound, the nature or cause of the disease, severity of the topoisomerase inhibitors include camptotheca derivatives disease, the age, weight, and/or health of the patient to be 25 and edotecarin. treated, and the judgment of the prescribing physician. An The term “treating” or “treatment” of any disease or disor appropriate amount in any given instance can be ascertained der refers to arresting orameliorating a disease, disorder, or at by those skilled in the art or capable of determination by least one of the clinical symptoms of a disease or disorder, routine experimentation. The terms “therapeutically effective reducing the risk of acquiring a disease, disorder, or at least amount” and “prophylactically effective amount” are used 30 one of the clinical symptoms of a disease or disorder, reduc interchangeably. A therapeutically effective amount can also ing the development of a disease, disorder or at least one of the mean a dose that has been recommended or approved by any clinical symptoms of the disease or disorder, or reducing the of the various regulatory or advisory organizations in the risk of developing a disease or disorder or at least one of the medical or pharmaceutical arts (e.g., FDA, AMA) or by the clinical symptoms of a disease or disorder. “Treating” or manufacturer or supplier. 35 “treatment” also refers to inhibiting the disease or disorder, The term “therapeutically effective dose' refers to a dose either physically, (e.g., stabilization of a discernible symp that provides effective treatment of a disease or disorder in a tom), physiologically, (e.g., stabilization of a physical param patient. A therapeutically effective dose can vary from com eter), or both, and to inhibiting at least one physical parameter pound to compound, and from patient to patient, and can which may or may not be discernible to the patient. In certain depend upon factors such as the condition of the patient and 40 embodiments, “treating or “treatment” refers to delaying the the route of delivery. A therapeutically effective dose can be onset of the disease or disorder or at least one or more symp determined in accordance with routine pharmacological pro toms thereof in a patient which may be exposed to or predis cedures known to those skilled in the art. posed to a disease or disorder even though that patient does The term “thiol-reactive group’ as used herein refers to a not yet experience or display symptoms of the disease or functional group capable of reacting with a thiol group to 45 disorder. form a thiol ether bond, a disulfide bond, or a thiourea bond. The term “tubulysin derivative” refers to a compound hav Examples of a thiol-reactive group include a vinyl group ing the basic structural scaffold: (-CH=CH-), a haloalkyl group, a haloacetyl group, and an isocyanate group (-NCO). In certain embodiments a thiol reactive group is selected from Formula (A'al), Formula 50 (A'a2). Formula (Aa3), and Formula (A'a4):

(Aa1) 55

60 and is potentially effective in treating cancer. Tubulysin derivatives depolymerize microtubules and induce mitotic (Aa2) arrest. Examples of tubulysin derivatives include tubulysin A, tubulysin B, and tubulysin D. The term "tyrosine kinase inhibitor” refers to a compound 65 that inhibits the enzyme tyrosine kinase, which regulates cellular activity through the phosphorylation of the amino acid tyrosine of a signaling protein. Tyrosine kinase inhibitors US 9,295,731 B2 21 22 are effective in treating chronic myelogenous leukemia and stituted C. heteroalkyl, Co aryl, Substituted Co are being for the treatment of other cancers. Examples of aryl, Coheteroaryl, and substituted Coheteroaryl; tyrosine kinase inhibitors include afatinib. bosutinib, caner D is selected from a therapeutic agent having at least one tinib, crizotinib, dacomitinib, dasatinib, dovitinib, erlotinib, primary amine group, a therapeutic agent having at least fostamatinib, gefitinib, imatinib, intedanib, lapatinib, lini one secondary amine group, a therapeutic agent having fanib, masitinib, motesanib, neratinib, nilotinib, paZopanib, at least one hydroxy group, and a therapeutic agent Saracatinib, Selumetinib, tellatinib, tipifarnib, Vandetanib, and having at least one thiol group; and Vatalanib. n5 is an integer selected from 1 to 20. The term “vinca alkaloid derivative' refers to a compound In certain embodiments of a compound of Formula (II), isolated or derived from a plant Madagascar Periwinkle or a 10 each A' is the same, and in certain embodiments, at least some structural analog thereof that inhibits microtubule polymer A' are different. ization. Examples of Vinca alkaloid derivatives include In certain embodiments of a compound of Formula (II), desacetylvinblastine hydrazide, vinblastine, Vincristine, Vin eachX' is the same, and in certain embodiments, at least some destine, Vinflunine, and vinorelbine. X' are different. Compounds are named using Symyx Draw 3.3, Symyx 15 In certain embodiments of a compound of Formula (II), Solutions Inc., 2010. eachX is the same, and in certain embodiments, at least some Reference is now made in detail to certain embodiments of X’ are different. In certain embodiments of a compound of Formula (II), compounds, compositions, and methods. The disclosed each X is the same, and in certain embodiments, at least some embodiments are not intended to be limiting of the claims. To X are different. the contrary, the claims are intended to coverall alternatives, In certain embodiments of a compound of Formula (II), modifications, and equivalents. each R' is the same, and in certain embodiments, at least some Cleavable Conjugates R" are different. 25 In certain embodiments of a compound of Formula (II), In certain embodiments, a compound provided by the eachR is the same, and in certain embodiments, at least some present disclosure has the structure of Formula (II): Rare different. In certain embodiments of a compound of Formula (II), each group within the brackets is the same, and in certain (II) 30 embodiments, at least one group within the brackets is differ O R1 R2 O ent. In certain embodiments of a compound of Formula (II), Al Nx11 x Yx3 -l O X O each A' is independently a moiety selected from Formula 5 (A'al), Formula (A'a2). Formula (Aa3), Formula (A'b1), 35 Formula (A'b2), Formula (A'c1), Formula (A'c2), Formula or a salt thereof, wherein: (A'c3). Formula (Ad1), Formula (Ad2). Formula (Ad3), each A' is independently selected from a thiol-reactive Formula (A'el), Formula (A' fl), and Formula (A"f2): group, an amine-reactive group, an avidin-binding group, a photoreactive group, an alkyne-reactive group, and an azide-reactive group; 40 (Aa1) each X and X is independently selected from a covalent R8 O bond, Co alkanediyl. Substituted Co alkanediyl. Co heteroalkanediyl. Substituted Co. heteroal kanediyl, C-2 cycloalkanediyl. Substituted C. cycloalkanediyl, C. heterocycloalkanediyl. Substi 45 R9 y tuted C. heterocycloalkanediyl, Cao alkanecycloal O kanediyl. Substituted Coalkanecycloalkanediyl, Cao (Aa2) heteroalkanecycloalkanediyl. Substituted Cao het eroalkanecycloalkanediyl, Co arenediyl. Substituted Co-oarenediyl. Co-o heteroarenediyl. Substituted C-20 50 heteroarenediyl, C-2 alkanearenediyl. Substituted NX C7-20 alkanearenediyl, C-20 heteroalkanearenediyl. (Aa3) Substituted Co. heteroalkanearenediyl, and (CH)—(CH-CH O), (CH), , wherein: 55 tsu', each n1 and n3 is independently an integer selected from (Ab1) 0 to 5; and O each n2 is independently an integer selected from 1 to 25; each X* is independently selected from a covalent bond, 60 O— —S , —N , —N-, - N=N , —N–C , SO— —SO , —SON.— —SS—, —C(O)—, (Ab2) C(O)O-, - C(O)N , —C(O)S –C(O)N O R8 R9 O

- NC(O)N , and - NC(S)N: 65 L2 ls, O each R" and R is independently selected from hydrogen, C. alkyl, Substituted C. alkyl, C. heteroalkyl, Sub US 9,295,731 B2 23 24 -continued -continued (Af2)

10

(Ac2) wherein: each R and R is independently selected from hydro gen, C. alkyl, Substituted C. alkyl, C. het 15 eroalkyl, Substituted Cheteroalkyl, Co aryl. Sub stituted Co aryl, Co heteroaryl, and Substituted Coheteroaryl; each E is selected from F, Cl, Br, and I; and each L is selected from halogen, N-hydroxysuccinim idyl, substituted N-hydroxysuccinimidyl, phenol-yl, substituted phenol-yl, hydroxybenzotriazolyl, substi tuted hydroxybenzotriazolyl, imidazolyl, and substi tuted imidazolyl. 25 In certain embodiments of a compound of Formula (II), each A' is independently selected from a thiol-reactive group, each A' is independently selected from an amine-reactive group, each A' is independently selected from an avidin binding group, and in certain embodiments, each A' is inde 30 pendently selected from a photoreactive group, each A' is (Ad1) independently selected from an alkyne-reactive group, and each A' is independently selected from an azide-reactive group. In certain embodiments of a compound of Formula (II), 35 each A' is a thiol-reactive group and is independently selected from Formula (A'al), Formula (A'a2), and Formula (Aa3). In certain embodiments of a compound of Formula (II), each A' is an amine-reactive group and is independently selected from Formula (Ab1) and Formula (A'b2). In certain (Ad2) 40 embodiments of Formula (II), each A' is an avidin-binding group and is independently selected from Formula (A'c1), Formula (Ac2), and Formula (A'c3). In certain embodi ments of a compound of Formula (II), each A' is a photore active group and is independently selected from Formula 45 (Ad1), Formula (Ad2), and Formula (Ad3). In certain embodiments of a compound of Formula (II), each A' is an alkyne-reactive group of Formula (A"e1). In certain embodi (Ad3) ments of a compound of Formula (II), each A' is an azide reactive group and is independently selected from Formula 50 (A' fl) and Formula (A" f2). In certain embodiments of a compound of Formula (II), each A' is Formula (Aa1). In certain embodiments of a compound of Formula (II), each A' is Formula (A'al), wherein each R and R is independently selected from 55 hydrogen, C alkyl and Ces aryl; in certain embodiments, (Ale1) each RandR is independently selected from hydrogen, C. alkyl, and Caryl; and in certain embodiments, each R and R is hydrogen. In certain embodiments of a compound of Formula (II), 60 each A' is Formula (A'a2). In certain embodiments of a compound of Formula (II), each A' is Formula (Aa2), wherein each R is selected from hydrogen and Cia alkyl; and (Alf1) in certain embodiments, each R is selected from hydrogen and methyl. 65 In certain embodiments of a compound of Formula (II), each A' is Formula (Aa3). In certain embodiments of a compound of Formula (II), each A' is Formula (Aa3), US 9,295,731 B2 25 26 wherein each E is selected from F, Cl, Br, and I; and in certain In certain embodiments of a compound of Formula (II), embodiments each E is selected from Brand I. each X" is selected from Co alkanediyl, Co cycloal In certain embodiments of a compound of Formula (II), kanediyl, Co arenediyl, and C-2 alkanearenediyl. In cer each A' is Formula (Ab1). In certain embodiments of a tain embodiments of a compound of Formula (II), each X is compound of Formula (II) wherein each A' is Formula Selected from Co alkanediyl, Co cycloalkanediyl, Co (Ab1), each L is selected from Cl, N-hydroxysuccinimidyl, arenediyl, and C- alkanearenediyl. Sulfo-N hydroxysuccinimidyl, 4-nitrophenol-yl, pentafluo In certain embodiments of a compound of Formula (II), rophenol-yl 4-methylsulfonylphenol-yl, and trichlorophe each X is a covalent bond. nol-yl. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), 10 each A' is Formula (Ab2). In certain embodiments of a each X is a covalent bond. compound of Formula (II) wherein each A' is Formula In certain embodiments of a compound of Formula (II), (Ab2), each RandR is independently selected from hydro each X and X is independently selected from a covalent gen, Calkyl, and Cesaryl, and each L is selected from Cl, bond, methane-diyl, ethane-1,2-diyl, propane-1,3-diyl. N-hydroxysuccinimidyl, sulfo-N hydroxysuccinimidyl, 4-ni 15 butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6-diyl, heptane trophenol-yl, pentafluorophenol-yl 4-methylsulfonylphe 1,7-diyl octane-1,8-diyl. nonane-1,9-diyl, decane-1,10-diyl. nol-yl, and trichlorophenol-yl. In certain embodiments of a undecane-1,11-diyl, dodecane-1,12-diyl, hexadecane-1,16 compound of Formula (II) wherein each A' is Formula diyl, benzene -diyl, methylbenzene-diyl, propylbenzene (Ab2), each RandR is independently selected from hydro diyl, methylcyclohexane-diyl, and —(CH)—(CH2— gen, methyl ethyl, propyl, isopropyl, and phenyl, and each Lif CH2—O) (CH) , wherein each n1 and n3 is is selected from Cl, N-hydroxysuccinimidyl, sulfo-N hydrox independently an integer selected from 0 to 3 and each n2 is ySuccinimidyl, 4-nitrophenol-yl, pentafluorophenol-yl, independently an integer selected from 1 to 20. 4-methylsulfonylphenol-yl, and trichlorophenol-yl. In cer In certain embodiments of a compound of Formula (II), tain embodiments of a compound of Formula (II) wherein each X and X is independently selected from a covalent each A' is Formula (A'b2), each RandR is hydrogen, and 25 bond, methane-diyl, ethane-1,2-diyl, propane-1,3-diyl, pen each L is selected from Cl, N-hydroxysuccinimidyl, sulfo-N tane-1,5-diyl 4-methylcyclohexane-diyl, benzene-1,2-diyl. hydroxySuccinimidyl, 4-nitrophenol-yl, pentafluorophenol benzene-1,3-diyl, and benzene-1,4-diyl. y1, 4-methylsulfonylphenol-yl, and trichlorophenol-yl. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each X is a covalent bond; and each X is selected from a each A' is Formula (A'c1). In certain embodiments of a 30 covalent bond, methane-diyl ethane-1,2-diyl, propane-1,3- compound of Formula (II), each A' is Formula (A'c2). In diyl, pentane-1,5-diyl 4-methylcyclohexane-diyl, benzene certain embodiments of a compound of Formula (II), each A' 1,2-diyl, benzene-1,3-diyl, and benzene-1,4-diyl. is Formula (A'c3). In certain embodiments of a compound of In certain embodiments of a compound of Formula (II), Formula (II), each A' is Formula (A'd1). In certain embodi each X is ethane-1,2-diyl; and each X is selected from a ments of a compound of Formula (II), each A' is Formula 35 covalent bond, methane-diyl ethane-1,2-diyl, propane-1,3- (A'd2). In certain embodiments of a compound of Formula diyl, pentane-1,5-diyl 4-methylcyclohexane-diyl, benzene (II), each A' is Formula (A'd3). In certain embodiments of a 1,2-diyl, benzene-1,3-diyl, and benzene-1,4-diyl. compound of Formula (II), each A' is Formula (A"e1). In In certain embodiments of a compound of Formula (II), certain embodiments of a compound of Formula (II), each A' each X* is pentane-1,5-diyl; and each X is selected from a is Formula (A'fl). In certain embodiments of a compound of 40 covalent bond, methane-diyl ethane-1,2-diyl, propane-1,3- Formula (II), each A' is Formula (A" f2). diyl, pentane-1,5-diyl 4-methylcyclohexane-diyl, benzene In certain embodiments of a compound of Formula (II), 1,2-diyl, benzene-1,3-diyl, and benzene-1,4-diyl. each X and X is independently selected from a covalent In certain embodiments of a compound of Formula (II), bond, Co alkanediyl. Substituted Co alkanediyl, C-12 eachX' is 4-methylcyclohexane-diyl; and X is selected from cycloalkanediyl. Substituted C-2 cycloalkanediyl. Substi 45 a covalent bond, methane-diyl ethane-1,2-diyl, propane-1,3- tuted C. heterocycloalkanediyl. Cao arenediyl. Substi diyl, pentane-1,5-diyl 4-methylcyclohexane-diyl, benzene tuted Coarenediyl. Czo alkanearenediyl. Substituted Czo 1,2-diyl, benzene-1,3-diyl, and benzene-1,4-diyl. alkanearenediyl, and (CH2)—(CH2—CH2—O), In certain embodiments of a compound of Formula (II) (CH), , wherein each n1 and n3 is independently an inte wherein each X and/or X is —(CH)—(CH CH ger selected from 0 to 5 and each n2 is independently an 50 O) (CH), , each n1 and n3 is independently an integer integer selected from 1 to 25. selected from 0 to 5, an integer selected from 0 to 4, an integer In certain embodiments of a compound of Formula (II), selected from 0 to 3, and in certain embodiments, an integer each X and X is independently selected from a covalent selected from 0 to 2. In certain embodiments of a compound bond, C- alkanediyl. Substituted C- alkanediyl, C. of Formula (II) wherein each X and/or X is —(CH),— cycloalkanediyl, benzene-diyl, substituted benzene-diyl. 55 (CH2—CH2—O), (CH), , each n2 is independently C, benzenealkane-diyl, and —(CH)—(CH-CH an integer selected from 1 to 25, an integer selected from 1 to O), (CH), , wherein each n1 and n3 is independently 20, an integer selected from 1 to 15, an integer selected from an integer selected from 0 to 5 and each n2 is independently 1 to 10, an integer selected from 1 to 5, an integer selected an integer selected from 1 to 20. from 1 to 4, and in certain embodiments, an integer selected In certain embodiments of a compound of Formula (II), 60 from 1 to 3. each X and X is independently selected from a covalent In certain embodiments of a compound of Formula (II), bond, Cs alkanediyl. Substituted Cls alkanediyl, Cs each n1 and n3 is independently an integer selected from 0 to cycloalkanediyl, benzene-diyl, substituted benzene-diyl. 5, in certain embodiments, an integer selected from 0 to 4, and C- benzenealkane-diyl, and —(CH2)—(CH2—CH2— in certain embodiments, an integer selected from 0 to 3; and O), (CH), , wherein each n1 and n3 is independently 65 each n2 is independently an integer selected from 1 to 25, in an integer selected from 0 to 3 and each n2 is independently certain embodiments an integer selected from 1 to 20, in an integer selected from 1 to 10. certain embodiments an integer selected from 1 to 15, in US 9,295,731 B2 27 28 certain embodiments an integer selected from 1 to 10, and in In certain embodiments of a compound of Formula (II), D certain embodiments an integer selected from 1 to 5. is selected from aminopterin, folitix.orin, methotrexate, pem In certain embodiments of a compound of Formula (II), etrexed, pralatrexate, raltitrexed, pelitrexol, talotrexin, each X is selected from a covalent bond, -O-, -S , deoxycoformycin, cladribine, clofarabine, fludarabine, N— —N—, N—N— —N=C —SO— —SO , thioguanine, mercaptopurine, berubicin, daunorubicin, doxo SON ... —SS , —C(O)— —C(O)O , —C(O)N , rubicin, epirubicin, idarubicin, amrubicin, pirarubicin, Zoru —C(O)S C(O)N N—, OP(O)(OH)O-, - OC(O) bicin, mitoxantrone, banoxantrone, ledoxantrone, nortopix O - OC(O)N - NC(O)N , and NC(S)N. In certain antrone, pixantrone, piroxantrone, Sabarubicin, topixantrone, embodiments of a compound of Formula (II), each X* is monomethyl auristatin E. monomethyl auristatin F. monom selected from a covalent bond, —O— —S— —SO—, 10 ethyl dolastatin 10, dolastatin 15, belotecan, atiratecan, SO —SON ... —SS , —C(O)— —C(O)O—, camptothecin, exatecan, irinotecan, namitecan, rubitecan, C(O)N , —C(O)N N=, OC(O)N , and - NC(O) topotecan, demecolcine, duocarmycin A, duocarmycin SA, N—. In certain embodiments of a compound of Formula (II), duocarmycin B, duocarmycin B1, abbeymycin, anthramycin, each X is selected from a covalent bond, SO , centanamycin, chicamycin, mazethramycin, porothramycin —SON , and —C(O)N . 15 A, porothramycin B, Sibanomycin, Sibiromycin, trabectedin, In certain embodiments of a compound of Formula (II), calicheamicin Y1. calicheamicin T, esperamicin A1, espe each RandR is independently selected from hydrogen, C. ramicin C, esperamicin D, dynemicin A, dynemicin H, dyne alkyl, Substituted Calkyl, Cheteroalkyl, Substituted C micin M, dynemicin N, dynemicin O. dynemicin P. dynemi heteroalkyl, Coo aryl, Substituted Co-o aryl, Co-o het cin Q, dynemicin S. neocarzinostatin chromophore, eroaryl, and Substituted Coheteroaryl. uncialamycin, 21-aminoepothilone B, eribulin, hemiasterlin, In certain embodiments of a compound of Formula (II), HTI-286, kahalatide F, elsamitrucin, lucanthone, melphalan, each RandR is independently selected from hydrogen, C. mitoguaZone, nimustine, procarbazine, dacarbazine, amsa alkyl, Substituted C. alkyl, and Co aryl. In certain crine, 5-amino-4-oxo-pentanoic acid, methyl 5-amino-4- embodiments of a compound of Formula (II), R' and Rare oXo-pentanoate, actinomycin D, 7-aminoactinomycin D, independently selected from hydrogen, methyl, ethyl, propyl. 25 bleomycin, mitomycin, staurosporine, desacetylvinblastine isopropyl, and phenyl. In certain embodiments of a com hydrazide, vinblastine, Vincristine, Vindestine, Vinflunine, pound of Formula (II), each R' and R is independently Vinorelbine, afatinib, apilimod, balamapimod, barasertib, selected from hydrogen and methyl. In certain embodiments bosutinib, canertinib, cevipabulin, crizotinib, dacomitinib, of a compound of Formula (II), each R' and R is indepen dasatinib, denibulin, dilimapimod, dinaciclib, dovitinib, duta dently selected from hydrogen and ethyl. In certain embodi 30 catib, duVoglustat, edotecarin, elisidepsin, entinostat, epetiri ments of a compound of Formula (II), each R' and R is mod, erlotinib, fingolimod, fostamatinib, gefitinib, goloti independently selected from hydrogen and propyl. In certain mod, gusperimus, imatinib, imiquimod, intedanib, ispinesib. embodiments of a compound of Formula (II), each R" and R' lapatinib, lenalidomide, linifanib, litronesib, loSmapimod, is independently selected from hydrogen and isopropyl. In metesind, mocetinostat, motesanib, masitinib, myriocin, certain embodiments of a compound of Formula (II), each R' 35 neratinib, nilotinib, odanacatib, ombrabulin, pamapimod, and R is independently selected from hydrogen and phenyl. panobinostat, paZopanib, plerixafor, pomalidomide, In certain embodiments of a compound of Formula (II), raZupenem, residuimod, Sabarubicin, Saracatinib, Seliciclib, each R' and R is hydrogen. In certain embodiments of a Selumetinib, Sotirimod, squalamine, tacedinaline, tallabostat, compound of Formula (II), each R" is hydrogen and each R taltobulin, tellatinib, tipifarnib, toZasertib, vandetanib, vata is methyl. In certain embodiments of a compound of Formula 40 lanib, Veliparib, Voreloxin, alvespimycin, amikacin, ampho (II), each R" is hydrogen and each R is ethyl. In certain tericin B, arbekacin, astromicin, bacitracin, balofloxacin, embodiments of a compound of Formula (II), each R" is bederocin, bekanamycin, besifloxacin, brodimoprim, ciprof hydrogen and each R is propyl. In certain embodiments of a loxacin, clinafloxacin, colistin, daptomycin, dibekacin, compound of Formula (II), each R" is hydrogen and each R enoxacin, framycetin, garenoxacin, gatifloxacin, gemifloxa is isopropyl. In certain embodiments of a compound of For 45 cin, gentamicin, gentamicin, grepafloxacin, hamycin, hexeti mula (II), each R" is hydrogen and each R is phenyl. dine, hygromycin B, ibacitabine, iclaprim, isepamicin, kana In certain embodiments of a compound of Formula (II), D mycin, lomefloxacin, lucimycin, lymecycline, mepartricin, is selected from a therapeutic agent having at least one pri moxifloxacin, natamycin, nemonoxacin, neomycin B, neo mary amine group, a therapeutic agent having at least one mycin C, netilmicin, norfloxacin, nystatin, omadacycline, secondary amine group, a therapeutic agent having at least 50 oritavancin, paromomycin, paZufloxacin, perimycin A, peri one hydroxy group, and a therapeutic agent having at least mycin B, perimycin C, pipemidic acid, polymyxin B, puro one thiol group. mycin, radeZolid, retaspimycin, ribostamycin, rimocidin, In certain embodiments of a compound of Formula (II), D Sisomicin, sitafloxacin, sparfloxacin, spectinomycin, strepto is selected from a folic acid analog, a purine analog, a pyri mycin, Sulfacetamide, Sulfadiazine, Sulfadimethoxine, Sul midine analog, an anthracycline derivative, a dolastatin 55 fadimidine, Sulfafurazole, Sulfalene, SulfamaZone, Sulfam derivative, a camptotheca derivative, an ectainascidin deriva erazine, sulfamethizole, Sulfamethoxazole, tive, a colchicine derivative, a duocarmycin derivative, an Sulfamethoxypyridazine, Sulfametomidine, Sulfametoxydi enediyne derivative, an epothilone derivative, a halichondrin azine, Sulfametrole, Sulfamoxole, Sulfanilamide, Sulfaperin, derivative, akahalalide derivative, a streptomyces, a tubulysin Sulfaphenazole, Sulfapyridine, Sulfathiazole, Sulfathiourea, derivative, a Vinca alkaloid, an antifolate, a hemiasterlin, a 60 Sulfisomidine, teicoplanin, telavancin, tanespimycin, tema cathepsin K inhibitor, dipeptidyl peptidase IV inhibitor, heat floxacin, tetroXoprim, tigecycline, tobramycin, toSufloxacin, shock protein 90 (Hsp90) inhibitor, a histone deacetylase trimethoprim, trimethoprim, trovafloxacin, tyrothricin, uli inhibitor, an immunomodulator, an aurora kinase inhibitor, a floxacin, Valnemulin, Vancomycin, Verdamicin, and Zabof cyclin-dependent kinase inhibitor, tyrosine kinase inhibitor, loxacin. kinesin-related motor protein Eg5 inhibitor, kinesin spindle 65 In certain embodiments of a compound of Formula (II), n5 protein inhibitor, microtubule interference compounds, is an integer selected from 1 to 20, from 1 to 5, and in certain topoisomerase inhibitor, and an antibiotic. embodiments, n5 is an integer selected from 1 to 3. In certain US 9,295,731 B2 29 30 embodiments, n5 is 1, n5 is 2, n5 is 3, and in certain embodi at least one primary amine group and a therapeutic agent ments, n5 is 4. In certain embodiments of a compound of having at least one secondary amine group; and n5 is selected Formula (II), n5 is selected from 1, 2, 3, and 4. from 1 to 4. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (A'al), wherein each Rand each A' is a moiety of Formula (A'al), wherein each Rand R is independently selected from hydrogen, C. alkyl, and R is hydrogen; each X is pentane-1,5-diyl; each X* is a Cao aryl; each X and X is independently selected from a covalent bond; each X is a covalent bond; each R" is hydro covalent bond, methane-diyl ethane-1,2-diyl, propane-1,3- gen; each R is methyl; D is selected from a therapeutic agent diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6-diyl, hep having at least one primary amine group and a therapeutic 10 agent having at least one secondary amine group; and n5 is tane-1,7-diyl octane-1,8-diyl. nonane-1,9-diyl. decane-1,10 selected from 1 to 4. diyl, undecane-1,11-diyl, dodecane-1,12-diyl, hexadecane-1, In certain embodiments of a compound of Formula (II), 16-diyl, benzene-diyl, methylbenzene-diyl, propylbenzene each A' is a moiety of Formula (A'al), wherein each Rand diyl, methylcyclohexane-diyl, and —(CH)—(CH2— R is hydrogen; each X is pentane-1,5-diyl; each X* is a CH-O), (CH), , wherein each n1 and n3 is 15 covalent bond; each X is a covalent bond; each R' and R is independently an integer selected from 0 to 5, and each n2 is hydrogen; D is selected from a therapeutic agent having at independently an integer selected from 1 to 25; each X is least one primary amine group and a therapeutic agent having independently selected from a covalent bond, —O— —S , at least one secondary amine group; and n5 is selected from 1 SO— —SO —SON.— —SS , —C(O)— —C(O) to 4. O—, C(O)N , C(O)N N=, OC(O)N , and In certain embodiments of a compound of Formula (II), —NC(O)N ; each RandR is independently selected from each A' is a moiety of Formula (A'al), wherein each Rand hydrogen, C alkyl, and Ces aryl; D is selected from a R is hydrogen; each X" is benzene-1,4-diyl; each X* is a therapeutic agent having at least one primary amine group, a covalent bond; each X is propane-1,3-diyl; each RandR is therapeutic agent having at least one secondary amine group, hydrogen; D is selected from a therapeutic agent having at a therapeutic agent having at least one hydroxy group, and a 25 least one primary amine group and a therapeutic agent having therapeutic agent having at least one thiol group; and n5 is at least one secondary amine group; and n5 is selected from 1 selected from 1 to 20. to 4. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (A'al), wherein each Rand each A' is a moiety of Formula (A'al), wherein each Rand R is independently selected from hydrogen and methyl; each 30 R is hydrogen; each X* is -(CH2)-(CH X' and X is independently selected from a covalent bond, CH2—O) (CH) , wherein n1 is 0, n2 is 2, and n3 is 2: methane-diyl ethane-1,2-diyl, propane-1,3-diyl, butane-1,4- each X* is a covalent bond; each X is a covalent bond; each diyl, pentane-1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl. R" and R is hydrogen; D is selected from a therapeutic agent octane-1,8-diyl. nonane-1,9-diyl. decane-1,10-diyl, unde having at least one primary amine group and a therapeutic cane-1,11-diyl, dodecane-1,12-diyl, hexadecane-1,16-diyl. 35 agent having at least one secondary amine group; and n5 is benzene-diyl, methylbenzene-diyl, propylbenzene-diyl. selected from 1 to 4. methylcyclohexane-diyl, and —(CH2)—(CH2—CH2— In certain embodiments of a compound of Formula (II), O), (CH), , wherein each n1 and n3 is independently each A' is a moiety of Formula (A'al), wherein each Rand an integer selected from 0 to 3, and each n2 is independently R is hydrogen; each X is pentane-1,5-diyl; each X is an integer selected from 1 to 20; each X* is selected from a 40 —C(O)N ; each X is pentane-1,5-diyl; each R" is hydro covalent bond, —O— —SO ... —SON.—, —C(O)—, gen; each R is methyl; D is selected from a therapeutic agent —C(O)C)—, and —C(O)N ; each R" and R is indepen having at least one primary amine group and a therapeutic dently selected from hydrogen, Calkyl, and Csaryl; D is agent having at least one secondary amine group; and n5 is selected from a therapeutic agent having at least one primary selected from 1 to 4. amine group and a therapeutic agent having at least one 45 In certain embodiments of a compound of Formula (II), secondary amine group; and n5 is selected from 1 to 5. each A' is a moiety of Formula (A'al), wherein each Rand In certain embodiments of a compound of Formula (II), R is hydrogen; each X* is 4-methylcyclohexane-diyl; each each A' is a moiety of Formula (A'al), wherein each Rand X is C(O)N ; each X is pentane-1,5-diyl; each R' is R is hydrogen; each X is pentane-1,5-diyl; each X* is a hydrogen; each R is methyl; D is selected from a therapeutic covalent bond; each X is a covalent bond; each R' and R is 50 agent having at least one primary amine group and a thera hydrogen; D is selected from a therapeutic agent having at peutic agent having at least one secondary amine group; and least one primary amine group and a therapeutic agent having n5 is selected from 1 to 4. at least one secondary amine group; and n5 is selected from 1 In certain embodiments of a compound of Formula (II), to 4. each A' is a moiety of Formula (Aa2), wherein each R is In certain embodiments of a compound of Formula (II), 55 selected from hydrogen, C. alkyl, and Cao aryl; each X' each A' is a moiety of Formula (A'al), wherein each Rand and X is independently selected from a covalent bond, meth R is hydrogen; each X" is benzene-1,3-diyl; each X is a ane-diyl ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl. covalent bond; each X is a covalent bond; each R' and R is pentane-1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl octane hydrogen; D is selected from a therapeutic agent having at 1,8-diyl. nonane-1,9-diyl. decane-1,10-diyl, undecane-1,11 least one primary amine group and a therapeutic agent having 60 diyl, dodecane-1,12-diyl, hexadecane-1,16-diyl, benzene at least one secondary amine group; and n5 is selected from 1 diyl. methylbenzene-diyl. propylbenzene-diyl. to 4. methylcyclohexane-diyl. and —(CH2)—(CH2— In certain embodiments of a compound of Formula (II), CH-O) (CH), , wherein each n1 and n3 is indepen each A' is a moiety of Formula (A'al), wherein each Rand dently an integer selected from 0 to 5, and each n2 is inde R is hydrogen; each X is 4-methylcyclohexane-diyl; each 65 pendently an integer selected from 1 to 25; each X* is selected X is a covalent bond; each X is a covalent bond; each R" and from a covalent bond, —O— —S— —SO— —SO , R is hydrogen; D is selected from a therapeutic agent having SON ... —SS , —C(O)— —C(O)O , —C(O)N , US 9,295,731 B2 31 32 C(O)N N=, OC(O)N , and NC(O)N-; each R' amine group and a therapeutic agent having at least one and R is independently selected from hydrogen, C. alkyl, secondary amine group; and n5 is selected from 1 to 4. and Cesaryl; D is selected from a therapeutic agent having at In certain embodiments of a compound of Formula (II), least one primary amine group, a therapeutic agent having at each A' is a moiety of Formula (A'a2), wherein each R is least one secondary amine group, a therapeutic agent having methyl; each X is a covalent bond; each X* is C(O)N : at least one hydroxy group, and a therapeutic agent having at each X is 4-methylcyclohexane-diyl; each R' and R is least one thiol group; and n5 is selected from 1 to 20. hydrogen; D is selected from a therapeutic agent having at In certain embodiments of a compound of Formula (II), least one primary amine group and a therapeutic agent having each A' is a moiety of Formula (A'a2), each wherein R is at least one secondary amine group; and n5 is selected from 1 selected from hydrogen and methyl; each X and X is inde 10 to 4. pendently selected from a covalent bond, methane-diyl. In certain embodiments of a compound of Formula (II), ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane each A' is a moiety of Formula (Aa2), wherein each R is 1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. methyl; each X* is a covalent bond; each X is C(O)N : nonane-1,9-diyl. decane-1,10-diyl, undecane-1,11-diyl. each X is pentane-1,5-diyl; each R" is hydrogen; each R is 15 methyl; D is selected from a therapeutic agent having at least dodecane-1,12-diyl, hexadecane-1,16-diyl, benzene -diyl. one primary amine group and a therapeutic agent having at methylbenzene-diyl, propylbenzene-diyl, methylcyclohex least one secondary amine group; and n5 is selected from 1 to ane-diyl, and —(CH)—(CH-CH O), (CH), , 4. wherein each n1 and n3 is independently an integer selected In certain embodiments of a compound of Formula (II), from 0 to 3, and each n2 is independently an integer selected each A' is a moiety of Formula (A'a2), wherein each R is from 1 to 20; each X* is selected from a covalent bond, methyl; each X is a covalent bond; each X* is C(O)N : O— —SO , —SON , —C(O)— —C(O)O—, and each X is 4-methylcyclohexane-diyl; each R" is hydrogen; —C(O)N ; each R" and R is independently selected from each R is methyl; D is selected from a therapeutic agent hydrogen, C alkyl, and Ces aryl; D is selected from a having at least one primary amine group and a therapeutic therapeutic agent having at least one primary amine group 25 agent having at least one secondary amine group; and n5 is and a therapeutic agent having at least one secondary amine selected from 1 to 4. group; and n5 is selected from 1 to 5. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (Aa2), wherein each R is each A' is a moiety of Formula (A'a2), wherein each R is methyl; each X* is a covalent bond; each X is C(O)O : hydrogen; each X* is a covalent bond; each X* is a covalent 30 each X is pentane-1,5-diyl; each RandR is hydrogen; D is bond; each X is a covalent bond; each RandR is hydrogen; selected from a therapeutic agent having at least one primary D is selected from a therapeutic agent having at least one amine group and a therapeutic agent having at least one primary amine group and a therapeutic agent having at least secondary amine group; and n5 is selected from 1 to 4. one secondary amine group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), 35 each A' is a moiety of (Aa3), wherein each E is selected from each A' is a moiety of Formula (A'a2), wherein each R is F, Cl, Br, and I; eachX' and X is independently selected from methyl; each X" is a covalent bond; each X* is a covalent a covalent bond, methane-diyl ethane-1,2-diyl, propane-1,3- bond; each X is a covalent bond; each RandR is hydrogen; diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6-diyl, hep D is selected from a therapeutic agent having at least one tane-1,7-diyl octane-1,8-diyl. nonane-1,9-diyl. decane-1,10 primary amine group and a therapeutic agent having at least 40 diyl, undecane-1,11-diyl, dodecane-1,12-diyl, hexadecane-1, one secondary amine group; and n5 is selected from 1 to 4. 16-diyl, benzene-diyl, methylbenzene-diyl, propylbenzene In certain embodiments of a compound of Formula (II), diyl, methylcyclohexane-diyl, and —(CH)—(CH each A' is a moiety of Formula (A'a2), wherein each R is CH2—O) (CH) , wherein each n1 and n3 is hydrogen; each X is a covalent bond; each X is SO : independently an integer selected from 0 to 5, and each n2 is each X is methane-diyl; each R' and R is hydrogen; D is 45 independently an integer selected from 1 to 25; each X is selected from a therapeutic agent having at least one primary selected from a covalent bond, —O— —S— —SO—, amine group and a therapeutic agent having at least one SO. , —SON ... —SS , —C(O)—, —C(O)C) , secondary amine group; and n5 is selected from 1 to 4. C(O)N , —C(O)N N—, OC(O)N , and - NC(O) In certain embodiments of a compound of Formula (II), N. ; each R" and R is independently selected from hydro each A' is a moiety of Formula (A'a2), wherein each R is 50 gen, Calkyl, and Cesaryl; D is selected from a therapeutic hydrogen; each X is a covalent bond; each X is SO : agent having at least one primary amine group, a therapeutic each X is ethane-1,2-diyl; each R" and R is hydrogen; D is agent having at least one secondary amine group, a therapeu selected from a therapeutic agent having at least one primary tic agent having at least one hydroxy group, and a therapeutic amine group and a therapeutic agent having at least one agent having at least one thiol group; and n5 is selected from secondary amine group; and n5 is selected from 1 to 4. 55 1 to 20. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (A'a2), wherein each R is each A' is a moiety of (Aa3), wherein each E is selected from hydrogen; each X* is a covalent bond; each X* is SO.N. ; F, Cl, Br, and I; eachX' and X is independently selected from each X is pentane-1,5-diyl; each R" and R is hydrogen; D is a covalent bond, methane-diyl ethane-1,2-diyl, propane-1,3- selected from a therapeutic agent having at least one primary 60 diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6-diyl, hep amine group and a therapeutic agent having at least one tane-1,7-diyl octane-1,8-diyl. nonane-1,9-diyl. decane-1,10 secondary amine group; and n5 is selected from 1 to 4. diyl, undecane-1,11-diyl, dodecane-1,12-diyl, hexadecane-1, In certain embodiments of a compound of Formula (II), 16-diyl, benzene-diyl, methylbenzene-diyl, propylbenzene each A' is a moiety of Formula (A'a2), wherein each R is diyl, methylcyclohexane-diyl, and —(CH)—(CH2— methyl; each X* is a covalent bond; each X is —C(O)N : 65 CH2—O) (CH) , wherein each n1 and n3 is each X is pentane-1,5-diyl; each R" and R is hydrogen; D is independently an integer selected from 0 to 3, and each n2 is selected from a therapeutic agent having at least one primary independently an integer selected from 1 to 20; each X is US 9,295,731 B2 33 34 selected from a covalent bond, —O— —SO ,—SON.— diyl, benzene-diyl, methylbenzene-diyl, propylbenzene-diyl. C(O)—, —C(O)C)—, and —C(O)N ; each R" and R is methylcyclohexane-diyl. and —(CH)—(CH independently selected from hydrogen, C alkyl, and Ces CH2—O) (CH) , wherein each n1 and n3 is indepen aryl; D is selected from a therapeutic agent having at least one dently an integer selected from 0 to 3, and each n2 is inde primary amine group and a therapeutic agent having at least pendently an integer selected from 1 to 20; each X* is selected one secondary amine group; and n5 is selected from 1 to 5. from a covalent bond, —O— —SO , —SON.— In certain embodiments of a compound of Formula (II), C(O) , —C(O)O , and C(O)N ; each R' and R is each A' is a moiety of Formula (Aa3), wherein each E is Br; independently selected from hydrogen, C alkyl, and Ces each X is a covalent bond; each X* is a covalent bond; each aryl; D is selected from a therapeutic agent having at least one X is a covalent bond; each R" is hydrogen; each R is methyl: 10 D is selected from a therapeutic agent having at least one primary amine group and a therapeutic agent having at least primary amine group and a therapeutic agent having at least one secondary amine group; and n5 is selected from 1 to 5. one secondary amine group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (A'b1), wherein each L is each A' is a moiety of Formula (Aa3), wherein each E is Br; 15 N-hydroxysuccinimidyl; each X* is ethane-1,2-diyl; each X each X* is a covalent bond; each X* is C(O)N ; each X is is a covalent bond; each X is a covalent bond; each RandR pentane-1,5-diyl; each R" is hydrogen; each R is methyl; Dis is hydrogen; D is selected from a therapeutic agent having at selected from a therapeutic agent having at least one primary least one primary amine group and a therapeutic agent having amine group and a therapeutic agent having at least one at least one secondary amine group; and n5 is selected from 1 secondary amine group; and n5 is selected from 1 to 4. to 4. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (Aa3), wherein each E is Br; each A' is a moiety of (A'b2), wherein each L is selected each X* is a covalent bond; each X* is C(O)N ; each X is from halogen, N-hydroxysuccinimidyl, substituted N -hy 4-methylcyclohexane-diyl; each R' is hydrogen; each R is droxysuccinimidyl, phenol-yl, and Substituted phenol-yl, and methyl; D is selected from a therapeutic agent having at least 25 each RandR is independently selected from hydrogen, Ca one primary amine group and a therapeutic agent having at alkyl, and Co aryl; each X and X is independently least one secondary amine group; and n5 is selected from 1 to selected from a covalent bond, methane-diyl, ethane-1,2-diyl. 4. propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane In certain embodiments of a compound of Formula (II), 1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl. each A' is a moiety of Formula (Aa3), wherein each E is Br; 30 decane-1,10-diyl, undecane-1,11-diyl, dodecane-1,12-diyl. each X* is a covalent bond; each X* is C(O)O ; each X is hexadecane-1,16-diy1, benzene-diyl, methylbenzene -diyl. pentane-1,5-diyl; each R" is hydrogen; each R is methyl: Dis propylbenzene-diyl. methylcyclohexane-diyl. and selected from a therapeutic agent having at least one primary (CH)—(CH-CH O), (CH), , wherein each amine group and a therapeutic agent having at least one n1 and n3 is independently an integer selected from 0 to 5, and secondary amine group; and n5 is selected from 1 to 4. 35 each n2 is independently an integer selected from 1 to 25; In certain embodiments of a compound of Formula (II), each X is selected from a covalent bond, -O-, -S , each A' is a moiety of (A'b1), wherein each L is selected from halogen, N-hydroxysuccinimidyl, substituted N -hy droxysuccinimidyl, phenol-yl, and Substituted phenol-yl; - NC(O)N ; each RandR is independently selected from each X and X is independently selected from a covalent 40 hydrogen, C alkyl, and Cs aryl; D is selected from a bond, methane-diyl, ethane-1,2-diyl, propane-1,3-diyl. therapeutic agent having at least one primary amine group, a butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6-diyl, heptane therapeutic agent having at least one secondary amine group, 1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl, decane-1,10-diyl. a therapeutic agent having at least one hydroxy group, and a undecane-1,11-diyl, dodecane-1,12-diyl, hexadecane-1,16 therapeutic agent having at least one thiol group; and n5 is diyl, benzene-diyl, methylbenzene-diyl, propylbenzene-diyl. 45 selected from 1 to 20. methylcyclohexane-diyl. and —(CH2)—(CH2— In certain embodiments of a compound of Formula (II), CH-O), (CH), , wherein each n1 and n3 is indepen each A' is a moiety of (Ab2), wherein each L is selected dently an integer selected from 0 to 5, and each n2 is inde from halogen, N-hydroxysuccinimidyl, substituted N -hy pendently an integer selected from 1 to 25; each X* is selected droxysuccinimidyl, phenol-yl, and Substituted phenol-yl, and 50 each RandR is independently selected from hydrogen, Ca alkyl, and Cao aryl; each X and X is independently selected from a covalent bond, methane-diyl, ethane-1,2-diyl. and R is independently selected from hydrogen, C. alkyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane and Csaryl; D is selected from a therapeutic agent having at 1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl. least one primary amine group, a therapeutic agent having at 55 decane-1,10-diyl, undecane-1,11-diyl, dodecane-1,12-diyl. least one secondary amine group, a therapeutic agent having hexadecane-1,16-diy1, benzene-diyl, methylbenzene -diyl. at least one hydroxy group, and a therapeutic agent having at propylbenzene-diyl. methylcyclohexane-diyl. and least one thiol group; and n5 is selected from 1 to 20. (CH2)—(CH2—CH2—O) (CH), , wherein each In certain embodiments of a compound of Formula (II), n1 and n3 is independently an integer selected from 0 to 3, and each A' is a moiety of (A'b1), wherein each L is selected 60 each n2 is independently an integer selected from 1 to 20; from halogen, N-hydroxysuccinimidyl, substituted N -hy each X is selected from a covalent bond, -O-, -SO , droxysuccinimidyl, phenol-yl, and Substituted phenol-yl; SON , —C(O)— —C(O)O—, and —C(O)N ; each each X and X is independently selected from a covalent R" and R is independently selected from hydrogen, Ca bond, methane-diyl, ethane-1,2-diyl, propane-1,3-diyl. alkyl, and Ces aryl; D is selected from a therapeutic agent butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6-diyl, heptane 65 having at least one primary amine group and a therapeutic 1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl, decane-1,10-diyl. agent having at least one secondary amine group; and n5 is undecane-1,11-diyl, dodecane-1,12-diyl, hexadecane-1,16 selected from 1 to 5. US 9,295,731 B2 35 36 In certain embodiments of a compound of Formula (II), 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben each A' is a moiety of Formula (Ab2), wherein each L is Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and N-hydroxysuccinimidyl, and each R and R is hydrogen; (CH2)—(CH2—CH2—O) (CH), , wherein each each X" is ethane-1,2-diyl; each X* is a covalent bond; each n1 and n3 is independently an integer selected from 0 to 5, and X is a covalent bond; each R" and R is hydrogen; D is each n2 is independently an integer selected from 1 to 25; selected from a therapeutic agent having at least one primary each X is selected from a covalent bond, -O-, -S , amine group and a therapeutic agent having at least one SO , —SO. , —SON ... —SS , —C(O) , —C(O) secondary amine group; and n5 is selected from 1 to 4. O—, C(O)N , C(O)N N=, OC(O)N , and In certain embodiments of a compound of Formula (II), NC(O)N ; each RandR is independently selected from each A' is a moiety of (A'c1); each X and X is indepen 10 hydrogen, C alkyl, and Ces aryl; D is selected from a dently selected from a covalent bond, methane-diyl ethane therapeutic agent having at least one primary amine group, a 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. therapeutic agent having at least one secondary amine group, hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane a therapeutic agent having at least one hydroxy group, and a 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, therapeutic agent having at least one thiol group; and n5 is 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben 15 selected from 1 to 20. Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and In certain embodiments of a compound of Formula (II), (CH2), —(CH2—CH2—O) (CH), , wherein each each A' is a moiety of (Ac2); each X and X is indepen n1 and n3 is independently an integer selected from 0 to 5, and dently selected from a covalent bond, methane-diyl ethane each n2 is independently an integer selected from 1 to 25; 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. each X is selected from a covalent bond, -O-, -S , hexane-1,6-diyl, heptane-1,7-diyl octane-1,8-diyl. nonane SO ,—SO. , —SON ... —SS , —C(O)— —C(O) 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, O—, C(O)N , C(O)N N=, OC(O)N , and 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben —NC(O)N ; each RandR is independently selected from Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and hydrogen, C alkyl, and Ces aryl; D is selected from a (CH2)—(CH2—CH2—O) (CH), , wherein each therapeutic agent having at least one primary amine group, a 25 n1 and n3 is independently an integer selected from 0 to 3, and therapeutic agent having at least one secondary amine group, each n2 is independently an integer selected from 1 to 20; a therapeutic agent having at least one hydroxy group, and a each X is selected from a covalent bond, -O-, -SO , therapeutic agent having at least one thiol group; and n5 is SON , —C(O)— —C(O)C)—, and —C(O)N ; each selected from 1 to 20. R" and R is independently selected from hydrogen, Ca In certain embodiments of a compound of Formula (II), 30 alkyl, and Cs aryl; D is selected from a therapeutic agent each A' is a moiety of (A'c1); each X and X is indepen having at least one primary amine group and a therapeutic dently selected from a covalent bond, methane-diyl, ethane agent having at least one secondary amine group; and n5 is 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. selected from 1 to 5. hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane In certain embodiments of a compound of Formula (II), 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, 35 each A' is a moiety of Formula (Ac2); each X is butane-1, 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben 4-diyl; each X* is a covalent bond; each X is a covalent bond; Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and each R" is hydrogen; each R is methyl; D is selected from a (CH2), —(CH2—CH2—O) (CH), , wherein each therapeutic agent having at least one primary amine group n1 and n3 is independently an integer selected from 0 to 3, and and a therapeutic agent having at least one secondary amine each n2 is independently an integer selected from 1 to 20; 40 group; and n5 is selected from 1 to 4. each X is selected from a covalent bond, -O-, -SO , In certain embodiments of a compound of Formula (II), SON , —C(O)—, —C(O)O , and —C(O)N ; each each A' is a moiety of Formula (Ac2); each X is butane-1, R" and R is independently selected from hydrogen, Ca 4-diyl; each X* is C(O)N ; each X is pentane-1,5-diyl: alkyl, and Ces aryl; D is selected from a therapeutic agent each R' is hydrogen; each R is hydrogen; D is selected from having at least one primary amine group and a therapeutic 45 a therapeutic agent having at least one primary amine group agent having at least one secondary amine group; and n5 is and a therapeutic agent having at least one secondary amine selected from 1 to 5. group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (A'c1); each X is butane-1, each A' is a moiety of Formula (Ac2); each X is butane-1, 4-diyl; each X* is a covalent bond; each X is a covalent bond; 50 4-diyl; each X* is C(O)N ; each X is 4-methylcyclohex each R" is hydrogen; each R is methyl; D is selected from a ane-diyl; each R" is hydrogen; each R is methyl; Dis selected therapeutic agent having at least one primary amine group from a therapeutic agent having at least one primary amine and a therapeutic agent having at least one secondary amine group and a therapeutic agent having at least one secondary group; and n5 is selected from 1 to 5. amine group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), 55 In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (A'c1); each X is butane-1, each A' is a moiety of (A'c3); each X and X is indepen 4-diyl; each X* is —C(O)N ; each X is pentane-1,5-diyl: dently selected from a covalent bond, methane-diyl ethane each R" and R is hydrogen; D is selected from a therapeutic 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. agent having at least one primary amine group and a thera hexane-1,6-diyl, heptane-1,7-diyl octane-1,8-diyl. nonane peutic agent having at least one secondary amine group; and 60 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, n5 is selected from 1 to 5. 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben In certain embodiments of a compound of Formula (II), Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and each A' is a moiety of (Ac2); each X and X is indepen (CH)—(CH-CH O), (CH), , wherein each dently selected from a covalent bond, methane-diyl ethane n1 and n3 is independently an integer selected from 0 to 5, and 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. 65 each n2 is independently an integer selected from 1 to 25; hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane each X is selected from a covalent bond, -O-, -S , 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, SO— —SO —SON.— —SS , —C(O)— —C(O) US 9,295,731 B2 37 38 O-, - C(O)N –C(O)N N—, OC(O)N , and a therapeutic agent having at least one hydroxy group, and a —NC(O)N ; each RandR is independently selected from therapeutic agent having at least one thiol group; and n5 is hydrogen, C alkyl, and Ces aryl; D is selected from a selected from 1 to 20. therapeutic agent having at least one primary amine group, a In certain embodiments of a compound of Formula (II), therapeutic agent having at least one secondary amine group, each A' is a moiety of (Ad1); each X and X is indepen a therapeutic agent having at least one hydroxy group, and a dently selected from a covalent bond, methane-diyl ethane therapeutic agent having at least one thiol group; and n5 is 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. selected from 1 to 20. hexane-1,6-diyl, heptane-1,7-diyl octane-1,8-diyl. nonane In certain embodiments of a compound of Formula (II), 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, each A' is a moiety of (A'c3); each X and X is indepen 10 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben dently selected from a covalent bond, methane-diyl ethane Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. (CH2)—(CH2—CH2—O) (CH), , wherein each hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane n1 and n3 is independently an integer selected from 0 to 3, and 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, each n2 is independently an integer selected from 1 to 20; 15 each X is selected from a covalent bond, -O-, -SO , 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben SON , —C(O)— —C(O)C)—, and —C(O)N ; each Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and R" and R is independently selected from hydrogen, Ca (CH),—(CH-CH O), (CH), , wherein each alkyl, and Ces aryl; D is selected from a therapeutic agent n1 and n3 is independently an integer selected from 0 to 3, and having at least one primary amine group and a therapeutic each n2 is independently an integer selected from 1 to 20; agent having at least one secondary amine group; and n5 is each X is selected from a covalent bond, -O-, -SO , selected from 1 to 5. SON , —C(O)— —C(O)O—, and —C(O)N ; each In certain embodiments of a compound of Formula (II), R" and R is independently selected from hydrogen, Ca each A' is a moiety of Formula (Ad1); each X* is propane alkyl, and Ces aryl; D is selected from a therapeutic agent 1,3-diyl; each X* is a covalent bond; each X is a covalent having at least one primary amine group and a therapeutic 25 bond; each R" is hydrogen; each R is hydrogen; D is selected agent having at least one secondary amine group; and n5 is from a therapeutic agent having at least one primary amine selected from 1 to 5. group and a therapeutic agent having at least one secondary In certain embodiments of a compound of Formula (II), amine group; and n5 is selected from 1 to 4. each A' is a moiety of Formula (A'c3); each X is butane-1, In certain embodiments of a compound of Formula (II), 30 each A' is a moiety of Formula (Ad1); each X* is propane 4-diyl; each X* is a covalent bond; each X is a covalent bond; 1,3-diyl; each X is C(O)N ; each X is pentane-1,5-diyl: each R" is hydrogen; each R is methyl; D is selected from a each R" is hydrogen; each R is hydrogen; D is selected from therapeutic agent having at least one primary amine group a therapeutic agent having at least one primary amine group and a therapeutic agent having at least one secondary amine and a therapeutic agent having at least one secondary amine group; and n5 is selected from 1 to 4. 35 group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (A'c3); each X is butane-1, each A' is a moiety of Formula (Ad1); each X is propane 4-diyl; each X* is —C(O)N ; each X is pentane-1,5-diyl: 1,3-diyl; each X* is —C(O)N ; each X is 4-methylcyclo each R" is hydrogen; each R is hydrogen; D is selected from hexane-diyl; each R' is hydrogen; each R is methyl; D is a therapeutic agent having at least one primary amine group 40 selected from a therapeutic agent having at least one primary and a therapeutic agent having at least one secondary amine amine group and a therapeutic agent having at least one group; and n5 is selected from 1 to 4. secondary amine group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (A'c3); each X is butane-1, each A' is a moiety of (A'd2); each X and X is indepen 4-diyl; each X* is C(O)N ; each X is 4-methylcyclohex 45 dently selected from a covalent bond, methane-diyl ethane ane-diyl; each R" is hydrogen; each R is methyl; Dis selected 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. from a therapeutic agent having at least one primary amine hexane-1,6-diyl, heptane-1,7-diyl octane-1,8-diyl. nonane group and a therapeutic agent having at least one secondary 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, amine group; and n5 is selected from 1 to 4. 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben In certain embodiments of a compound of Formula (II), 50 Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and each A' is a moiety of (Ad1); each X and X is indepen (CH2)—(CH2—CH2—O) (CH), , wherein each dently selected from a covalent bond, methane-diyl ethane n1 and n3 is independently an integer selected from 0 to 5, and 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. each n2 is independently an integer selected from 1 to 25; hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane each X is selected from a covalent bond, -O-, -S , 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, 55 SO— —SO —SON.— —SS , —C(O)— —C(O) 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben O—, C(O)N , C(O)N N=, OC(O)N , and Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and - NC(O)N ; each RandR is independently selected from (CH2), —(CH2—CH2—O) (CH), , wherein each hydrogen, C alkyl, and Ces aryl; D is selected from a n1 and n3 is independently an integer selected from 0 to 5, and therapeutic agent having at least one primary amine group, a each n2 is independently an integer selected from 1 to 25; 60 therapeutic agent having at least one secondary amine group, each X is selected from a covalent bond, -O-, -S , a therapeutic agent having at least one hydroxy group, and a SO— —SO —SON.— —SS , —C(O)— —C(O) therapeutic agent having at least one thiol group; and n5 is O—, C(O)N , C(O)N N=, OC(O)N , and selected from 1 to 20. —NC(O)N ; each RandR is independently selected from In certain embodiments of a compound of Formula (II), hydrogen, C alkyl, and Ces aryl; D is selected from a 65 each A' is a moiety of (A'd2); each X and X is indepen therapeutic agent having at least one primary amine group, a dently selected from a covalent bond, methane-diyl ethane therapeutic agent having at least one secondary amine group, 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. US 9,295,731 B2 39 40 hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane each X is selected from a covalent bond, -O-, -SO , 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, SON , —C(O)— —C(O)C)—, and —C(O)N ; each 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben R" and R is independently selected from hydrogen, Ca Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and alkyl, and Ces aryl; D is selected from a therapeutic agent (CH2), —(CH2—CH2—O) (CH), , wherein each having at least one primary amine group and a therapeutic n1 and n3 is independently an integer selected from 0 to 3, and agent having at least one secondary amine group; and n5 is each n2 is independently an integer selected from 1 to 20; selected from 1 to 5. each X is selected from a covalent bond, -O-, -SO , In certain embodiments of a compound of Formula (II), SON , —C(O)—, —C(O)O , and —C(O)N ; each each A' is a moiety of Formula (Ad3); each X* is a covalent R" and R is independently selected from hydrogen, Ca 10 bond; each X is a covalent bond; each X is a covalent bond; alkyl, and Ces aryl; D is selected from a therapeutic agent each R' is hydrogen; each R is hydrogen; D is selected from having at least one primary amine group and a therapeutic a therapeutic agent having at least one primary amine group agent having at least one secondary amine group; and n5 is and a therapeutic agent having at least one secondary amine selected from 1 to 5. group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), 15 In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (Ad2); each X* is ethane-1, each A' is a moiety of Formula (Ad3); each X* is a covalent 2-diyl; each X* is a covalent bond; each X is a covalent bond; bond; each X* is C(O)N ; each X is pentane-1,5-diyl: each R' is hydrogen; each R is hydrogen; D is selected from each R' is hydrogen; each R is hydrogen; D is selected from a therapeutic agent having at least one primary amine group a therapeutic agent having at least one primary amine group and a therapeutic agent having at least one secondary amine and a therapeutic agent having at least one secondary amine group; and n5 is selected from 1 to 4. group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (Ad2); each X* is ethane-1, each A' is a moiety of Formula (Ad3); each X* is a covalent 2-diyl; each X* is —C(O)N ; each X is pentane-1,5-diyl: bond; each X is —C(O)N ; each X is 4-methylcyclohex each R' is hydrogen; each R is hydrogen; D is selected from 25 ane-diyl; each R" is hydrogen; each R is methyl; Dis selected a therapeutic agent having at least one primary amine group from a therapeutic agent having at least one primary amine and a therapeutic agent having at least one secondary amine group and a therapeutic agent having at least one secondary group; and n5 is selected from 1 to 4. amine group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (Ad2); each X* is ethane-1, 30 each A' is a moiety of (A"e1); each X and X is indepen 2-diyl; each X* is C(O)N ; each X is 4-methylcyclohex dently selected from a covalent bond, methane-diyl ethane ane-diyl; each R" is hydrogen; each R is hydrogen; D is 1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. selected from a therapeutic agent having at least one primary hexane-1,6-diyl, heptane-1,7-diyl octane-1,8-diyl. nonane amine group and a therapeutic agent having at least one 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, secondary amine group; and n5 is selected from 1 to 4. 35 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben In certain embodiments of a compound of Formula (II), Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and each A' is a moiety of (Ad3); each X and X is indepen (CH2)—(CH2—CH2—O) (CH), , wherein each dently selected from a covalent bond, methane-diyl ethane n1 and n3 is independently an integer selected from 0 to 5, and 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. each n2 is independently an integer selected from 1 to 25; hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane 40 each X is selected from a covalent bond, -O-, -S , 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, SO— —SO —SON.— —SS , —C(O)— —C(O) 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben O—, C(O)N , C(O)N N=, OC(O)N , and Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and - NC(O)N ; each RandR is independently selected from (CH2), —(CH2—CH2—O) (CH), , wherein each hydrogen, C alkyl, and Ces aryl; D is selected from a n1 and n3 is independently an integer selected from 0 to 5, and 45 therapeutic agent having at least one primary amine group, a each n2 is independently an integer selected from 1 to 25; therapeutic agent having at least one secondary amine group, each X is selected from a covalent bond, -O-, -S , a therapeutic agent having at least one hydroxy group, and a SO— —SO —SON.— —SS , —C(O)— —C(O) therapeutic agent having at least one thiol group; and n5 is O—, C(O)N , C(O)N N=, OC(O)N , and selected from 1 to 20. —NC(O)N ; each RandR is independently selected from 50 In certain embodiments of a compound of Formula (II), hydrogen, C alkyl, and Ces aryl; D is selected from a each A' is a moiety of (A'el); each X and X is indepen therapeutic agent having at least one primary amine group, a dently selected from a covalent bond, methane-diyl ethane therapeutic agent having at least one secondary amine group, 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. a therapeutic agent having at least one hydroxy group, and a hexane-1,6-diyl, heptane-1,7-diyl octane-1,8-diyl. nonane therapeutic agent having at least one thiol group; and n5 is 55 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, selected from 1 to 20. 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben In certain embodiments of a compound of Formula (II), Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and each A' is a moiety of (Ad3); each X and X is indepen (CH2)—(CH2—CH2—O) (CH), , wherein each dently selected from a covalent bond, methane-diyl ethane n1 and n3 is independently an integer selected from 0 to 3, and 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. 60 each n2 is independently an integer selected from 1 to 20; hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane each X is selected from a covalent bond, -O-, -SO , 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, SON , —C(O)— —C(O)O—, and —C(O)N ; each 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben R" and R is independently selected from hydrogen, Ca Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and alkyl, and Ces aryl; D is selected from a therapeutic agent (CH2), —(CH2—CH2—O) (CH), , wherein each 65 having at least one primary amine group and a therapeutic n1 and n3 is independently an integer selected from 0 to 3, and agent having at least one secondary amine group; and n5 is each n2 is independently an integer selected from 1 to 20; selected from 1 to 5. US 9,295,731 B2 41 42 In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (A"e1); each X is pentane each A' is a moiety of Formula (A'f1); each X is methane 1,5-diyl; each X* is a covalent bond; each X is a covalent diyl; each X* is a covalent bond; each X is a covalent bond; bond; each R" is hydrogen; each R is hydrogen; D is selected each R' is hydrogen; each R is hydrogen; D is selected from from a therapeutic agent having at least one primary amine a therapeutic agent having at least one primary amine group group and a therapeutic agent having at least one secondary and a therapeutic agent having at least one secondary amine amine group; and n5 is selected from 1 to 4. group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (A"e1); each X is pentane each A' is a moiety of Formula (A' fl); each X is a covalent 1,5-diyl; each X is C(O)N ; each X is pentane-1,5-diyl: 10 bond; each X* is C(O)N ; each X is pentane-1,5-diyl: each R' is hydrogen; each R is hydrogen; D is selected from each R" is hydrogen; each R is methyl; D is selected from a a therapeutic agent having at least one primary amine group therapeutic agent having at least one primary amine group and a therapeutic agent having at least one secondary amine and a therapeutic agent having at least one secondary amine group; and n5 is selected from 1 to 4. group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), 15 In certain embodiments of a compound of Formula (II), each A' is a moiety of Formula (A"e1); each X is pentane each A' is a moiety of Formula (A' fl); each X is a covalent 1,5-diyl; each X* is C(O)N ; each X is 4-methylcyclo bond; each X is —C(O)N ; each X is 4-methylcyclohex hexane-diyl; each R' is hydrogen; each R is methyl; D is ane-diyl; each R" is hydrogen; each R is methyl; Dis selected selected from a therapeutic agent having at least one primary from a therapeutic agent having at least one primary amine amine group and a therapeutic agent having at least one group and a therapeutic agent having at least one secondary secondary amine group; and n5 is selected from 1 to 4. amine group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), In certain embodiments of a compound of Formula (II), each A' is a moiety of (A' fl); each X and X is indepen each A' is a moiety of Formula (A'f1); each X is pentane-1, dently selected from a covalent bond, methane-diyl ethane 5-diyl; each X* is C(O)N ; each X is pentane-1,5-diyl; 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. 25 each R" is hydrogen; each R is methyl; D is selected from a hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane therapeutic agent having at least one primary amine group 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, and a therapeutic agent having at least one secondary amine 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben group; and n5 is selected from 1 to 4. Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and In certain embodiments of a compound of Formula (II), (CH),—(CH-CH O), (CH), , wherein each 30 each A' is a moiety of (A" f2); each X and X is indepen n1 and n3 is independently an integer selected from 0 to 5, and dently selected from a covalent bond, methane-diyl ethane each n2 is independently an integer selected from 1 to 25: 1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. each X is selected from a covalent bond, -O-, -S , hexane-1,6-diyl, heptane-1,7-diyl octane-1,8-diyl. nonane SO— —SO —SON.— —SS , —C(O)— —C(O) 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, O—, C(O)N , C(O)N N=, OC(O)N , and 35 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben —NC(O)N ; each RandR is independently selected from Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and hydrogen, C alkyl, and Ces aryl; D is selected from a (CH2)—(CH2—CH2—O) (CH), , wherein each therapeutic agent having at least one primary amine group, a n1 and n3 is independently an integer selected from 0 to 5, and therapeutic agent having at least one secondary amine group, each n2 is independently an integer selected from 1 to 25; a therapeutic agent having at least one hydroxy group, and a 40 each X is selected from a covalent bond, -O-, -S , therapeutic agent having at least one thiol group; and n5 is SO— —SO —SON.— —SS , —C(O)— —C(O) selected from 1 to 20. O—, C(O)N , C(O)N N=, OC(O)N , and In certain embodiments of a compound of Formula (II), - NC(O)N ; each RandR is independently selected from each A' is a moiety of (A' fl); each X and X is indepen hydrogen, C alkyl, and Ces aryl; D is selected from a dently selected from a covalent bond, methane-diyl ethane 45 therapeutic agent having at least one primary amine group, a 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. therapeutic agent having at least one secondary amine group, hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane a therapeutic agent having at least one hydroxy group, and a 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, therapeutic agent having at least one thiol group; and n5 is 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben selected from 1 to 20. Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and 50 In certain embodiments of a compound of Formula (II), (CH2), —(CH2—CH2—O) (CH), , wherein each each A' is a moiety of (A'f2); each X and X is indepen n1 and n3 is independently an integer selected from 0 to 3, and dently selected from a covalent bond, methane-diyl ethane each n2 is independently an integer selected from 1 to 20; 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. each X is selected from a covalent bond, -O-, -SO , hexane-1,6-diyl, heptane-1,7-diyl octane-1,8-diyl. nonane SON , —C(O)— —C(O)O—, and —C(O)N ; each 55 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, R" and R is independently selected from hydrogen, Ca 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben alkyl, and Ces aryl; D is selected from a therapeutic agent Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and having at least one primary amine group and a therapeutic (CH2)—(CH2—CH2—O) (CH), , wherein each agent having at least one secondary amine group; and n5 is n1 and n3 is independently an integer selected from 0 to 3, and selected from 1 to 5. 60 each n2 is independently an integer selected from 1 to 20; In certain embodiments of a compound of Formula (II), each X is selected from a covalent bond, -O-, -SO , each A' is a moiety of Formula (A'f1); each X" is a covalent SON , —C(O)— —C(O)O—, and —C(O)N ; each bond; each X* is a covalent bond; each X is a covalent bond; R" and R is independently selected from hydrogen, Ca each R' is hydrogen; each R is hydrogen; D is selected from alkyl, and Ces aryl; D is selected from a therapeutic agent a therapeutic agent having at least one primary amine group 65 having at least one primary amine group and a therapeutic and a therapeutic agent having at least one secondary amine agent having at least one secondary amine group; and n5 is group; and n5 is selected from 1 to 4. selected from 1 to 5. US 9,295,731 B2 43 44 In certain embodiments of a compound of Formula (II), diyl; each X* is —C(O)N ; each X is 4-methylcyclohex each A' is a moiety of Formula (A'f2); each X is methane ane-diyl; each R" is hydrogen; each R is methyl; D is selected diyl; each X* is a covalent bond; each X is a covalent bond; from a therapeutic agent having at least one primary amine each R" is hydrogen; each R is hydrogen; D is selected from group and a therapeutic agent having at least one secondary a therapeutic agent having at least one primary amine group and a therapeutic agent having at least one secondary amine amine group; and n5 is selected from 1 to 4. group; and n5 is selected from 1 to 4. In certain embodiments of a compound of Formula (II), the In certain embodiments of a compound of Formula (II), compound is selected from a compound of Formula (II-a). each A' is a moiety of Formula (A'f2); each X* is methane Formula (II-b), Formula (II-c). Formula (II-d), Formula (II diyl; each X is C(O)N ; each X is pentane-1,5-diyl: e), Formula (II-f). Formula (II-g), Formula (II-h), Formula each R" is hydrogen; each R is methyl; D is selected from a 10 (II-i), Formula (II-). Formula (II-k). Formula (II-1), Formula therapeutic agent having at least one primary amine group (II-m). Formula (II-n). Formula (II-o), Formula (II-p), For and a therapeutic agent having at least one secondary amine mula (II-q). Formula (II-r), Formula (II-s). Formula (II-t), group; and n5 is selected from 1 to 4. Formula (II-u), Formula (II-V). Formula (II-w), Formula (II In certain embodiments of a compound of Formula (II), X). Formula (II-y), Formula (II-Z). Formula (II-aa). Formula each A' is a moiety of Formula (A'f2); each X* is methane (II-ab), and Formula (II-ac), or a salt of any of the foregoing:

(II-a)

(II-b)

US 9,295,731 B2 45 46 -continued

(II-c)

(II-d)

US 9,295,731 B2 51 52 -continued (II-m) (II-n)

(II-o)

(II-p) US 9,295,731 B2 53 54 -continued

(II-q)

(II-r)

OH

(II-s)

US 9,295,731 B2

-continued (II-Z) (II-aa)

(II-ab)

(II-ac)

HN O O NH

N N O O f O O N N1 O O l O O 1No N O O W H N

O

60 In certain embodiments, a compound of Formula (II) is a In certain embodiments, a compound of Formula (II) is a compound of Formula (I-a) or a salt thereof. compound of Formula (II-d) or a salt thereof. In certain embodiments, a compound of Formula (II) is a In certain embodiments, a compound of Formula (II) is a compound of Formula (II-b) or a salt thereof. is compound of Formula (II-e) or a salt thereof. In certain embodiments, a compound of Formula (II) is a In certain embodiments, a compound of Formula (II) is a compound of Formula (II-c) or a salt thereof. compound of Formula (II-f) or a salt thereof. US 9,295,731 B2 59 60 In certain embodiments, a compound of Formula (II) is a each A is independently selected from a covalent bond, an compound of Formula (II-g) or a salt thereof. amide group, an avidin-binding group, a carbamate In certain embodiments, a compound of Formula (II) is a group, a carbonyl group, a disulfide group, an ester compound of Formula (II-h) or a salt thereof. group, a hydraZone group, an imine group, a Succinim In certain embodiments, a compound of Formula (II) is a ide group, a Sulfonamide group, a Sulfone group, a Sul compound of Formula (II-i) or a salt thereof. foxide group, a thioether group, a triazole group, and a In certain embodiments, a compound of Formula (II) is a urea group: compound of Formula (II-) or a salt thereof. each X and X is independently selected from a covalent In certain embodiments, a compound of Formula (II) is a bond, Co alkanediyl. Substituted Co alkanediyl. compound of Formula (II-k) or a salt thereof. 10 Co heteroalkanediyl. Substituted Co. heteroal In certain embodiments, a compound of Formula (II) is a kanediyl. C. cycloalkanediyl. Substituted C compound of Formula (II-1) or a salt thereof. cycloalkanediyl, C. heterocycloalkanediyl. Substi In certain embodiments, a compound of Formula (II) is a tuted C. heterocycloalkanediyl, Cao alkanecycloal compound of Formula (II-m) or a salt thereof. 15 kanediyl. Substituted Cao alkanecycloalkanediyl, Cao In certain embodiments, a compound of Formula (II) is a heteroalkanecycloalkanediyl. Substituted Cao het compound of Formula (II-n) or a salt thereof. eroalkanecycloalkanediyl, Co arenediyl. Substituted In certain embodiments, a compound of Formula (II) is a Coarenediyl. Coheteroarenediyl. Substituted Co compound of Formula (II-o) or a salt thereof. heteroarenediyl, C-2 alkanearenediyl. Substituted In certain embodiments, a compound of Formula (II) is a C7-20 alkanearenediyl, C-20 heteroalkanearenediyl. compound of Formula (II-p) or a salt thereof. Substituted Co heteroalkanearenediyl, and In certain embodiments, a compound of Formula (II) is a (CH2)—(CH2—CH2—O) (CH), , compound of Formula (II-q) or a salt thereof. wherein: In certain embodiments, a compound of Formula (II) is a each n1 and n3 is independently an integer selected from compound of Formula (II-r) or a salt thereof. 25 0 to 5; and In certain embodiments, a compound of Formula (II) is a each n2 is independently an integer selected from 1 to compound of Formula (II-s) or a salt thereof. 25; In certain embodiments, a compound of Formula (II) is a each X* is independently selected from a covalent bond, compound of Formula (II-t) or a salt thereof. O— —S——N , —N-, - N=N , —N–C , In certain embodiments, a compound of Formula (II) is a 30 SO— —SO. , —SON— —SS—, —C(O)—, compound of Formula (II-u) or a salt thereof. C(O)O-, - C(O)N , —C(O)S –C(O)N In certain embodiments, a compound of Formula (II) is a N=, OP(O)(OH)O OC(O)C OC(O)N , compound of Formula (II-V) or a salt thereof. - NC(O)N , and - NC(S)N: In certain embodiments, a compound of Formula (II) is a each R' and R is independently selected from hydrogen, compound of Formula (II-w) or a salt thereof. 35 C. alkyl, Substituted C. alkyl, C. heteroalkyl, Sub In certain embodiments, a compound of Formula (II) is a stituted C. heteroalkyl, Co aryl, Substituted Co compound of Formula (II-X) or a salt thereof. aryl, Coheteroaryl, and Substituted Coheteroaryl; In certain embodiments, a compound of Formula (II) is a each D is independently selected from therapeutic agent compound of Formula (II-y) or a salt thereof. having at least one primary amine group, a therapeutic In certain embodiments, a compound of Formula (II) is a 40 agent having at least one secondary amine group, a compound of Formula (II-Z) or a salt thereof. therapeutic agent having at least one hydroxy group, and In certain embodiments, a compound of Formula (II) is a a therapeutic agent having at least one thiol group; and compound of Formula (II-aa) or a salt thereof. nó is an integer selected from 1 to 20. In certain embodiments, a compound of Formula (II) is a In certain embodiments of a compound of Formula (III), Q compound of Formula (II-ab) or a salt thereof. 45 is a ligand having at least one thiol group, in certain embodi In certain embodiments, a compound of Formula (II) is a ments at least one primary amine group, in certain embodi compound of Formula (II-ac) or a salt thereof. ments at least one secondary amine group, in certain embodi In certain embodiments, conjugates have the structure of ments at least one biotin-binding group, in certain Formula (III): embodiments at least one photoreactive group, in certain 50 embodiments at least one alkyne group, and in certain embodiments at least one azide group. (III) In certain embodiments of a compound of Formula (III), O R1 R2 O the ligand is selected from a peptide, a protein, a pegylated protein, a fusion protein, an antibody, an antibody fragment, Q Y 1N11AS -XS Nx ls O X O ul D. 55 a polymer, a copolymer, and a lipid. In certain embodiments of a compound of Formula (III), the ligand having at least one thiol group is selected from a a pharmaceutically acceptable salt thereof, wherein: peptide, a protein, a pegylated protein, a fusion protein, an Q is selected from a ligand having at least one thiol group, antibody, an antibody fragment, a polymer, a copolymer, and a ligand having at least one primary amine group, a 60 a lipid. ligand having at least one secondary amine group, a In certain embodiments of a compound of Formula (III), ligand having at least one biotin-binding group, a ligand the ligand having at least one primary amine group is selected having at least one photoreactive group, a ligand having from a peptide, a protein, a pegylated protein, a fusion pro at least one alkyne group, and aligandhaving at least one tein, an antibody, an antibody fragment, a polymer, a copoly azide group: 65 mer, and a lipid. each Y is independently selected from a covalent bond and In certain embodiments of a compound of Formula (III), a hydrogen bond; the ligand having at least one secondary amine group is US 9,295,731 B2 61 62 selected from a peptide, a protein, a pegylated protein, a detumomab, dorlixizumab, drozitumab, ecromeximab, ecu fusion protein, an antibody, an antibody fragment, a polymer, lizumab, edrecolomab, efalizumab, efungumab, ensituX a copolymer, and a lipid. imab, efalizumab, elotuZumab, elsilimomab, enlimomab, In certain embodiments of a compound of Formula (III), enlimomab pegol, epitumomab, epitumomab, epitumomab the ligand having at least one biotin-binding group is selected 5 cituxetan, epratuZumab, erlizumab, ertumaxomab, etaner from a peptide and a protein. cept, etaracizumab, exbivirumab, fanolesomab, faralimo In certain embodiments of a compound of Formula (III), mab, farletuZumab, felvizumab, fezakinumab, figitumumab, fontolizumab, foralumab, foravirumab, fresolimumab, galix the ligand having at least one biotin-binding group is selected imab, ganitumab, gantenerumab, gantenerumab, gavilimo from avidin, deglycosylated avidin, Streptavidin, and strep mab, gemtuzumab, girentuximab, glembatumumab, goli Tactin. 10 mumab, gomiliximab, ibalizumab, ibritumomab, In certain embodiments of a compound of Formula (III), ibritumomab tiuXetan, igovomab, imciromab, infliximab, the ligand having at least one primary photoreactive group is inolimomab, inotuZumab, intetumumab, ipilimumab, itoli selected from a peptide, a protein, a pegylated protein, a Zumab, keliximab, labetuzumab, lebrikizumab, lemaleso fusion protein, an antibody, an antibody fragment, a polymer, mab, lenercept, lerdelimumab, lexatumumab, libivirumab, a copolymer, and a lipid. 15 lintuZumab, lorVotuZumab, lucatumumab, lumiliximab, In certain embodiments of a compound of Formula (III), mapatumumab, maslimomab, matuZumab, mavrilimumab, the ligand having at least one primary alkyne group is selected mepolizumab, metelimumab, milatuZumab, minretumomab, from a peptide, a protein, a pegylated protein, a fusion pro mirococept, mitumomab, morolimumab, motavizumab, tein, an antibody, an antibody fragment, a polymer, a copoly moxetumomab, moxetumomab pasudotox, muromonab mer, and a lipid. CD3, nacolomab, nacolomab tafenatox, naptumomab, nap In certain embodiments of a compound of Formula (III), tumomab estafenatox, natalizumab, nebacumab, necitu the ligand having at least one primary azide group is selected mumab, nerelimomab, nimotuZumab, ocrelizumab, from a peptide, a protein, a pegylated protein, a fusion pro odulimomab. ofatumumab, olaratumab, olokiZumab, omali tein, an antibody, an antibody fragment, a polymer, a copoly Zumab, oportuzumab, oportuzumab monatox, Oregovomab, mer, and a lipid. 25 otelixiZumab, oxelumab, pagibaximab, palivizumab, panitu In certain embodiments of a compound of Formula (III), mumab, panobacumab, pascolizumab, pegsunercept, pertu the ligand is a peptide and is selected from glutathione, Zumab, pexelizumab, pintumomab, priliximab, pritumumab, racotumomab, rafivirumab, ramucirumab, ranibizumab, raxi camosine, and pantetheline. bacumab, regavirumab, reslizumab, rilonacept, rilotu In certain embodiments of a compound of Formula (III), mumab, rituximab, robatumumab, roledumab, rontalizumab, the ligand is a protein and is selected from bovine serum 30 rovelizumab, ruplizumab, Samalizumab, Satumomab, secuki albumin, human serum albumin, and lactosaminated human numab. Sevirumab, sibrotuZumab, Sifalimumab, siltuximab, serum albumin. Siplizumab, SolaneZumab, SontuZumab, Sotatercept, stamu In certain embodiments of a compound of Formula (III), lumab, Sulesomab, Suvizumab, tacatuZumab, tadocizumab, the ligand is a protein and is selected from avidin, deglyco talizumab, taneZumab, taplitumomab, taplitumomab paptoX, Sylated avidin, streptavidin, and strep-Tactin. 35 tefibazumab, tellimomab, tellimomab aritox, teneliximab, In certain embodiments of a compound of Formula (III), tenatumomab, tepliZumab, teprotumumab, ticilimumab, tig the ligand is a protein and is selected from interferon alfa, atuZumab, tocilizumab, toralizumab, to situmomab, traloki interferon alfa-2a, interferon alfa-2b, interferon beta, inter numab, trastuzumab, tremelimumab, tucotuZumab, tucotu feron beta-1a, interferon beta-1b, interferon gamma, and Zumab celmoleukin, tuvirumab, urtoxazumab, ustekinumab, interferon gamma-1b. 40 vapaliximab, vedolizumab, VeltuZumab, Vepalimomab, In certain embodiments of a compound of Formula (III), Visilizumab, Volociximab, Votumumab, Zalutumumab, Zano the ligand is a pegylated protein and is selected from pegy limumab, Ziralimumab, Zolimomab, Zolimomab aritoX. lated interferon alfa, pegylated interferon alfa-2a, pegylated In certain embodiments of a compound of Formula (III), interferon alfa-2b, pegylated interferon beta, pegylated inter the ligand is a polymer and is selected from polyethylene feron beta-1a, pegylated interferon beta-1b, pegylated inter 45 glycol (PEG), monomethyl polyethylene glycol (MPEG), feron gamma, and pegylated interferon gamma-1b. polypropylene glycol (PPG), polystyrene, and polylactide. In certain embodiments of a compound of Formula (III), In certain embodiments of a compound of Formula (III), the ligand is an antibody and is selected from a humanized the ligand is a copolymer and is selected from N-(2-hydrox monoclonal antibody, a murine monoclonal antibody, a chi ypropyl)methacrylamide (HPMA) copolymer and poly(sty meric monoclonal antibody, and a human monoclonal anti 50 rene-co-maleic acid). body. In certain embodiments of a compound of Formula (III), In certain embodiments of a compound of Formula (III), the ligand is MPEG having a number average molecular the ligand is an antibody and is selected from abagovomab, weight from about 100 Daltons to about 60,000 Daltons. In abatacept, abciximab, adalimumab, adecatumumab, afelimo certain embodiments of a compound of Formula (III), the mab, alefacept, aflibercept, afutuZumab, alacizumab pegol, 55 ligand is MPEG having a number average molecular weight alefacept, alemtuzumab, anakinra, anatumomab, anrukin from about 1,000 Daltons to about 40,000 Daltons. In certain Zumab, apolizumab, arcitumomab, aselizumab, atacicept, embodiments of a compound of Formula (III), the ligand is atorolimumab, baminercept, bapineuZumab, basiliximab, MPEG having a number average molecular weight from bavituximab, bectumomab, belatacept, belimumab, benrali about 1,000 Daltons to about 12,500 Daltons. Zumab, bertilimumab, besilesomab, bevacizumab, biciro 60 In certain embodiments of a compound of Formula (III), mab, bifarcept, blinatumomab, brentuximab, briakinumab, the ligand is a lipid and is selected from Cso alkyl. briobacept, canakinumab, catumaxomab, cantuzumab, In certain embodiments of a compound of Formula (III), cedelizumab, certolizumab, certolizumab pegol, cetuximab, each Y is the same, and in certain embodiments, at least some cintredekin, cintredekin besudotox, citatuZumab, citatu Y are different. Zumab bogatoX, cixutumumab, clenoliximab, clivatuZumab, 65 In certain embodiments of a compound of Formula (III), conatumumab, dacetuZumab, dacliximab, daclizumab, dalo each A is the same, and in certain embodiments, at least some tuZumab, daratumumab, denileukin diftitox, denosumab, A are different. US 9,295,731 B2 63 64 In certain embodiments of a compound of Formula (III), -continued eachX' is the same, and in certain embodiments, at least some (Ab2) X' are different. O R8 R9 O In certain embodiments of a compound of Formula (III), each X is the same, and in certain embodiments, at least some X’ are different. X ls-X. -x

In certain embodiments of a compound of Formula (III), (Ac1) each X is the same, and in certain embodiments, at least some X are different. 10 In certain embodiments of a compound of Formula (III), each R' is the same, and in certain embodiments, at least some R" are different. In certain embodiments of a compound of Formula (III), each R is the same, and in certain embodiments, at least some 15 Rare different. In certain embodiments of a compound of Formula (III), (Ac2) each D is the same, and in certain embodiments, at least some Dare different. In certain embodiments of a compound of Formula (III), each group within the bracket is the same, and in certain embodiments, at least one group within the brackets is differ ent.

In certain embodiments of a compound of Formula (III), 25 each Y is independently selected from a covalent bond and a hydrogen bond. (Ac3) In certain embodiments of a compound of Formula (III), each Y is a covalent bond to the at least one thiol group. In certain embodiments of a compound of Formula (III), each Y 30 is a covalent bond to the at least one primary amine group. In certain embodiments of a compound of Formula (III), each Y is a covalent bond to the at least one secondary amine group. "..." In certain embodiments of a compound of Formula (III), each O Y is a covalent bond to the at least one photoreactive group. In 35 certain embodiments of a compound of Formula (III), each Y X (Af1) is a covalent bond to the at least one triazole group. In certain embodiments of a compound of Formula (III), each Y is a N-N covalent bond to the at least one Succinimide group. In certain k \ and embodiments of a compound of Formula (III), each Y is a 40 hydrogen bond to the at least one biotin-binding group. In certain embodiments of a compound of Formula (III), % each A is independently selected from a covalent bond, an (Af2) amide group, an avidin-binding group, a carbamate group, a carbonyl group, a disulfide group, an estergroup, a hydrazone 45 group, an imine group, a Succinimide group, a Sulfonamide group, a Sulfone group, a Sulfoxide group, a thioether group, a triazole group, and a urea group. In certain embodiments of a compound of Formula (III), each A is independently selected from a covalent bond, 50 OC(O)N , NC(O)N , —SO. , —SON ... —S , —SS , a moiety of Formula (Aa1), a moiety of Formula (Ab2), a moiety of Formula (Ac1), a moiety of Formula (Ac2), a moiety of Formula (Ac3), a moiety of Formula 55 (Af1), and a moiety of Formula (Af2): wherein each R and R is independently selected from hydrogen, C alkyl, Substituted C alkyl, C. het eroalkyl, Substituted C. heteroalkyl, Co aryl, Substi (Aa1) 60 tuted Cao aryl, Coheteroaryl, and Substituted Co heteroaryl. In certain embodiments of a compound of Formula (III), each A is a thioether group and is independently selected from - S - and a moiety of Formula (Aa1). 65 In certain embodiments of a compound of Formula (III), each A is an amide group and is independently selected from —C(O)N and a moiety of Formula (Ab2). US 9,295,731 B2 65 66 In certain embodiments of a compound of Formula (III), In certain embodiments of a compound of Formula (III), each A is an avidin-binding group and is independently each X is a covalent bond. selected from a moiety of Formula (Ac1), a moiety of For In certain embodiments of a compound of Formula (III), mula (Ac2), and a moiety of Formula (Ac3). each X and X is independently selected from a covalent In certain embodiments of a compound of Formula (III), bond, methane-diyl, ethane-1,2-diyl, propane-1,3-diyl. each A is a triazole group and is independently selected from butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6-diyl, heptane a moiety of Formula (Afl) and a moiety of Formula (Af2). 1,7-diyl octane-1,8-diyl. nonane-1,9-diyl, decane-1,10-diyl. In certain embodiments of a compound of Formula (III), undecane-1,11-diyl, dodecane-1,12-diyl, hexadecane-1,16 each A is Formula (Aa1). In certain embodiments of a diyl, benzene-diyl, methylbenzene-diyl, propylbenzene-diyl. compound of Formula (III), each A is Formula (Aa1), 10 methylcyclohexane-diyl. and —(CH2)—(CH2— wherein each R and R is independently selected from CH2—O) (CH) , wherein each n1 and n3 is indepen hydrogen, C alkyl and Cesaryl; in certain embodiments, dently an integer selected from 0 to 3 and each n2 is indepen each RandR is independently selected from hydrogen, C dently an integer selected from 1 to 20. alkyl, and Caryl; and in certain embodiments, each R and In certain embodiments of a compound of Formula (III), R is hydrogen. 15 each X and X is independently selected from a covalent In certain embodiments of a compound of Formula (III), bond, methane-diyl, ethane-1,2-diyl, propane-1,3-diyl, pen each A is Formula (Ab2). In certain embodiments of a tane-1,5-diyl 4-methylcyclohexane-diyl, benzene-1,2-diyl. compound of Formula (III) wherein each A is Formula benzene-1,3-diyl, and benzene-1,4-diyl. (Ab2), each RandR is independently selected from hydro In certain embodiments of a compound of Formula (III), gen, C alkyl, and Cs aryl. In certain embodiments of a each X is a covalent bond; and each X is selected from a compound of Formula (III) wherein each A is Formula covalent bond, methane-diyl ethane-1,2-diyl, propane-1,3- (Ab2), each RandR is independently selected from hydro diyl, pentane-1,5-diyl 4-methylcyclohexane-diyl, benzene gen, methyl, ethyl, propyl, isopropyl, and phenyl. In certain 1,2-diyl, benzene-1,3-diyl, and benzene-1,4-diyl. embodiments of a compound of Formula (III) wherein each In certain embodiments of a compound of Formula (III), A is Formula (Ab2), each RandR is hydrogen. 25 each X is ethane-1,2-diyl; and each X is selected from a In certain embodiments of a compound of Formula (III), covalent bond, methane-diyl ethane-1,2-diyl, propane-1,3- each A is Formula (Ac1). In certain embodiments of a diyl, pentane-1,5-diyl 4-methylcyclohexane-diyl, benzene compound of Formula (III), each A is Formula (Ac2). In 1,2-diyl, benzene-1,3-diyl, and benzene-1,4-diyl. certain embodiments of a compound of Formula (III), each A In certain embodiments of a compound of Formula (III), is Formula (Ac3). In certain embodiments of a compound of 30 each X* is pentane-1,5-diyl; and each X is selected from a Formula (III), each A is Formula (Af1). In certain embodi covalent bond, methane-diyl ethane-1,2-diyl, propane-1,3- ments of a compound of Formula (III), each A is Formula diyl, pentane-1,5-diyl 4-methylcyclohexane-diyl, benzene (Af2). 1,2-diyl, benzene-1,3-diyl, and benzene-1,4-diyl. In certain embodiments of a compound of Formula (III), In certain embodiments of a compound of Formula (III), each X and X is independently selected from a covalent 35 each X* is 4-methylcyclohexane-diyl; and X is selected from bond, Co alkanediyl. Substituted Co alkanediyl, C-12 a covalent bond, methane-diyl ethane-1,2-diyl, propane-1,3- cycloalkanediyl. Substituted C-2 cycloalkanediyl. Substi diyl, pentane-1,5-diyl 4-methylcyclohexane-diyl, benzene tuted C. heterocycloalkanediyl. Cao arenediyl. Substi 1,2-diyl, benzene-1,3-diyl, and benzene-1,4-diyl. tuted C-2 arenediyl, C7-20 alkanearenediyl. Substituted C7-20 In certain embodiments of a compound of Formula (III) alkanearenediyl, and (CH)—(CH-CH-O), 40 wherein each X and/or X is —(CH)—(CH (CH), , wherein each n1 and n3 is independently an inte CH2—O) (CH) , each n1 and n3 is independently an ger selected from 0 to 5 and each n2 is independently an integer selected from 0 to 5, an integer selected from 0 to 4, an integer selected from 1 to 25. integer selected from 0 to 3, and in certain embodiments, an In certain embodiments of a compound of Formula (III), integer selected from 0 to 2. In certain embodiments of a each X and X is independently selected from a covalent 45 compound of Formula (III) wherein each X and/or X is bond, C. alkanediyl. Substituted C- alkanediyl. C. (CH2)—(CH2—CH2—O) (CH), , each n2 is cycloalkanediyl, benzene-diyl, substituted benzene-diyl. independently an integer selected from 1 to 25, an integer C- benzenealkane-diyl, and —(CH2)—(CH2—CH2— selected from 1 to 20, an integer selected from 1 to 15, an O), (CH), , wherein each n1 and n3 is independently integer selected from 1 to 10, an integer selected from 1 to 5, an integer selected from 0 to 5 and each n2 is independently 50 an integer selected from 1 to 4, and in certain embodiments, an integer selected from 1 to 20. an integer selected from 1 to 3. In certain embodiments of a compound of Formula (III), In certain embodiments of a compound of Formula (III), each X and X is independently selected from a covalent each n1 and n3 is independently an integer selected from 0 to bond, Cs alkanediyl. Substituted Cls alkanediyl, Cs 5, in certain embodiments, an integer selected from 0 to 4, and cycloalkanediyl, benzene-diyl, substituted benzene-diyl. 55 in certain embodiments, an integer selected from 0 to 3; and C, benzenealkane-diyl, and —(CH)—(CH-CH each n2 is independently an integer selected from 1 to 25, in O), (CH), , wherein each n1 and n3 is independently certain embodiments an integer selected from 1 to 20, in an integer selected from 0 to 3 and each n2 is independently certain embodiments an integer selected from 1 to 15, in an integer selected from 1 to 10. certain embodiments an integer selected from 1 to 10, and in In certain embodiments of a compound of Formula (III), 60 certain embodiments an integer selected from 1 to 5. each X* is selected from Co alkanediyl, Co cycloal In certain embodiments of a compound of Formula (III), kanediyl, Co arenediyl, and C-2 alkanearenediyl. In cer each X is selected from a covalent bond, -O-, -S , tain embodiments of a compound of Formula (III), each X is N—, N=, N=N , N=C , —SO , —SO , Selected from C-20 alkanediyl, C-2 ocycloalkanediyl. Colo SON ... —SS , —C(O)— —C(O)O , —C(O)N , arenediyl, and C7 alkanearenediyl. 65 - C(O)S C(O)N N—, OP(O)(OH)O-, - OC(O) In certain embodiments of a compound of Formula (III), O - OC(O)N - NC(O)N , and NC(S)N. In certain each X" is a covalent bond. embodiments of a compound of Formula (III), each X is US 9,295,731 B2 67 68 selected from a covalent bond, —O— —S— —SO—, Sabarubicin, topixantrone, monomethylauristatin E. monom SO. , —SON ... —SS , —C(O)— —C(O)C) , ethyl auristatin F, monomethyl dolastatin 10, dolastatin 15, C(O)N , —C(O)N N=, OC(O)N , and - NC(O) belotecan, atiratecan, camptothecin, exatecan, irinotecan, N—. In certain embodiments of a compound of Formula (III), namitecan, rubitecan, topotecan, demecolcine, duocarmycin each X is selected from a covalent bond, SO , A, duocarmycin SA, duocarmycin B, duocarmycin B1, —SON , and —C(O)N . abbeymycin, anthramycin, centanamycin, chicamycin, In certain embodiments of a compound of Formula (III), mazethramycin, porothramycin A, porothramycin B, Sibano each RandR is independently selected from hydrogen, C. mycin, Sibiromycin, trabectedin, calicheamicin Y1. cali alkyl, Substituted Calkyl, Cheteroalkyl, Substituted C. cheamicin T, esperamicin A1, esperamicin C, esperamicin D, heteroalkyl, Coo aryl, Substituted Co-o aryl, Co-o het 10 dynemicin A, dynemicin H, dynemicin M, dynemicin N. eroaryl, and Substituted Coheteroaryl. dynemicin O. dynemicin P. dynemicin Q, dynemicin S. neo In certain embodiments of a compound of Formula (III), carzinostatin chromophore, uncialamycin, 21-aminoe each RandR is independently selected from hydrogen, C. pothilone B, eribulin, hemiasterlin, HTI-286, kahalatide F. alkyl, Substituted C. alkyl, and Co aryl. In certain elsamitrucin, lucanthone, melphalan, mitoguaZone, nimus embodiments of a compound of Formula (III), R' and Rare 15 tine, procarbazine, dacarbazine, amsacrine, 5-amino-4-oxo independently selected from hydrogen, methyl, ethyl, propyl. pentanoic acid, methyl 5-amino-4-oxo-pentanoate, actino isopropyl, and phenyl. In certain embodiments of a com mycin D, 7-aminoactinomycin D, bleomycin, mitomycin, pound of Formula (III), each R' and R is independently staurosporine, desacetylvinblastine hydrazide, vinblastine, selected from hydrogen and methyl. In certain embodiments Vincristine, vindestine, Vinflunine, vinorelbine, afatinib, of a compound of Formula (III), each R' and R is indepen apilimod, balamapimod, barasertib, boSutinib, canertinib, dently selected from hydrogen and ethyl. In certain embodi cevipabulin, crizotinib, dacomitinib, dasatinib, denibulin, dil ments of a compound of Formula (III), each R" and R is mapimod, dinaciclib, dovitinib, dutacatib, duVoglustat, edot independently selected from hydrogen and propyl. In certain ecarin, elisidepsin, entinostat, epetirimod, erlotinib, fingoli embodiments of a compound of Formula (III), each RandR mod, fostamatinib, gefitinib, golotimod, gusperimus, is independently selected from hydrogen and isopropyl. In 25 imatinib, imiquimod, intedanib, ispinesib, lapatinib, lenali certain embodiments of a compound of Formula (III), each R' domide, linifanib, litronesib, loSmapimod, metesind, moceti and R is independently selected from hydrogen and phenyl. nostat, motesanib, masitinib, myriocin, neratinib, nilotinib, In certain embodiments of a compound of Formula (III), odanacatib, ombrabulin, pamapimod, panobinostat, paZo each R' and R is hydrogen. In certain embodiments of a panib, plerixafor, pomalidomide, raZupenem, residuimod, compound of Formula (III), each R" is hydrogen and each R 30 Sabarubicin, Saracatinib, Seliciclib, Selumetinib, Sotirimod, is methyl. In certain embodiments of a compound of Formula squalamine, tacedinaline, tallabostat, taltobulin, tellatinib, tipi (III), each R" is hydrogen and each R is ethyl. In certain farnib, toZasertib, vandetanib, Vatalanib, veliparib, Voreloxin, embodiments of a compound of Formula (III), each R' is alvespimycin, amikacin, amphotericin B, arbekacin, astromi hydrogen and each R is propyl. In certain embodiments of a cin, bacitracin, balofloxacin, bederocin, bekanamycin, besi compound of Formula (III), each R" is hydrogen and each R 35 floxacin, brodimoprim, ciprofloxacin, clinafloxacin, colistin, is isopropyl. In certain embodiments of a compound of For daptomycin, dibekacin, enoxacin, framycetin, garenoxacin, mula (III), each R" is hydrogen and each R is phenyl. gatifloxacin, gemifloxacin, gentamicin, gentamicin, grepa In certain embodiments of a compound of Formula (III), floxacin, hamycin, hexetidine, hygromycin B, ibacitabine, each D is independently selected from a therapeutic agent iclaprim, isepamicin, kanamycin, lomefloxacin, lucimycin, having at least one primary amine group, a therapeutic agent 40 lymecycline, mepartricin, moxifloxacin, natamycin, nemon having at least one secondary amine group, a therapeutic oxacin, neomycin B, neomycin C, netilmicin, norfloxacin, agent having at least one hydroxy group, and a therapeutic nystatin, omadacycline, oritavancin, paromomycin, paZu agent having at least one thiol group. floxacin, perimycin A, perimycin B, perimycin C, pipemidic In certain embodiments of a compound of Formula (III), acid, polymyxin B, puromycin, radeZolid, retaspimycin, each D is independently selected from a folic acid analog, a 45 ribostamycin, rimocidin, Sisomicin, sitafloxacin, sparfloxa purine analog, a pyrimidine analog, an anthracycline deriva cin, spectinomycin, Streptomycin, Sulfacetamide, Sulfadiaz tive, a dolastatin derivative, a camptotheca derivative, an ine, Sulfadimethoxine, Sulfadimidine, Sulfafurazole, Sul ectainascidin derivative, a colchicine derivative, a duocarmy falene, SulfamaZone, Sulfamerazine, Sulfamethizole, cin derivative, an enediyne derivative, an epothilone deriva Sulfamethoxazole, Sulfamethoxypyridazine, Sulfametomi tive, a halichondrin derivative, a kahalalide derivative, a 50 dine, Sulfametoxydiazine, Sulfametrole, Sulfamoxole, Sulfa streptomyces, a tubulysin derivative, a Vinca alkaloid, an anti nilamide, Sulfaperin, Sulfaphenazole, Sulfapyridine, Sul folate, a hemiasterlin, a cathepsin Kinhibitor, dipeptidyl pep fathiazole, Sulfathiourea, Sulfisomidine, teicoplanin, tidase IV inhibitor, heat shock protein 90 (Hsp90) inhibitor, a telavancin, tanespimycin, temafloxacin, tetroXoprim, tigecy histone deacetylase inhibitor, an immunomodulator, an cline, tobramycin, to Sufloxacin, trimethoprim, trimethoprim, aurora kinase inhibitor, a cyclin-dependent kinase inhibitor, 55 trovafloxacin, tyrothricin, ulifloxacin, valnemulin, Vancomy tyrosine kinase inhibitor, kinesin-related motor protein Eg5 cin, Verdamicin, and Zabofloxacin. inhibitor, kinesin spindle protein inhibitor, microtubule inter In certain embodiments of a compound of Formula (III), né ference compounds, topoisomerase inhibitor, and an antibi is an integer selected from 1 to 20; in certain embodiments, an otic. integer selected from 1 to 10; in certain embodiments, an In certain embodiments of a compound of Formula (III), 60 integer selected from 1 to 4; and in certain embodiments, né each D is independently selected from aminopterin, folitix is 1, 2, 3, or 4. In certain embodiments, né is 1, né is 2, né is orin, methotrexate, pemetrexed, pralatrexate, raltitrexed, 3, and in certain embodiments né is 4. pelitrexol, talotrexin, deoxycoformycin, cladribine, clofara In certain embodiments of a compound of Formula (III), Q bine, fludarabine, thioguanine, mercaptopurine, berubicin, is a ligand selected from a Cso alkyl, a polyethylene glycol, daunorubicin, doxorubicin, epirubicin, idarubicin, amrubi 65 an avidin, an albumin, an antibody, a polymer, and a glass cin, pirarubicin, Zorubicin, mitoxantrone, banoxantrone, (SiO) substrate; each Y is independently selected from a ledoxantrone, nortopixantrone, pixantrone, piroXantrone, covalent bond and a hydrogen bond; each A is independently US 9,295,731 B2 69 70 selected from a covalent bond, —C(O)N , —C(O)O—, O— —SO , —SON , —C(O)— —C(O)O—, and C—N –C(O)N N=C OC(O)N-, - NC(O) —C(O)N ; each R" and R is independently selected from N— —SO ... —SON ... —S , —SS—, a moiety of For- hydrogen and methyl; each D is selected from a therapeutic mula (Aa1), a moiety of Formula (Ab2), a moiety of For agent having at least one primary amine group and a thera mula (Ac1), a moiety of Formula (Ac2), a moiety of 5 peutic agent having at least one secondary amine group; and Formula (Ac3), a moiety of Formula (Afl), and a moiety of nó is selected from 1 to 5. Formula (Af2); each X and X is independently selected In certain embodiments of a compound of Formula (III), Q from a covalent bond, methane-diyl ethane-1,2-diyl, pro is a Cso alkyl; each Y is a covalent bond; each A is a moiety pane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6- of Formula (Aa1); each X and X is independently selected diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl. 10 from a covalent bond, methane-diyl ethane-1,2-diyl, pro decane-1,10-diyl, undecane-1,11-diyl, dodecane-1,12-diyl. pane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6- hexadecane-1,16-diyl, benzene-diyl, methylbenzene-diyl. diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl. propylbenzene-diyl. methylcyclohexane-diyl. and decane-1,10-diyl, undecane-1,11-diyl, dodecane-1,12-diyl. (CH),—(CH-CH O), (CH), , wherein each hexadecane-1,16-diy1, benzene-diyl, methylbenzene-diyl. n1 and n3 is independently an integer selected from 0 to 5, and 15 each n2 is independently an integer selected from 1 to 25; propylbenzene-diyl. methylcyclohexane-diyl. and each X* is independently selected from a covalent bond, (CH2)—(CH2—CH2—O) (CH), , wherein each O— —S— —SO— —SO , —SON ... —SS—, n1 and n3 is independently an integer selected from 0 to 3, and C(O) , C(O)O , C(O)N , C(O)N N=, each n2 is independently an integer selected from 1 to 20; —OC(O)N , and NC(O)N ; each R' and R is indepen- 20 each X is selected from a covalent bond, -O-, -SO , dently selected from hydrogen, Calkyl, and Cesaryl; each SON , —C(O)— —C(O)C)—, and —C(O)N ; each D is selected from a therapeutic agent having at least one R" and R is independently selected from hydrogen and primary amine group, a therapeutic agent having at least one methyl; each D is selected from a therapeutic agent having at secondary amine group, a therapeutic agent having at least least one primary amine group and a therapeutic agent having one hydroxy group, and a therapeutic agent having at least 25 at least one secondary amine group; and né is selected from 1 one thiol group; and né is selected from 1 to 20. to 5. In certain embodiments of a compound of Formula (III), Q In certain embodiments of a compound of Formula (III), Q is a ligand selected from a Cso alkyl, an MPEG, a human is a Cso alkyl; each Y is a covalent bond; each A is a moiety serum albumin, an antibody, a polymer, and a glass (SiO2) of Formula (Aa1), wherein each RandR is hydrogen; each substrate; each Y is independently selected from a covalent 30 X' is pentane-1,5-diyl; each X* is a covalent bond; each X is bond and a hydrogen bond; each A is independently selected a covalent bond; each R" and R is hydrogen; each D is from a covalent bond, —C(O)N —SO —SON . independently selected from a therapeutic agent having at —S-, -SS , a moiety of Formula (Aa1), a moiety of least one primary amine group and a therapeutic agent having Formula (Ab2), a moiety of Formula (Ac1), a moiety of at least one secondary amine group; and né is selected from 1 Formula (Ac2), a moiety of Formula (Ac3), a moiety of 35 to 4. Formula (Af1), and amoiety of Formula (Af2); eachX' and In certain embodiments of a compound of Formula (III), Q X is independently selected from a covalent bond, methane is a Cs2 alkyl; each Y is a covalent bond; each A is a moiety diyl ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pen of Formula (Aa1), wherein each RandR is hydrogen; each tane-1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl, octane-1.8- X' is 4-methylcyclohexane-diyl; each X* is a covalent bond; diyl. nonane-1,9-diyl. decane-1,10-diyl, undecane-1,11-diyl. 40 each X is a covalent bond; each R" and R is hydrogen; each dodecane-1,12-diyl, hexadecane-1,16-diyl, benzene-diyl. D is selected from a therapeutic agent having at least one methylbenzene-diyl, propylbenzene-diyl, methylcyclohex primary amine group and a therapeutic agent having at least ane-diyl, and —(CH)—(CH2—CH2—O) (CH), , one secondary amine group; and né is selected from 1 to 4. wherein each n1 and n3 is independently an integer selected In certain embodiments of a compound of Formula (III), Q from 0 to 3, and each n2 is independently an integer selected 45 is a Cso alkyl; each Y is a covalent bond; each A is a moiety from 1 to 20; each X* is selected from a covalent bond, of Formula (Aa1), wherein each RandR is hydrogen; each O— —SO , —SON , —C(O)— —C(O)C)—, and X' is pentane-1,5-diyl; each X* is —C(O)N ; each X is —C(O)N ; each R' and R is independently selected from pentane-1,5-diyl; each R" and R is hydrogen; each D is hydrogen, methyl, ethyl, propyl, isopropyl, and phenyl; each selected from a therapeutic agent having at least one primary D is selected from a therapeutic agent having at least one 50 amine group and a therapeutic agent having at least one primary amine group and a therapeutic agent having at least secondary amine group; and né is selected from 1 to 4. one secondary amine group; and né is selected from 1 to 20. In certain embodiments of a compound of Formula (III), Q In certain embodiments of a compound of Formula (III), Q is a Cs2 alkyl; each Y is a covalent bond; each A is a moiety is a Cso alkyl; each Y is a covalent bond; each A is inde of Formula (Af1); each X and X is independently selected pendently selected from a covalent bond, —C(O)N , 55 from a covalent bond, methane-diyl ethane-1,2-diyl, pro —SO. , —S , a moiety of Formula (Aa1), a moiety of pane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6- Formula (Af1), and amoiety of Formula (Af2); eachX' and diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl. X is independently selected from a covalent bond, methane decane-1,10-diyl, undecane-1,11-diyl, dodecane-1,12-diyl. diyl ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pen hexadecane-1,16-diy1, benzene-diyl, methylbenzene-diyl. tane-1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl, octane-1.8- 60 propylbenzene-diyl. methylcyclohexane-diyl. and diyl. nonane-1,9-diyl. decane-1,10-diyl, undecane-1,11-diyl. (CH)—(CH-CH O), (CH), , wherein each dodecane-1,12-diyl, hexadecane-1,16-diyl, benzene-diyl. n1 and n3 is independently an integer selected from 0 to 3, and methylbenzene-diyl, propylbenzene-diyl, methylcyclohex each n2 is independently an integer selected from 1 to 20; ane-diyl, and —(CH)—(CH2—CH2—O) (CH), , each X is selected from a covalent bond, -O-, -SO , wherein each n1 and n3 is independently an integer selected 65 SON , —C(O)— —C(O)O—, and —C(O)N ; each from 0 to 3, and each n2 is independently an integer selected R" and R is independently selected from hydrogen and from 1 to 20; each X* is selected from a covalent bond, methyl; each D is selected from a therapeutic agent having at US 9,295,731 B2 71 72 least one primary amine group and a therapeutic agent having moiety of Formula (Af1), and a moiety of Formula (Af2): at least one secondary amine group; and né is selected from 1 each X and X is independently selected from a covalent to 5. bond, methane-diyl, ethane-1,2-diyl, propane-1,3-diyl. In certain embodiments of a compound of Formula (III), Q butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6-diyl, heptane is a Cso alkyl; each Y is a covalent bond; each A is a moiety 1,7-diyl octane-1,8-diyl. nonane-1,9-diyl, decane-1,10-diyl. of Formula (Af1); each X is pentane-1,5-diyl; each X is a undecane-1,11-diyl, dodecane-1,12-diyl, hexadecane-1,16 covalent bond; each X is a covalent bond; each R' and R is diyl, benzene-diyl, methylbenzene-diyl, propylbenzene-diyl. hydrogen; each D is independently selected from a therapeu methylcyclohexane-diyl. and —(CH2)—(CH2— tic agent having at least one primary amine group and a CH-O) (CH), , wherein each n1 and n3 is indepen therapeutic agent having at least one secondary amine group; 10 dently an integer selected from 0 to 3, and each n2 is inde and né is selected from 1 to 4. pendently an integer selected from 1 to 20; each X* is selected In certain embodiments of a compound of Formula (III), Q from a covalent bond, —O— —SO , —SON.— is a Cs2 alkyl; each Y is a covalent bond; each A is a moiety C(O) , —C(O)O , and C(O)N ; each R' and R is of Formula (Af1); each X is 4-methylcyclohexane-diyl; independently selected from hydrogen and methyl; D is each X* is a covalent bond; each X is a covalent bond; each 15 selected from a therapeutic agent having at least one primary R" and R is hydrogen; each D is selected from a therapeutic amine group and a therapeutic agent having at least one agent having at least one primary amine group and a thera secondary amine group; and né is selected from 1 to 5. peutic agent having at least one secondary amine group; and In certain embodiments of a compound of Formula (III), Q nó is selected from 1 to 4. is an MPEG having a number average molecular weight from In certain embodiments of a compound of Formula (III), Q about 10,000 to 60,000 daltons; each Y is a covalent bond; is a Cso alkyl; each Y is a covalent bond; each A is a moiety each A is a moiety of Formula (Aa1); each X and X is of Formula (Af1); each X" is pentane-1,5-diyl; each X* is independently selected from a covalent bond, methane-diyl. —C(O)N ; each X is pentane-1,5-diyl; each R" and R is ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane hydrogen; each D is selected from a therapeutic agent having 1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. at least one primary amine group and a therapeutic agent 25 nonane-1,9-diyl. decane-1,10-diyl, undecane-1,11-diyl. having at least one secondary amine group; and né is selected dodecane-1,12-diyl, hexadecane-1,16-diyl, benzene-diyl. from 1 to 4. methylbenzene-diyl, propylbenzene-diyl, methylcyclohex In certain embodiments of a compound of Formula (III), Q ane-diyl, and —(CH)—(CH-CH O), (CH), , is a Cs2 alkyl; each Y is a covalent bond; each A is a moiety wherein each n1 and n3 is independently an integer selected of Formula (Af2); each X and X is independently selected 30 from 0 to 3, and each n2 is independently an integer selected from a covalent bond, methane-diyl ethane-1,2-diyl, pro from 1 to 20; each X* is selected from a covalent bond, pane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6- O —SO —SON , —C(O)— —C(O)O—, and diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl. —C(O)N ; each R" and R is independently selected from decane-1,10-diyl, undecane-1,11-diyl, dodecane-1,12-diyl. hydrogen and methyl; each D is selected from a therapeutic hexadecane-1,16-diyl, benzene-diyl, methylbenzene-diyl. 35 agent having at least one primary amine group and a thera propylbenzene-diyl. methylcyclohexane-diyl. and peutic agent having at least one secondary amine group; and (CH2), —(CH2—CH2—O) (CH), , wherein each nó is selected from 1 to 5. n1 and n3 is independently an integer selected from 0 to 3, and In certain embodiments of a compound of Formula (III), Q each n2 is independently an integer selected from 1 to 20; is an MPEG having a number average molecular weight from each X is selected from a covalent bond, -O-, -SO , 40 about 10,000 to 60,000 daltons; each Y is a covalent bond; SON , —C(O)— —C(O)O—, and —C(O)N ; each each A is a moiety of Formula (Aa1), wherein each Rand R" and R is independently selected from hydrogen and R is hydrogen; each X is pentane-1,5-diyl; each X* is a methyl; each D is selected from a therapeutic agent having at covalent bond; each X is a covalent bond; each R' and R is least one primary amine group and a therapeutic agent having hydrogen; each D is independently selected from a therapeu at least one secondary amine group; and né is selected from 1 45 tic agent having at least one primary amine group and a to 5. therapeutic agent having at least one secondary amine group; In certain embodiments of a compound of Formula (III), Q and né is selected from 1 to 4. is a Cs2 alkyl; each Y is a covalent bond; each A is a moiety In certain embodiments of a compound of Formula (III), Q of Formula (Af2); each X* is methane-diyl; each X* is a is an MPEG having a number average molecular weight from covalent bond; each X is a covalent bond; each R' and R is 50 about 10,000 to 60,000 daltons; each Y is a covalent bond; hydrogen; each D is independently selected from a therapeu each A is a moiety of Formula (Aa1), wherein each Rand tic agent having at least one primary amine group and a R is hydrogen; each X* is 4-methylcyclohexane-diyl; each therapeutic agent having at least one secondary amine group; X is a covalent bond; each X is a covalent bond; each R" and and né is selected from 1 to 4. R is hydrogen; each D is selected from a therapeutic agent In certain embodiments of a compound of Formula (III), Q 55 having at least one primary amine group and a therapeutic is a Cso alkyl; each Y is a covalent bond; each A is a moiety agent having at least one secondary amine group; and né is of Formula (Af2); each X is methane-diyl; each X is selected from 1 to 4. —C(O)N ; each X is pentane-1,5-diyl; each R' and R is In certain embodiments of a compound of Formula (III), Q hydrogen; each D is selected from a therapeutic agent having is an MPEG having a number average molecular weight from at least one primary amine group and a therapeutic agent 60 about 10,000 to 60,000 daltons; each Y is a covalent bond; having at least one secondary amine group; and né is selected each A is a moiety of Formula (Aa1), wherein each Rand from 1 to 4. R is hydrogen; each X is pentane-1,5-diyl; each X is In certain embodiments of a compound of Formula (III), Q —C(O)N ; each X is pentane-1,5-diyl; each R" and R is is an MPEG having a number average molecular weight from hydrogen; each D is selected from a therapeutic agent having about 10,000 to 60,000 daltons; each Y is a covalent bond; 65 at least one primary amine group and a therapeutic agent each A is independently selected from a covalent bond, having at least one secondary amine group; and né is selected —C(O)N , -SO. , S: , a moiety of Formula (Aa1), a from 1 to 4. US 9,295,731 B2 73 74 In certain embodiments of a compound of Formula (III), Q In certain embodiments of a compound of Formula (III), Q is an MPEG having a number average molecular weight from is an MPEG having a number average molecular weight from about 10,000 to 60,000 daltons; each Y is a covalent bond; about 10,000 to 60,000 daltons; each Y is a covalent bond; each A is a moiety of Formula (Afl); each X and X is each A is a moiety of Formula (Af2); each X* is methane independently selected from a covalent bond, methane-diyl. diyl; each X* is a covalent bond; each X is a covalent bond; ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane each RandR is hydrogen; each Dis independently selected 1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. from a therapeutic agent having at least one primary amine nonane-1,9-diyl. decane-1,10-diyl, undecane-1,11-diyl. group and a therapeutic agent having at least one secondary dodecane-1,12-diyl, hexadecane-1,16-diyl, benzene-diyl. amine group; and né is selected from 1 to 4. methylbenzene-diyl, propylbenzene-diyl, methylcyclohex 10 ane-diyl, and —(CH)—(CH2—CH2—O) (CH), , In certain embodiments of a compound of Formula (III), Q wherein each n1 and n3 is independently an integer selected is an MPEG having a number average molecular weight from from 0 to 3, and each n2 is independently an integer selected about 10,000 to 60,000 daltons; each Y is a covalent bond; from 1 to 20; each X* is selected from a covalent bond, each A is a moiety of Formula (Af2); each X* is methane O— —SO , —SON , —C(O)— —C(O)O—, and 15 diyl; each X* is —C(O)N ; each X is pentane-1,5-diyl; —C(O)N ; each R" and R is independently selected from each R" and R is hydrogen; each D is selected from a thera hydrogen and methyl; each D is selected from a therapeutic peutic agent having at least one primary amine group and a agent having at least one primary amine group and a thera therapeutic agent having at least one secondary amine group; peutic agent having at least one secondary amine group; and and né is selected from 1 to 4. nó is selected from 1 to 5. In certain embodiments of a compound of Formula (III), Q In certain embodiments of a compound of Formula (III), Q is an avidin; each Y is a hydrogen bond; each A is indepen is an MPEG having a number average molecular weight from dently selected from a moiety of Formula (Ac1), a moiety of about 10,000 to 60,000 daltons; each Y is a covalent bond; Formula (Ac2), and a moiety of Formula (Ac3); each X' each A is a moiety of Formula (Af1); each X is pentane-1, and X is independently selected from a covalent bond, meth 5-diyl; each X* is a covalent bond; each X is a covalent bond; 25 ane-diyl ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl. each RandR is hydrogen; each Dis independently selected pentane-1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl octane from a therapeutic agent having at least one primary amine 1,8-diyl. nonane-1,9-diyl. decane-1,10-diyl, undecane-1,11 group and a therapeutic agent having at least one secondary diyl, dodecane-1,12-diyl, hexadecane-1,16-diyl, benzene amine group; and né is selected from 1 to 4. diyl. methylbenzene-diyl. propylbenzene-diyl. In certain embodiments of a compound of Formula (III), Q 30 methylcyclohexane-diyl. and —(CH)—(CH is an MPEG having a number average molecular weight from CH2—O) (CH) , wherein each n1 and n3 is indepen about 10,000 to 60,000 daltons; each Y is a covalent bond; dently an integer selected from 0 to 3, and each n2 is inde each A is a moiety of Formula (Af1); each X* is 4-methyl pendently an integer selected from 1 to 20; each X* is selected cyclohexane-diyl; each X is a covalent bond; each X is a from a covalent bond, —O— —SO , —SON.— covalent bond; each RandR is hydrogen; each D is selected 35 C(O) , —C(O)C)—, and —C(O)N ; each R" and R is from a therapeutic agent having at least one primary amine independently selected from hydrogen and methyl; each Dis group and a therapeutic agent having at least one secondary selected from a therapeutic agent having at least one primary amine group; and né is selected from 1 to 4. amine group and a therapeutic agent having at least one In certain embodiments of a compound of Formula (III), Q secondary amine group; and né is selected from 1 to 4. is an MPEG having a number average molecular weight from 40 In certain embodiments of a compound of Formula (III), Q about 10,000 to 60,000 daltons; each Y is a covalent bond; is an avidin; each Y is a hydrogen bond; each A is indepen each A is a moiety of Formula (Afl); each X is pentane-1, dently selected from a moiety of Formula (Ac1), a moiety of 5-diyl; each X* is —C(O)N ; each X is pentane-1,5-diyl; Formula (Ac2), and a moiety of Formula (Ac3); each X is each R" and R is hydrogen; each D is selected from a thera butane-1,4-diyl; each X is selected from a covalent bond and peutic agent having at least one primary amine group and a 45 —C(O)N ; each X is selected from a covalent bond, pen therapeutic agent having at least one secondary amine group; tane-1,5-diyl, and (CH2)—(CH2—CH2—O), and né is selected from 1 to 4. (CH), , wherein each n1 and n3 is independently an inte In certain embodiments of a compound of Formula (III), Q ger selected from 0 to 3, and each n2 is independently an is an MPEG having a number average molecular weight from integer selected from 1 to 10; each R" and R is hydrogen; about 10,000 to 60,000 daltons; each Y is a covalent bond; 50 each D is independently selected from a therapeutic agent each A is a moiety of Formula (Af2); each X and X is having at least one primary amine group and a therapeutic independently selected from a covalent bond, methane-diyl. agent having at least one secondary amine group; and né is ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane selected from 1 to 4. 1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. In certain embodiments of a compound of Formula (III), Q nonane-1,9-diyl. decane-1,10-diyl, undecane-1,11-diyl. 55 is an avidin; each Y is a hydrogen bond; each A is a moiety of dodecane-1,12-diyl, hexadecane-1,16-diyl, benzene-diyl. Formula (Ac1); each X is butane-1,4-diyl; each X* is methylbenzene-diyl, propylbenzene-diyl, methylcyclohex —C(O)N ; each X is pentane-1,5-diyl; each R' and R is ane-diyl, and —(CH)—(CH2—CH2—O) (CH), , hydrogen; each D is independently selected from a therapeu wherein each n1 and n3 is independently an integer selected tic agent having at least one primary amine group and a from 0 to 3, and each n2 is independently an integer selected 60 therapeutic agent having at least one secondary amine group; from 1 to 20; each X* is selected from a covalent bond, and né is selected from 1 to 4. O— —SO , —SON , —C(O)— —C(O)O—, and In certain embodiments of a compound of Formula (III), Q —C(O)N ; each R" and R is independently selected from is an avidin; eachY is a hydrogenbond; each A is a moiety of hydrogen and methyl; each D is selected from a therapeutic Formula (Ac2); each X" is butane-1,4-diyl; each X is agent having at least one primary amine group and a thera 65 —C(O)N ; each X is pentane-1,5-diyl; each R' and R is peutic agent having at least one secondary amine group; and hydrogen; each D is independently selected from a therapeu nó is selected from 1 to 5. tic agent having at least one primary amine group and a US 9,295,731 B2 75 76 therapeutic agent having at least one secondary amine group; is a moiety of Formula (Aa1), wherein each R and R is and né is selected from 1 to 4. hydrogen; each X is pentane-1,5-diyl; each X* is C(O) In certain embodiments of a compound of Formula (III), Q N. ; each X is pentane-1,5-diyl; each RandR is hydrogen; is an avidin; each Y is a hydrogen bond; each A is a moiety of each Dis selected from a therapeutic agent having at least one Formula (Ac3); each X is butane-1,4-diyl; each X* is 5 primary amine group and a therapeutic agent having at least —C(O)N ; each X is pentane-1,5-diyl; each R' and R is one secondary amine group; and né is 1. hydrogen; each D is independently selected from a therapeu In certain embodiments of a compound of Formula (III), Q tic agent having at least one primary amine group and a is a human serum albumin; each Y is a covalent bond; each A therapeutic agent having at least one secondary amine group; is a moiety of Formula (Af1); each X and X is indepen and né is selected from 1 to 4. 10 dently selected from a covalent bond, methane-diyl ethane In certain embodiments of a compound of Formula (III), Q 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. is a human serum albumin; each Y is a covalent bond; each A hexane-1,6-diyl, heptane-1,7-diyl octane-1,8-diyl. nonane is independently selected from a covalent bond, —C(O)N 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, —SO. , —S , a moiety of Formula (Aa1), a moiety of 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben Formula (Af1), and amoiety of Formula (Af2); eachX' and 15 X is independently selected from a covalent bond, methane Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and diyl ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pen (CH2)—(CH2—CH2—O) (CH), , wherein each tane-1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl, octane-1.8- n1 and n3 is independently an integer selected from 0 to 3, and diyl. nonane-1,9-diyl. decane-1,10-diyl, undecane-1,11-diyl. each n2 is independently an integer selected from 1 to 20; dodecane-1,12-diyl, hexadecane-1,16-diyl, benzene-diyl. each X is selected from a covalent bond, -O-, -SO , methylbenzene-diyl, propylbenzene-diyl, methylcyclohex SON , —C(O)— —C(O)C)—, and —C(O)N ; each ane-diyl, and —(CH)—(CH2—CH2—O) (CH), , R" and R is independently selected from hydrogen and wherein each n1 and n3 is independently an integer selected methyl; each D is selected from a therapeutic agent having at from 0 to 3, and each n2 is independently an integer selected least one primary amine group and a therapeutic agent having from 1 to 20; each X* is selected from a covalent bond, 25 at least one secondary amine group; and né is 1. O— —SO , —SON , —C(O)— —C(O)O—, and In certain embodiments of a compound of Formula (III), Q —C(O)N ; each R' and R is independently selected from is a human serum albumin; each Y is a covalent bond; each A hydrogen and methyl; each D is selected from a therapeutic is a moiety of Formula (Afl); each X* is pentane-1,5-diyl; agent having at least one primary amine group and a thera each X* is a covalent bond; each X is a covalent bond; each peutic agent having at least one secondary amine group; and 30 R" and R is hydrogen; each Dis independently selected from nó is 1. a therapeutic agent having at least one primary amine group In certain embodiments of a compound of Formula (III), Q and a therapeutic agent having at least one secondary amine is a human serum albumin; each Y is a covalent bond; each A group; and né is 1. is a moiety of Formula (Aa1); each X and X is indepen In certain embodiments of a compound of Formula (III), Q dently selected from a covalent bond, methane-diyl ethane 35 is a human serum albumin; each Y is a covalent bond; each A 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. is a moiety of Formula (Af1); each X is 4-methylcyclohex hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane ane-diyl; each X is a covalent bond; each X is a covalent 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, bond; each R" and R is hydrogen; each D is selected from a 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben therapeutic agent having at least one primary amine group Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and 40 and a therapeutic agent having at least one secondary amine (CH2), —(CH2—CH2—O) (CH), , wherein each group; and né is 1. n1 and n3 is independently an integer selected from 0 to 3, and In certain embodiments of a compound of Formula (III), Q each n2 is independently an integer selected from 1 to 20; is a human serum albumin; each Y is a covalent bond; each A each X is selected from a covalent bond, -O-, -SO , is a moiety of Formula (Af1); each X* is pentane-1,5-diyl: SON , —C(O)— —C(O)O—, and —C(O)N ; each 45 each X* is C(O)N ; each X is pentane-1,5-diyl; each R' R" and R is independently selected from hydrogen and and R is hydrogen; each D is selected from a therapeutic methyl; each D is selected from a therapeutic agent having at agent having at least one primary amine group and a thera least one primary amine group and a therapeutic agent having peutic agent having at least one secondary amine group; and at least one secondary amine group; and né is 1. nó is 1. In certain embodiments of a compound of Formula (III), Q 50 In certain embodiments of a compound of Formula (III), Q is a human serum albumin; each Y is a covalent bond; each A is an antibody; each Y is a covalent bond; each A is indepen is a moiety of Formula (Aa1), wherein each R and R is dently selected from a covalent bond, —C(O)N —SO , hydrogen; each X" is pentane-1,5-diyl; each X is a covalent —S , a moiety of Formula (Aa1), a moiety of Formula bond; each X is a covalent bond; each RandR is hydrogen; (Af1), and a moiety of Formula (Af2); each X and X is each D is independently selected from a therapeutic agent 55 independently selected from a covalent bond, methane-diyl. having at least one primary amine group and a therapeutic ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane agent having at least one secondary amine group; and né is 1. 1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. In certain embodiments of a compound of Formula (III), Q nonane-1,9-diyl. decane-1,10-diyl, undecane-1,11-diyl. is a human serum albumin; each Y is a covalent bond; each A dodecane-1,12-diyl, hexadecane-1,16-diyl, benzene-diyl. is a moiety of Formula (Aa1), wherein each R and R is 60 methylbenzene-diyl, propylbenzene-diyl, methylcyclohex hydrogen; each X* is 4-methylcyclohexane-diyl; each X* is a ane-diyl, and —(CH)—(CH-CH O), (CH), , covalent bond; each X is a covalent bond; each R' and R is wherein each n1 and n3 is independently an integer selected hydrogen; each D is selected from a therapeutic agent having from 0 to 3, and each n2 is independently an integer selected at least one primary amine group and a therapeutic agent from 1 to 20; each X* is selected from a covalent bond, having at least one secondary amine group; and né is 1. 65 O— —SO , —SON , —C(O)— —C(O)O—, and In certain embodiments of a compound of Formula (III), Q —C(O)N ; each R" and R is independently selected from is a human serum albumin; each Y is a covalent bond; each A hydrogen and methyl; each D is selected from a therapeutic US 9,295,731 B2 77 78 agent having at least one primary amine group and a thera In certain embodiments of a compound of Formula (III), Q peutic agent having at least one secondary amine group; and is an antibody; each Y is a covalent bond; each A is a moiety nó is selected from 1 to 5. of Formula (Af1); each X is pentane-1,5-diyl; each X is a In certain embodiments of a compound of Formula (III), Q covalent bond; each X is a covalent bond; each R' and R is is an antibody; each Y is a covalent bond; each A is a moiety hydrogen; each D is independently selected from a therapeu of Formula (Aa1); each X and X is independently selected tic agent having at least one primary amine group and a from a covalent bond, methane-diyl ethane-1,2-diyl, pro therapeutic agent having at least one secondary amine group; pane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6- and né is selected from 1 to 5. diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl. In certain embodiments of a compound of Formula (III), Q 10 is an antibody; each Y is a covalent bond; each A is a moiety decane-1,10-diyl, undecane-1,11-diyl, dodecane-1,12-diyl. of Formula (Af1); each X is 4-methylcyclohexane-diyl: hexadecane-1,16-diyl, benzene-diyl, methylbenzene-diyl. each X* is a covalent bond; each X is a covalent bond; each propylbenzene-diyl. methylcyclohexane-diyl. and R" and R is hydrogen; each D is selected from a therapeutic (CH2), —(CH2—CH2—O) (CH), , wherein each agent having at least one primary amine group and a thera n1 and n3 is independently an integer selected from 0 to 3, and 15 peutic agent having at least one secondary amine group; and each n2 is independently an integer selected from 1 to 20; nó is selected from 1 to 5. each X is selected from a covalent bond, -O-, -SO , In certain embodiments of a compound of Formula (III), Q SON , —C(O)—, —C(O)O , and —C(O)N ; each is an antibody; each Y is a covalent bond; each A is a moiety R" and R is independently selected from hydrogen and of Formula (Afl); each X* is pentane-1,5-diyl; each X* is methyl; each D is selected from a therapeutic agent having at —C(O)N ; each X is pentane-1,5-diyl; each R' and R is least one primary amine group and a therapeutic agent having hydrogen; each D is selected from a therapeutic agent having at least one secondary amine group; and né is selected from 1 at least one primary amine group and a therapeutic agent to 5. having at least one secondary amine group; and né is selected In certain embodiments of a compound of Formula (III), Q from 1 to 5. is an antibody; each Y is a covalent bond; each A is a moiety 25 In certain embodiments of a compound of Formula (III), Q of Formula (Aa1), wherein each RandR is hydrogen; each is an antibody; each Y is a covalent bond; each A is a moiety X' is pentane-1,5-diyl; each X is a covalent bond; each X is of Formula (Af2); each X and X is independently selected a covalent bond; each R" and R is hydrogen; each D is from a covalent bond, methane-diyl ethane-1,2-diyl, pro independently selected from a therapeutic agent having at pane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6- least one primary amine group and a therapeutic agent having 30 diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl. at least one secondary amine group; and né is selected from 1 decane-1,10-diyl, undecane-1,11-diyl, dodecane-1,12-diyl. to 5. hexadecane-1,16-diyl, benzene-diyl, methylbenzene-diyl. In certain embodiments of a compound of Formula (III), Q propylbenzene-diyl. methylcyclohexane-diyl. and is an antibody; each Y is a covalent bond; each A is a moiety (CH2)—(CH2—CH2—O) (CH), , wherein each of Formula (Aa1), wherein each RandR is hydrogen; each 35 n1 and n3 is independently an integer selected from 0 to 3, and X' is 4-methylcyclohexane-diyl; each X* is a covalent bond; each n2 is independently an integer selected from 1 to 20; each X is a covalent bond; each R' and R is hydrogen; each each X is selected from a covalent bond, -O-, -SO , D is selected from a therapeutic agent having at least one SON , —C(O)— —C(O)O—, and —C(O)N ; each primary amine group and a therapeutic agent having at least R" and R is independently selected from hydrogen and one secondary amine group; and né is selected from 1 to 5. 40 methyl; each D is selected from a therapeutic agent having at In certain embodiments of a compound of Formula (III), Q least one primary amine group and a therapeutic agent having is an antibody; each Y is a covalent bond; each A is a moiety at least one secondary amine group; and né is selected from 1 of Formula (Aa1), wherein each RandR is hydrogen; each to 5. X' is pentane-1,5-diyl; each X* is —C(O)N ; each X is In certain embodiments of a compound of Formula (III), Q pentane-1,5-diyl; each R" and R is hydrogen; each D is 45 is an antibody; each Y is a covalent bond; each A is a moiety selected from a therapeutic agent having at least one primary of Formula (Af2); each X* is methane-diyl; each X* is a amine group and a therapeutic agent having at least one covalent bond; each X is a covalent bond; each R' and R is secondary amine group; and né is selected from 1 to 5. hydrogen; each D is independently selected from a therapeu In certain embodiments of a compound of Formula (III), Q tic agent having at least one primary amine group and a is an antibody; each Y is a covalent bond; each A is a moiety 50 therapeutic agent having at least one secondary amine group; of Formula (Afl); each X and X is independently selected and né is selected from 1 to 5. from a covalent bond, methane-diyl ethane-1,2-diyl, pro In certain embodiments of a compound of Formula (III), Q pane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6- is an antibody; each Y is a covalent bond; each A is a moiety diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl. of Formula (Af2); each X is methane-diyl; each X* is decane-1,10-diyl, undecane-1,11-diyl, dodecane-1,12-diyl. 55 —C(O)N ; each X is pentane-1,5-diyl; each R' and R is hexadecane-1,16-diyl, benzene-diyl, methylbenzene-diyl. hydrogen; each D is selected from a therapeutic agent having propylbenzene-diyl. methylcyclohexane-diyl. and at least one primary amine group and a therapeutic agent (CH2), —(CH2—CH2—O) (CH), , wherein each having at least one secondary amine group; and né is selected n1 and n3 is independently an integer selected from 0 to 3, and from 1 to 5. each n2 is independently an integer selected from 1 to 20; 60 In certain embodiments of a compound of Formula (III), Q each X is selected from a covalent bond, -O-, -SO , is a polystyrene; each Y is a covalent bond; each A is inde SON , —C(O)— —C(O)O—, and —C(O)N ; each pendently selected from a covalent bond, —C(O)N . R" and R is independently selected from hydrogen and —SO. , —S , a moiety of Formula (Aa1), a moiety of methyl; each D is selected from a therapeutic agent having at Formula (Af1), and a moiety of Formula (Af2); eachX' and least one primary amine group and a therapeutic agent having 65 X is independently selected from a covalent bond, methane at least one secondary amine group; and né is selected from 1 diyl ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pen to 5. tane-1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl, octane-1.8- US 9,295,731 B2 79 80 diyl. nonane-1,9-diyl. decane-1,10-diyl, undecane-1,11-diyl. n1 and n3 is independently an integer selected from 0 to 3, and dodecane-1,12-diyl, hexadecane-1,16-diyl, benzene-diyl. each n2 is independently an integer selected from 1 to 20; methylbenzene-diyl, propylbenzene-diyl, methylcyclohex each X is selected from a covalent bond, -O-, -SO , ane-diyl, and —(CH)—(CH2—CH2—O) (CH), , SON , —C(O)— —C(O)O—, and —C(O)N ; each wherein each n1 and n3 is independently an integer selected R" and R is independently selected from hydrogen and from 0 to 3, and each n2 is independently an integer selected methyl; each D is selected from a therapeutic agent having at from 1 to 20; each X* is selected from a covalent bond, least one primary amine group and a therapeutic agent having O— —SO , —SON , —C(O)— —C(O)O—, and at least one secondary amine group; and né is selected from 1 —C(O)N ; each R" and R is independently selected from to 20. hydrogen and methyl; each D is selected from a therapeutic 10 agent having at least one primary amine group and a thera In certain embodiments of a compound of Formula (III), Q peutic agent having at least one secondary amine group; and is a polystyrene; each Y is a covalent bond; each A is a moiety nó is selected from 1 to 20. of Formula (Af1); each X is pentane-1,5-diyl; each X is a In certain embodiments of a compound of Formula (III), Q covalent bond; each X is a covalent bond; each R' and R is is a polystyrene; each Y is a covalent bond; each A is a moiety 15 hydrogen; each D is independently selected from a therapeu of Formula (Aa1); each X and X is independently selected tic agent having at least one primary amine group and a from a covalent bond, methane-diyl ethane-1,2-diyl, pro therapeutic agent having at least one secondary amine group; pane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6- and né is selected from 1 to 20. diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl. In certain embodiments of a compound of Formula (III), Q decane-1,10-diyl, undecane-1,11-diyl, dodecane-1,12-diyl. is a polystyrene; eachY is a covalent bond; each A is a moiety hexadecane-1,16-diyl, benzene-diyl, methylbenzene-diyl. of Formula (Af1); each X is 4-methylcyclohexane-diyl; propylbenzene-diyl. methylcyclohexane-diyl. and each X* is a covalent bond; each X is a covalent bond; each (CH),—(CH-CH O), (CH), , wherein each R" and R is hydrogen; each D is selected from a therapeutic n1 and n3 is independently an integer selected from 0 to 3, and agent having at least one primary amine group and a thera each n2 is independently an integer selected from 1 to 20; 25 peutic agent having at least one secondary amine group; and each X is selected from a covalent bond, -O-, -SO , nó is selected from 1 to 20. SON , —C(O)— —C(O)O—, and —C(O)N ; each In certain embodiments of a compound of Formula (III), Q R" and R is independently selected from hydrogen and is a polystyrene; eachY is a covalent bond; each A is a moiety methyl; each D is selected from a therapeutic agent having at of Formula (Afl); each X* is pentane-1,5-diyl; each X* is least one primary amine group and a therapeutic agent having 30 —C(O)N ; each X is pentane-1,5-diyl; each R" and R is at least one secondary amine group; and né is selected from 1 hydrogen; each D is selected from a therapeutic agent having to 20. at least one primary amine group and a therapeutic agent In certain embodiments of a compound of Formula (III), Q having at least one secondary amine group; and né is selected is a polystyrene; each Y is a covalent bond; each A is a moiety from 1 to 20. of Formula (Aa1), wherein each RandR is hydrogen; each 35 X' is pentane-1,5-diyl; each X is a covalent bond; each X is In certain embodiments of a compound of Formula (III), Q a covalent bond; each R" and R is hydrogen; each D is is a polystyrene; eachY is a covalent bond; each A is a moiety independently selected from a therapeutic agent having at of Formula (Af2); each X and X is independently selected least one primary amine group and a therapeutic agent having from a covalent bond, methane-diyl ethane-1,2-diyl, pro at least one secondary amine group; and né is selected from 1 40 pane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6- to 20. diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl. In certain embodiments of a compound of Formula (III), Q decane-1,10-diyl, undecane-1,11-diyl, dodecane-1,12-diyl. is a polystyrene; each Y is a covalent bond; each A is a moiety hexadecane-1,16-diy1, benzene-diyl, methylbenzene-diyl. of Formula (Aa1), wherein each RandR is hydrogen; each propylbenzene-diyl. methylcyclohexane-diyl. and X' is 4-methylcyclohexane-diyl; each X* is a covalent bond; 45 (CH2)—(CH2—CH2—O) (CH), , wherein each each X is a covalent bond; each R' and R is hydrogen; each n1 and n3 is independently an integer selected from 0 to 3, and D is selected from a therapeutic agent having at least one each n2 is independently an integer selected from 1 to 20; primary amine group and a therapeutic agent having at least each X is selected from a covalent bond, -O-, -SO , one secondary amine group; and né is selected from 1 to 20. SON , —C(O)— —C(O)C)—, and —C(O)N ; each In certain embodiments of a compound of Formula (III), Q 50 R" and R is independently selected from hydrogen and is a polystyrene; each Y is a covalent bond; each A is a moiety methyl; each D is selected from a therapeutic agent having at of Formula (Aa1), wherein each RandR is hydrogen; each least one primary amine group and a therapeutic agent having X' is pentane-1,5-diyl; each X* is —C(O)N ; each X is at least one secondary amine group; and né is selected from 1 pentane-1,5-diyl; each R" and R is hydrogen; each D is to 20. selected from a therapeutic agent having at least one primary 55 In certain embodiments of a compound of Formula (III), Q amine group and a therapeutic agent having at least one is a polystyrene; each Y is a covalent bond; each A is a moiety secondary amine group; and né is selected from 1 to 20. of Formula (Af2); each X* is methane-diyl; each X* is a In certain embodiments of a compound of Formula (III), Q covalent bond; each X is a covalent bond; each R' and R is is a polystyrene; each Y is a covalent bond; each A is a moiety hydrogen; each D is independently selected from a therapeu of Formula (Afl); each X and X is independently selected 60 tic agent having at least one primary amine group and a from a covalent bond, methane-diyl ethane-1,2-diyl, pro therapeutic agent having at least one secondary amine group; pane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6- and né is selected from 1 to 20. diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane-1,9-diyl. In certain embodiments of a compound of Formula (III), Q decane-1,10-diyl, undecane-1,11-diyl, dodecane-1,12-diyl. is a polystyrene; eachY is a covalent bond; each A is a moiety hexadecane-1,16-diyl, benzene-diyl, methylbenzene-diyl. 65 of Formula (Af2); each X is methane-diyl; each X is propylbenzene-diyl. methylcyclohexane-diyl. and —C(O)N ; each X is pentane-1,5-diyl; each R" and R is (CH2), —(CH2—CH2—O) (CH), , wherein each hydrogen; each D is selected from a therapeutic agent having US 9,295,731 B2 81 82 at least one primary amine group and a therapeutic agent primary amine group and a therapeutic agent having at least having at least one secondary amine group; and né is selected one secondary amine group; and né is selected from 1 to 20. from 1 to 20. In certain embodiments of a compound of Formula (III), Q In certain embodiments of a compound of Formula (III), Q is a glass (SiO.) substrate; each Y is a covalent bond; each A is a glass (SiO) substrate; each Y is a covalent bond; each A 5 is a moiety of Formula (Af1); each X and X is indepen is independently selected from a covalent bond, —C(O)N dently selected from a covalent bond, methane-diyl ethane —SO. , —S , a moiety of Formula (Aa1), a moiety of 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. Formula (Af1), and amoiety of Formula (Af2); eachX' and hexane-1,6-diyl, heptane-1,7-diyl octane-1,8-diyl. nonane X is independently selected from a covalent bond, methane 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, diyl ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pen 10 tane-1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl, octane-1.8- 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben diyl. nonane-1,9-diyl. decane-1,10-diyl, undecane-1,11-diyl. Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and dodecane-1,12-diyl, hexadecane-1,16-diyl, benzene-diyl. (CH2)—(CH2—CH2—O) (CH), , wherein each methylbenzene-diyl, propylbenzene-diyl, methylcyclohex n1 and n3 is independently an integer selected from 0 to 3, and ane-diyl, and —(CH)—(CH2—CH2—O) (CH), , 15 each n2 is independently an integer selected from 1 to 20; wherein each n1 and n3 is independently an integer selected each X is selected from a covalent bond, -O-, -SO , from 0 to 3, and each n2 is independently an integer selected SON , —C(O)— —C(O)O—, and —C(O)N ; each from 1 to 20; each X* is selected from a covalent bond, R" and R is independently selected from hydrogen and O— —SO , —SON , —C(O)— —C(O)O—, and methyl; each D is selected from a therapeutic agent having at —C(O)N ; each R' and R is independently selected from least one primary amine group and a therapeutic agent having hydrogen and methyl; each D is selected from a therapeutic at least one secondary amine group; and né is selected from 1 agent having at least one primary amine group and a thera to 20. peutic agent having at least one secondary amine group; and In certain embodiments of a compound of Formula (III), Q nó is selected from 1 to 20. is a glass (SiO) substrate; each Y is a covalent bond; each A In certain embodiments of a compound of Formula (III), Q 25 is a moiety of Formula (Af1); each X* is pentane-1,5-diyl: is a glass (SiO) substrate; each Y is a covalent bond; each A each X* is a covalent bond; each X is a covalent bond; each is a moiety of Formula (Aa1); each X and X is indepen R" and R is hydrogen; each Dis independently selected from dently selected from a covalent bond, methane-diyl ethane a therapeutic agent having at least one primary amine group 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. and a therapeutic agent having at least one secondary amine hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl. nonane 30 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, group; and né is selected from 1 to 20. 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben In certain embodiments of a compound of Formula (III), Q Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and is a glass (SiO) substrate; each Y is a covalent bond; each A (CH2), —(CH2—CH2—O) (CH), , wherein each is a moiety of Formula (Af1); each X is 4-methylcyclohex n1 and n3 is independently an integer selected from 0 to 3, and 35 ane-diyl; each X is a covalent bond; each X is a covalent each n2 is independently an integer selected from 1 to 20; bond; each R' and R is hydrogen; each D is selected from a each X is selected from a covalent bond, -O-, -SO , therapeutic agent having at least one primary amine group SON , —C(O)— —C(O)O—, and —C(O)N ; each and a therapeutic agent having at least one secondary amine R" and R is independently selected from hydrogen and group; and né is selected from 1 to 20. methyl; each D is selected from a therapeutic agent having at 40 In certain embodiments of a compound of Formula (III), Q least one primary amine group and a therapeutic agent having is a glass (SiO.) substrate; each Y is a covalent bond; each A at least one secondary amine group; and né is selected from 1 is a moiety of Formula (Afl); each X* is pentane-1,5-diyl; to 20. each X* is C(O)N ; each X is pentane-1,5-diyl; each R' In certain embodiments of a compound of Formula (III), Q and R is hydrogen; each D is selected from a therapeutic is a glass (SiO) substrate; each Y is a covalent bond; each A 45 agent having at least one primary amine group and a thera is a moiety of Formula (Aa1), wherein each R and R is peutic agent having at least one secondary amine group; and hydrogen; each X* is pentane-1,5-diyl; each X* is a covalent nó is selected from 1 to 20. bond; each X is a covalent bond; each RandR is hydrogen; In certain embodiments of a compound of Formula (III), Q each D is independently selected from a therapeutic agent is a glass (SiO.) substrate; each Y is a covalent bond; each A having at least one primary amine group and a therapeutic 50 is a moiety of Formula (Af2); each X and X is indepen agent having at least one secondary amine group; and né is dently selected from a covalent bond, methane-diyl ethane selected from 1 to 20. 1.2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl. In certain embodiments of a compound of Formula (III), Q hexane-1,6-diyl, heptane-1,7-diyl octane-1,8-diyl. nonane is a glass (SiO) substrate; each Y is a covalent bond; each A 1.9-diyl, decane-1,10-diyl, undecane-1,11-diyl, dodecane-1, is a moiety of Formula (Aa1), wherein each R and R is 55 hydrogen; each X* is 4-methylcyclohexane-diyl; each X* is a 12-diyl, hexadecane-1,16-diyl, benzene-diyl, methylben covalent bond; each X is a covalent bond; each R' and R is Zene-diyl, propylbenzene-diyl, methylcyclohexane-diyl, and hydrogen; each D is selected from a therapeutic agent having (CH2)—(CH2—CH2—O) (CH), , wherein each at least one primary amine group and a therapeutic agent n1 and n3 is independently an integer selected from 0 to 3, and having at least one secondary amine group; and né is selected 60 each n2 is independently an integer selected from 1 to 20; from 1 to 20. each X is selected from a covalent bond, -O-, -SO , In certain embodiments of a compound of Formula (III), Q SON , —C(O)— —C(O)O—, and —C(O)N ; each is a glass (SiO) substrate; each Y is a covalent bond; each A R" and R is independently selected from hydrogen and is a moiety of Formula (Aa1), wherein each R and R is methyl; each D is selected from a therapeutic agent having at hydrogen; each X* is pentane-1,5-diyl; each X* is C(O) 65 least one primary amine group and a therapeutic agent having N—; each X is pentane-1,5-diyl; each RandR is hydrogen; at least one secondary amine group; and né is selected from 1 each Dis selected from a therapeutic agent having at least one to 20. US 9,295,731 B2 83 84 In certain embodiments of a compound of Formula (III), Q having at least one primary amine group and a therapeutic is a glass (SiO) substrate; each Y is a covalent bond; each A agent having at least one secondary amine group; and né is is a moiety of Formula (Af2); each X is methane-diyl; each selected from 1 to 20. X is a covalent bond; each X is a covalent bond; each R" and The compound of claim 48, wherein the compound is R’ is hydrogen; each D is independently selected from a selected from a compound of Formula (III-a). Formula (III b), Formula (III-c). Formula (III-d), Formula (III-e), Formula therapeutic agent having at least one primary amine group (III-f). Formula (III-g). Formula (III-h). Formula (III-i). For and a therapeutic agent having at least one secondary amine mula (III-). Formula (III-k). Formula (III-1), Formula (III-m), group; and né is selected from 1 to 20. Formula (III-n), Formula (III-o), Formula (III-p). Formula In certain embodiments of a compound of Formula (III), Q 10 (III-q). Formula (III-r). Formula (III-s), Formula (III-t), For is a glass (SiO.) substrate; each Y is a covalent bond; each A mula (III-u). Formula (III-III). Formula (III-w). Formula (III is a moiety of Formula (Af2); each X* is methane-diyl; each X). Formula (III-y), Formula (III-Z). Formula (III-aa). For X is —C(O)N ; each X is pentane-1,5-diyl; each R" and mula (III-ab), and Formula (III-ac), or a pharmaceutically R is hydrogen; each D is selected from a therapeutic agent acceptable salt of any of the foregoing:

(III-a)

Human Serum S Albumin

O

(III-b) Trastuzumab S O O

N 1N

I-10 US 9,295,731 B2 85 86 -continued (III-c) Trastuzumab S O

I-10

(III-d)

Immunoglobulin

1-0 US 9,295,731 B2 87 88 -continued (III-e) O OH

H OH N

O O O O Rituximab S O l 1No N N r O N21 N

HN lsN NH2 I-10

(III-f) OH O O Rituximab S O ul N o1 No N y N - O

6H I-10

(III-g)

I-10 US 9,295,731 B2 89 90 -continued (III-h) O

Trastuzumab S O O ulO -x o1 No N H N

O

O H H S H Null N - N

O 10 O On O 1-0 (III-i)

O

Trastuzumab S O O lO NullO -x O 1\o N N

N O N OH

O

O H s H N N Y N

O E O O O O 1. N 1-0

(III-) O Brentuximab S O

N

N N

1-0 US 9,295,731 B2 91 92 -continued (III-k) OH

(III-I)

Bevacizumab S O US 9,295,731 B2 93 94 -continued (III-m) HO O O O O l o1 No N1 O NH N O S \ a III / O HN O V

- O O HN / O HO (III-n)

O Trastuzumab S O

N

O

- I-10 (III-o)

Glembatumumab S O

O

I-10 US 9,295,731 B2 95 96 -continued (III-p)

No OH -O O in a

S O I OH O E OH

O S

I-10

(III-q)

N

O Milatuzumab

6H

I-10

(III-r)

Trastuzumab

I-10 US 9,295,731 B2 97 98 -continued (III-s)

O O O O Trastuzumab S O ls H 1No N '

N

O

I-10 (III-t)

O

N OH H

Trastuzumab S O N- Null O O

I-10

(III-u) HO O O Rituximab S O ls o1 No N H N OH

O I-10

(III-v) OH OH O O Alemtuzumab S O ul 21 o1 No N H OH O N HO O

O I-10 US 9,295,731 B2 99 100 -continued (III-w)

O Bivatuzumab S O O

N

O s'

I-10

(III-X) O O Lintuzumab S O l o1 No NH O H N N

O

I-10

(III-y) o1 1n-'N O

O O NN Bevacizumab S O ul 2 o1 No N N

N

O

Šs I-10

(III-Z) O

HN-( o O 3. O O H N Citatuzumab S O ls 1No N

N H O

O I-10 US 9,295,731 B2 101 102 -continued (III-aa) in-(O O

O O H Oportuzumab S O ul o1 No N

N H O

O I-10 (III-ab)

N NH . N O O NN O O N1 O O O O

N N O 1. O O O O N

HN O O NH N O O l N N1 21 Trastuzumab S O O O O O

I-10

(III-ac)

Human Serum Albumin I-10.

In certain embodiments of a compound of Formula (III), In certain embodiments of a compound of Formula (III), the compound has the structure of Formula (III-a) or a phar- the compound has the structure of Formula (III-d) or a phar maceutically acceptable salt thereof. maceutically acceptable salt thereof. In certain embodiments of a compound of Formula (III), In certain embodiments of a compound of Formula (III), the compound has the structure of Formula (III-b) or a phar- the compound has the structure of Formula (III-e) or a phar maceutically acceptable salt thereof. maceutically acceptable salt thereof. In certain embodiments of a compound of Formula (III), 65. In certain embodiments of a compound of Formula (III), the compound has the structure of Formula (III-c) or a phar- the compound has the structure of Formula (III-f) or a phar maceutically acceptable salt thereof. maceutically acceptable salt thereof. US 9,295,731 B2 103 104 In certain embodiments of a compound of Formula (III), pound, are labile under mild basic conditions. In certain the compound has the structure of Formula (III-g) or a phar embodiments, a conjugate of Formula (II) or Formula (III) maceutically acceptable salt thereof. can be cleaved from a primary or secondary amine-containing In certain embodiments of a compound of Formula (III), compound from about pH 7.4 to about pH 10, from about pH the compound has the structure of Formula (III-h) or a phar 7.4 to about pH 9.0, and in certain embodiments from about maceutically acceptable salt thereof. pH 7.4 to about pH 8.5. In certain embodiments of a compound of Formula (III), the compound has the structure of Formula (III-i) or a phar Method of Synthesis of Cleavable Drug Conjugates maceutically acceptable salt thereof. In certain embodiments of a compound of Formula (III), 10 the compound has the structure of Formula (III-) or a phar Cleavable crosslinkers, cleavable crosslinking intermedi maceutically acceptable salt thereof. ates, and cleavable conjugates provided by the present dis In certain embodiments of a compound of Formula (III), closure may be synthesized by methods known to those the compound has the structure of Formula (III-k) or a phar skilled in the art. For example, methods of synthesizing maceutically acceptable salt thereof. 15 related acyloxyalkyl carbamate compounds are described by In certain embodiments of a compound of Formula (III), Alexander, U.S. Pat. No. 4,760,057; Alexander, U.S. Pat. No. the compound has the structure of Formula (III-1) or a phar 4,916,230; Lund, U.S. Pat. No. 5,401,868: Alexander, U.S. maceutically acceptable salt thereof. Pat. No. 5,466,811: Alexander, U.S. Pat. No. 5,684,018; Gal In certain embodiments of a compound of Formula (III), lop, U.S. Pat. No. 7,227,028, and Singh, PCT Publication No. the compound has the structure of Formula (III-m) or a phar WO 2005097760. maceutically acceptable salt thereof. In certain embodiments of a compound of Formula (III), In certain embodiments, methods of synthesizing conju the compound has the structure of Formula (III-n) or a phar gates of Formula (II) or Formula (III) comprise (i) mixing a maceutically acceptable salt thereof. Solution containing a compound having at least one primary In certain embodiments of a compound of Formula (III), amine group and/or at least one secondary amine group the compound has the structure of Formula (III-o) or a phar 25 (D-NH) and a cleavable crosslinker of Formula (I) in dim maceutically acceptable salt thereof. ethylformamide (DMF), dimethyl sulfoxide (DMSO) or an In certain embodiments of a compound of Formula (III), aqueous buffer (pH 7.0-8.5) such as phosphate buffered saline the compound has the structure of Formula (III-p) or a phar (PBS) for at least 30 minutes at room temperature to obtain maceutically acceptable salt thereof. the corresponding conjugate of Formula (II); and (ii) mixing In certain embodiments of a compound of Formula (III), 30 a solution containing the conjugate of Formula (II) and a the compound has the structure of Formula (III-q) or a phar ligand having at least one thiol group (Q-SH) in dimethylfor maceutically acceptable salt thereof. mamide (DMF), dimethyl sulfoxide (DMSO) or an aqueous In certain embodiments of a compound of Formula (III), buffer (pH 7.0-8.5) such as phosphate buffered saline (PBS) the compound has the structure of Formula (III-r) or a phar for at least 30 minutes at room temperature to obtain the maceutically acceptable salt thereof. 35 corresponding conjugates of Formula (III) as shown in In certain embodiments of a compound of Formula (III), Scheme A. the compound has the structure of Formula (III-s) or a phar maceutically acceptable salt thereof. In certain embodiments of a compound of Formula (III), the compound has the structure of Formula (III-t) or a phar Scheme A maceutically acceptable salt thereof. 40 O O In certain embodiments of a compound of Formula (III), the compound has the structure of Formula (III-u) or a phar A X2 ls X ls -- maceutically acceptable salt thereof. Nx1 Nx3 O O Ll In certain embodiments of a compound of Formula (III), (I) the compound has the structure of Formula (III-V) or a phar maceutically acceptable salt thereof. 45 L' is a leaving group such as a halogen In certain embodiments of a compound of Formula (III), or N-hydroxysuccinimidyl the compound has the structure of Formula (III-w) or a phar D-NH).5 maceutically acceptable salt thereof. DMSO or DMF In certain embodiments of a compound of Formula (III), or PBS Buffer the compound has the structure of Formula (III-X) or a phar 50 O R1 R2 O maceutically acceptable salt thereof. A. Y X2 ls X -- In certain embodiments of a compound of Formula (III), Nx1 Nx3 O O D the compound has the structure of Formula (III-y) or a phar 5 maceutically acceptable salt thereof. (II) In certain embodiments of a compound of Formula (III), 55 Q-SH) the compound has the structure of Formula (III-Z) or a phar DMSO or DMF maceutically acceptable salt thereof. or PBS Buffer In certain embodiments of a compound of Formula (III), O R1 R2 O the compound has the structure of Formula (III-aa) or a phar maceutically acceptable salt thereof. Q A2 -X ls X l In certain embodiments of a compound of Formula (III), 60 1.Nx11N31 No O D the compound has the structure of Formula (III-ab) or a phar maceutically acceptable salt thereof. (III) In certain embodiments of a compound of Formula (III), the compound has the structure of Formula (III-ac) or a phar maceutically acceptable salt thereof. 65 In certain embodiments, methods of synthesizing cleav In certain embodiments, cleavable crosslinkers, when able drug conjugates of Formula (II) and Formula (III) are coupled to a primary or secondary amine-containing com shown in Schemes 1 to 6. US 9,295,731 B2 105 106

Scheme 1 O s1 O

C ---O C Base C ---O S

la 1b

O O O

O 1b O 1N ls 1. / OH --TEA / O O S -e-N-Hydroxysuccinimide N heat N Peracetic acid, AcOH

O O 1c 1d

O O O O ls doxorubicin 1No O1 N -e-DIEA W DMF N O

O 1e

O O 1N l

O

1

Scheme 2

1e -- mitoxantrone

DMSO or DMF or PBS Buffer US 9,295,731 B2 107 108 -continued O O

N O

O O-r O JO \ O O N O O N O

Scheme 3 1 + MPEG(40K Daltons)-SH - > ACN, DMSO or DMF/ Aqueous Buffer (pH 7.0 to 9.0)

O O O MPEG(40K Daltons)-S O ls 1No N

N O

O

Scheme 4

1 + > *rsE -e-ACN, DMSO or DMF/ O Aqueous Buffer Nshi (pH 7.0 to 9.0) US 9,295,731 B2 109 110 -continued

Scheme 5 1 + L-Glutathione (reduced) -- ACN, DMSO or DMF/ Aqueous Buffer (pH 7.0 to 9.0)

O Glutathione-S O l 1No N

N O

O

5

Scheme 6 1 + Human Serum Albumin-SH -- DMSO or DMF, Aqueous Buffer (pH 7.0 to 9.0) US 9,295,731 B2 111 112

-continued

Human Serum Albumin-S O ~1.

Example of therapeutic agents having a primary amine gatifloxacin, gemifloxacin, gentamicin, gentamicin, grepa group and/or a secondary amine group include aminopterin, floxacin, hamycin, hexetidine, hygromycin B, ibacitabine, folitiXorin, methotrexate, pemetrexed, pralatrexate, raltitr 25 iclaprim, isepamicin, kanamycin, lomefloxacin, lucimycin, exed, pelitrexol, talotrexin, deoxycoformycin, cladribine, lymecycline, mepartricin, moxifloxacin, natamycin, nemon clofarabine, fludarabine, thioguanine, mercaptopurine, beru oxacin, neomycin B, neomycin C, netilmicin, norfloxacin, bicin, daunorubicin, doxorubicin, epirubicin, idarubicin, nystatin, omadacycline, oritavancin, paromomycin, paZu amrubicin, pirarubicin, Zorubicin, mitoxantrone, banox floxacin, perimycin A, perimycin B, perimycin C, pipemidic antrone, ledoxantrone, nortopixantrone, pixantrone, piroX 30 acid, polymyxin B, puromycin, radeZolid, retaspimycin, antrone, Sabarubicin, topixantrone, monomethyl auristatin E, ribostamycin, rimocidin, sisomicin, sitafloxacin, sparfloxa monomethylauristatin F. monomethyldolastatin 10, dolasta cin, spectinomycin, Streptomycin, Sulfacetamide, Sulfadiaz tin 15, belotecan, atiratecan, camptothecin, exatecan, irinote ine, Sulfadimethoxine, Sulfadimidine, Sulfafurazole, Sul can, namitecan, rubitecan, topotecan, demecolcine, duocar 35 falene, SulfamaZone, Sulfamerazine, Sulfamethizole, mycin A, duocarmycin SA, duocarmycin B, duocarmycin B1, Sulfamethoxazole, Sulfamethoxypyridazine, Sulfametomi abbeymycin, anthramycin, centanamycin, chicamycin, dine, Sulfametoxydiazine, Sulfametrole, Sulfamoxole, Sulfa mazethramycin, porothramycin A, porothramycin B, Sibano nilamide, Sulfaperin, Sulfaphenazole, Sulfapyridine, Sul mycin, Sibiromycin, trabectedin, calicheamicin Y1. cali fathiazole, Sulfathiourea, Sulfisomidine, teicoplanin, cheamicin T, esperamicin A1, esperamicin C, esperamicin D, 40 telavancin, tanespimycin, temafloxacin, tetroXoprim, tigecy dynemicin A, dynemicin H, dynemicin M, dynemicin N. cline, tobramycin, to Sufloxacin, trimethoprim, trimethoprim, dynemicin O. dynemicin P. dynemicin Q, dynemicin S. neo trovafloxacin, tyrothricin, ulifloxacin, valnemulin, Vancomy carzinostatin chromophore, uncialamycin, 21-aminoe cin, Verdamicin, and Zabofloxacin. Other therapeutic agents pothilone B, eribulin, hemiasterlin, HTI-286, kahalatide F. having a primary amine group and/or a secondary amine elsamitrucin, lucanthone, melphalan, mitoguaZone, nimus 45 group are described in various compendia, such as, for tine, procarbazine, dacarbazine, amsacrine, 5-amino-4-oxo example, the Merck Index, 14th Edition, 2006; and the Phy pentanoic acid, methyl 5-amino-4-oxo-pentanoate, actino sicians Desk Reference, 64th Edition, 2009. mycin D, 7-aminoactinomycin D, bleomycin, mitomycin, An example of a therapeutic agent having at least one thiol staurosporine, desacetylvinblastine hydrazide, vinblastine, group is captopril. Vincristine, vindestine, Vinflunine, vinorelbine, afatinib, 50 Examples of therapeutic agents having at least one hydroxy apilimod, balamapimod, barasertib, boSutinib, canertinib, group include doxorubicin and camptothecin. cevipabulin, crizotinib, dacomitinib, dasatinib, denibulin, dil mapimod, dinaciclib, dovitinib, dutacatib, duVoglustat, edot Pharmaceutical Compositions Comprising ecarin, elisidepsin, entinostat, epetirimod, erlotinib, fingoli Conjugates mod, fostamatinib, gefitinib, golotimod, gusperimus, 55 imatinib, imiquimod, intedanib, ispinesib, lapatinib, lenali Certain embodiments provided by the present disclosure domide, linifanib, litronesib, loSmapimod, metesind, moceti relate to pharmaceutical compositions, formulations, routes nostat, motesanib, masitinib, myriocin, neratinib, nilotinib, of administration and doses of conjugates of Formula (II) or odanacatib, ombrabulin, pamapimod, panobinostat, paZo Formula (III). panib, plerixafor, pomalidomide, raZupenem, residuimod, 60 Conjugates of Formula (II) or Formula (III) may beformu Sabarubicin, Saracatinib, Seliciclib, Selumetinib, Sotirimod, lated with one or more suitable pharmaceutically acceptable squalamine, tacedinaline, tallabostat, taltobulin, tellatinib, tipi vehicles, diluents, and/or excipients, which are well known, farnib, toZasertib, vandetanib, Vatalanib, veliparib, Voreloxin, and can be determined, by one of skill in the art. alvespimycin, amikacin, amphotericin B, arbekacin, astromi In certain embodiments, when the route of administration cin, bacitracin, balofloxacin, bederocin, bekanamycin, besi 65 is parenteral (e.g. intravenous, intramuscular, intravenous, floxacin, brodimoprim, ciprofloxacin, clinafloxacin, colistin, Subcutaneous or interperitoneal), the vehicle or excipient or daptomycin, dibekacin, enoxacin, framycetin, garenoxacin, excipient mixture can be a solvent or a dispersive medium US 9,295,731 B2 113 114 containing, for example, various polar or nonpolar solvents, embodiments, from about 1 mg to about 1,000 mg of a con suitable mixtures thereof or oils. Examples of vehicles or jugate of Formula (II) or Formula (III). excipients include, but are not limited to, pharmaceutically acceptable solvents, dispersive agents or media, coatings, Therapeutic Use antimicrobial agents, iso- or hypo- or hypertonic agents, absorption modifying agents. Moreover, other or Supplemen A conjugate of Formula (II), a conjugate of Formula (III), tary active ingredients can also be incorporated into the com or a pharmaceutical composition of any of the foregoing, can position. be administered to a mammal. Such as a human, to treat a In certain embodiments, a pharmaceutical composition cancer, an autoimmune disease, or an infectious disease. 10 In certain embodiments, methods of treating a cancer in a includes carriers and excipients such as buffers, carbohy patient comprise administering to a patient in need of Such drates, mannitol, proteins, polypeptides or amino acids Such treatment a conjugate of Formula (II), a conjugate of Formula as glycine, antioxidants, bacteriostats, chelating agents, Sus (III), or a pharmaceutical composition of any of the forego pending agents, thickening agents and/or preservatives, ing, wherein D is therapeutically effective for treating the water, oils including but not limited to those of petroleum, 15 cancer. In certain embodiments, the cancer is selected from animal, vegetable or synthetic origin, Such as peanut oil, adrenal cortical cancer, anal cancer, aplastic anemia, bile duct Soybean oil, mineral oil, sesame oil and the like, Saline Solu cancer, bladder cancer, bone cancer, bone metastasis, a brain tions, aqueous dextrose and glycerol solutions, flavoring cancer, a central nervous system (CNS) cancer, a peripheral agents, coloring agents, detackifiers and other acceptable nervous system (PNS) cancer, breast cancer, cervical cancer, additives, adjuvants, or binders, other pharmaceutically childhood Non-Hodgkin’s lymphoma, colon and rectum can acceptable auxiliary Substances as required to approximate cer, endometrial cancer, esophagus cancer, Ewing's family of physiological conditions, such as pH buffering agents, tonic tumors, eye cancer, gallbladder cancer, a gastrointestinal car ity adjusting agents, emulsifying agents, wetting agents and cinoid tumor, a gastrointestinal stromal tumor, gestational the like. Examples of excipients include, but are not limited trophoblastic disease, hairy cell leukemia, Hodgkin’s lym to, starch, glucose, lactose. Sucrose, gelatin, malt, rice, flour, 25 phoma, Kaposi's sarcoma, kidney cancer, laryngeal and chalk, silica gel, Sodium Stearate, glycerol monostearate, talc, hypopharyngeal cancer, acute lymphocytic leukemia, acute Sodium chloride, dried skim milk, glycerol, propylene, gly myeloid leukemia, children's leukemia, chronic lymphocytic col, water, ethanol and the like. In certain embodiments, a leukemia, chronic myeloid leukemia, liver cancer, lung can pharmaceutical preparation is Substantially free of preserva cer, a lung carcinoid tumor, Non-Hodgkin’s lymphoma, male tives. In certain embodiments, a pharmaceutical preparation 30 breast cancer, malignant mesothelioma, multiple myeloma, contains at least one preservative. myelodysplastic syndrome, myeloproliferative disorders, nasal cavity and paranasal cancer, nasopharyngeal cancer, Administration neuroblastoma, oral cavity and oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile can In certain embodiments, conjugates of Formula (II) and 35 cer, pituitary tumor, prostate cancer, retinoblastoma, rhab Formula (III) may be administered or applied singly, in com domyosarcoma, salivary gland cancer, a sarcoma, melanoma bination with one or more pharmaceutically active agents, skin cancer, non-melanoma skin cancers, stomach cancer, including another conjugate provided by the present disclo testicular cancer, thymus cancer, thyroid cancer, uterine can SUC. cer, transitional cell carcinoma, vaginal cancer, Vulvar cancer, Conjugates of Formula (II) and Formula (III) and compo 40 mesothelioma, squamous cell or epidermoid carcinoma, sitions thereofmay be administered by any appropriate route. bronchialadenoma, choriocarcinoma, head and neck cancers, In certain embodiments, conjugates of Formula (II) and For teratocarcinoma, and Waldenstrom's macroglobulinemia. In mula (III) and compositions thereof may be administered certain embodiments, the cancer is selected from acute lym parenterally, for example, by infusion or bolus injection. phocytic leukemia, acute myeloid leukemia, chronic lympho 45 cytic leukemia, chronic myeloid leukemia, multiple Dosage myeloma, breast cancer, prostate cancer, lung cancer, kidney cancer, liver cancer, and colon cancer. Pharmaceutical compositions provided by the present dis In certain embodiments, methods of treating an autoim closure comprise a conjugate of Formula (II) or Formula (III) mune disease in a patient comprise administering to a patient and a pharmaceutically acceptable vehicle. In certain 50 in need of Such treatment a conjugate of Formula (II), a embodiments, a compound of Formula (II) or Formula (III) is conjugate of Formula (III), or a pharmaceutical composition present in a therapeutically effective amount, i.e., in an of any of the foregoing, wherein D is therapeutically effective amount effective to achieve therapeutic benefit in a patient for treating the autoimmune disease. In certain embodiments, having a cancer, an autoimmune disease, or an infectious the autoimmune disease is selected from alopecia areata, disease. The amount effective for a particular treatment will 55 ankylosing spondylitis, Chagas disease, chronic obstructive depend, at least in part, on the disease or diseases being pulmonary disease, Crohns Disease, dermatomyositis, diabe treated, the condition of the patient, the severity of the dis tes mellitus type 1, endometriosis, Goodpasture’s syndrome, ease, the formulation, and the route of administration, as well Graves disease, Guillain-Barré syndrome, Hashimoto's dis as other factors known to those of skill in the art. In vitro or in ease, Hidradenitis Suppurativa, Kawasaki disease, IgA neph Vivo assays can optionally be employed to help identify opti 60 ropathy, idiopathic thrombocytopenic purpura, interstitial mal doses and dosing regimens. cystitis, lupus erythematosus, lupus nephritis, mixed connec In certain embodiments, a therapeutically effective amount tive tissue disease, morphea, multiple Sclerosis, Myasthenia of a conjugate of Formula (II) or Formula (III) sufficient to gravis, carcolepsy, meuromyotonia, Pemphigus Vulgaris, treat a cancer, an autoimmune disease, or an infectious dis Pernicious anaemia, psoriasis, psoriatic arthritis, polymyosi ease in a patient is from about 1 mg to about 10,000 mg of a 65 tis, primary biliary cirrhosis, relapsing polychondritis, rheu conjugate of Formula (II) or Formula (III); in certain embodi matoid arthritis, sarcoidosis, Schizophrenia, Scleroderma, ments, from about 1 mg to about 5,000 mg; and in certain Sjögren's syndrome, temporal arteritis, ulcerative colitis, US 9,295,731 B2 115 116 vasculitis, and Wegener's granulomatosis. In certain embodi pH 7.4 to about pH 10, from about pH 7.4 to about pH 9.0, and ments, the autoimmune disease is selected from lupus erythe in certain embodiments from about pH 7.4 to about pH 8.5. matosus, psoriasis and multiple Sclerosis. In certain embodi EXAMPLES ments, the autoimmune disease is multiple Sclerosis and the multiple Sclerosis is selected from relapsing remitting mul The following examples describe in detail methods of syn tiple Sclerosis, secondary progressive multiple Sclerosis, pri thesizing crosslinkers, intermediates, and conjugates pro mary progressive multiple Sclerosis, and progressive relaps vided by the present disclosure, assays for characterizing ing multiple Sclerosis. crosslinkers and conjugates provided by the present disclo Sure, and methods of using crosslinkers and conjugates pro In certain embodiments, methods of treating an infectious 10 vided by the present disclosure. It will be apparent to those disease in a patient comprise administering to a patient in skilled in the art that many modifications, both to materials need of Such treatment a conjugate of Formula (II), a conju and methods, may be practiced without departing from the gate of Formula (III), or a pharmaceutical composition of any Scope of the disclosure. of the foregoing, wherein D is therapeutically effective for treating the infectious disease. In certain embodiments, the 15 infectious disease is selected from a fungal disease, a bacte Example 1 rial disease, and a viral disease. In certain embodiments, the infectious disease is a viral disease and the viral disease is Synthesis of Doxorubicin Conjugate (1)

O O

O 1No l N f N O

O selected from Hepatitis A, Hepatitis B. Hepatitis C, and 40 A mixture of 1-chloromethyl chloroformate (1.0 eq) and autoimmune deficiency disorder (AIDS). tetrabutylammonium bisulfate (0.20 eq) in dichloromethane In certain embodiments, a conjugate of Formula (II) or was cooled to 0°C. with an ice-water bath. A 21% solution of Formula (III) or a pharmaceutical composition of any of the sodium methanethiolate (1.0 eq) in water was added. The foregoing may be administered to a patient together with at 45 bilayer was stirred at 0°C. for 1 h and then at room tempera least one second therapeutic agent. In certain embodiments, ture for 16 h. The reaction mixture was diluted with dichlo the at least one second therapeutic agent is selected from an romethane, washed with water, Saturated bicarbonate solu antitumor alkylating agent, an antitumor antimetabolite, an tion and brine, then dried over NaSO, filtered and antitumor antibiotic, a plant-derived antitumor agent, an anti concentrated in vacuo to provide compound (1b) as a color tumor organoplatinum compound, an antitumor campthote 50 less liquid. cinderivative, an antitumor tyrosine kinase inhibitor, a mono A mixture of 4-(2,5-dioxopyrrol-1-yl)methylcyclohex clonal antibody, an interferon, a biological response modifier, anecarboxylic acid (1c), compound (1b) (1.0 eq), and triethy a hormonal anti-tumor agent, an angiogenesis inhibitor, a lamine (1.0 eq) was stirred at 70° C. for 24 h. The reaction differentiating agent, and a pharmaceutically acceptable salt mixture was diluted with ethyl acetate, and washed with of any of the foregoing. In certain embodiments, the at least 55 water, Saturated bicarbonate solution and brine. The organic one second therapeutic agent is selected from paclitaxel, doc phase was separated, dried over NaSO filtered and concen trated in vacuo to yield the crude product. The crude material etaxel, cyclophosphamide, ribavirin, and bortezomib. was then purified by silica gel column chromatography using A second therapeutic agent can be administered prior to, ethyl acetate and hexanes to yield compound (1 d). concomitant with, or Subsequent to administering a conjugate 60 To a solution of compound (1 d) (1.0 eq) in dichlo of Formula (II) or Formula (III) or pharmaceutical composi romethane (10 mL) was added N-hydroxysuccinimide (NHS) tion of any of the foregoing. (3.0 eq) and the reaction mixture cooled to 0°C. A solution of The conjugate compound of Formula (II) or Formula (III) 32% (v/v) peracetic acid in acetic acid (3.0 eq) was added can be cleaved under mild basic conditions and/or by dropwise over a period of 10 mins, and the solution was esterases in Vivo to release the pharmaceutical agent. In cer 65 stirred at 0°C. for 3 hand then at room temperature overnight. tain embodiments, the conjugate Formula (II) or Formula The reaction mixture was then diluted with dichloromethane (III) can release the pharmaceutical agent in vivo from about and washed with water, Saturated Sodium bicarbonate solu US 9,295,731 B2 117 118 tion and brine, then dried over NaSO filtered and concen Performance Liquid Chromatography (HPLC) using aceto trated in vacuo to give the crude product. The crude material nitrile/aqueous buffer as eluent to provide compound (1). was then purified by silica gel column chromatography using ethyl acetate and hexanes to yield compound (1e). A mixture of compound (1e) (25.25 mg) and doxorubicin HCl (1.0 eq) and TEA (1.0 eq) in DMF (0.40 mL) was stirred Example 2 at room temperature for 1 h and then concentrated under vacuum. The crude was purified by reverse-phase C-18 High Synthesis of Mitoxantrone Conjugate (2)

O O N1 r l ico,

O O

A mixture of compound (1e) (4.0 eq), mitoxantrone (1.0 35 eq), and TEA (5.0 eq) in acetonitrile/aqueous buffer is stirred at room temperature for 3 h and then concentrated under vacuum. The crude product is purified by reverse-phase C-18 High Performance Liquid Chromatography (HPLC) using acetonitrile/aqueous buffer as eluent to provide compound 40 (2). Example 3 Synthesis of MPEG-Maleimido-Doxorubicin Conjugate (3)

O

O O O MPEG(40K Daltons)-S O ul 1No N

N O

O US 9,295,731 B2 119 120 A mixture of compound (1) and MPEG-SH (average MW 40K Daltons) in DMSO/aqueous buffer is stirred at room temperature for 1 hand then concentrated under vacuum. The crude product is purified using size-exclusion chromatogra phy to provide compound (3). 5 Example 4 Synthesis of Boc-Cysteine-Maleimido-Doxorubicin Conjugate (4)

O O

S N O

> O ul N O 6

O

35 A mixture of compound (1) (5.81 mg) and Boc-cysteine (1.23 mg) inacetonitrile/aqueous potassium phosphate buffer (1.6 mL, pH 7.4) was stirred at room temperature for 1 h to provide compound (4). Mass found: M+Na: 1080. Example 5 40 Synthesis of L-Glutathione-Maleimido-Doxorubicin Conjugate (5)

O O Glutathione-S O l o1 No N

N O

O US 9,295,731 B2 121 122 A mixture of compound (1) (8.37 mg) and L-glutathione Liver Homogenate: Compounds (1-500 uM) are incubated (reduced) (9.94 mg) in acetonitrile/aqueous potassium phos with rat or human liver S9 at 0.5 mg protein/mL in the pres phate buffer (2 mL, pH 7.4) was stirred at room temperature ence of 1 mMNADPH at pH 7.4 and at 37°C. for 1 hour to 10 for 3 h to provide compound (5). Mass found: M+Na: 1167. days. Samples are obtained at Zero. 1 hour, and up to 10 days post-addition and are immediately quenched with methanol Example 6 or acetonitrile to prevent further conversion. Quenched samples are immediately analyzed or are frozen and main Synthesis of Human Serum tained at -80° C. prior to analysis. Samples are analyzed by Albumin-Maleimido-Doxorubicin Conjugate (6) LC, LC/MS or LC/MS/MS.

O O O O

O O O Human Serum Albumin S O ul o1 No N

N O

O

A mixture of compound (1) and human serum albumin in Caco-2Cell Homogenate: Caco-2 cells are grown in flasks DMSO/aqueous potassium phosphate buffer was stirred at over 21 days. Cells are then rinsed/scraped into ice-cold 10 room temperature for 3 h to provide compound (6). mM sodium phosphate/0.15 M potassium chloride, pH 7.4. 35 Cells are lysed by Sonication at 4°C. using a probe Sonicator Example 7 and centrifuged at 9,000xg for 20 min at 4°C. Aliquots of the Chemical Stability resulting Supernatant (Caco-2 cell homogenate S9 fraction) are transferred into 0.5 mL vials and stored at -80°C. prior to For chemical stability studies, buffers are prepared at pH 40 analysis. For stability studies, compounds (5 LM) are incu 2.0, pH 7.4 and pH 8.0. Compounds (1-500 uM) are incubated bated with Caco-2 S9 (0.5 mg protein/mL) at pH 7.4 and 37° with buffers at 37° C. for 1 h in a temperature-controlled C. for 1 hour to 10 days. Samples are obtained at Zero, 1 hour, HPLC autosampler. Samples for analysis are extracted at Zero and up to 10 days post-addition and are immediately time and 1 h post-addition. Samples are analyzed by HPLC, quenched with methanol or acetonitrile to prevent further liquid chromatography-mass spectrometry (LC/MS), or by 45 conversion. Quenched samples are immediately analyzed or liquid chromatography-tandem mass spectrometry (LC/MS/ are frozen and maintained at -80° C. prior to analysis. MS). Samples are analyzed by LC, LC/MS or LC/MS/MS. Pancreatin: Compounds (5uM) are incubated with porcine Example 8 pancreatin (10 mg/mL in pH 7.5 buffer) at 37°C. for 24 hour. 50 Samples are obtained at Zero, 1 hour, and up to 10 days Metabolic Stability/Drug Release post-addition and are immediately quenched with methanol or acetonitrile to prevent further conversion. Quenched Plasma Stability: Compounds (1-500 uM) are incubated samples are immediately analyzed or are frozen and main with 10-100% mouse, rat or human serum or plasma at 37°C. tained at -80° C. prior to analysis. Samples are analyzed by for 1 hour to 10 days. Samples are obtained at Zero, 1 hour, 55 LC, LC/MS or LC/MS/MS. and up to 10 days post-addition and are immediately quenched with methanol or acetonitrile to prevent further Example 9 conversion. Quenched samples are immediately analyzed or are frozen and maintained at -80° C. prior to analysis. In Vitro Cytotoxicity Samples are analyzed by LC, LC/MS or LC/MS/MS. 60 Compound 4 (5 uL of a 10 mM DMSO solution) was The effect of compounds provided by the present disclo incubated with a 50% rat serum (Aleken Biologics) in potas Sure on cancer cell growth and viability is measured in a panel sium phosphate buffer (pH 7.4) solution (195uL). After 1 of human cancer cells grown in vitro. All cancer cell lines are hour, the sample was quenched with acetonitrile (600 uL), obtained, for example, from the American Tissue Type Col centrifuged at 20,000xg, and the supernatant analyzed by LC 65 lection (ATTC). Cells are maintained in the following media: at 220 nm. The ratio of doxorubicin released to compound 4 TUR, H69AR, Calu-6, ME180, and PC3 cells are grown in was 10:90. RPMI medium containing 4.5 g/L glucose, 10% FBS, 4 mM US 9,295,731 B2 123 124 L-glutamine, and 1 mM sodium pyruvate); and HT29 cells are What is claimed is: grown in DMEM with 4.5 g/L glucose, 10% FBS, and 6 1. A compound of Formula (II): mM/L-glutamine. All cells are grown at 37°C. in 5% CO and passaged bi-weekly. For cytoxicity studies, cells are seeded into 96-well clear (II) bottom microtiter plates at the following densities: TUR O R1 R2 O (7,500 cells/well), HT29 (5,000 cells/well), H69 AR (10,000 2 cells/well), Calu-6 (15,000 cells/well), and ME180 (2,500 AS-XSX X ls O X O cells/well). Cells are maintained at 37°C. with 5% CO, for 24 5 hours prior to addition of a test compound. Each condition is 10 measured in triplicate. For compounds dissolved in DMSO, or a salt thereof, wherein: the final DMSO concentration in each well and in the non each A' is independently a moiety selected from Formula drug treated control cells is constant and below 1% in all (A'al), Formula (Aa2). Formula (Aa3 ), formula studies. The number of viable cells is determined after 24, 48, (Ab1), Formula (A 2), Formula (A'c1), Formula and 72 hours using Alamar blue fluorescence. Briefly, Alamar 15 (Ac2 ), Formula (A'c3). Formula (Ad1), Formula blue (10 mM) is added to each well for 4 hours, and the (A'd2). Formula (A'd3), Formula (A"e1). Formula A' fl number of viable cells estimated by measuring the fluores and Formula (A'f2): cence of reduced Alamar blue (530 nm excitation, 590 nm emission). To correct for background fluorescence, cells are treated with 100% DMSO to eliminate all viable cells. The (Aa1) effect of a test compound on cell growth is determined by R8 O calculating the fraction of live cells treated with the test com pound compared to untreated cells: (F1 drug treated Fl DMSO killed)/(F1 untreated F1 DMSO killed). The half maximal growth inhibition value (GI50) is calculated using 25 R9 y commercial curve-fitting Software (e.g., Prism). O Example 10 (Aa2)

Efficacy in Xenograph Model 30 Tumor xenograft studies are performed using nude athy R8 mic CD-1 mice (for example, from Charles River Laborato (Aa3) ries). Human cancer cells (5x10 PC3, 5x10 HT29, 107 Calu-6, and 5x10 ME 180) are implanted in the hind flank of 35 nude mice. Tumor growth is measured using calipers and tumor Volume calculated using the formula: (width (Ab1) 2x(length/2)). Tumor Volume is measured 3 days per week. O Animal weight is measured 3 days per week to assess com pound toxicity. 40 L2 Compounds are tested for in vivo tumor growth inhibition as follows. Animals bearing tumors (100-200 mg) are sorted into groups (5-8 animals) with similar average tumor mass (Ab2) per group. Drug doses are administered by intraperitoneal (IP) injection (0.5 mL). Test compounds are dissolved in 45 phosphate buffered saline (PBS) with the concentration adjusted for animal weight. Actual drug concentrations in formulated doses are verified by quantitative nitrogen detec (A'c1) tion. Animals are dosed every 7 days for 21 days in most studies. In several studies, animals are dosed twice weekly 50 (days 0, 4, 7, 10, 14, etc.). Tumor Volume and animal weight are measured on days 0, 2, 4, 7, 9, 11, 14, 16, 18, 21, 23, 25, and 28. At the end of the study, animal blood is removed for analysis of liver and kidney function (BUN, creatine, AST and ALT: analytical measurements performed by Quality 55 Clinical Labs). Tumor volume at the end of the study is calculated as a percentage of the tumor Volume on day 0. The (Ac2) percent tumor growth inhibition is obtained from the ratio of the percentage tumor Volume increase in drug treated animals to the percentage tumor Volume increase in Saline treated 60 animals. Finally, it should be noted that there are alternative ways of implementing the disclosures contained herein. Accordingly, the present embodiments are to be considered as illustrative and not restrictive, and the claims are not to be limited to the 65 details given herein, but may be modified within the scope and equivalents thereof. US 9,295,731 B2 125 126 -continued tuted C-2 arenediyl. Coheteroarenediyl. Substituted Co heteroarenediyl, C, alkanearenediyl. Substi tuted C-2 alkanearenediyl. Co. heteroalkanearene diyl. Substituted Co. heteroalkanearenediyl, and y v. (A'c3) —(CH2)—(CH-CH-O) (CH), , wherein: each n1 and n3 is independently an integer selected from 0 to 5; and each n2 is independently an integer selected from 1 to " 25; O (Ad1) 10 each X* is independently selected from a covalent bond,

O O O 15 each R" and R is independently selected from hydrogen, N 7 C. alkyl, Substituted C. alkyl, C. heteroalkyl, Sub stituted C. heteroalkyl, Co aryl, Substituted Co (Ad2) aryl, Coheteroaryl, and Substituted Coheteroaryl; D is selected from aminopterin, folitix.orin. methotrexate, pemetrexed, ralatrexate, raltitrexed, pelitrexol, talotr exin. deoxycoformycin, cladribine, clofarabine, flu N.--N darabine, thioguanine, mercaptopurine, berubicin (Ad3) daunorubicin, doxorubicin, epirubicin, idarubicin, 25 amrubicin, pirarubicin, Zorubicin mitoxantrone, banox antrone, ledoxantrone, nortoixantrone, pixantrone, piroXantrone, Sabarubicin, topixantrone, monomethyl, auristatin E. monomethyl, auristatin F, monomethyl, dolastatin, 10, dolastatin 15, belotecan, atiratecan, 30 camptopthecin, exatecan, irinotecan, namitecan, rubite (Ale1) can, topotecan, demecolcine, duocarmycin A, duocar mycin SA, duocarmycin B, duocarmycin Blabbeymy cin, anthramycin, centanamycin, chicamycin, X mazethramycin, porothrarmycin A, porothramycin B, (Alf1) 35 Sibanomycin, Sibiromycm, trabectedin, calicheamicin Y1. calicheamicin T, esperamicin Al, esperamicin C, H o , and esperamicin D, dynemicin A, dynemicin H, dynemicin M, dynemicin N, dynemicin O. dynemicin p, dynemicin (Alf2) Q, dynemicin S, neocarzinostatin chromophore, 40 uncialamycin, 21-aminoepothilone B, eribulin, hemi asterlin, HTI-286, kahalatide F, elsamitrucin, lucan p-- s thone, melphalan, mitoguaZone, nimustine, procarba Zine, dacarbazine, amsacrine 5-amino-4-oxo-pentanoic acid. methyl 5-amino-4-oxo-pentanoate, actimomycin 45 D. 7-aminoactinomycin D, bleomycin, mitomycin, Stau rosporine, desacetylvinblastine hydrazide, vinblastine, Vincristine, vindesine, Vinflunine, vinorelbine, afatinib, apilimbd, balamanimod, barasertib. bosutinib, ceaner wherein: tinib, cevipabulin, crizotinib, dacomitinib, dasatinib, each R and R is independently selected from hydro 50 denibulin. dilimapimod, dinaciclib, dovitiriib, dutacatib, gen, methyl, ethyl, n-propyl, isopropyl, phenyl, and duVoglustat, edotecarin, elisidepsin, entinostat, epetiri benzyl: mod, erlotinib, fingolimod, fostamatinib, gefitinib, golo each E is selected from F, Cl, Br, and I; and timod, gusperimus, imatinib, imiquimod, intedanib, each L is selected from halogen, N-hydroxysuccinim ispinesib, lapatinib, lenalidomide, linifanib, litronesib, idyl, substituted N-hydroxvsuccinimidyl, phenol-yl, 55 loSmapimod, metesind, mocetinostat, motesanib, masi substituted phenol-yl, hydroxybenzotriazolyl, substi tinib, myriocin, neratinib, nilotinib, odanacatib, tuted hydroxybenzotriazolyl, imidazolyl, and substi ombrabulin, pamapimod, panobinostat, paZopanib, tuted imidazolyl: plerixafor, pomalidomide, raZupenem, residuimod, each X and X is independently selected from a covalent Sabarubicin, Saracatinib, Seliciclib, Selumetinib, Sotiri bond, Co alkanediyl. Substituted Co alkanediyl. 60 mod, squalamine, tacedinaline, tallabostat, taltobulin, Co heteroalkanediyl. Substituted Co heteroal telatinib, tipifarnib, toZasertib, Vandetanib, Vatalanib, kanediyl, C-2 cycloalkanediyl. Substituted C. Veliparib, Voreloxin, alvespimycin, amikacin, amphot cycloalkanediyl. C. heterocycloalkanediyl. Substi ericin B, arbekacin, astromicin, bacitracin, balofloxacin, tuted C" '' heterocycloalkanediyl, Cao alkanecy bederocin, bekanamycin, besifloxacin, brodimoprim, cloalkanediyl. Substituted Cao alkanecycloalkanediyl. 65 ciprofloxacin, clinafloxacin, colistin, daptomycin, Cao heteroalkanecycloalkanediyl. Substituted Cao dibekacin, enoxacin, framycetin, garenoxacin, gati heteroalkanecycloalkanediyl, Co arenediyl. Substi floxacin, gemifloxacin, gentamicin, gentamicin, grepa US 9,295,731 B2 127 128 floxacin, hamycin, hexetidine, hygromycin B, ibacitab -continued ine, iclaprim, isepamicin, kanamycin, lomefloxacin, lucimycin, lymecycline, mepartricin, moxifloxacin, (Ab1b) natamycin, nemonoxacin, neomycin B, neomycin C, netilmicin, norfloxacin, nystatin, omadacycline, orita vancin, paromomycin, paZufloxacin, perimycin A, peri mycin B, perimycin C, pipemidic acid, polymyxin B, puromycin, radeZolid, retaspimycin, ribostamycin, rimocidin, Sisomicin, sitafloxacin, sparfloxacin, specti (Ab1c) nomycin, Streptomycin, Sulfacetamide, Sulfadiazine, 10 sulfadimethoxine, sulfadimidine, sulfafurazole, sul falene, SulfamaZone, Sulfamerazine, Sulfamethizole, Sulfamethoxazole, Sulfamethoxypyridazine, Sulfameto midine, Sulfametoxydiazine, Sulfametrole, Sulfamoxole, Sulfanilamide, Sulfaperin, Sulfaphenazole, Sulfapyri 15 dine, Sulfathiazole, Sulfathiourea, Sulfisomidine, teico planin, telavancin, tanespimycin, temafloxacin, tetroX (Ab2a) oprim, tigecycline, tobramycin, to Sufloxacin, O trimethoprim, trimethoprim, trovafloxacin, tyrothricin, ulifloxacin, valnemulin, Vancomycin, Verdamicin, and Zabofloxacin; and O n5 is an interger selected from 1 to 20. (Ab2b) 2. The compound of claim 1, wherein each A' is indepen 25 dently a moiety selected from Formula (A'ala), Formula (A' O a2a). Formula (Aa2b), Formula (Aa3a). Formula (Aa3b). Formula (Ab 1a). Formula (Ab1b). Formula (Ab1c), For mula (Ab2a). Formula (Ab2b), Formula (A''2c), Formula cy (A'c I). Formula (Ac2). Formula (A'c3). Formula (Ad1), 30 Formula (A'd2), Formula (A'd3), Formula (A'c1), Formula (Ab2c) (A'fl), and Formula (Af2): O

(A'ala) 35 O

/ y 40 O (Aa2a) N-X s 45 (Aa2b) (Ac2) 50

(Aa3a) cry 55 (Aa3ba)

ry s 60 (Ab1a) O HN NH

65 US 9,295,731 B2 129 130 -continued (II-A-2). Formula (II-A-3). Formula (II-A-4), Formula (II-A- (Ad1) 5), Formula (II-A-6). Formula (II-A-7), Formula (II-A-8), Formula (II-A-9), and Formula (II-A-10):

O O O (II-A-1) O S / W O O (Ad2) 10 Nulls 1No ls N1 D,

O (II-A-2) O X 15 (Ad3) N-N-N's D, O O O (Ale1) (II-A-3) O W O O X. 25 (Alf1) N--- o1 No ul N1 D, i-- and O 30 (II-A-4) (Af2) O O O 1No ul N1 D 35 ( \,

O 5 (II-A-5) 40 3. The compound of claim 1. wherein each X" and X is O independently selected from a covalent bond, Coal / O O kanediyl, C-20 cycloalkanediyl. Co-o arenediyl, C7-20 alka nearenediyl, and (CH2)—(CH2—CH2—O). N 1No ul-DN1 (CH), , wherein each n1 and n3 is independently an inte 45 ger selected from 0 to 5 and each n2 is independently an O integer selected from 1 to 25. 4. The compound of claim 1, wherein each X" and X is (II-A-6) independently selected from a covalent bond, methane-diyl. O ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane 50 1,5-diyl, hexane-1,6-diyl, benzene-diyl, methylbenzene-diyl. ethylbenzene-diyl, propylbenzcine-diyl, methylcyclohexane ( diyl. (CH2) -(CH2—CH2—O)—, (CH.) (CH CH-O) (CH)—, and -(CH2) (CH-CH O), O O N (CH2) . 55 O N1 r ND. 5. The compound of claim 1, wherein each X* is selected from a covalent bond, —O— —S— —SO— —SO , O O (II-A-7) O 6. The compound of claim 1, wherein each. X is selected 60 from a covalent bond, —O — —SONH , —SS , C(O) = C(O)O , and –C(O)NH 7. The compound of claim 1, wherein each R" and R is independently selected from hydrogen, methyl, ethyl, propyl. ( S. N-s-s isopropyl, and phenyl. 65 O O O 8. The compound of claim 1, wherein the compound is selected from the compound of Formula (II-A-1), Formula US 9,295,731 B2 131 132 -continued -continued (II-A-8) (II-B-5) O O ( \, o1 No ulN1

O Ouluo1 No ul N1 10 (II-A-9) O (II-B-6) / O O O N ls 15 o1 No N1 o1 No lsN1 O

and

(II-A-10) O (II-B-7) 25 O O t 1No ul N1 D, O O--- 30 " O (II-B-8) 9. The compound of claim 1, wherein the compound is O O O N N1 ND. selected from the compound of Formula (II-B-1), Formula 35 (II-B-2), Formula (II-B-3), Formula (II-B-4), Formula (II-B- O 5), Formula (II-B-6). Formula (II-B-7), Formula (II-B-8), O Formula (II-B-9). Formula (II-B-10). Formula (II-B-1 1), Formula (II-13-1 and Formula (II-B-13): O O O 40 N / (II-B-1) O O (II-B-9) O O ls 45 N- N1 Sar-s-s ulN1 NEN (II-B-2) O O (II-B-10) O O 50 claus 1No ul N1 D, ul-D (II-B-3) O O O Cr'so1 No N1 N 55 O ul-D (II-B-11) N1 ~~ N1 O O O O N--- l D, (II-B-4) 60 1No N1 O O O (II-B-12) (t O r O n1 O ~~ ul N1 D, O O O O 65 O ------" " US 9,295,731 B2 133 134 -continued 10. The compound of claim 1, wherein the compound is (II-B-13) selected from Formula (II-a). Formula (II-b), Formula (II-c), O Formula (II-d), Formula (II-e), Formula (II-f), Formula (II-g), )- s Formula (II-h). Formula (II-i). Formula (II-j), Formula (II-k), O \-o Formula (II-l). Formula (II-m). Formula (II-n), Formula (II X- o). Formula (II-p). Formula (II-q). Formula (II-r), Formula O O O Y. (II-s), Formula (II-t), Formula (II-u). Formula (II-V). Formula o (II-w). Formula (II-X). Formula (II-y), Formula (II-Z). For mula (II-aa). Formula (II-ab), and Formula (II-ac), or a salt of any of the foregoing:

(II-a)

(II-b)

(II-c) US 9,295,731 B2 135 136 -continued

(II-d) OH O r O \ Cluu COO W --- ? r f O N

(II-e)

OH US 9,295,731 B2 137 138 -continued (II-f) OH

OH (II-g) cryO

O H sus N S NH 19 O ON O (II-h)

O H N N

19 O O-N O (II-i) O O O ry o1 No lulu-NullsN N H i H O NoH US 9,295,731 B2 139 140 -continued (II-)

(II-k)

(II-I) US 9,295,731 B2 141 142 -continued (II-m) O O

N

(II-n)

O O

N

(II-o) US 9,295,731 B2 143 144 -continued

(II-q)

(II-r)

US 9,295,731 B2 149 150 -continued

(II-aa)

(II-ab)

O O N1

O O N , and O O O N

(II-ac)

k k k k k