Compounds targeting fibroblast-activation protein and methods of use thereof

Abstract

The present disclosure provides, in certain embodiments, compounds chelated or covalently bound to therapeutic or diagnostic radionuclides, such as a chelated or covalently bound complexes of Formula (I) as described herein and their methods of use in the treatment of a disease (e.g., a disease described herein) or imaging.

Description

Attorney Docket No. 00020.026.1801COMPOUNDS TARGETING FIBROBLAST-ACTIVATION PROTEIN AND METHODS OF USE THEREOF CROSS REFERENCE

[0001] This application claims the benefit of and priority to U. S. Provisional Application No.63 / 746,755, filed on January 17. 2025, the contents of which are incorporated herein by reference in their entirety.SUMMARY

[0002] The present disclosure provides, in certain embodiments, compounds such as complexes that are chelated or covalently bound to therapeutic or diagnostic radionuclides (e.g., a chelated complex represented by Formula (I)) and their methods of use in the treatment of a disease (e.g., a disease described herein) and / or imaging. Also described herein, in certain embodiments, are non-chelated compounds of the complexes described herein (e.g., a compound represented by Formula (I)) and compounds represented by Formula (II).

[0003] In certain embodiments, provided herein are complexes represented by Formula (I):D-L-RAIL-EFormula (I)or a pharmaceutically acceptable salt thereof, wherein:D is a ligand that targets fibroblast activation protein (FAP);L is a linker;LEis a peptide linker comprising at least one proline;E is a secondary effector moiety; andRAis a ligand chelated or covalently linked to a therapeutic radionuclide or a diagnostic radionuclide.

[0004] In certain embodiments, provided herein are complexes chelated or covalently linked to a radionuclide, comprising a linker represented by:1#583961Attorney Docket No. 00020.026.1801wherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl, 6-membered heteroaryl, and C3-6 cycloalkyl;n is an integer from 1 to 6,q is an integer from 1 to 20;p is 0 or 1; andLEis a peptide linker comprising at least one proline.

[0005] In certain embodiments, provided herein are compounds represented by Formula (II):D-L-RLEFormula (II)or a pharmaceutically acceptable salt thereof, wherein:D is a ligand that targets fibroblast activation protein (FAP);L is a linker;LEis a peptide linker comprising at least one proline;E is a secondary effector moiety; andRLis a ligand.

[0006] In certain embodiments, provided herein are compounds, comprising a linker represented by:#583961Attorney Docket No. 00020.026.1801wherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl. 6-membered heteroaryl, and C3-6 cycloalkyln is an integer from 1 to 6,q is an integer from 1 to 20;p is 0 or 1; andLEis a peptide linker comprising at least one proline.

[0007] In certain embodiments, provided herein are pharmaceutical compositions comprising a compound described herein (e.g., a complex represented by Formula (I) or compound represented by Formula (I)), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0008] In certain embodiments, disclosed herein are methods of treating a disease (e.g., a disease described herein) in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein (e.g., a complex represented by Formula (I)).

[0009] Additionally disclosed herein, in certain embodiments, are methods of imaging a cancer-associated fibroblast in a patient in need thereof, comprising administering to the patient an effective amount of a compound described herein (e.g., a complex represented by Formula (I)), and identifying the location of the cancer-associated fibroblast in the patient.BRIEF DESCRIPTION OF THE DRAWINGS177

[0010] FIG. 1 depicts a radiochromatogram of Lu-Compound 1.

[0011] FIG. 2 depicts SPECT / CT scans at 3 hours, 24 hours, 48 hours, 96 hours, and 192 hours post injection of177Lu-Compound 1 using Balb / c mice bearing U87MG tumors.177

[0012] FIG. 3A shows binding results of Lu-Compound 1 in an HT1080-hFAP 177xenograft model. FIG. 3B shows specific binding results of Lu-Compound 1 in an HT1080-hFAP xenograft model.177

[0013] FIG. 4A shows biodistribution of Lu-Compound 1 across various regions in 177male naive rats. FIG. 4B shows biodistribution of Lu-Compound 1 across various regions in female naive rats.3#583961Attorney Docket No. 00020.026.1801

[0014] FIG. 5 shows dosimetry results of Lu-Compound 1 across various regions in male (M) and female (F) naive rats.177

[0015] FIG. 6A shows dose recovery results of Lu-Compound 1 in female naive 177rats. FIG. 6B shows dose recovery7results of Lu-Compound 1 in male naive rats.177

[0016] FIG. 7A shows biodistribution of Lu-Compound 1 in HT1080-hFAP tumor 177bearing rats 1 hour after dosing of Lu-Compound 1. FIG. 7B shows biodistribution of Lu-Compound 1 in HT1080-hFAP tumor bearing rats 24 hours after dosing of Lu- 177Compound 1. FIG. 7C shows biodistribution of Lu-Compound 1 in HT1080-hFAP tumor 177bearing rats 48 hours after dosing of Lu-Compound 1. FIG. 7D shows biodistribution of 177 177 Lu-Compound 1 in HT1080-hFAP tumor bearing rats 120 hours after dosing of Lu- Compound 1.177

[0017] FIG. 8 shows dosimetry results of Lu-Compound 1 across various regions in male (M) and female (F) HT1080-hFAP tumor bearing rats.DETAILED DESCRIPTIONDefinitions

[0018] As used herein the specification, “a” or "an" may mean one or more. As used herein, when used in conjunction with the word "comprising", the words “a” or "an" may mean one or more than one. As used herein "another” may mean at least a second or more. Still further, the terms ’‘having”, “including”, “containing” and “comprising” are interchangeable and one of skill in the art is cognizant that these terms are open ended terms. Some embodiments of the disclosure may consist of or consist essentially of one or more elements, method steps, and / or methods of the disclosure. It is contemplated that any method, compound, or composition described herein can be implemented with respect to any other method, compound, or composition described herein.

[0019] " About" and "approximately" shall generally mean an acceptable degree of error for the quantity measured given the nature or precision of the measurements. Exemplary degrees of error are within 20 percent (%), typically, within 10%, and more typically, within 5% of a given value or range of values.

[0020] As used herein, “alkyl” refers to a radical of a straight-chain or branched saturated hydrocarbon group, e.g, having 1 to 20 carbon atoms (“C1-20 alkyl”). In some 4#583961Attorney Docket No. 00020.026.1801embodiments, an alkyl group has 1 to 10 carbon atoms (“C1-10 alkyl”). In some embodiments, an alkyl group has 1 to 9 carbon atoms (“C1-9 alkyl”). In some embodiments, an alkyl group has 1 to 8 carbon atoms (“C1-8 alkyl”). In some embodiments, an alkyl group has 1 to 7 carbon atoms (“C1-7 alkyl”). In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C1-6 alkyl”). In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C1-5 alkyl”). In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C1-4 alkyl”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C1-3 alkyl”). In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C1-2 alkyl”). In some embodiments, an alkyl group has 1 carbon atom (“C1 alkyl”). Examples of C1-6 alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, hexyl, and the like. In some embodiments, “alkyl” is unsubstituted or substituted with one or more substituents as described herein.

[0021] As used herein, “cycloalkyl” refers to a monovalent saturated cyclic, bicyclic, or bridged cyclic (e.g., adamantyl) hydrocarbon group of 3-12, 3-10, 3-8, 4-8, or 4-6 carbons ("C3-10cycloalkyl") derived from a cycloalkane. Exemplary cycloalkyl groups include, but are not limited to, cyclohexanes, cyclopentanes, cyclobutanes and cyclopropanes. A some embodiments, a cycloalkyl group is a monovalent saturated cyclic hydrocarbon group of 3-6 carbons (“C3-6 cycloalkyl”).

[0022] As used herein, “halogen” refers to -F, -Cl, -Br, or -I.

[0023] As used herein, “heterocyclyl” refers to a radical of a 3- to 10- membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“3-10 membered heterocyclyl”). In some embodiments, in heterocyclyl groups that contain one or more nitrogen atoms, the point of attachment is a carbon or nitrogen atom, as valency permits. In some embodiments, a heterocyclyl group is either be monocyclic (“monocyclic heterocyclyl”) or a fused, bridged or spiro ring system such as a bicyclic system (“bicyclic heterocyclyl”), and is saturated or is partially unsaturated. In some embodiments, heterocyclyl bicyclic ring systems include one or more heteroatoms in one or both rings. “Heterocyclyl” also includes ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more cycloalkyl groups wherein the point of attachment is either on the cycloalkyl or heterocyclyl ring, or ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more phenyl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heterocyclyl ring system. The terms “heterocycle,” “heterocyclyl,” “heterocyclyl ring,” “heterocyclic group,” “heterocyclic 5#583961Attorney Docket No. 00020.026.1801moiety,” and “heterocyclic radical,” are used interchangeably. In some embodiments, a heterocyclyl group is a 3-6 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“3 to 6-membered heterocyclyl”). In some embodiments, a heterocyclyl group is a 6 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“6-membered heterocyclyl”).

[0024] As used herein, “heteroaryl” refers to a radical of a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 electrons shared in a cyclic array) having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur (“5-10 membered heteroaryl”). In some embodiments, in heteroaryl groups that contain one or more nitrogen atoms, the point of attachment is a carbon or nitrogen atom, as valency permits. In some embodiments, a heteroaryl group is a 6 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“6-membered heteroaryl”).

[0025] Exemplary 6-membered heteroary l groups containing one heteroatom include, without limitation, pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, without limitation, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6- membered heteroaryl groups containing three or four heteroatoms include, without limitation, triazinyl and tetrazinyl, respectively.

[0026] It will be appreciated that the compounds, as described herein, may be substituted with any number of substituents or functional moieties. In general, the term “substituted” whether preceded by the term “optionally” or not, and substituents contained in formulas of this disclosure, refer to the replacement of hydrogen radicals in a given structure with the radical of a specified substituent. When more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position.

[0027] Unless otherwise indicated, any heteroatom with unsatisfied valences is assumed to have hydrogen atoms sufficient to satisfy the valences.

[0028] Certain compounds of the present disclosure may exist in particular geometric or stereoisomeric forms. The present disclosure contemplates all such compounds, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (d)-isomers, (l)-isomers, the 6#583961Attorney Docket No. 00020.026.1801racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the disclosure. Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this disclosure.

[0029] All stereoisomers of the present compounds (for example, those which may exist due to asymmetric carbons on various substituents), including enantiomeric forms and diastereomeric forms, are contemplated within the scope of this disclosure. Individual stereoisomers of the compounds of the disclosure may, for example, be substantially free of other isomers (e.g., as a pure or substantially pure optical isomer having a specified activity), or may be admixed, for example, as racemates or with all other, or other selected, stereoisomers. The chiral centers of the present disclosure may have the S or R configuration as defined by the International Union of Pure and Applied Chemistry (IUPAC) 1974 Recommendations. The racemic forms can be resolved by physical methods, such as, for example, fractional cry stallization, separation or crystallization of diastereomeric derivatives, or separation by chiral column chromatography. The individual optical isomers can be obtained from the racemates by any suitable method, including without limitation, conventional methods, such as, for example, salt formation with an optically active acid followed by crystallization.

[0030] All configurational isomers of the compounds of the present disclosure are contemplated, either in admixture or in pure or substantially pure form. The definition of compounds of the present disclosure embraces both cis (Z) and trans (E) alkene isomers, as well as cis and trans isomers of cyclic hydrocarbon or heterocyclic rings.

[0031] Isomeric mixtures containing any of a variety of isomer ratios may be utilized in accordance with the present disclosure. For example, where only two isomers are combined, mixtures containing 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99: 1, or 100:0 isomer ratios are all contemplated by the present disclosure.

[0032] The present disclosure also includes isotopically-labeled compounds, which are identical to the compounds disclosed herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, and chlorine, such as2H,3H,13C,11C,14C,15N,18O,170,31P,32P,35S,18F, and36C1, respectively. Compounds of the present disclosure, or a stereoisomer, tautomer, or pharmaceutically acceptable salt or solvate thereof, which contain the7#583961Attorney Docket No. 00020.026.1801aforementioned isotopes and / or other isotopes of other atoms are within the scope of this disclosure.

[0033] As used herein, “pharmaceutically acceptable salt’’ refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al., describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of this disclosure include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2–hydroxy–ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropi onate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(Ci-4alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate.

[0034] As used herein, “pharmaceutically acceptable excipient” refers to any substance in a pharmaceutical formulation other than an active pharmaceutical ingredient(s). Exemplary pharmaceutical excipients include those that aid the manufacturing process; protect, support or enhance stability: increase bioavailability; or increase patient8#583961Attorney Docket No. 00020.026.1801acceptability. They may also assist in product identification or enhance the overall safety or function of the product during storage or use.

[0035] As used herein, a '‘subject” to which administration is contemplated includes, but is not limited to, humans (i.e., a male or female of any age group, e.g., a pediatric subject (e.g., infant, child, adolescent) or adult subject (e.g., young adult, middle-aged adult or senior adult)) and / or a non-human animal, e.g.. a mammal such as primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human animal. The terms “human,” “patient,” “subject,” and “individual” are used interchangeably herein. None of these terms require the active supervision of medical personnel.

[0036] Disease, disorder, and condition are used interchangeably herein.

[0037] As used herein, and unless otherwise specified, the terms “treat,” “treating” and “treatment” contemplate an action that occurs while a subject is suffering from the specified disease, disorder or condition, which reduces the severity of the disease, disorder or condition, or reverses or slows the progression of the disease, disorder or condition (also “therapeutic treatment”).

[0038] In general, the “effective amount” of a compound refers to an amount sufficient to elicit the desired biological response. As will be appreciated by those of ordinary skill in this art. the effective amount of a compound of the disclosure may vary depending on such factors as the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, weight, health, and condition of the subject. A “therapeutically effective amount” of a compound is an amount sufficient to provide a therapeutic benefit in the treatment of a disease, disorder or condition, or to delay or minimize one or more symptoms associated with the disease, disorder or condition. A therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapies, which provides a therapeutic benefit in the treatment of the disease, disorder or condition. The term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms or causes of disease or condition, or enhances the therapeutic efficacy of another therapeutic agent. A “prophylactically effective amount” of a compound is an amount sufficient to prevent a disease, disorder or condition, or one or more sy mptoms associated with the disease, disorder or condition, or prevent its recurrence. A prophylactically effective amount of a compound means an amount of a therapeutic agent, alone or in combination with other agents, which 9#583961Attorney Docket No. 00020.026.1801provides a prophylactic benefit in the prevention of the disease, disorder or condition. The term "prophy lactically effective amount” can encompass an amount that improves overall prophylaxis or enhances the prophylactic efficacy of another prophylactic agent. A “prophylactic treatment” contemplates an action that occurs before a subject begins to suffer from the specified disease, disorder or condition.

[0039] As used herein, “177Lu-Compound 1” refers to a chelated complex represented by the formula:Compounds

[0040] In certain embodiments, provided herein are complexes represented by Formula (1):D-L-RAILEEFormula (I)or a pharmaceutically acceptable salt thereof, wherein:D is a ligand that targets fibroblast activation protein (FAP);L is a linker;LEis a peptide linker comprising at least one proline;E is a secondary effector moiety; and10#583961Attorney Docket No. 00020.026.1801RAis a ligand chelated or covalently linked to a therapeutic radionuclide or a diagnostic radionuclide.

[0041] In some embodiments, D is represented by Formula X-B:T is substituted or unsubstituted methylene (-CH2-), substituted or unsubstituted amino (-NH-), -O-, or -S- (e.g., wherein the substitution of T is C1-C3 alkyl, haloalkyl, or halo);J is C(RJ)0-3, wherein each RJis independently H or alkyl, or two or more RJare taken together to form oxo;R1and R2are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-. -C=C-C(O)aryl, -C=C-S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, -CONH2, and 5-tetrazolyl;R3and R4are independently selected from the group consisting of -H, -OH, F, Cl, Br, I, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl;R5, R6, R7, and R8are independently selected from the group consisting of H, alkyl, and halo; andR9, R10, andR11are independently selected from the group consisting of H,-Ci-ealkyl, -Ci-ehaloalkyl, -O-Ci-ealkyl, -S-C1-6alkyl, F, Cl, Br, and I.

[0042] In some embodiments, D is represented by Formula X-C:wherein:R21is -CN or#583961Attorney Docket No. 00020.026.1801oR22and R23are independently selected from the group consisting of H, C1-6 alkyl, and halo; andR24is C1-6 alkyl;Q is -N(R25)(C(R26)(R27))t- or -O(C(R28)(R29))UC(O)-;R25is selected from H and C1-6 alkyl; andeach R26, R27, R28, and R29is independently selected from H and Ci-6alkyl; each r and s is independently 1 or 2; andeach t and u is independently 0 or 1.

[0043] In some embodiments, D is represented by:R41is selected from the group consisting of -H, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=C-C(O)aryl, -C=C-S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, and 5-tetrazolyl,R42, R43a, R43band R44are independently selected from the group consisting of -H, -OH, halogen, -Ci-6alkyl, -O-Ci-6alkyl, and -S-Ci-6alkyl,R45is selected from the group consisting of -H, -CH2OH, and -CH(CH3)2,R46, R47, and R48are each independently selected from the group consisting of -H, -OH, oxo, halogen, CF3, -Ci-6alkyl, -O-Ci-6alkyl, and -S-Ci-6alkyl, -NR49R50, -OR51, -Het2, and -Ar2; each of -Ci-6alkyl being optionally substituted with from 1 to 3 substituents selected from -OH and halogen;R49, R50, and R51are each independently selected from the group consisting of -H, -OH, oxo, halogen, CF3, -Ci-6alkyl, -O-Ci-6alkyl, -S-Ci-6alkyl, and -Ar3.Ar2and Ar3are each independently a 5- or 6-membered aromatic monocycle optionally comprising 1 or 2 heteroatoms selected from O, N, and S; each of Ar2and Ar312#583961Attorney Docket No. 00020.026.1801being optionally and independently substituted with from 1 to 3 substituents selected from -NR52R53, -Ci-6alkyl, -O-Ci-6alkyl, and -S-Ci-6alkyl,R52and R53are each independently selected from the group consisting of -H, -OH, CF3, -Ci-6alkyl, -O-Ci-6alkyl, and -S-Ci-6alkyl,Het2 is a 5- or 6-membered non-aromatic monocycle optionally comprising 1 or 2 heteroatoms selected from O, N and S; Het2 being optionally substituted with from 1 to 3 substituents selected from -NR54R55, -Ci-6alkyl, -O-Ci-6alkyl, and -S-Ci-6alkyl,R54and R55are each independently selected from the group consisting of -H, -OH, halogen, CF3, -Ci-6alkyl, -O-Ci-6alkyl, and -S-Ci-6alkyl,the fragment:represents a 5- to 10-membered N-containing aromatic or non-aromatic mono- or bicyclic heterocycle, said heterocycle optionally further comprising 1 to 3 heteroatoms selected from O, N, and S; andJ is selected from the group consisting of a bond, -C1-3alkyl, -C1-3alkyl-NH-, C=O, and -O-.

[0044] In some embodiments, D is represented by:Fwherein,R30is -H, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=C-C(O)aryl, -C=C-S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, or 5-tetrazolyl;R31is H or CH3; andAr1is substituted phenyl, pyridyl, chloropyridyl, or quinolinyl.

[0045] In some embodiments, D is represented by:13#583961Attorney Docket No. 00020.026.1801wherein:each m is independently 0 to 3.14#583961Attorney Docket No. 00020.026.1801

[0046] In some embodiments, L is represented by:Ywherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl, 6-membered heteroaryl, and C3-6 cycloalkyl;L1is selected from a covalent bond and a linker; andL2is a linker,whereina is the point of attachment to D;P is the point of attachment to RL; andY is the point of attachment to LE.

[0047] In some embodiments, L is represented by:wherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl, 6-membered heteroaryl, and C3-6 cycloalkyl;n is an integer from 1 to 6,q is an integer from 1 to 20; andp is 0 or 1;wherein:a is the point of attachment to D;is the point of attachment to RL; andY is the point of attachment to LE.

[0048] In some embodiments, ring A is 3 to 6-membered heterocyclyl.

[0049] In some embodiments, ring A is selected from the group consisting of:#583961Attorney Docket No. 00020.026.1801wherein \| / represents the point of attachment to L1, and e represents the point of attachment to L2.

[0050] In some embodiments, LEis represented by:θ-(X1)m-LC-μwherein:each X1is an amino acid;Lcis absent or a cleavable linker; andm is an integer from 1 to 3;provided that at least one X1is proline; andwherein 0 represents the point of attachment to L and p represents the point of attachment to E.

[0051] In some embodiments, each X1is independently selected from the group consisting of absent, proline, glycine, alanine, threonine, and serine, provided that at least one X1is proline.

[0052] In some embodiments, each X1is independently selected from the group consisting of proline and glycine, provided that at least one X1is proline. In some embodiments, each X1is independently selected from the group consisting of proline and glycine, provided that at least one X1is proline, and m is 2.

[0053] In some embodiments, Lcis absent.

[0054] In some embodiments, Lcis a cleavable linker represented by:

[0055] In some embodiments, the ligand in RAis represented by:#583961Attorney Docket No. 00020.026.1801or17#583961Attorney Docket No. 00020.026.1801

[0056] In some embodiments, RAis represented by:, wherein M is a therapeutic radionuclide or a diagnostic radionuclide.

[0057] In some embodiments, E is an albumin binding ligand.

[0058] In some embodiments, E comprises a cleavable linker.

[0059] In some embodiments, E is represented by:#583961Attorney Docket No. 00020.026.1801wherein each of X and Y is independently selected from the group consisting of halogen and C1-6 alkyl, and n is an integer from 1 to 20. In some embodiments, X is halogen. In some embodiments, X is I.

[0060] In certain embodiments, provided herein are complexes chelated or covalently linked to a radionuclide, comprising a linker represented by:wherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl, 6-membered heteroaryl, and C3-6 cycloalkyl;n is an integer from 1 to 6,q is an integer from 1 to 20;p is 0 or 1; andLEis a peptide linker comprising at least one proline.

[0061] In some embodiments, LEis represented by:θ-(X1)m-LC-μwherein:each X1is an amino acid;Lcis absent or a cleavable linker; andm is an integer from 1 to 3;provided that at least one X1is proline; andwherein 0 represents the point of attachment to L and p represents the point of attachment to E.

[0062] In some embodiments, each X1is independently selected from the group consisting of absent, proline, glycine, alanine, threonine, and serine, provided that at least one X1is proline.

[0063] In some embodiments, each X1is independently selected from the group consisting of proline and glycine, provided that at least one X1is proline. In some embodiments, each X1is independently selected from the group consisting of proline and glycine, provided that at least one X1is proline, and m is 2.

[0064] In some embodiments, Lcis absent.#583961Attorney Docket No. 00020.026.1801

[0065] In some embodiments, Lcis a cleavable linker represented by:

[0066] In some embodiments, the complex is a complex of a compound selected from the group consisting of:#583961Attorney Docket No. 00020.026.1801#583961Attorney Docket No. 00020.026.1801or a pharmaceutically acceptable salt thereof, and a therapeutic radionuclide or a diagnostic radionuclide.

[0067] In some embodiments, the therapeutic radionuclide is selected from the group consisting of177Lu,90Y,135Tb,161Tb,225Ac,67Cu,223Ra,186Re,188Re,131I,212Pb,212BI,213BI, and211At.

[0068] In some embodiments, the therapeutic radionuclide is177Lu.

[0069] In some embodiments, the diagnostic radionuclide is selected from the group consisting of "mTc,mIn,18F,66Ga,67Ga,68Ga,1241,125I.44Sc.47Sc,64Cu,52Mn.55Co.86Y,89Zr,149Tb,152Tb, and203Pb.

[0070] In certain embodiments, provided herein are compounds represented by Formula (II):22#583961Attorney Docket No. 00020.026.1801Formula (II)or a pharmaceutically acceptable salt thereof, wherein:D is a ligand that targets fibroblast activation protein (FAP);L is a linker;LEis a peptide linker comprising at least one proline;E is a secondary effector moiety; andRLis a ligand.

[0071] In some embodiments, D is represented by Formula X-B:T is substituted or unsubstituted methylene (-CH2-), substituted or unsubstituted amino (-NH-), -O-, or -S- (e.g., wherein the substitution of T is C1-C3 alkyd, haloalkyl, or halo);J is C(RJ)0-3, wherein each RJis independently H or alkyl, or two or more RJare taken together to form oxo;R1and R2are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aiyl-, -C=C-C(O)aryl, -C=C-S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, -CONH2, and 5-tetrazolyl;R3and R4are independently selected from the group consisting of -H, -OH, F, CL Br, I, -Ci ealkyl, -O-Ci-6alkyl, and -S-Ci ealkyl;R5, R6, R7, and R8are independently selected from the group consisting of H, alkyl, and halo; and23#583961Attorney Docket No. 00020.026.1801R9, R10, andR11are independently selected from the group consisting of H,-Ci-ealkyl, -Ci-6haloalkyl, -O-Ci-6alkyl, -S-C1-6alkyl, F, Cl, Br, and I.

[0072] In some embodiments, D is represented by Formula X-C:(X-C)wherein:R21is -CN oreach r and s is independently 1 or 2;R22and R23are independently selected from the group consisting of H, C1-6 alkyl, and halo; andR24is C1-6 alkyl;Q is -N(R25)(C(R26)(R27))t- or -O(C(R28)(R29))uC(O)-;R25is selected from H and C1-6 alkyl; andeach R26, R27, R28, and R29is independently selected from H and C1-6 alkyl; and each t and u is independently 0 or 1.

[0073] In some embodiments, D is represented by:wherein,R30is -H, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=C-C(O)aryl, -C €-, S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, or 5-tetrazolyl;R31is H or CH3; andAr1is substituted phenyl, pyridyl, chloropyridyl, or quinolinyl.

[0074] In some embodiments, D is represented by:#583961Attorney Docket No. 00020.026.1801R41is selected from the group consisting of -H, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=C-C(O)aryl, -C=C-S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, and 5-tetrazolyl, R42j^43a j^43ban(j 44are eachInc|epenc|ent|y selected from the group consisting of -H, -OH, halogen, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl,R45is selected from the group consisting of -H, -CH2OH, and -CH(CH3)2,R46anj j^48are eacindependently selected from the group consisting of -H, -OH, oxo, halogen, CF3, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl, -NR49R50, -OR51, - Het2, and -Arc; each of -Ci-ealkyl being optionally substituted with from 1 to 3 substituents selected from -OH and halogen;R49anj:> Iare eacindependently selected from the group consisting of -H, -OH, oxo. halogen, CF3, -Ci-ealkyl. -O-Ci-ealkyl. -S-Ci-ealkyl, and -An.Arc and An are each independently a 5- or 6-membered aromatic monocycle optionally comprising 1 or 2 heteroatoms selected from O, N, and S; each of Arc and Arc being optionally and independently substituted with from 1 to 3 substituents selected from -NR52R53. -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl,R52and R53are each independently selected from the group consisting of -H, -OH, CF3, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl,Het2 is a 5- or 6-membered non-aromatic monocycle optionally comprising 1 or 2 heteroatoms selected from O, N and S; Het2 being optionally substituted with from 1 to 3 substituents selected from -NR54R55, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl,R34and R55are each independently selected from the group consisting of -H, -OH, halogen, CFs, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl,the fragment:25#583961Attorney Docket No. 00020.026.1801represents a 5- to 10-membered N-containing aromatic or non-aromatic mono- or bicyclic heterocycle, said heterocycle optionally further comprising 1 to 3 heteroatoms selected from O, N, and S; andJ is selected from the group consisting of a bond, -Ci salkyl, -Ci salkyl-NH-, C=O, and -O-.

[0075] In some embodiments, D is represented by:F F26#583961Attorney Docket No. 00020.026.1801wherein:each m is independently 0 to 6.

[0076] In some embodiments, L is represented by:wherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl, 6-membered heteroaryl, and C3-6 cycloalkyl;L1is selected from a covalent bond and a linker; andL2is a linker,whereina is the point of attachment to D;P is the point of attachment to RL; andy is the point of attachment to LE.

[0077] In some embodiments, L is represented by:27#583961Attorney Docket No. 00020.026.1801wherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl, 6-membered heteroaryl, and C3-6 cycloalkyl;n is an integer from 1 to 6,q is an integer from 1 to 20; andp is 0 or 1;wherein:a is the point of attachment to D;P is the point of attachment to RL; andy is the point of attachment to LE.

[0078] In some embodiments, ring A is 3 to 6-membered heterocyclyl.

[0079] In some embodiments, ring A is selected from the group consisting of:wherein \| / represents the point of attachment to L1, and s represents the point of attachment to L2.

[0080] In some embodiments, LEis represented by:wherein each X1is an amino acid;Lcis absent or a cleavable linker; andm is an integer from 1 to 3;provided that at least one X1is proline; andwherein 0 represents the point of attachment to L and p represents the point of attachment to E.#583961Attorney Docket No. 00020.026.1801

[0081] In some embodiments, each X1is independently selected from the group consisting of absent, proline, glycine, alanine, threonine, and serine, provided that at least one X1is proline.

[0082] In some embodiments, each X1is independently selected from the group consisting of proline and glycine, provided that at least one X1is proline. In some embodiments, each X1is independently selected from the group consisting of proline and glycine, provided that at least one X1is proline, and m is 2.

[0083] In some embodiments, Lcis absent.

[0084] In some embodiments, Lcis a cleavable linker represented by:

[0085] In some embodiments, RLis capable of chelating or covalently binding to a therapeutic radionuclide or a diagnostic radionuclide.

[0086] In some embodiments, RLis represented by:29#583961Attorney Docket No. 00020.026.1801or30#583961Attorney Docket No. 00020.026.1801

[0087] In some embodiments, RLis represented by:

[0088] In some embodiments, E is an albumin binding ligand.

[0089] In some embodiments, E comprises a cleavable linker.

[0090] In some embodiments, E is represented by:#583961Attorney Docket No. 00020.026.1801wherein each of X and Y is independently selected from the group consisting of halogen and C1-6 alkyl, and n is an integer from 1 to 20. In some embodiments, X is halogen. In some embodiments, X is I.

[0091] In certain embodiments, provided herein are compounds, comprising a linker represented by:wherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl, 6-membered heteroaryl, and C3-6 cycloalkyl;n is an integer from 1 to 6,q is an integer from 1 to 20;p is 0 or 1; andLEis a peptide linker comprising at least one proline.

[0092] In some embodiments, LEis represented by:θ-(X1)m-LC-μwherein:each X1is an amino acid;Lcis absent or a cleavable linker; andm is an integer from 1 to 3;provided that at least one X1is proline; andwherein 0 represents the point of attachment to L and p represents the point of attachment to E.

[0093] In some embodiments, each X1is independently selected from the group consisting of absent, proline, glycine, alanine, threonine, and serine provided that at least one X1is proline.

[0094] In some embodiments, each X1is independently selected from the group consisting of proline and glycine, provided that at least one X1is proline. In some#583961Attorney Docket No. 00020.026.1801embodiments, each X1is independently selected from the group consisting of proline and glycine, provided that at least one X1is proline, and m is 2.

[0095] In some embodiments, Lcis absent.

[0096] In some embodiments, Lcis a cleavable linker represented by:ci $

[0097] In some embodiments, the compound is a compound selected from the group consisting of: A bCompound Structurenumber1c2Ay AFF cxy HN O?H3 / ^. A?HNA^SN \ _QXVOA MAUAA4}£w Vv“$? A; y A U5^ ^33#583961Attorney Docket No. 00020.026.1801#583961Attorney Docket No. 00020.026.1801Pharmaceutical Compositions

[0098] Compounds provided in accordance with the present disclosure, in certain embodiments, are administered in the form of pharmaceutical compositions. This disclosure therefore provides, in certain embodiments, pharmaceutical compositions that comprise, as the active ingredient, one or more of the compounds described, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, including inert solid diluents and fillers, diluents, including sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers and adjuvants. In certain embodiments, the pharmaceutical compositions are administered in combination with other therapeutic agents.

[0099] Provided herein, in certain embodiments, is a pharmaceutical composition, comprising: a) a compound disclosed herein, or a pharmaceutically acceptable salt thereof: and b) a pharmaceutically acceptable excipient.35#583961Attorney Docket No. 00020.026.1801[000100] In certain embodiments, the pharmaceutical compositions are administered in a single dose by any of the accepted modes of administration of agents having similar utilities, including rectal, buccal, intranasal and transdermal routes, by intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, as an inhalant, or via an impregnated or coated device such as a stent, for example, or an artery-inserted cylindrical polymer. In certain embodiments, the pharmaceutical compositions are administered in multiple doses.[000101] One mode for administration is parenteral, for example by injection.Administration by injection comprises, in some embodiments, aqueous or oil suspensions, or emulsions, with sesame oil, corn oil. cottonseed oil, or peanut oil. as well as elixirs, mannitol, dextrose, or a sterile aqueous solution, and similar pharmaceutical vehicles. Aqueous solutions in saline are also conventionally used for injection. In some embodiments, compositions suitable for injection comprise ethanol, glycerol, propylene glycol, liquid polyethylene glycol, or the like (and suitable mixtures thereof), cyclodextrin derivatives, or vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, or the like.[000102] Sterile injectable solutions are prepared by incorporating a compound according to the present disclosure in the required amount in the appropriate solvent with various other ingredients as enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.[000103] Oral administration is another route for administration of compounds in accordance with the disclosure. Oral administration includes, for example, capsule or enteric coated tablets, or the like. In making the pharmaceutical compositions that include at least one compound described herein, the active ingredient is, in some embodiments, diluted by an excipient and / or enclosed within such a carrier that can be in the form of a capsule, sachet, paper or other container. When the excipient serves as a diluent, it can be in the form of a 36#583961Attorney Docket No. 00020.026.1801solid, semi-solid, or liquid material (as above), which acts as a vehicle, carrier or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments comprising the active compound, soft and hard gelatin capsules, sterile injectable solutions, or sterile packaged powders.[000104] The compositions of the disclosure can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient by employing procedures known in the art. Controlled release drug delivery systems for oral administration include osmotic pump systems and dissolutional systems containing polymer-coated reservoirs or drug-polymer matrix formulations. Another formulation for use in the methods of the present disclosure employs transdermal delivery devices ("patches"). Such transdermal patches may be used to provide continuous or discontinuous infusion of the compounds of the present disclosure in controlled amounts. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents.[000105] The compositions are preferably formulated in a unit dosage form. The term "unit dosage forms" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient (e.g., a tablet, capsule, or ampoule). The compounds are generally administered in a pharmaceutically effective amount. It will be understood, however, that the amount of the compound actually administered usually will be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered and its relative activity, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.[000106] For preparing solid compositions such as tablets, the principal active ingredient is mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound of the present disclosure. When referring to these preformulation compositions as homogeneous, it is meant that the active ingredient is dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules.[000107] In certain embodiments, the tablets or pills of the present disclosure are coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action, or to protect from the acid conditions of the stomach. For example, the tablet or pill 37#583961Attorney Docket No. 00020.026.1801can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer that serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.[000108] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. In some embodiments, the compositions are administered by the oral or nasal respiratory route for local or systemic effect. Compositions in preferably pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizing device or the nebulizing device may be attached to a facemask tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered, preferably orally or nasally, from devices that deliver the formulation in an appropriate manner.Methods of Use[000109] In certain embodiments, provided herein are methods of treating cancer in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein (e.g., a complex of Formula (I) described herein or a compound of Formula (I) or (II) described herein, or a pharmaceutically acceptable salt thereof). In certain embodiments, the cancer is selected from the group consisting of breast cancer, pancreatic cancer, mesothelioma, esophageal cancer, cholangiocellular carcinoma, head and neck cancer, carcinoma of unknown primary origin, brain metastases, sarcoma, salivary’ gland carcinoma, lung cancer, prostate cancer, thymus cancer, ovarian cancer, desmoid, chordoma, colorectal cancer, anal carcinoma, neuroendocrine tumors, small intestine cancer, medullary’ thyroid cancer, cervical cancer, hepatocellular carcinoma, gastric cancer, adenoid cystic cancer, pheochromocytoma, differentiated thyroid cancer, insulinoma, renal cell cancer, gastric cancer, renal cancer, bone metastases, lymph node metastases, lung metastases, pleura, bladder cancer, myeloma, lymphoma, brain cancer, endometrial cancer, testicular cancer, neuroblastoma, astrocytoma, meningioma, rectal cancer, cervical cancer, stomach cancer, skin cancer, thyroid cancer, and / or liver cancer.[000110] In certain embodiments, provided herein are methods of treating a disease mediated by cancer-associated fibroblasts in a patient in need thereof, comprising38#583961Attorney Docket No. 00020.026.1801administering to the patient a therapeutically effective amount of a compound described herein (e.g., a complex of Formula (I) described herein or a compound of Formula (I) or (II) described herein, or a pharmaceutically acceptable salt thereof,). In certain embodiments, the disease is cancer. In certain embodiments, the cancer is selected from the group consisting of breast cancer, pancreatic cancer, mesothelioma, esophageal cancer, cholangiocellular carcinoma, head and neck cancer, carcinoma of unknown primary origin, brain metastases, sarcoma, salivary gland carcinoma, lung cancer, prostate cancer, thymus cancer, ovarian cancer, desmoid, chordoma, colorectal cancer, anal carcinoma, neuroendocrine tumors, small intestine cancer, medullary thyroid cancer, cervical cancer, hepatocellular carcinoma, gastric cancer, adenoid cystic cancer, pheochromocytoma, differentiated thyroid cancer, insulinoma, renal cell cancer, gastric cancer, renal cancer, bone metastases, lymph node metastases, lung metastases, pleura, bladder cancer, myeloma, lymphoma, brain cancer, endometrial cancer, testicular cancer, neuroblastoma, astrocytoma, meningioma, rectal cancer, cervical cancer, stomach cancer, skin cancer, thyroid cancer, and / or liver cancer.[000111] In certain embodiments, provided herein are methods of imaging a cancer-associated fibroblast in a patient in need thereof, comprising administering to the patient an effective amount of a compound described herein (e.g., a complex of Formula (I) described herein or a compound of Formula (I) or (II) described herein, or a pharmaceutically acceptable salt thereof), and identifying the location of the cancer-associated fibroblast in the patient.[000112] In certain embodiments, provided herein are method of imaging activated fibroblasts in a patient with idiopathic pulmonary fibrosis, cirrhosis or liver fibrosis, Crohn’s disease, rheumatoid arthritis, ulcerative colitis, sclerosis, cardiac fibrosis, and / or kidney fibrosis in need thereof, comprising administering to the patient an effective amount of a compound described herein (e.g., a complex of Formula (I) described herein or a compound of Formula (I) or (II) described herein, or a pharmaceutically acceptable salt thereof), and identifying the location of activated fibroblasts in the patient for noninvasive diagnosis.[000113] In certain embodiments, provided herein are compounds (e.g., a complex of Formula (I) or a compound of Formula (I) or (II) described herein, or a pharmaceutically acceptable salt thereof), for use as a medicament.[000114] In certain embodiments, provided herein are compounds (e.g., a complex of Formula (I) or a compound of Formula (I) or (II) described herein, or a pharmaceutically acceptable salt thereof) for use in treating cancer in a patient in need thereof, comprising 39#583961Attorney Docket No. 00020.026.1801administering to the patient a therapeutically effective amount of the compound described herein (e.g., a complex of Formula (I) described herein or a compound of Formula (I) or (II) described herein, or a pharmaceutically acceptable salt thereof). In certain embodiments, the cancer is selected from the group consisting of breast cancer, pancreatic cancer, mesothelioma, esophageal cancer, cholangiocellular carcinoma, head and neck cancer, carcinoma of unknown primary origin, brain metastases, sarcoma, salivary gland carcinoma, lung cancer, prostate cancer, thymus cancer, ovarian cancer, desmoid, chordoma, colorectal cancer, anal carcinoma, neuroendocrine tumors, small intestine cancer, medullary thyroid cancer, cervical cancer, hepatocellular carcinoma, gastric cancer, adenoid cystic cancer, pheochromocytoma, differentiated thyroid cancer, insulinoma, renal cell cancer, gastric cancer, renal cancer, bone metastases, lymph node metastases, lung metastases, pleura, bladder cancer, myeloma, lymphoma, brain cancer, endometrial cancer, testicular cancer, neuroblastoma, astrocytoma, meningioma, rectal cancer, cervical cancer, stomach cancer, skin cancer, thyroid cancer, and / or liver cancer.[000115] In certain embodiments, provided herein are compounds (e.g.. a complex of Formula (I) or a compound of Formula (I) or (II) described herein, or a pharmaceutically acceptable salt thereof) for use in treating a disease mediated by cancer-associated fibroblasts in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of the compound described herein (e.g., a complex of Formula (I) described herein or a compound of Formula (I) or (II) described herein, or a pharmaceutically acceptable salt thereof). In certain embodiments, the disease is cancer. In certain embodiments, the cancer is selected from the group consisting of breast cancer, pancreatic cancer, mesothelioma, esophageal cancer, cholangiocellular carcinoma, head and neck cancer, carcinoma of unknown primary origin, brain metastases, sarcoma, salivary gland carcinoma, lung cancer, prostate cancer, thymus cancer, ovarian cancer, desmoid, chordoma, colorectal cancer, anal carcinoma, neuroendocrine tumors, small intestine cancer, medullary thyroid cancer, cervical cancer, hepatocellular carcinoma, gastric cancer, adenoid cystic cancer, pheochromocytoma, differentiated thyroid cancer, insulinoma, renal cell cancer, gastric cancer, renal cancer, bone metastases, lymph node metastases, lung metastases, pleura, bladder cancer, myeloma, lymphoma, brain cancer, endometrial cancer, testicular cancer, neuroblastoma, astrocytoma, meningioma, rectal cancer, cervical cancer, stomach cancer, skin cancer, thyroid cancer, and / or liver cancer.[000116] In certain embodiments, provided herein are compounds (e.g.. a complex of Formula (I) or a compound of Formula (I) or (II) described herein, or a pharmaceutically 40#583961Attorney Docket No. 00020.026.1801acceptable salt thereof) for use in imaging a cancer-associated fibroblast in a patient in need thereof, comprising administering to the patient an effective amount of the compound described herein (e.g., a complex of Formula (I) described herein or a compound of Formula (I) or (II) described herein, or a pharmaceutically acceptable salt thereof), and identifying the location of the cancer-associated fibroblast in the patient.[000117] In certain embodiments, provided herein are compounds (e.g.. a complex of Formula (I) or a compound of Formula (I) or (II) described herein, or a pharmaceutically acceptable salt thereof) for use in imaging activated fibroblasts in a patient with idiopathic pulmonary fibrosis, cirrhosis or liver fibrosis, Crohn’s disease, rheumatoid arthritis, ulcerative colitis, sclerosis, cardiac fibrosis, and / or kidney fibrosis in need thereof, comprising administering to the patient an effective amount of the compound described herein (e.g., a complex of Formula (I) described herein or a compound of Formula (I) or (II) described herein, or a pharmaceutically acceptable salt thereof), or a pharmaceutically acceptable salt thereof, and identifying the location of activated fibroblasts in the patient for noninvasive diagnosis.Dosage Regimens[000118] Dosage regimens may be adjusted to provide the optimum desired response. The skilled artisan would appreciate, based upon the disclosure provided herein, that the dose and dosing regimen is adjusted in accordance with methods well-known in the therapeutic arts. That is, the maximum tolerable dose can be readily established, and the effective amount providing a detectable therapeutic benefit to a patient may also be determined, as can the temporal requirements for administering each agent to provide a detectable therapeutic benefit to the patient. Accordingly, while certain dose and administration regimens are exemplified herein, these examples in no way limit the dose and administration regimen that may be provided to a patient in practicing the present disclosure.[000119] It is to be noted that dosage values may vary with the type and severity of the condition to be alleviated and may include single or multiple doses. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that dosage ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition. For example, doses may be adjusted based on pharmacokinetic or pharmacodynamic parameters, which may include clinical effects such as toxic effects and / or 41#583961Attorney Docket No. 00020.026.1801laboratory values. Thus, the present disclosure encompasses intra-patient dose-escalation as determined by the skilled artisan. Determining appropriate dosages and regimens for administration of the chemotherapeutic agent are well-known in the relevant art and would be understood to be encompassed by the skilled artisan once provided the teachings disclosed herein.[000120] The amount of the compound of the disclosure administered will be dependent on the subject being treated, the severity of the disorder or condition, the rate of administration, the disposition of the compound and the discretion of the prescribing physician. However, an effective dosage is in the range of about 0.001 to about 100 mg per kg body weight per day. In some instances, dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases still larger doses may be employed without causing any harmful side effect, provided that such larger doses are first divided into several small doses for administration throughout the day.Kits[000121] In another aspect provided herein are kits, comprising materials useful for the treatment or prevention of the diseases and disorders described above. In some embodiments, the kit comprises a container comprising a compound described herein (e.g., a complex of Formula (I) or compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof). In some embodiments, the kit further comprises a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, blister pack, etc. The container can be formed from a variety of materials such as glass or plastic. The container can hold a compound described herein (e.g., a complex of Formula (I) or a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof) or a composition thereof which is effective for treating or preventing the condition and can have a sterile access port (for example, the container can be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). At least one active agent in the composition is a compound described herein (e g., a complex of Formula (I) or a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof) or a pharmaceutically acceptable salt thereof. The label or package insert indicates that the composition is used for treating the condition of choice. The label or package insert can also indicate that the composition can be used to treat other disorders. Alternatively, or additionally, the article of manufacture can further comprise a second container comprising a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI),42#583961Attorney Docket No. 00020.026.1801phosphate-buffered saline, Ringer’s solution and dextrose solution. It can further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.[000122] The kit can further comprise directions for the administration of a compound described herein (e.g., a complex of Formula (I) or a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof) or a pharmaceutically acceptable salt thereof and, if present, the second pharmaceutical formulation. For example, if the kit comprises a first composition comprising a compound described herein (e.g., a complex of Formula (I) or a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof) or a pharmaceutically acceptable salt thereof and a second pharmaceutical formulation, the kit can further comprise directions for the simultaneous, sequential or separate administration of the first and second pharmaceutical compositions to a patient in need thereof.[000123] In another embodiment, the kits are suitable for the delivery of solid oral forms of a compound described herein (e.g., a complex of Formula (I) or a compound of Formula (I) or (II). or a pharmaceutically acceptable salt thereof) or a pharmaceutically acceptable salt thereof, such as tablets or capsules. Such a kit preferably includes a number of unit dosages. Such kits can include a card having the dosages oriented in the order of their intended use. An example of such a kit is a blister pack. Blister packs are well known in the packaging industry and are widely used for packaging pharmaceutical unit dosage forms. If desired, a memory aid can be provided, for example in the form of numbers, letters, or other markings or with a calendar insert, designating the days in the treatment schedule in which the dosages can be administered.[000124] In some embodiments, a kit comprises (a) a first container with a compound described herein (e.g., a complex of Formula (I) or a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof) or a pharmaceutically acceptable salt thereof contained therein; and optionally (b) a second container with a second pharmaceutical formulation contained therein. Alternatively, or additionally, the kit can further comprise a third container comprising a pharmaceutically-acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer’s solution and dextrose solution. It can further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.[000125] In some embodiments wherein the kit comprises a compound described herein (e.g., a complex of Formula (I) or a compound of Formula (I) or (II), or a pharmaceutically acceptable salt thereof) or a pharmaceutically acceptable salt thereof and a second therapeutic ’ 43#583961Attorney Docket No. 00020.026.1801agent, the kit can comprise a container for containing the separate compositions such as a divided bottle or a divided foil packet, however, the separate compositions can also be contained within a single, undivided container. Typically, the kit comprises directions for the administration of the separate components. The kit form is particularly advantageous when the separate components are preferably administered in different dosage forms (e.g., oral and parenteral), are administered at different dosage intervals, or when titration of the individual components of the combination is desired by the prescribing physician.EXAMPLESSelected Abbreviations[000126] ACN: acetonitrile; DCM: dichloromethane; DIPEA: diisopropylethylamine; DMAP: 4-dimethylaminopyridine; DMF: dimethylformamide; DMSO: dimethylsulfoxide; ESI: electrospray ionization; EtOAc: ethyl acetate; EtsN: triethylamine; HATU: hexafluorophosphate azabenzotriazole tetramethyl uronium; HPLC: high-performance liquid chromatography; h: hours; LC-MS: liquid chromatography-mass spectrometry; MeOH: methanol; 2-MeTHF: 2-methyltetrahydrofuran; min: minutes; MS: mass spectrometry;MTBE: methyl tert-butyl ether NMR: nuclear magnetic resonance; RT: room temperature; t-BuOH: tert-butyl alcohol; TFA: trifluoroacetic acid; THF: tetrahydrofuran; TLC: thin layer chromatography; T3P: propylphosphonic anhydride; XPhos: 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl; XPhos Pd G4: (-4-3)-[Dicyclohexyl[2',4',6'-tris(1-methylethyl)[1,1'-biphenyl]-2-yl]phosphine](methanesulfonato-κ)[2'-(methylamino-κ)[1,1'-biphenyl]-2-yl-κ]palladium.General synthetic methods[000127] The following methods were used in the exemplified Examples, except where noted otherwise.[000128] Purification of intermediates and final products was carried out via either normal or reverse phase chromatography. Normal phase chromatography was carried out using prepacked SiO₂ cartridges eluting with either gradients of hexane and ethyl acetate or DCM and MeOH. Reverse phase was carried out using Cl 8 column with UV 215 nm to 254 nm, eluting with gradients of solvents A (water with 0.1%TFA) and Solvent B (Acetonitrile) or solvents A: water and Solvent B: Acetonitrile. Reverse phase preparative HPLC was carried out using GEMINI C18 (250*21.2*)5u, AGILENT C18 (50*50mm)5u, Shim-pack C18 (250*20.0mm) 5u and Luna C18 (250*80mm lOp) with UV detection at 215& 254 nm.44#583961Attorney Docket No. 00020.026.1801LC-MS / HPLC Employed in characterization of examples[000129] LC-MS was performed on Waters (Acquity H Class plus coupled with SQ Detector 2,) column: Acquity BEH Cl 8 (2.1*50*mml.7p), Mobile phase A:0.01% Formic acid in water, Mobile phase B: Acetonitrile, Diluent; water: acetonitrile (9: 1), UV 215 nm, injection volume:1 µL, Run time: 6.6 min.[000130] Mass detector Parameters: Ionisation mode: ESI, Capillary’ voltage: 2.0 kV, Cone voltage: 35v, Desolation temp: 450°C[000131] HPLC was performed on Waters (e2695 with PDA Detector) column: Inertsil ODS 3V (250X4.6 mm5p), Mobile phase A:0.1%TFA in water, Mobile phase B:Acetonitrile: Water (90:10), Diluent; water: acetonitrile (9:1), UV 215 nm, injection volume:7 µL, Run time: 25 minPrep HPLC employed for purification[000132] Purification was performed using Shimadzu Nexera Prep with FRC-40 fraction collector; using with methods (A-H)Method-A:Column Kinetex C18 (250*21.2mm 5p)Mobile phase-A 0.1% TF A in WaterMobile phase-B Acetonitrile: water (80:20)Gradient (T / %B) 0 / 20,10 / 40,15 / 45,20 / 95Flow Rate 13 mL / minUV Detection 215nm & 254nmSolubility Acetonitrile: water (1:9)Method-B:Column Agilent Cl 8 (50*50mm 5p) + Kinetex C18(250*21.2mm 5p)Mobile phase-A 0.1% TFA in WaterMobile phase-B Acetonitrile: water (80:20)Gradient (T / %B) 0 / 25.12 / 38,25 / 38,25.1 / 98Flow Rate 9 mL / minUV Detection 215nm & 254nmSolubility Acetonitrile: water (1:9)Method-C:Column Gemini C18 (250*21.2mm 5p)Mobile phase-A 0.1% TF A in WaterMobile phase-B Acetonitrile: water (80:20)Gradient (T / %B) 0 / 10.1 / 10,15 / 50,18 / 50,18.05 / 9845#583961Attorney Docket No. 00020.026.1801Flow Rate 14 mL / minUV Detection 215nm & 254nmSolubility Acetonitrile: water (1:9)Method-D:Column XBridge C18 (250*19 mm 5p)Mobile phase-A 0.1% TF A in WaterMobile phase-B Acetonitrile: water (80:20)Gradient (T / %B) 0 / 5,2 / 5,10 / 38,20 / 38,21 / 96Flow Rate 13 mL / minUV Detection 215nm & 254nmSolubility Acetonitril e: water( 1:9)Method-E:Column Shim-pack C18 (250*20 mm 5p)Mobile phase-A 0.1% TFA in WaterMobile phase-B Acetonitrile: water (80:20)Gradient (T / %B) 0 / 30, 2 / 30, 20 / 80, 20.1 / 98Flow Rate 14 mL / minUV Detection 215nm & 254nmSolubility Acetonitrile: water (1:9)Method-F:Column name Luna C18 (250*80 mm 10 p)Mobile phase-A 0.1% TFA in WaterMobile phase-B 0.1% Acetonitrile: Water (80:20)Method (%B) 0 / 30, 2 / 30, 25 / 60, 26 / 90, 31 / 30,36 / 30Flow Rate 180 mL / minSolubility Acetonitrile: water (1:9)NMR Employed in characterization of examples[000133]NMR spectra were obtained with Brucker spectrometers operating at frequencies as follows: 'H NMR: 400 MHz. Spectra data are reported in the format:Chemical shift (multiplicity, number of hydrogens. Chemical shifts are specified in ppm downfield of a tetramethyl silane internal standard (5 units, TMS=0 ppm) and reference to solvent peaks, which in ¹HNMR appear at 2.49 ppm for DMSO-de. sample quantity 2 to 3 mg, solvents quantity 0.6 mL.#583961Attorney Docket No. 00020.026.1801Example 1: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 1)[000134] To a solution of 4-(4-iodophenyl)butanoic acid 1 (30 g, 0.103 mol) in DMF (60 mL, 2 Vol) at 0°C was added DIPEA (27 mL, 0.155 mol) followed by HATU (39.3 g, 0.103 mol). Reaction mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl 4-(aminomethyl)benzoate 2 (21.4 g, 0.103 mol). Resultant reaction mixture was stirred at room temperature for 4 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (400 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 400 mL). Combined organic layer was washed with water (400 mL). The separated organic layer 47#583961Attorney Docket No. 00020.026.1801was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (36.1 g, crude) as a brown liquid. MS (ESI) m / z: 480.1 [M+H]+Step-2: Synthesis of 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid (4):[000135] TFA (160 mL, 4 vol) was added to a solution of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (40 g, 0.083 mol) in DCM (200 mL, 5 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 5 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (200 mL) to afford 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid 4 (31 g, yield: 88%) as a brown solid. MS (ESI) m / z: 424.42 [M+H]+Step-3: Synthesis of tert-butyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycinate[000136] To a solution of 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid 4 (10 g, 23.64 mmol) in DMF (100 mL, 10 Vol) at 0°C, was added DIPEA (12.3 mL, 70.92 mmol ) followed by HATU (13.4 g, 35.46 mmol). The reaction was stirred at room temperature for 30 min, followed by addition of tert-butyl glycinate 5 (3.5 mL, 26.00 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min. Precipitated solid was filtered and wash with hexane (2* 100 mL) to afford tert-butyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycinate 6 (10 g, yield: 83 %) as a white solid. MS (ESI) m / z:538.28 [M+H]+.Step-4: Synthesis of (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine (7):48#583961Attorney Docket No. 00020.026.1801[000137] TFA (18 mL, 3 vol) was added to a solution of tert-butyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycinate 6 (6 g, 0.011 mol) in DCM (60 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 5 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (100 mL) to afford (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine 7 (4.5 g, yield: 86%) as a white solid. MS (ESI) m / z: 481.13 [M+H]+Step-5: Synthesis of (S)-2-( ( <(9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-( < S)-l-((4-( 4-( 4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid (8):[000138] To a solution of (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine 7 (10 g, 20.83 mmol) in DMF (10 mL, 10 Vol) at 0°C was added DIPEA (5.4 mL, 31.25 mmol) followed by addition of HATU (8.7 g. 22.91 mmol). The reaction was stirred at room temperature for 10 min and added (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid Int#1 (8.1 g, 18.75 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted with cold water (500 mL) and stirred for 15 min. Obtained white solid was dissolve in 10%MeOH: DCM (50 mL), added diethyl ether (250 mL) to afford (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid 8 (9.5 g, yield: 52%) as a white solid. MS (ESI) m / z: 900.45 (M+H)+.Step-6: Synthesis of tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate (9):49#583961Attorney Docket No. 00020.026.1801[000139] To a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid 8 (9.2 g. 10.5 mmol) in DMF (90 mL, 10 Vol) at 0 °C. was added DIPEA (3.66 mL, 23.00 mmol ) followed by HATU (4.38 g, 11.5 mmol). The mixture was stirred at 0°C for 30 min, and added tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate Int#5A (3.64 g, 11.51 mmol). Resultant reaction mixture was stirred at room temperature for 2h. Upon completion of the reaction, reaction mixture was diluted with cold water (500 mL) and stirred for 15 min. Obtained white solid was dissolve in 10%MeOH: DCM (50 mL), added diethyl ether (250 mL) to afford tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-y l)methoxy)carbonyl)amino)-4-((S)- 1 -((4-((4-(4-iodopheny 1) butanamido) methyl) benzoyl) glycyl) pyrrolidine-2-carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate 9 (10.2 g, yield: 83%) as a white solid. MS (ESI) m / z: 1198.69 [M+H]+.Step-7: Synthesis of (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin-[000140] TFA (40 mL, 4 vol) was added to a solution of tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy )carbony l)amino)-4-((S)- 1 -((4-((4-(4- 50#583961Attorney Docket No. 00020.026.1801iodophenyl)butanamido)methyl)benzoyl)glycyl) pyrrolidine-2 -carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate 9 (10.0 g, 8.34 mmol) in DCM (100 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (400 mL) to afford (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-1-(4-(isoindolin-4-ylmethyl)piperidin-1-yl)-1-oxobutan-2-yl)carbamate 10 (8.0 g, yield: 87%) as an off white solid. MS (ESI) m / z: 1098.94 [M+H]+Step-8: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-1-yl)-4-((S)-1-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-1-oxobutan-2-yl)carbamate (11):[000141] To a solution of CD A-5 (3.32 g, 11.05 mmol) in DMF (20 mL, 10Vol) at 0°C was added DIPEA (2.96 mL, 17.0 mmol) followed by addition of HATU (3.55 g, 9.35 mmol). The mixture was stirred at room temperature for 10 min followed by addition of (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate 10 (9.34 g, 8.504 mmol). Resultant reaction was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min. Obtained white solid was further triturated with diethyl ether (100 mL) to afford (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-1-yl)-4-((S)-1-((4-((4-(4- 51#583961Attorney Docket No. 00020.026.1801iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 11 (8.0 g, yield: 62%) as an off white solid. MS (ESI) m / z: 1382.19 (M+H)+. Step-9: Synthesis of(S)-N-((S)-3-amino-4-(4-( (2-(2-( ( 3S.5S)-5-( (S)-2-cyano-4, 4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)-1-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide (12):[000142] Diethyl amine (0.44 mL, 4.2 mmol) was added to a solution of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl) butanamido) methyl)benzoyl) glycyl) pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 11 (7.3 g, 5.28 mmol) in DMF (73 mL, 10 Vol) at room temperature. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with MTBE ( 730 mL) and obtained solid was filtered and washed with EtOAc (30 mL) to afford (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-1-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-1-yl)-4-oxobutyl)-1-((4-((4-(4-iodophenyl) butanamido) methyl) benzoyl)glycyl)pyrrolidine-2-carboxamide 12 (4.9 g, yield: 79%) as a white solid. MS (ESI) m / z: 1159.78 (M+H).Step-10: Synthesis of2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidme-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4, 7, 10-tetraazacyclododecane-l,4, 7-triyl)triacetic acid (compound 1)52#583961Attorney Docket No. 00020.026.1801[000143] To a solution of (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-1-yl)-4-oxobutyl)-1-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide 12 (4.78 g, 4.12 mmol) in acetonitrile (48 mL, 10 Vol), 10% MeOH: DCM (48 mL) at room temperature, was added DIPEA (6.57 mL, 37.7 mmol) followed by 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid DOTA NHS ester (3.55 g, 7.08 mmol). Resultant reaction was stirred at room temperature for Ih. Upon completion of the reaction, reaction was concentrated under reduced pressure to afford crude. Crude was triturated with EtOAc (50 mL), resulted material was purified by grace reverse 53#583961Attorney Docket No. 00020.026.1801phase, eluted at 45-50% CH₃CN: H₂O. Required fractions were concentrated under reduced pressure and further lyophilized to afford 2,2'.2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l -yl)-4-((S)-l -((4-((4-(4-iodophenyl) butanamido) methyl)benzoy l)gly cy l)py rrolidine-2-carboxamido)- 1 -oxobutan-2-y l)amino)-2-oxoethyl)- 1,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 1) (2.4 g, yield: 37%) as a white solid. MS (ESI) m / z:1546.15 [M+H]+. ‘H-NMR (400 MHz. DMSO-d6): 5 8.57 (brs, 1H), 8.39 (brs, 1H), 8.31-8.29 (m, 1H), 8.05-8.04 (m, 1H), 7.85-7.80 (m, 2H), 7.65-7.62 (m, 2H), 7.30 (d, J= 6.0 Hz, 2H), 7.22-7.15 (m, 2 H), 7.05-7.00 (m, 3H), 5.11-5.00 (m, 1H), 4.90-4.80 (m, 2H), 4.72-4.60 (m, 3H), 4.50-4.20 (m, 4H), 4.20-4.00 (m, 4H), 3.95-3.80 (m, 2H), 3.70-3.60 (m, 2H), 3.60-3.50 (m, 8H), 3.20-3.10 (m, 4H), 3.10-2.90 (m, 13H), 2.85-2.75 (m, 8H). 2.45-2.40 (m, 4H), 2.30-2.25 (m, 2H), 2.15-2.10 (m, 3H), 1.95-1.90 (m, 1H), 1.75-1.70 (m, 6H), 1.60-1.50 (m, 3H), 1.20-0.90 (m, 2H).Step-1 A: Synthesis of tert-butyl 4-bromoisoindoline-2-carboxylate (2A):[000144] To a solution of 4-bromoisoindoline 1A (1 g, 0.005 mol) in 2-MeTHF (10 mL, 10 Vol) at 0 °C was added Et₃N (2.9 mL, 0.02 mol) and (Boc)2O (1.47 mL, 0.0067 mol).Reaction mixture was stirred at room temperature for 2h. Upon completion of the reaction, reaction was diluted with EtOAc (30 mL). water (30 mL) and stirred for 15 min. Organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 30 mL).Combined organic layer was washed with water (30 mL). Separated organic layer was dried over anhydrous sodium sulphate, fdtered and concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 5% EtOAc in pet ether as an eluent. Required fraction were concentrated under reduced pressure 54#583961Attorney Docket No. 00020.026.1801to afford tert-buty l 4-bromoisoindoline-2-carboxylate 2A (1.14 g, yield: 76%) as a white solid.Step-2A: Synthesis of tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate (4A):[000145] To a solution of tert-butyl 4-bromoisoindoline-2-carboxylate 2A (500 mg, 1.67 mmol) in DMF:H₂O (9:1, 10 vol) was added 4-((4.4.5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)methylene)piperidine 3A (561.5 mg, 2.51 mmol) followed by Cs₂CO₃ (1.63 g, 5.03 mmol). The reaction mixture was degassed with argon for 15 min followed by addition of XPhos Pd G4 (80 mg, 0.67 mmol), resultant reaction mixture was stirred at 100°C for 16 h. Upon completion of the reaction, reaction mixture was filtered through celite bed washed with ethyl acetate (20 mL), filtrate was diluted with water (30 mL). The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 30 mL). Combined organic layers was washed with water (30 mL). Separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 10% MeOH in DCM. Required fractions were concentrated under reduced pressure to afford tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate 4A (0.415 g, yield: 79%) as a brown solid. MS (ESI) m / z: 315.1[M+H]+Step-3A: Synthesis of lert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylale (Inl45A)#583961Attorney Docket No. 00020.026.1801[000146] To a solution of tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate 4A (680 mg, 2.162 mmol) in MeOH (3 mL) was added 10% Pd / C (680 mg, wt / wt). Resultant reaction mixture was stirred at room temperature for 16 h under hydrogen pressure (30 psi). Upon completion of the reaction, reaction mixture was filtered through celite bed, washed with MeOH (20 mL) and filtrate was concentrated under reduced pressure to get tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate (Int#5A) (523 mg, crude) as an off white solid. MS (ESI) m / z: 317.09 [M+H]+Step-1B: Synthesis of 4-((4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methylene)piperidine[000147] To a solution of tert-butyl 4-((4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methylene)piperidine-1-carboxylate 1B (1 g, 0.003 mmol) in DCM (10 mL, 10 vol), was added 4N HC1 in 1,4-dioxane (6 mL, 6 vol) at 0 °C. Reaction was stirred at room temperature for Ih. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude (1.3 g). Crude was triturated with diethyl ether to afford 4-((4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methylene)piperidine 3A (635 mg, yield: 92%) as an off white solid. MS (ESI) m / z: 224 [M+H]+Synthesis of Int-1:Step-1A: 1-(tert-butyl) 2-(2,5-dioxopyrrolidin-1-yl) (S)-pyrrolidine-1,2-dicarboxylate (3A)56#583961Attorney Docket No. 00020.026.1801[000148] To a solution of (tert-butoxycarbonyl)-L-proline 1A (20 g, 93.00 mmol) in DMF (200 mL, 10 Vol) at 0 °C, was added DIPEA (32 mL. 186.04 mmol) followed by HBTU (38 g, 102.32 mmol). The mixture was stirred at room temperature for 30 min, followed by addition of 1 -hydroxypyrrolidine-2, 5-dione 2A (16 g, 139.50 mmol). Resultant reaction was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min. Precipitated solid was filter, wash with hexane (2* 100 mL) to afford 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-l-yl) (S)-pyrrolidine-l,2-dicarboxylate 3 A (22 g, yield: 75 %) as a white solid.Step-2A: (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)butanoic acid (5A)[000149] To a solution of 1 -(tert-butyl) 2-(2.5 -di oxo py nolidin- 1-yl) (S)-pyrrolidine-l,2-dicarboxylate 3A (10 g, 32.05 mmol) in DCM (100 mL, 10 Vol) at 0 °C, was added DIPEA (11 mL, 64.10 mmol) followed by (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-aminobutanoic acid 4A (10.8 g, 32.05 mmol). Resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was diluted with water (200 mL) and extract with 10% MeOH: DCM (2* 100 mL). Finally organic layers was separated and dried over Na₂SO₄ and concentrated to obtained (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)butanoic acid 5A (15 g, yield: 87%) as a brown gummy. MS (ESI) m / z: 538.8 [M+H]+Step-3A: (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid (Int-1)[000150] TFA (30 mL, 3 vol) was added to a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)butanoic acid (10 g, 0.018 mol) in DCM (100 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (100 mL) to afford (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid Int-1) (5 g, yield: 64%) as an off white solid. MS (ESI) m / z: 438.47 [M+H]+Example 2: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4- 57#583961Attorney Docket No. 00020.026.1801iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 2).Step-1: Synthesis of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate (3):[000151] To a solution of 4-(4-iodophenyl)butanoic acid 1 (30 g, 0.103 mol) in DMF (60 mL, 2 Vol) at 0°C was added DIPEA (27 mL, 0.155 mol) followed by HATU (39.3 g, 0.103 mol). The reaction mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl 4-(aminomethyl)benzoate 2 (21.4 g, 0.103 mol). Resultant reaction mixture was stirred at room temperature for 4 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (200 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 400 mL). Combined organic layer was washed with water (400 mL). The separated organic layer was dried over anhydrous sodium sulphate, fdtered and concentrated under reduced pressure to afford tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (36.1 g, crude) as a brown liquid. MS (ESI) m / z: 480.1 [M+H]+Step-2: Synthesis of 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid (4):58#583961Attorney Docket No. 00020.026.1801[000152] TFA (160 mL, 4 vol) was added to a solution of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (40 g, 0.083 mol) in DCM (200 mL, 5 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 5 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude, crude was triturated with diethyl ether (200 mL) to afford 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid 4 (31 g, yield: 88%) as a brown solid. MS (ESI) m / z: 424.42 [M+H]+Step-3: Synthesis of tert-butyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycinate[000153] To a solution of 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid (4) (10 g, 23.64 mmol) in DMF (100 mL, 10 Vol) at 0°C, was added DIPEA (12.3 mL, 70.92 mmol ) followed by HATU (13.4 g, 35.46 mmol). The mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl glycinate 5 (3.5 mL, 26.00 mmol). Resultant reaction was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min, precipitated solid was filtered and wash with hexane (2* 100 mL) to afford tert-butyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycinate 6 (10 g, yield: 83 %) as a white solid. MS (ESI) m / z: 538.28 [M+H]+.Step-4: Synthesis of (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine (7):#583961Attorney Docket No. 00020.026.1801[000154] TFA (18 mL, 3 vol) was added to a solution of tert-butyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycinate 6 (6 g, 0.011 mol) in DCM (60 mL, 10 Vol) at 0 °C. Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (100 mL) to afford (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine 7 (4.5 g, yield: 86%) as a white solid. MS (ESI)m / z: 481.13 [M+H]+Step-5: Synthesis of (S)-2-( ( f9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-( ( 4-( 4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid (8):[000155] To a solution of (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine 7 (2.5 g, 5.20 mmol) in DMF (25 mL, 10 Vol) at 0°C was added DIPEA (1.3 mL, 7.81 mmol) followed by addition of HATU (2.17 g, 5.72 mmol). The reaction mixture was stirred at room temperature for 10 min followed by addition of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid Int#l (2.27 g, 5.20 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted with cold water (100 mL) and stirred for 15 min, obtained solid was dissolve in 10% MeOH: DCM (20 mL) then triturated with diethyl ether (100 mL) to afford (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid 8 (3.6 g, yield: 78%) as a white solid. MS (ESI) m / z: 900.47 [M+H]+.Step-6: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-(3-((4-((2-((S)-2-cyano-4,4-difl uoropyrrolidin- 1 -yl) -2-oxoethyl) carbamoyl) quinolin-6- 60#583961Attorney Docket No. 00020.026.1801yl)(methyl)amino)propyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (9):[000156] To a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l- ((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2- carboxamido)butanoic acid 8 (1.6 g, 1.78 mmol) in DMF (16 mL, 10 Vol) at 0°C, was added DIPEA (0.92 mL, 5.34 mmol) followed by HATU (1.35 g, 3.56 mmol). Reaction mixture was stirred at room temperature for 10 min, followed by addition of (S)-N-(2-(2-cyano-4,4- difluoropyrrolidin- 1 -yl)-2-oxoethyl)-6-(methyl(3-(piperazin-l -yl)propyl)amino)quinoline-4- carboxamide Int-1 (1.3 g, 2.66 mmol). Resultant reaction mixture was stirred at room temperature for 2h. Upon completion of the reaction, reaction mixture was diluted with cold water (100 mL) and stirred for 15 min. Obtained white solid was dissolve in 10%MeOH: DCM (20 mL) then triturated with DEE (100 mL) to afford (9H-fluoren-9-yl)methyl ((S)-l-(4-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-l-yl)-2- oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 9 (1.5 g, yield: 65 %) as a reddish gummy. MS (ESI) m / z:1382 (M+H)+.Step-7: Synthesis of 6-((3-(4-((S)-2-amino-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperazin-l-yl)propyl)(methyl)amino)-N-(2-((S)-2-cyano-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)quinoline-4-carboxamide (10):[000157] Diethyl amine (4.8 mL, 4 Vol) was added to a solution of (9H-fluoren-9-yl)methyl ((S)-l-(4-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-l-yl)-2-#583961Attorney Docket No. 00020.026.1801oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl) butanamido) methyl) benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 9 (1.5 g, 1.08 mmol) in acetonitrile (15 mL, 10 Vol) at room temperature. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude. Crude material was triturated with cold EtOAc (10 mL) followed by diethyl ether (10 mL) to afford 6-((3-(4-((S)-2-amino-4-((S)-l-((4-((4-(4-iodophenyl) butanamido) methyl) benzoyl) glycyl) pyrrolidine-2-carboxamido)butanoyl)piperazin-l-yl)propyl)(methyl)amino)-N-(2-((S)-2-cyano-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)quinoline-4-carboxamide 10 (0.9 g, yield: 72%) as a white solid. MS (ESI) m / z: 1160 (M+H)+.Step-8: Synthesis of 2.2', 2"-( 10-(2-( ( (S)-l-( 4-(3-( (4-((2-( (S)-2-cyano-4, 4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazin-1-yl)-4-((S)-1-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-1-oxobutan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid[000158] To a solution of 6-((3-(4-((S)-2-amino-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperazin-l-yl)propyl)(methyl)amino)-N-(2-((S)-2-cyano-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)quinoline-4-carboxamide (10) (200 mg, 0.17 mmol) in 62#583961Attorney Docket No. 00020.026.1801acetonitrile (4 mL, 10 Vol) at room temperature, was added DIPEA (0.06 mL, 0.34 mmol) followed by 2,2'.2"-( 10-(2-((2.5 -dioxopyrrolidin- 1 -y l)oxy )-2-oxoethy 1)- 1,4,7, 10-tetraazacyclododecane-l,4,7-triyl)triacetic acid DOTA NHS ester (130 mg, 0.25 mmol). The resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. The crude material was triturated with EtOAc (20 mL) then obtained material was purified by grace reverse phase, eluted at 45-50% CH₃CN: H₂O. Required fractions were concentrated under reduced pressure and further lyophilized to afford 2,2',2"-(10-(2-(((S)-l-(4-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-y l)(methyl)amino)propy l)piperazin- 1 -y 1) -4 - ((S ) - 1 -((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7, 10-tetraazacyclododecane- 1,4, 7-triy l)tri acetic acid (compound 2) (20 mg, yield: 8.8%) as a white solid. MS (ESI) m / z: 1546.2[M+H]+. ‘H-NMR (400 MHz, DMSO-ds): 59.2-8.9 (brs, 1H), 8.7-8.6 (brs, 1H), 8.56 (d, J = 4.4 Hz, 1H), 8.4-8.2 (m, 3H), 7.9-7.80 (m, 3H), 7.7-7.62 (m. 2H), 7.55-7.45 (m. 1H), 7.40-7.32 (m. 4H), 7.0-6.9 (m, 2H), 5.13-5.11 (m, 1H). 4.7-4.6 (m. 1H), 4.4-4.0 (m, 10H). 3.70-3.50 (m, 1 OH), 3.50-3.30 (m. 15H), 3.20-3.1 (m, 2H), 3.1-2.9 (m, 12H), 2.8-2.70 (m, 5H), 2.30-2.25 (m, 4H), 2.2-2.10 (m, 2H), 1.2-1.90 (m, 1H), 1.9-1.6 (m, 10H).Synthesis of Int-1[000159] To a solution of 6-bromoquinoline-4-carboxylic acid 1 (10 g, 39.7 mmol), in t-BuOH (37.6 mL. 10 vol), at 0° C was added DMAP (5.8 g, 47.6 mmol) followed by Boc anhydride (18.2 mL, 79.36 mmol). Resultant reaction mixture was stirred at room63#583961Attorney Docket No. 00020.026.1801temperature for 16 h. Progress of reaction was monitored by TLC. Upon completion of reaction, the reaction mixture was concentrated under reduced pressure to afford crude. Crude was purified by silica gel column chromatography (100-200 mesh), eluted with 10% MeOH in DCM to afford / e / 7- butyl 6-bromoquinoline-4-carboxylate 2 (8.5 g) as a blue solid. MS (ESI) m / z: 308.2 [M+H]+.Step-2B: Synthesis of tert-butyl 6-((3-(4-(tert-butoxycarbonyl)piperazin-l-yl)propyl) (methyl)amino)quinoline-4-carboxylate ( 4)[000160] To a solution of tert-butyl 6-bromoquinoline-4-carboxylate 2 (2 g, 6.51 mmol), in 1,4-dioxane (60 mL, 10 Vol), at room temperature was added tert-butyl 4-(3-(methylamino)propyl)piperazine-l -carboxylate 3 (3 g, 11.7 mmol), CS2CO3 (4.2 g, 13 mmol) and Xphos (310 mg, 0.65 mmol) and Pd2(dba)3 (298 mg, 0.32 mmol). Resultant reaction mixture was stirred at 90 °C for 16 h. Progress of reaction was monitored by TLC. Upon completion of reaction, the reaction mixture was concentrated under reduced pressure to afford crude. Crude was purified by silica gel column chromatography (100-200 mesh) eluted at 5% MeOH in DCM and compound contained fractions were concentrated under reduced pressure to afford tert-butyl 6-((3-(4-(tert-butoxycarbonyl)piperazin-l-yl)propyl)(methyl)amino)quinoline-4-carboxylate (4) (2 g, Yield: 64%) as a red gummy. MS (ESI) m / z: 485.3 [(M+H]+.Step-3B: Synthesis of 6-(methyl(3-(piperazin-l-yl)propyl)amino)quinoline-4-carboxylic acid(5)[000161] To a solution of tert-butyl 6-((3-(4-(tert-butoxycarbonyl)piperazin-l-yl)propyl)(methyl)amino)quinoline-4-carboxylate (4) (3.5 g, 7.23 mmol) in TFA (35 mL, 10 vol). Resultant reaction mixture was stirred at 0 °C to room temperature for 4 h. Progress of reaction was monitored by TLC. Upon completion of reaction, the reaction mixture was 64#583961Attorney Docket No. 00020.026.1801concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (80 mL) and 10% MeOH in DCM (10 mL) to afford 6-(methyl(3-(piperazin-l-yl)propyl)amino)quinoline-4-carboxylic acid (5) (4 g, Crude) as a reddish gummy. MS (ESI) m / z: 329 [M+H]⁺Step-4B: Synthesis of 6-((3-(4-(tert-butoxycarbonyl)piperazin-l-yl)propyl) (methyl) amino) quinoline-4-carboxylic acid ( 6)[000162] To a solution 6-(methyl(3-(piperazin-l-yl)propyl)amino)quinoline-4-carboxylic acid (5) (3.5 g. 10.6 mmol) in THF (35 mL. 20 vol), at 0 °C was added NaHCO₃ (4.4 g, 3.6 mmol) followed by Boc anhydride (3.6 mL, 16 mmol). Resultant reaction mixture was stirred at room temperature for 3 h. Progress of reaction was monitored by TLC. Upon completion of reaction, the reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (25 mL) and 10% MeOH in DCM (5 mL) to afford 6-((3-(4-(tert-butoxycarbonyl)piperazin-l-yl)propyl)(methyl)amino)quinoline-4-carboxylic acid (6) (4 g) as a red gummy. MS (ESI) m / z: 429 [M+H]+.Step-5B: Synthesis of tert-butyl (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylate (7)[000163] To a solution of 6-((3-(4-(tert-butoxycarbonyl)piperazin-l-yl)propyl)(methyl)amino)quinoline-4-carboxylic acid (6) (1.5 g, 3.5 mmol) in DMF (15 mL, 10 Vol), at 0 °C was added DIPEA (1.8 mL, 10.5 mmol) followed by (S)-pyrrolidine-2-carbonitrile (Int-3) (1.3 g, 7 mmol) stirred for 15 min, and added HATU (2.6 g, 2 mmol). Resultant reaction mixture was stirred at room temperature for 30 min. Progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was poured into ice water (50 mL), compound was extracted with 10% MeOH in DCM (2 x 30 mL), dried over sodium sulphate, concentrated under reduced pressure to afforded crude. Crude was 65#583961Attorney Docket No. 00020.026.1801purified by silica gel column chromatography (100-200 mesh) eluted at 5% MeOH in DCM and compound contained fractions were concentrated under reduced pressure to afford tertbutyl (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-l -carboxylate (7) (1.3 g, yield: 65%) as a red solid compound. MS (ESI) m / z: 600.3 [M+H]+.Step-6B: Synthesis of (S)-N-(2-(2-cyano-4.4-difluoropyrrolidin-l-yl)-2-oxoethyl)-6-(methyl(3-(piperazin-l-yl)propyl)amino)quinoline-4-carboxamide (Int-1)[000164] To a solution of tert-butyl (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-l-carboxylate (7) (2 g, 3.33 mmol) in TFA (10 mL, 10 vol). Resultant reaction mixture was stirred at room temperature for 2 h. Progress of reaction was monitored by TLC. Upon completion of reaction, the reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (80 mL) and 10% MeOH in DCM (10 mL) to afford (S)-N-(2-(2-cyano-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)-6-(methyl(3-(piperazin-l-yl) propyl) amino) quinoline-4-carboxamide (Int-1) (1.2 g, Yield: 75%) as a reddish solid. MS (ESI) m / z: 500 [M+H]’.Synthesis of Int-3:Step-1 & 2: Synthesis of tert-butyl (S)-(2-(2-cyano-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamate (4)66#583961Attorney Docket No. 00020.026.1801[000165] To a solution of (tert-butoxycarbonyl)glycine 2 (5.3 g, 28.57 mmol) in THF (20 mL, 10 Vol) at 0 °C was added T3P 50% in EtOAc (13.6 g. 42.85 mmol) stirred for 10 min and added (S)-4,4-difluoropyrrolidine-2-carboxamide hydrochloride 1 (5 g, 28.54 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of starting material and formation of intermediate by TLC, again added T3P (6.8 mL) and the reaction mixture was heated to 60 °C for 16 h. Progress of the reaction was monitored by TLC. Reaction mixture was diluted with ethyl acetate (100 mL) and ice water (50 mL), organic layer was separated and the aqueous layer was extracted with ethyl acetate (2 *25 mL). Organic layer separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford crude. Crude was purified by silica gel column chromatography (60-100 mesh) using 30% ethyl acetate in hexane as an eluent. Required fractions were concentrated under reduced pressure to afford tert-butyl (S)-(2-(2-cyano-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamate (4) (4 g, crude) as a colour less thick liquid. MS (ESI) m / z: 290.08 (M+H)+.Step-3: Synthesis of (S)-4,4-difluoro-l-glycylpyrrolidine-2-carbonitrile (5):[000166] To a solution of tert-butyl (S)-(2-(2-cyano-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamate (4) (2 g, 6.9 mmol) in DCM (20 mL, 10 vol) at 0 °C, was added TFA (8 mL, 4 vol). Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude. Crude was triturated with diethyl ether to afford (S)-4,4-difluoro-l-glycylpyrrolidine-2-carbonitrile (5) (1.5 g) as a gummy. MS (ESI) m / z: 190.2 [M+H]+.Example 3: Synthesis of 2,2’,2"-(10-(2-(((S)-l-(4-(2-((4-((2-((S)-2-(2-((3,4-dimethoxybenzyl)amino)-2-oxoacetyl)-4,4-difluoropyrrolidin-l-yl)-2- 67#583961Attorney Docket No. 00020.026.1801oxoethyl)carbamoyl)quinolin-6-yl)oxy)acetyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl) butanamido) methyl) benzoyl) glycyl)pyrrolidine-2-carboxamido)-1-oxobutan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (compound 3)Step-1: Synthesis of benzyl (2-((2S)-2-(2-((3,4-dimethoxybenzyl)amino)-l-hydroxy-2-oxoethyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamate (4):[000167] To a solution tert-butyl (S)-4,4-difluoro-2-formylpyrrolidine-l -carboxylate (1) (2 g, 8.15 mmol) in DCM (20 mL. 10 Vol) at room temp was added ((benzyloxy) carbonyl)glycine (2) (1.7 g 8.15 mmol) and 4-(isocy anomethyl)- 1,2-dimethoxy benzene (3) (1.5 g, 8.510 mmol). Reaction mixture was stirred at room temperature for 3 h, followed by addition of TFA (10 mL 5 vol) and reaction was stirred at room temperature for 1 h. Upon completion starting material, reaction mixture was concentrated under vacuum, residue was re-dissolved in DCM (20 mL) and added Et₃N (25 mL, 5 vol). Resultant reaction mixture was stirred at room temperature for Ih. Progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was diluted with DCM (50 mL), water (50 mL) and stirred for 15 min, aqueous layer was extracted with DCM (2 * 50 mL), combined68#583961Attorney Docket No. 00020.026.1801organic layers was washed with water (50 mL). The organic phase separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by column chromatography using silica gel (100-200 mesh), eluted in 50% EtOAc in hexane, collected fraction were concentrated under reduced pressure to afford benzyl(2-((2S)-2-(2-((3,4-dimethoxybenzyl)amino)-1-hydroxy-2-oxoethyl)-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamate (4) (1.3 g. yield: 29.5%) as a red gummy. MS (ESI) m / z: 522.3 [M+H]+.Step-2: Synthesis of 2-((S)-4,4-difluoro-l-glycylpyrrolidin-2-yl)-N-(3,4-dimethoxybenzyl)-2-[000168] To a solution of benzyl (2-((2S)-2-(2-((3,4-dimethoxybenzyl)amino)-l-hydroxy-2-oxoethyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamate (4) (3 g. 5.75 mmol) in MeOH (3 mL, 10 Vol) at room temperature, was added Pd / C (1.5 g, 50%w / w). Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was filtered, filtrate was concentrated under reduced pressure to afford 2-((S)-4,4-difluoro-l-glycylpyrrolidin-2-yl)-N-(3,4-dimethoxybenzyl)-2-hydroxy acetamide (5) (2 g, yield: 90%) as a brown gummy. MS (ESI) m / z: 388.4 [M+H]+.Step-3: Synthesis of tert-butyl 4-(2-((4-((2-((2S)-2-(2-((3,4-dimethoxybenzyl)amino)-l-hydroxy-2-oxoethyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-#583961Attorney Docket No. 00020.026.1801[000169] To a solution of 6-(2-(4-(tert-butoxycarbonyl)piperazin-l-yl)-2-oxoethoxy)quinoline-4-carboxylic acid (Int#3B) (139 mg, 0.361mmol) in DMF (1.5 mL, 10 Vol) at 0°C, were added DIPEA (0.13mL, 0.72 mmol) followed by HATU (411 mg, 1.083 mmol). Reaction mixture was stirred at room temperature for 10 min, and added 2-((S)-4,4-difluoro-l-glycylpyrrolidin-2-yl)-N-(3,4-dimethoxybenzyl)-2-hydroxy acetamide (5) (150 mg, 0.361 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. Progress of the reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was quenched with ice cold water, solid obtained was filtered and re-dissolved in 10% MeOH in DCM, dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was triturated with diethyl ether (3* 20 mL) to afford tert-butyl 4-(2-((4-((2-((2S)-2-(2-((3,4-dimethoxybenzyl)amino)-l-hydroxy-2-oxoethyl)-4.4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)acetyl)piperazine-l-carboxylate (6) (90 mg, yield: 31 %) as a white solid. MS (ESI) m / z: 785.6 [M+H]+.Step-4: Synthesis ofN-(2-((2S)-2-(2-((3,4-dimethoxybenzyl)amino)-l-hydroxy-2-oxoethyl)-4.4-difluoropyrrolidin-l-yl)-2-oxoethyl)-6-(2-oxo-2-(piperidin-4-yl)ethoxy)quinoline-4-[000170] To a solution of tert-butyl 4-(2-((4-((2-((2S)-2-(2-((3,4-dimethoxy benzy l)amino)- 1 -hydroxy-2-oxoethyl)-4,4-difluoropyrrolidin- 1 -yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)acetyl)piperidine-l -carboxylate (6) (800 mg 0.893 mmol) in DCM (8mL, 10 Vol), at 0°C was added TFA ( 3.2 mL, 5vol ). Resultant reaction mixture was stirred at room temperature for 2 h. Progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude. Crude was triturated with diethyl ether (5 mL) to get N-(2-((2S)-2-(2-((3,4-dimethoxybenzyl)amino)-l -hydroxy -2-oxoethyl)-4,4-difluoropyrroli din-1 -yl)-2-oxoethyl)-6-(2-oxo-2-(piperidin-4-yl)ethoxy)quinoline-4-carboxamide (Int#2) (600 mg, yield: 86%) as a white solid. MS (ESI) m / z: 685.4 [M+H]+.70#583961Attorney Docket No. 00020.026.1801Step-5: Synthesis of (9H-fluoren-9-yl)methyl ((2S)-l-(4-(2-((4-((2-(2-(2-((3,4-dimethoxybenzyl)amino)-l-hydroxy-2-oxoethyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)acetyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)henzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-[000171] To a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid (Int#8) (300 mg, 0.33 mmol) in DMF (3 mL, 10 Vol), at 0°C was added DIPEA (0.12 mL, 0.66 mmol) followed by HATU (253 mg. 0.66 mmol). The reaction mixture was stirred for 10 min, and added N-(2-(2-(2-((3,4-dimethoxybenzyl)amino)ethyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)-6-(2-oxo-2-(piperazin-l-yl)ethoxy)quinoline-4-carboxamide (Int#2) ( 273 mg, 0.40 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. Progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was diluted with ice cold water (20 mL). stirred for 15 min, solid obtained was dissolved in 10% MeOH in DCM organic layer was washed with ice cold water (30 mL), dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was triturated with diethyl ether (2*20 mL ), dried under reduced pressure to afford (9H-fluoren-9-yl)methyl ((2S)-l-(4-(2-((4-((2-(2-(2-((3.4-dimethoxybenzyl)amino)-l-hydroxy-2-oxoethyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoy l)quinolin-6-yl)oxy)acetyl)piperazin- 1 -yl)-4-((S)- 1 -((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (7) (300 mg, yield: 67%) as a white solid. MS (ESI) m / z: 1507.8 [M+H]+. Step-6: Synthesis of (9H-fluoren-9-yl)methyl ((2S)-l-(4-(2-((4-((2-(2-(2-((3,4-dimethoxybenzyl)amino)-2-oxoacetyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)acetyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (8):71#583961Attorney Docket No. 00020.026.1801DMP, DCM0 °C to RT, 2 h Step-6[000172] To a solution of (9H-fluoren-9-yl)methyl ((2S)-l-(4-(2-((4-((2-(2-(2-((3,4-dimethoxy benzy l)amino)-l-hydroxy-2-oxoethyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)acetyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (7) (250 mg, 0.159 mmol) in DCM (5 mL, 20 Vol), followed by addition of DMP at 0°C (203 mg, 0.47 mmol), resultant reaction was stirred at room temperature for 2h. Progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was fdter through celite, concentrated under reduced pressure to afford crude. Crude triturated with diethyl ether (2*5 mL ) to afford (9H-fluoren-9-yl)methyl ((2S)-l-(4-(2-((4-((2-(2-(2-((3,4-dimethoxy benzy l)amino)-2-oxoacetyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoy l)quinolin-6-yl)oxy )acetyl)piperazin- 1 -yl)-4-((S)- 1 -((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (8) (150 mg, yield: 60 %) as a white solid. MS (ESI) m / z: 1265 [M+H]+.Step-7: Synthesis of 6-(2-(4-((S)-2-amino-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperazin-l-yl)-2-oxoethoxy)-N-(2-((S)-2-(2-((3,4- 72#583961Attorney Docket No. 00020.026.1801dimethoxybenzyl)amino)-2-oxoacetyl)-4,4-difluoropyrrolidm-l-yl)-2-oxoethyl)quinoline-4-[000173] To a solution of (9H-fluoren-9-yl)methyl ((2S)-l-(4-(2-((4-((2-(2-(2-((3,4-dimethoxybenzyl)amino)-2-oxoacetyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)acetyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (8) (270 mg, 0.17 mmol) in acetonitrile (1.3 mL) at 0°C was added 30% piperidine in DMF (7 mL, 10 vol), resultant reaction mixture was stirred at room temperature for 2 h. Progress of reaction was monitored by TLC, upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (2*20 mL) to afford 6-(2-(4-((S)-2-amino-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperazin-l-yl)-2-oxoethoxy)-N-(2-((S)-2-(2-((3,4-dimethoxybenzyl)amino)-2-oxoacetyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)quinoline-4-carboxamide (9) (200 mg, yield: 86% ) as a brown solid MS (ESI) m / z: 1342 [M+H]+.73#583961Attorney Docket No. 00020.026.1801Step-8: Synthesis of O-(tert-butyl) S-(2-(((S)-l-(4-(2-((4-((2-((S)-2-(2-((3,4-dlmethoxybenzyl)amino)-2-oxoacetyl)-4.4-difluoropyrrolidm-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)acetyl)piperidin-l-yl)-6-(4-(4-iodophenyl)butanamido)-l-oxohexan-2-yl)amino)-2-oxoethyl) carbonothioate (compound 3):[000174] To a solution of 6-(2-(4-((S)-2-amino-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperazin-l-yl)-2-oxoethoxy)-N-(2-((S)-2-(2-((3,4-dimethoxybenzyl)amino)-2-oxoacetyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)quinoline-4-carboxamide (9) (800 mg, 0.59 mmol) in acetonitrile (8 mL, 10 Vol) at room temperature, was added DIPEA (0.9 mL, 5.36 mmol) followed by 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid DOTA NHS ester (0.746 mg, 1.43 mmol). Resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was concentrated under reduced74#583961Attorney Docket No. 00020.026.1801pressure to afford crude. Crude was triturated with EtOAc (20 mL) and purified by prep-HPLC using below condition.Prep conditionColumn GEMINI C18 (250*21.5 mm) 5µ Buffer A:0.2%FA in H2O, B: 0.2% FA ACN: H2O (80:20)Gradient (T / %B) 0 / 35,15 / 55,17 / 55,17. l / 98@12mL / min Flow Rate 12 mL / minSolubility ACN+THF+H2O[000175] The required fractions were collected and lyophilized to afford O-(tert-butyl) S-(2-(((S)-l-(4-(2-((4-((2-((S)-2-(2-((3,4-dimethoxybenzyl)amino)-2-oxoacetyl)-4,4-difluoropyrrolidin-l-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)acetyl)piperidin-l-yl)-6-(4-(4-iodopheny 1) butanamido)- 1 -oxohexan-2-y l)amino)-2-oxoethy 1) carbonothioate (compound 3) (101 mg. yield: 10%) as a white solid. MS (ESI) m / z: 1729.16 [M+H]+.[000176] ¹H-NMR (400 MHz, DMSO-d6): δ 13-12 (brs,1H), 9.1-90 (brs, 1H), 8.85-8.82 (m, 1H), 8.7-8.6 (m, 1H), 8.5-8.2 (m, 3H), 8.1-8.0 (m, 1H), 7.9-7.8 (m, 3H), 7.7-7.6 (m, 2H), 7.5-7.4 (m, 2H), 7.4-7.3 (m, 2H), 7.0-6.5 (m, 5H), 5-4.5 (m, 3H), 4.5-4 (m, 11H), 3.7-3.65 (m, 6H), 3.65-3.6 (m. 2H), 3.6-3.5 (m, 11 H), 3.2-3. l(m. 4H), 3.1-2.9 (m, 16 H), 2.8-2.7 (m, 4H), 2.4-2.3 (m, 4H), 2.2-2.1 (m, 2H), 2-1.6 (m, 10 H)Intermediate synthesis (Int#3B)Step-1 A: Synthesis of 2-(4-(tert-butoxycarbonyl)piperazin-l-yl)-2-oxoethyl 6-(2-(4-(tert-butoxycarbonyl)piperazin-l-yl)-2-oxoethoxy)quinoline-4-carboxylate (Int#2B)#583961Attorney Docket No. 00020.026.1801[000177] To a solution of 6-hydroxyquinoline-4-carboxylic acid (SM-02) (3 g, 15.87 mmol) in DMF (30 mL, 10 vol) was added tert-butyl 4-(2-bromoacetyl)piperazine-l-carboxylate (SM-01) (12 g, 39.68 mmol) followed by addition of K.2CO3 (6.5 g, 47.6 mmol). Resultant reaction mixture was stirred at room temperature for 6 h. Progress of reaction was monitored by TLC.[000178] The reaction mixture was diluted with ice water (20 mL) and filtered the solid, washed with diethyl ether (20 mL) to afford 2-(4-(tert-butoxycarbonyl)piperazin-l-yl)-2-oxoethyl 6-(2-(4-(tert-butoxycarbonyl)piperazin-l-yl)-2-oxoethoxy)quinoline-4-carboxylate (Int#2B) (2.6 g, yield: 30% ) as a brown solid. MS (ESI) m / z: 642 [M+H]+.Step-2A: Synthesis of 6-(2-(4-(tert-butoxycarbonyl)piperazin-l-yl)-2-oxoethoxy)quinoline-4-[000179] To a solution of 2-(4-(tert-butoxycarbonyl)piperazin-l-yl)-2-oxoethyl 6-(2-(4-(tert-butoxycarbonyl)piperazin-l-yl)-2-oxoethoxy)quinoline-4-carboxylate (Int#2B)[000180] (7 g, 10.9 mmol) in THF (35 mL, 5Vol) and water (35 mL, 5Vol) was added KOH (1.2 g, 21.84 mmol). Resultant reaction mixture was stirred at 50°C for 2 h. Progress of reaction was monitored by TLC, upon completion of the reaction, reaction mixture was diluted with ice water (50 mL) and washed with ethyl acetate (50 mL). Separated aqueous layer was acidified with IN HC1, obtained solid was filtered and washed with diethyl ether (2*20 mL) to afford 6-(2-(4-(tert-butoxycarbonyl)piperazin-l-yl)-2-oxoethoxy)quinoline-4-carboxylic acid (Int#3B) (2 g, yield: 44% ) as a brown solid. MS (ESI) m / z: 416 [M+H]+.Example 4: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 4):76#583961Attorney Docket No. 00020.026.1801Step-1: Synthesis of (S)-2-( ( l(9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-( 4-( 4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)butanoic acid[000181] To a solution of 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid Int-2 (5 g, 0.011 mol) in DMF (50 mL, 10 Vol) at 0°C was added DIPEA (3 rnL, 0.017 mol) followed by HATU (4.9 g. 0.012 mol). The mixture w as stirred at room temperature for 30 min, and added (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-(L-prolyl)pyrrolidine-2-carboxamido)butanoic acid Int-1 (6.3 g, 0.011 mol). Resultant reaction mixture was stirred at room temperature for 2h. Upon completion of the reaction, reaction mixture w as diluted with EtOAc (50 mL), cold water (100 rnL) and stirred for 15 min. The organic layer w as separated, and aqueous layer was further extracted with EtOAc (2 x 400 mL). Combined organic layer was washed with water (400 rnL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford (S)-2- 77#583961Attorney Docket No. 00020.026.1801((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)butanoic acid (2) (6 g, Yield:54%) as a brown liquid. MS (ESI) m / z: 939.85 [M+H]+Step-2: Synthesis of tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4- ((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate (3):[000182] To a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)butanoic acid (2) (5 g, 0.005 mol) in DMF (50 mL, 10 Vol) at 0°C was added DIPEA (1.8 mL, 0.01 mol) followed by HATU (2.2 g, 0.005 mol, 1.1 eq). The mixture was stirred at room temperature for 15 min and added tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate (Int#5A) (1.6 g, 0.005 mol). Resultant reaction mixture was stirred at room temperature for 3h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (20 mL), ice cold w ater (50 mL) and stirred for 15 min. The aqueous layer was extracted with EtOAc (2 * 25 mL), combined organic layer was washed with ice cold water (2 * 25 mL). The organic phase separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate (3) (5.7 g, yield: 87%) as a white solid. MS (ESI) m / z: 1239.84 (M+H)+.Step-3: Synthesis of (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate (4)78#583961Attorney Docket No. 00020.026.1801[000183] TFA (22.8 mL, 4 Vol) was added to a solution of tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate (3) (5.7 g, 0.0046 mol) in DCM (57 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room for 2 h. Reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (30 mL), to afford (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate (4) (3.5 g, yield:67%) a brown solid. MS (ESI) m / z: 1138.16 (M+H)+.Step-4: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carb oxamido)-l-oxobutan-2-yl) carbamate (5):[000184] To a solution of CDA-5 (1.1 g, 0.003 mmol) in DMF (1.1 mL, 10 Vol) at 0°C was added DIPEA (1.3 mL, 0.007 mmol) followed by addition of HATU (1.16 g, 0.003 mmol). The mixture was stirred at room temperature for 30 min and added (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate 4 (3.5 g, 0.003 mmol). The resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with 10% MeOH in DCM (25 mL) and ice water (20 mL). organic layer was separated and the aqueous 79#583961Attorney Docket No. 00020.026.1801layer was extracted with 10% MeOH in DCM (2 *25 mL). Organic layer separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 5 (3.7g, yield: 86%) as an off white solid. MS (ESI) m / z:1421.99(M+H)+.Step-5: Synthesis of (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-1-yl)-4-oxobutyl)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamide (6)[000185] Diethyl amine (14 mL, 4 Vol) was added to a solution (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 5 (3.5 g, 0.002 mol) in acetonitrile (35 mL, 10 Vol) at room temperature. Resultant reaction mixture was stirred at room temperature for 4 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude. Crude material was triturated with cold EtOAc (10 mL) followed by diethyl ether (10 mL) to afford 80#583961Attorney Docket No. 00020.026.1801(S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamide 6 (2 g, yield: 69%) as a white solid. MS (ESI) m / z: 1200.84 (M+H)+.Step-6: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidme-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)-l-oxobutcin-2-yl)ammo)-2-oxoethyl)-l, 4, 7, 10-tetraazacyclododecane-l, 4, 7-triyl)triacetic acid (Compound 4)81#583961Attorney Docket No. 00020.026.1801[000186] To a solution of (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamide 6 (2 g, 0.00166 mol) in acetonitrile (20 mL, 10 Vol), at room temperature, was added DIPEA (2.6 mL, 0.015 mmol) followed by 2,2',2"-(10-(2-((2,5-dioxopyrrolidin- 1 -yl)oxy)-2-oxoethyl)- 1,4,7, 10-tetraazacyclododecane- 1,4,7-triyl)triacetic acid DOTA NHS ester (1.25 g, 0.0025 mol). The resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude (430 mg crude). The crude material was triturated with EtOAc (20 mL) and further purified by prep HPLC using the below condition. Prep conditionColumn GEMINI Cl 8 (250*21.5mm)5uBuffer A:0.2% FA IN WATER, B:0.2 % FA in ACN: H20(80:20)Gradient (T / %B) 0 / 35,15 / 50,22 / 50,22.1 / 98Flow Rate 11mL / minSolubility ACN+H2O[000187] The required fractions were collected and lyophilized to afford 2.2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane-1.4,7-triyl)triacetic acid (compound 4) (215.7 mg, yield: 8.1% ) as a white solid. MS (ESI) m / z: 1585.98 [M+H]+. 'H-NMR (400 MHz, DMSO-d6): 58.5-8.2 (brs, 3H), 8.1-8.0 (Brs, 1H), 7.61 (d, J = 8Hz, 2H), 7.5-7.4 (m, 1H), 7.1-7.3 (m, 5 H), 7.1-7.05 (m, 1H), 7.00 (d, J= 8 Hz, 2H), 5.12-5.09 (m, 1H), 4.9-4.8 (m, 2H), 4.7-4.6 (m, 4H), 4.5-4.4(m. 1 H), 4.4-4.3 (m. 7H), 4.0-3.5 (m, 6H), 3.1-3.4 (m, 7H), 3.0-2.9 (m. 13H), 2.9-2.7 (m, 8 H), 2.6-2.5 (m, 2H). 2.4-2.3 (m. 4H), 2.2-2.0 (m, 7H), 2.0-1.5 (m, 15H), 1.2-1 (m, 2H)Synthesis of Int-2:OStep-1: Synthesis of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate (3):82#583961Attorney Docket No. 00020.026.1801[000188] To a solution of 4-(4-iodophenyl)butanoic acid 1 (30 g, 0.103 mol) in DMF (60 mL, 2 Vol) at 0°C was added DIPEA (27 mL, 0.155 mol) followed by HATU (39.3 g, 0.103 mol). The reaction mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl 4-(aminomethyl)benzoate 2 (21.4 g, 0.103 mol). Resultant reaction mixture was stirred at room temperature for 4 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (400 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 400 mL). Combined organic layer was washed with water (400 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (36.1 g, crude) as a brown liquid. MS (ESI) m / z: 480.1 [M+H]+Step-2: Synthesis of 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid (Int-2):[000189] TFA (160 mL, 4 vol) was added to a solution of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (40 g, 0.083 mol) in DCM (200 mL, 5 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 5 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (200 mL) to afford 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid Int-2 (31 g, yield: 88%) as a brown solid. MS (ESI) m / z: 424.42 [M+H]+Synthesis of Int#3A:Step- 1 A: Synthesis of tert-butyl 4-bromoisoindoline-2-carboxylate (2A):83#583961Attorney Docket No. 00020.026.1801[000190] To a solution of 4-bromoisoindoline 1A (1 g, 0.005 mol) in 2-MeTHF (10 mL, 10 Vol) at 0°C was added Et₃N (2.9 mL, 0.02 mol) and (Boc)2O (1.47 mL, 0.0067 mol). The reaction mixture was stirred at room temperature for 2h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (30 mL), water (30 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 30 mL). Combined organic layer was washed with water (30 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 5% EtOAc in pet ether as an eluent. Required fraction were concentrated under reduced pressure to afford tert-butyl 4-bromoisoindoline-2-carboxylate 2A (1.14 g, yield: 76%) as a white solid.Step-2A: Synthesis of tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate (4A):[000191] To a solution of tert-butyl 4-bromoisoindoline-2-carboxylate 2A (500 mg, 1.67 mmol) in DMF:H₂O (9:1, 10 vol) was added 4-((4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)methylene)piperidine 3A (561.5 mg, 2.51 mmol) followed by Cs₂CO₃ (1.63 g, 5.03 mmol). The reaction mixture was degassed with argon for 15 min followed by addition of XPhos Pd G4 (80 mg, 0.67 mmol), resultant reaction mixture was stirred at 100°C for 16 h. Upon completion of the reaction, reaction mixture was filtered through celite bed washed with ethyl acetate (20 mL), filtrate was diluted with water (30 mL). The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 30 mL). Combined organic layers was washed with water (30 mL). The separated organic layer was dried over 84#583961Attorney Docket No. 00020.026.1801anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 10% MeOH in DCM as an eluant. Required fractions were concentrated under reduced pressure to afford tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate 4A (0.415 g, yield: 79%) as a brown solid. MS (ESI) m / z: 315.1[M+H]+Step-3A: Synthesis of tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate (Int#5A)[000192] To a solution of tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate 4A (680 mg, 2.162 mmol) in MeOH (10 mL) was added 10% Pd / C (680 mg, wt / wt). The resultant reaction mixture was stirred at room temperature for 16 h under hydrogen pressure (30 psi). Upon completion of the reaction, reaction mixture was filtered through celite bed, washed with MeOH (20 mL) and filtrate was concentrated under reduced pressure to get tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate (Int#5A) (523 mg, crude) as an off white solid. MS (ESI) m / z: 317.09 [M+H]+Step-IB: Synthesis of 4-( ( 4, 4, 5, 5-tetramethyl- 1, 3, 2-dioxaborolan-2-yl)methylene)piperidine (3A)[000193] To a solution of tert-butyl 4-((4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)methylene)piperidine-l -carboxylate IB (1 g, 0.003 mmol) in DCM (10 mL, 10 vol), was added 4M HC1 in 1,4-di oxane (6 mL, 6 vol) at 0°C. The reaction mixture was stirred at room temperature for 1h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude (1.3 g). Crude was triturated with diethyl ether to afford#583961Attorney Docket No. 00020.026.18014-((4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)methylene)piperidine 3A (635 mg, yield: 92%) as an off white solid. MS (ESI) m / z: 224 [M+H]+Example 5: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((2S,3R)-3-hydroxy-2-((S)-l-(4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)pyrrolidine-2-carboxamido)butanamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 5):Step-1: Synthesis of tert-butyl 4-(isoindolm-4-ylmethylene)piperidine-l -carboxylate (3):[000194] To a solution of 4-bromoisoindoline (1) (10 g, 42.7 mmol) in DMF:H₂O (9:2, 10 vol) at room temperature was added tert-butyl 4-((4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)methylene)piperidine-l -carboxylate (2) (16.5 g, 51.28 mmol) followed by CS2CO3 (41.6 g, 128.2 mmol). The reaction mixture was degassed with argon for 15 min followed by addition of XPhos Pd G4 (3.67 g, 4.27 mmol), resultant reaction mixture was stirred at 100°C for 5 h. Upon completion of the reaction, reaction mixture was filtered through celite bed washed with ethyl acetate (20 mL), filtrate was concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 5% MeOH in DCM as an eluant. Required fractions were concentrated under reduced pressure to 86#583961Attorney Docket No. 00020.026.1801afford tert-butyl 4-(isoindolin-4-ylmethylene)piperidine-l-carboxylate (3) (8 g. yield: 59%) as a brown solid. MS (ESI) m / z: 315.1 [M+H]Step-2: Synthesis tert-butyl 4-(isoindolin-4-ylmethyl)piperidine-l-carboxylate (4):[000195] To a solution of tert-butyl 4-(isoindolin-4-ylmethylene)piperidine-l-carboxylate (3) (8 g, 25.47 mmol) in MeOH (80 mL) was added 10% Pd / C (5 g, wt / wt). The resultant reaction mixture was stirred at room temperature for 16 h under hydrogen pressure (30 psi). Upon completion of the reaction, reaction mixture was filtered through celite bed, washed with MeOH (20 mL) and filtrate was concentrated under reduced pressure to get tertbutyl 4-(isoindolin-4-ylmethyl)piperidine-l -carboxylate (4) (5 g, yield: 62%) as a brown semi-solid. MS (ESI) m / z: 317.29 [M+H]+Step-3: Synthesis of tert-butyl 4-((2-(2-((3S,5S)-5-((S)-2-cyano-4.4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidine-l-ccirboxylate (5)[000196] To a solution of CDA-5 (5.7 g. 22.72 mmol) in DMF (50 mL. 10 Vol) at 0°C was added DIPEA (5.5 mL, 31 mmol) followed by addition of HATU (6 g, 15 mmol). The mixture was stirred at room temperature for 30 min and added tert-butyl 4-(isoindolin-4-ylmethyl)piperidine- 1 -carboxylate (4) (5 g, 16 mmol). Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was diluted with ice water (20 mL) solid w as filter and washed with diethyl ether (50 mL) to afford tertbutyl 4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidine-l-carboxylate (5) (6 g, yield: 63%) as a brown solid. MS (ESI) m / z: 600.2(M+H)+.87#583961Attorney Docket No. 00020.026.1801Step-4: Synthesis of (S)-4,4-difluoro-l-((2S,4S)-5-oxo-4-(2-oxo-2-(4-(piperidin-4-ylmethyl)isoindolin-2-yl)ethyl)pyrrolidine-2-carbonyl)pyrrolidme-2-carbonitrile (6):[000197] TFA (18 mL, 3 vol) was added to a solution of tert-butyl 4-((2-(2-((3S,5S)-5- ((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidine-l-carboxylate (5) (6 g, 10.0 mmol) in DCM (60 mL, 5 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (50 mL) to afford (S)-4,4-difluoro-l-((2S,4S)- 5-oxo-4-(2-oxo-2-(4-(piperidin-4-ylmethyl)isoindolin-2-yl)ethyl)pyrrolidine-2- carbonyl)pyrrolidine-2-carbonitrile (6) (4.5 g, yield: 90%) as a brown solid. MS (ESI) m / z: 500.3 [M+H]+Step-5: Synthesis of (9H-fluoren-9-yl)methyl tert-butyl ((S)-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutane-l,3-diyl)dicarbamate (7)[000198] To a solution of (S)-2-((((9H-fluoren-9-y l)methoxy)carbonyl)amino)-4-((tert- butoxycarbonyl)amino)butanoic acid (6) (5.2 g, 11.82 mmol) in DMF (59 mL, 10 Vol) at 0°C was added DIPEA (4.1 mL, 23.51 mmol) followed by addition ofHATU (4.9 g, 12.89 mmol). The mixture was stirred at room temperature for 30 min and (S)-4,4-difluoro-l- ((2S,4S)-5-oxo-4-(2-oxo-2-(4-(piperidin-4-ylmethyl)isoindolin-2-yl)ethyl)pyrrolidine-2- carbonyl)pyrrolidine-2-carbonitrile (6A) (5.9 g, 11.82 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was 88#583961Attorney Docket No. 00020.026.1801diluted with ice water (20 mL) solid was filtered and washed with diethyl ether (50 mL) to afford (9H-fluoren-9-yl)methyl tert-butyl ((S)-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutane-l,3-diyl)dicarbamate (7) (6 g, yield: 55%) as an off white solid. MS (ESI) m / z: 922.6(M+H)+.Step-6: Synthesis of (9H-fluoren-9-yl)methyl ((S)-4-amino-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate (8)[000199] TFA (32 mL, 3 vol) was added to solution of ((9H-fluoren-9-yl)methyl tertbutyl ((S)-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutane-l,3-diyl)dicarbamate (7) (9 g, 9.77 mmol) in DCM (90 mL, 5 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (200 mL) to afford (9H-fluoren-9-yl)methyl ((S)-4-amino-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrroli din-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate (8) (7 g, yield: 87% ) as a white solid. MS (ESI) m / z: 822.7 [M+H]+.Step-7: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin- l-yl)-4-((2S,3R)-3-hydroxy-2-((S)-l-(4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)pyrrolidine-2-carboxamido)butanamido)-l-oxobutan-2-yl)carbamate (9)89#583961Attorney Docket No. 00020.026.1801L-threonine Int-1 (2.2 g, 2.67 mmol) in DMF (22 mL, 10 Vol) at 0°C was added DIPEA (1.27 mL, 7.28 mmol) followed by addition of HATU (1.5 g, 2.93 mmol). The mixture was stirred at room temperature for 30 min and (9H-fluoren-9-yl)methyl ((S)-4-amino-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate (8) (3 g, 3.65 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted with 10% MeOH in DCM (25 mL) and ice water (20 mL), organic layer was separated and the aqueous layer was extracted with 10% MeOH in DCM (2 *25 mL). Organic layer separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-1-yl)-4-((2S,3R)-3-hydroxy-2-((S)-1-(4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)pyrrolidine-2-carboxamido)butanamido)-1-oxobutan-2-yl)carbamate (9) (3.1 g, yield: 59.6%) as an off white solid. MS (ESI) m / z: 822.7 (M+H)+.Step-8: Synthesis of (S)-N-((2S, 3R)-l-( ( (S)-3-amino-4-( 4-((2-(2-( (3S, 5S)-5-( (S)-2-cyano-4.4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)amino)-3-hydroxy-l -oxobutan-2-yl)-l-( 4-((4-( 4-iodophenyl)butanamido)methyl)benzoyl)pyrrolidine-2-carboxamide (10)[000201] Diethyl amine (11 mL, 4 Vol) was added to a solution of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2- 90#583961Attorney Docket No. 00020.026.1801oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((2S,3R)-3-hydroxy-2-((S)-l-(4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)pyrrolidine-2-carboxamido)butanamido)-l-oxobutan-2-yl)carbamate 9 (4.7 g, 3.29 mmol) in acetonitrile (47 mL, 10 Vol) at room temperature. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude. Crude material was triturated with cold EtOAc (10 mL) followed by diethyl ether (30 mL) to afford (S)-N-((2S,3R)-l-(((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperi din-1 -yl)-4-o\obutyl)amino)-34iydroxy-l-o\obutan-2-yl)-l-(4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)pyrrolidine-2-carboxamide 10 (3.2 g, yield: 88%) as a white solid. MS (ESI) m / z: 1203.8 (M+H)+.Step-9: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S.5S)-5-((S)-2-cyano-4.4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((2S,3R)-3-hydroxy-2-((S)-l-(4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)pyrrolidine-2-carboxamido)butanamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4.7, 10-tetraazacyclododecane-l.4.7 -triyl) triacetic acid (compound 5)[000202] To a solution of (S)-N-((2S,3R)-l-(((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)- 2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4- 91#583961Attorney Docket No. 00020.026.1801yl)methyl)piperidin-l-yl)-4-oxobutyl)amino)-3-hydroxy-l-oxobutan-2-yl)-l-(4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)pyrrolidine-2-carboxamide 10 (1.6 g, 1.33 mmol) in acetonitrile (160 mL, 10 Vol) at room temperature, was added DIPEA (2.1 mL, 12 mmol) followed by 2,2',2"-( 10-(2-((2,5 -dioxopy rrolidin- 1 -y l)oxy)-2-oxoethy 1)- 1,4,7, 10-tetraazacyclododecane-l,4,7-triyl)triacetic acid DOTA NHS ester (950 mg, 1.89 mmol). The resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. The crude material was triturated with EtOAc (20 mL) then obtained material was purified by grace reverse phase, eluted at 45-50% CH₃CN:H₂O. Required fractions were concentrated under reduced pressure and further lyophilized to afford 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4.4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((2S,3R)-3-hydroxy-2-((S)-l-(4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)pyrrolidine-2-carboxamido)butanamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 5) (210.7 mg, yield: 10%) as a white solid. MS (ESI) m / z: 1590.19 [M+H]+ 1H-NMR (400 MHz, DMSO-d6): 5 12-10 (brs, 1H), 8.5-8.3 (brs. 1H), 8.1-80 (brs. 1H), 7.61 (d. J = 8Hz, 2H), 7.5-7.05 (m, 7H), 7.01 (d, J= 8.4 Hz, 2H), 5.11-5.00 (m, 1H), 4.90-4.80 (m, 2H), 4.72-4.60 (m, 4H), 4.50-4.00 (m, 8H), 3.95-3.80 (m, 2H), 3.70-3.60 (m, 2H), 3.60-3.50 (m, 8H), 3.20-3.10 (m, 5H), 3.10-2.90 (m, 18H), 2.6-2.40 (m, 6H), 2.30-2.25 (m, 2H), 2.15-2.10 (m. 5H), 1.95-1.70 (m. 7H), 1.60-1.50 (m. 3H), 1.3-1.2 (m, 2H), 1.20-0.90 (m, 4H).Synthesis of Int-1:HO-WHO o '%-N *H0Step-1A: Synthesis of 1 -(tert-butyl) 2-(2.5-dioxopyrrolidin-l-yl) (S) -pyrrolidine- 1,2-dicarboxylate (2A)#583961Attorney Docket No. 00020.026.1801[000203] To a solution (tert-butoxycarbonyl)-L-proline 1A (10 g, 46.51 mmol) in DMF (100 mL. 10 Vol) at 0°C was added DIPEA (16.2 mL, 90.41 mmol) followed by addition of HBTU (19.3 g, 50.92 mmol). The mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted ice water (100 mL), solid was filtered and washed with diethyl ether (20 mL) to afford 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin- 1-yl) (S)-pyrrolidine-l,2-dicarboxylate (2A) (10.2 g, yield: 69%) as an off white solid. MS (ESI) m / z: 313.6 (M+H)+.Step-2A: Synthesis of tert-butyl (S)-2-(((2S,3R)-3-hydroxy-l-methoxy-l-oxobutan-2-yl)carbamoyl)pyrrolidine-l -carboxylate (4A)[000204] To a solution 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-l-yl) (S)-pyrrolidine-l,2-dicarboxylate (2A) (5 g, 16.02 mmol) in DCM (50 mL, 10 Vol) at 0°C was added DIPEA (1.27 mL, 31.5 mmol). The mixture was stirred at room temperature for 30 min and added methyl L-threoninate (3A) (2.7 g, 16.02 mmol). Resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was diluted with 10% MeOH in DCM (25 mL) and ice water (20 mL), organic layer was separated and the aqueous layer was extracted with 10% MeOH in DCM (2 *25 mL). Organic layer separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert-butyl (S)-2-(((2S,3R)-3-hydroxy-l-methoxy-l-oxobutan-2-yl)carbamoyl)pyrrolidine-l -carboxylate (4A) (3.1 g, yield: 59.6%) as a pale yellow liquid. MS (ESI) m / z: 331.9 (M+H)+.Step-3A: Synthesis of methyl L-prolyl-L-threoninate(5A)TFA. DCM, 0 °C, RT, 3h[000205] TFA (12 mL. 3 vol) was added to a solution of tert-butyl (S)-2-(((2S,3R)-3-hy droxy- 1 -methoxy- 1 -oxobutan-2-yl)carbamoyl)pyrrolidine- 1 -carboxylate 4A (4 g, 12.1293#583961Attorney Docket No. 00020.026.1801mmol) in DCM (40 mL, 10 Vol) at 0 °C. Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (50 mL) to afford methyl L-prolyl-L-threoninate (5A) (2.4 g, yield: 88%) as a paleyellow gummy solid. MS (ESI) m / z: 231.1 [M+H]+.Step-4A: Synthesis of methyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl-L-[000206] To a solution 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid Int-2 (2 g, 4.72 mmol) in DMF (20 mL, 10 Vol) at 0°C was added DIPEA (1.7 mL. 9.44 mmol) followed by addition of HATU (2.1 g, 5.66 mmol). The mixture was stirred at room temperature for 30 min and added methyl L-prolyl-L-threoninate (5A) (1 g, 4.72 mmol). Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was diluted with 10% MeOH in DCM (50 mL) and ice water (20 mL), organic layer was separated and the aqueous layer was extracted with 10% MeOH in DCM (2 *50 mL). Organic layer separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert-buty l methyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl-L-threoninate (7A) (2 g, yield: 68%) as a brown gummy solid. MS (ESI) m / z: 636.3 [M+H]+.Step-5 A: Synthesis of (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl-L-threonine[000207] To a solution methyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl-L-threoninate 7A (2 g, 3.1 mmol) in THF:MeOH:H₂O(2:4:4) (20 mL, 10 Vol) at 0°C 94#583961Attorney Docket No. 00020.026.1801was added LiOH.2H₂O (0.26 g, 6.2 mmol). Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was acidified with IN HC1 and diluted with 10% MeOH in DCM (25 mL) and ice water (20 mL), organic layer was separated and the aqueous layer was extracted with 10% MeOH in DCM (2 *25 mL).Organic layer separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl-L-threonine Int-1 (400 mg, yield: 21%) as an off white solid. MS (ESI) m / z: 622.2 (M+H)+Synthesis of Int-2:Step-1: Synthesis of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate (3):[000208] To a solution of 4-(4-iodophenyl)butanoic acid 1 (30 g, 0.103 mol) in DMF (60 mL, 2 Vol) at 0°C was added DIPEA (27 mL, 0.155 mol) followed by HATU (39.3 g, 0.103 mol). The reaction mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl 4-(aminomethyl)benzoate 2 (21.4 g. 0.103 mol). Resultant reaction mixture was stirred at room temperature for 4 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (400 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 400 mL). Combined organic layer was washed with water (400 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (36.1 g, crude) as a brown liquid. MS (ESI) m / z: 480.1 [M+H]+Step-2: Synthesis of 4-((4-(4-iodophenyl)bulanamido)methyl)benzoic acid (Inl-2):95#583961Attorney Docket No. 00020.026.1801[000209] TFA (160 mL, 4 vol) was added to a solution of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (40 g, 0.083 mol) in DCM (200 mL, 5 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 5 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (200 mL) to afford 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid Int-2 (31 g, yield: 88%) as a brown solid. MS (ESI) m / z: 424.42 [M+H]+Example 6: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-alanyl)pyi olidirie-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane- 1, 4, 7-triyl)triacetic acid (compound 6):Step-7: Synthesis of lerl-bulyl (S)-2-(((S)-3-((((9H-fluoren-9-yl) methoxy) carbonyl)amino)-4- ( 4-( (2-(2-(( 3S, 5S)-5-((S)-2-cyano-4, 4-difluoropyrrolidine-l -carbonyl)-2-oxopyrrolidiri-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)carbamoyl)pyrrolidine-l-carboxylate (11):96#583961Attorney Docket No. 00020.026.1801[000210] To a solution of (tert-butoxycarbonyl)-L-proline 10 (2.3 g, 10.95 mmol) in DMF (30 mL, 10 Vol) at 0°C, was added DIPEA (1 mL, 5.347 mmol) followed by HATU (1.5 g, 4.00 mmol). The mixture was stirred at room temperature for 10 min, and added (9H-fluoren-9-yl)methyl ((S)-4-amino-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate Int-1 (3 g, 3.65 mmol). Resultant reaction mixture was stirred at room temperature for 1.5 h. Upon completion of the reaction, reaction mixture was diluted with cold water (50 mL) and stirred for 15 min, obtained solid was dissolved in 10% MeOH in DCM, dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (20 mL), pentane (10 mL), concentrated under reduced pressure to afford tert-butyl (S)-2-(((S)-3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)carbamoyl)pyrrolidine-l -carboxy late 11 (3 g, yield: 81 %) as a brown solid. MS (ESI) m / z: 1019.6 (M+H)+.Step-8: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxo-4-((S)-pyrrolidine-2-carboxamido)butan-2-yl)carbamate (12):97#583961Attorney Docket No. 00020.026.1801[000211] To a solution of tert-butyl (S)-2-(((S)-3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)carbamoyl)pyrrolidine-l -carboxylate 11 (3 g, 2.94 mol) in DCM (60 mL, 20 Vol) at 0°C was added TFA (6 mL, 2 vol). Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction the reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (50 mL) to afford (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxo-4-((S)-pyrrolidine-2-carboxamido)butan-2-yl)carbamate 12 (2.7 g, yield: 99%) as an off white solid. MS (ESI) m / z: 919.7 (M+H)+.Step-9: ( (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-( ( tert-butoxycarbonyl)-L-alanyl)pyrrolidine-2-carboxamido)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidme-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate#583961Attorney Docket No. 00020.026.1801[000212] To a solution of ((tert-butoxycarbonyl)-L-alanine 12A (1.3 g, 7.35 mmol) in DMF (15 mL, 10 Vol) at 0°C, was added DIPEA (0.8 mL, 4.41 mmol) followed by COMU (1.1 g, 2.94 mmol). The mixture was stirred at room temperature for 10 min, and added (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxo-4-((S)-pyrrolidine-2-carboxamido)butan-2-yl)carbamate 12 (2.7 g. 2.94 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction the reaction mixture was diluted with ice water (50 mL) and filter the obtained solid. Solid was dissolved in 10% MeOH in DCM, dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether(20 mL), pentane (20 mL) concentrated under reduced pressure to afford 9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((tert-butoxycarbonyl)-L-alanyl)pyrrolidine-2-carboxamido)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate (13) (3 g, yield: 93 %) as a brown solid. MS (ESI) m / z: 1089.4 (M+H)+.Step-10: Synthesis of (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-(L-alanyl)pyrrolidine-2-carboxamido)-l-(4-((2-(2-((3S.5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate[000213] To a solution of (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((tert-butoxycarbonyl)- L-alanyl)pyrrolidine-2-carboxamido)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate 13 (3 g, 2.75 mol) in DCM (60 mL, 2099#583961Attorney Docket No. 00020.026.1801Vol) at 0°C was added TFA (6 mL, 2 vol). Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (50 mL) to afford 9H-fluoren-9-yl)methy 1 ((S)-4-((S)- 1 -(L-alany l)pyrrolidine-2-carboxamido)- 1 -(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate (14) (2.6 g, yield: 96%) as a brown solid. MS (ESI) m / z:990.96 (M+H)+.Step-11: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-( 4-( (2-(2-(( 3S, 5S)-5-( (S)-2-cyano-4, 4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-cilanyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (15):[000214] To a solution of (4-((4-(4-iodophenyl) butanamido) methyl)benzoic acid Int-2 (0.96 g, 2.27 mmol) in DMF (15 mL, 10 Vol) at 0°C, was added DIPEA (4.1 mL, 2.27mmol ) followed by COMU (0.664 g, 1.51 mmol). The mixture was stirred at room temperature for 10 min, and added (9H-fluoren-9-yl)methyl((S)-4-((S)-l-(L-alanyl)pyrrolidine-2-carboxamido)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate 14 (1.5 g, 1.51 mmol). Resultant reaction mixture was stirred at room temperature for 1.5 h. Upon completion of the reaction, the reaction mixture was dilute with cold water (50 mL) and 100#583961Attorney Docket No. 00020.026.1801stirred for 15 min, obtained solid was filtered, solid was dissolved in 10% MeOH in DCM, dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether, pentane concentrated under reduced pressure to afford 9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-alanyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 15 (1.6 g, yield: 80%) as a brown solid. MS (ESI) m / z: 1395.4 (M+H)+.Step-12: Synthesis of (S)-N-((S)-3-ammo-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-l)methyl)piperidin- 1-yl)-4-oxobutyl)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-lanyl)pyrrolidine- 2-carboxamide( 16):[000215] Diethyl amine (6 mL, 4 Vol) was added to a solution of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4.4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-alanyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 15 (1.5 g, 1.075 mmol) in acetonitrile (15 mL, 10 Vol) at room temperature. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the 101#583961Attorney Docket No. 00020.026.1801reaction, reaction mixture was concentrated under reduced pressure to get crude. Crude was triturated with cold EtOAc (40 mL) followed by diethyl ether (80 mL) to afford (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-alanyl)pyrrolidine-2-carboxamide 16 (1.2 g, yield: 98%) as a brown solid. MS (ESI) m / z:1173.1 (M+H)+Step-13: Synthesis of 2,2 2”-(10-(2-(((S)-l-(4-((2-(2-((3S.5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-alanyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l, 4, 7,10-tetraazacyclododecane-l, 4, 7-triyl)triacetic acid[000216] To a solution of (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-y l)methy l)piperidin- 1 -y l)-4-oxobutyl)- 1 -((4-((4-(4-iodophenyl)butanamido)methy l)benzoy 1)-L-alanyl)pyrrolidine-2-carboxamide 16 (1.2 g, 1.023 mmol) in acetonitrile (120mL, 10 Vol), at room temperature, was added DIPEA (1.6 mL, 9.207 mmol) followed by 2,2',2"-(10-(2-((2,5 -dioxopy rrolidin- 1 -y l)oxy)-2-oxoethy 1)- 1,4,7, 10-tetraazacy clododecane- 1,4,7-triyl)triacetic acid DOTA NHS ester (768 mg, 1.5345 mmol). Resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with EtOAc (20 mL) and further purified by prep HPLC using the below condition.Prep conditionColumn Gemini, C18(250*21.5mm)5pBuffer A:0.2% FA in Water, B:0.2 % FA in ACN: H2O (80:20)102#583961Attorney Docket No. 00020.026.1801Gradient (T / %B) 0 / 35,20 / 55,20.1 / 98Flow Rate 12 ml / minSolubility ACN+THF+H2O[000217] The required fractions were collected and lyophilized to afford 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindohn-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-alanyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)- 1,4,7, 10-tetraazacyclododecane-1,4,7-triyl)triaceticacid (compound 6) (225 mg, yield: 13%) as a white solid. MS (ESI) m / z: 1560.29 [M+H], ’H-NMR (400 MHz, DMSO-d6): 58.56-8.54 (d, J= 8 Hz,, 1H)., 8.38-8.4 (m. 1H), 8.28 (m, 2H). 8.07-8.05 (m, 1H), 7.80 (dd, J= 8Hz, 2H), 7.63-7.61 (d, J= 8 Hz,, 2H), 7.3-7.1 (m, 5 H), 7.01-6.99 (d, J= 8 Hz, 2H), 5.12-5.09 (m, 1H), 4.9-4.6 (m, 6H), 4.4-4.1 (m, 7H), 3.9-3.6 (m, 4H), 3.5-3.4 (m, 12H), 3.1-3.0(m, 14 H), 2.8-2.6 (m, 8H), 2.1-2.5 (m, 8H), 1.9-1.6 (m, 11H), 1.4-1.3 (m, 3H), 1.1-1 (m, 2H)Synthesis of Int-1:0%^NHStep-IA: Synthesis of 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-l-yl) (S) -pyrrolidine- 1,2-dicarboxylate (2A)CzBoc _..'B°cN HBTU, DIPEA, DMF / N O J.^OH - - '|f 0°C to RT,1 h 11 1 / o01A _ Step-1 A _ 2A[000218] To a solution (tert-butoxycarbonyl)-L-proline 1A (10 g, 46.51 mmol) in DMF (100 mL, 10 Vol) at 0°C was added DIPEA (16.2 rnL, 90.41 mmol) followed by addition of HBTU (19.3 g. 50.92 mmol). The mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted ice water (100 mL), solid was filtered and washed with diethyl ether (20 mL) to afford 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-1-yl) (S)-pyrrolidine-l,2-di carboxylate (2A) (10.2 g, yield: 69%) as an off white solid. MS (ESI) m / z: 313.6 (M+H)+.103#583961Attorney Docket No. 00020.026.1801Step-2A: Synthesis of tert-butyl (S)-2-(((2S,3R)-3-hydroxy-l-methoxy-l-oxobutan-2-yl)carbamoyl)pyrrolidine-l-carboxylate (4A)[000219] To a solution 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-l -yl) (S)-pyrrolidine-l,2-dicarboxylate (2A) (5 g, 16.02 mmol) in DCM (50 mL, 10 Vol) at 0°C was added DIPEA (1.27 mL, 31.5 mmol). The mixture was stirred at room temperature for 30 min and added methyl L-threoninate (3A) (2.7 g, 16.02 mmol). Resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was diluted with 10% MeOH in DCM (25 mL) and ice water (20 mL), organic layer was separated and the aqueous layer was extracted with 10% MeOH in DCM (2 *25 mL). Organic layer separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert-butyl (S)-2-(((2S,3R)-3-hydroxy-l-methoxy-l-oxobutan-2-yl)carbamoyl)pyrrolidine-l -carboxylate (4A) (3.1 g, yield: 59.6%) as a pale yellow liquid. MS (ESI) m / z: 331.9 (M+H)+.Step-3A: Synthesis of methyl L-prolyl-L-threoninate(5A)[000220] TFA (12 mL, 3 vol) was added to a solution of tert-butyl (S)-2-(((2S,3R)-3-hydroxy-l-methoxy-l-oxobutan-2-yl)carbamoyl)pyrrolidine-l-carboxylate 4A (4 g, 12.12 mmol) in DCM (40 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (50 mL) to afford methyl L-prolyl-L-threoninate (5A) (2.4 g, yield: 88%) as a paleyellow gummy solid. MS (ESI) m / z: 231.1 [M+H]+.104#583961Attorney Docket No. 00020.026.1801Step-4A: Synthesis of methyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl-L-[000221] To a solution 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid lnt-2 (2 g, 4.72 mmol) in DMF (20 rnL, 10 Vol) at 0°C was added DIPEA (1.7 mL, 9.44 mmol) followed by addition of HATU (2.1 g, 5.66 mmol). The mixture was stirred at room temperature for 30 min and added methyl L-prolyl-L-threoninate (5A) (1 g, 4.72 mmol). Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was diluted with 10% MeOH in DCM (50 mL) and ice water (20 mL), organic layer was separated and the aqueous layer was extracted with 10% MeOH in DCM (2 *50 mL). Organic layer separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert-butyl methyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl-L-threoninate (7 A) (2 g, yield: 68%) as a brown gummy solid. MS (ESI) m / z: 636.3 [M+H]+.Step-5A: Synthesis of (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl-L-threonine[000222] To a solution methyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl-L-threoninate 7A (2 g, 3.1 mmol) in THF:MeOH:H₂O(2:4:4) (20 mL, 10 Vol) at 0°C was added LiOH.2H2O (0.26 g, 6.2 mmol). Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was acidified with IN HC1 and diluted with 10% MeOH in DCM (25 mL) and ice water (20 rnL), organic layer was separated and the aqueous layer w as extracted with 10% MeOH in DCM (2 *25 mL).Organic layer separated was dried over anhydrous sodium sulphate, filtered and concentrated 105#583961Attorney Docket No. 00020.026.1801under reduced pressure to afford tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)-L-prolyl-L-threonine Int-1 (400 mg. yield: 21%) as an off white solid. MS (ESI) m / z: 622.2 (M+H)+.Synthesis of Int-2:Step-1: Synthesis of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate (3):[000223] To a solution of 4-(4-iodophenyl)butanoic acid 1 (30 g, 0.103 mol) in DMF (60 mL, 2 Vol) at 0°C was added DIPEA (27 mL, 0.155 mol) followed by HATU (39.3 g, 0.103 mol). The reaction mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl 4-(aminomethyl)benzoate 2 (21.4 g, 0.103 mol). Resultant reaction mixture was stirred at room temperature for 4 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (400 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 400 mL). Combined organic layer was washed with water (400 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (36.1 g, crude) as a brown liquid. MS (ESI) m / z: 480.1 [M+H]+Step-2: Synthesis of 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid (Int-2):[000224] TFA (160 mL, 4 vol) was added to a solution of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (40 g, 0.083 mol) in DCM (200 mL, 5 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 5 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude 106#583961Attorney Docket No. 00020.026.1801material, crude material was triturated with diethyl ether (200 mL) to afford 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid Int-2 (31 g, yield: 88%) as a brown solid. MS (ESI) m / z: 424.42 [M+H]+Example 7: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)gIycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacydododecane-l,4,7-triyl)triacetic acid (compound 7):Step-1: Synthesis of tert-butyl 4-((2-(naphthalen-2-yl)acetamido)methyl)benzoate (3):[000225] To a solution of 2-(naphthalen-2-yl)acetic acid 1 (5 g, 0.02 mol) in DMF (50 mL, 2 Vol) at 0 °C was added DIPEA (7 mL, 0.04 mol) followed by HATU (10.2 g, 0.02 mol). The mixture was stirred at room temperature for 30 min, and added tert-butyl 4-(aminomethyl)benzoate 2 (5.56 g, 0.02 mol). Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (400 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 100 mL). Combined organic layer was washed with water (400 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert- 107#583961Attorney Docket No. 00020.026.1801buty l 4-((2-(naphthalen-2-yl)acetamido)methyl)benzoate (3) (6.8 g, yields: 68%) as an off white solid. MS (ESI) m / z: 376.8 [M+H]+Step-2: Synthesis of4-((2-(naphthalen-2-yl)acetamido)methyl)benzoic acid (4):[000226] TFA (27.2 mL, 4 vol) was added to a solution of tert-butyl 4-((2-(naphthalen-2-yl)acetamido)methyl)benzoate 3 (6.8 g, 0.083 mol) in DCM (68 mL. 10 Vol) at 0°C.Resultant reaction mixture was stirred at room temperature for 2 h. Reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (200 mL) to afford 4-((2-(naphthalen-2-yl)acetamido)methyl)benzoic acid 4 (4.5 g, yield: 78%) as a off white solid. MS (ESI) m / z: 320.4 [M+H]+Step-3: Synthesis of tert-butyl (4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycinate[000227] To a solution of 4-((2-(naphthalen-2-yl)acetamido)methyl)benzoic acid 4 (1 g, 3.13 mmol) in DMF (10 mL, 10 Vol) at 0°C was added DIPEA (1.6 mL, 9 mmol) followed by HATU (1.78 g, 0.004 mol). The mixture was stirred at room temperature for 30 min, and added tert-butyl glycinate 4A (452 mg, 3.44 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (400 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 400 mL). Combined organic layer was washed with water (400 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tertbutyl (4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycinate (5) (900 mg, yield: 69%) as a white solid. MS (ESI) m / z: 431.2 [M-H]- Step-4: Synthesis of (4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycme (6):108#583961Attorney Docket No. 00020.026.1801[000228] TFA (3.6 mL, 4 vol) was added to a solution of tert-butyl (4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycinate (5) (900 mg, 2.078 mmol) in DCM (9 mL, 10 Vol) at 0 °C. Resultant reaction mixture was stirred at room temperature for 3 h. Reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (200 mL) to afford (4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycine (6) (450 mg, yield: 57%) as a white solid. MS (ESI) m / z: 377.1 [M+H]+Step-5: Synthesis of (S)-2-( ( f9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-( (S)-l-( ( 4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrroUdine-2-carboxamido)butanoic acid[000229] To a solution of (4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycine (6) (15 g, 39 mmol) in DMF (150 mL, 10 Vol) at 0°C, was added DIPEA (10 mL, 5 mmol ) followed by HATU (16.65 g, 43.8 mmol). The mixture was stirred at room temperature for 30 min, and added (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid (Int#6A) (17.4 g, 39 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. Reaction mixture was diluted with EtOAc (200 mL), cold water (200 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 150 mL). Combined organic layer was washed with water (150 mL). The separated organic layer was dried over anhydrous sodium sulphate. filtered and concentrated under reduced pressure to afford crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 50% Ethyl acetate in hexane as an eluant. Required fractions were concentrated under reduced pressure to afford (S)-2-((((9H-fluoren-9-y l)methoxy )carbony l)amino)-4-((S)- 1 -((4-((2-(naphthalen-2-109#583961Attorney Docket No. 00020.026.1801yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid (7) (10 g, yield: 40 %) as awhile solid. MS (ESI) m / z:796.7 (M+H)+.Step-6: Synthesis of tert-butyl 4-( ( l-((S)-2-( ( ((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4- ((S)-l-((4-(( 2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate (8)[000230] To a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid (7) (5 g, 6.28 mmol) in DMF (50 mL, 10 Vol) at 0°C was added DIPEA (2.19 mL, 13.1 mmol) followed by HATU (2.65 g, 6.97 mmol, 1.1 eq). The mixture was stirred at room temperature for 15 min and added tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate (Int#5A) (1.59 g. 5.02 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. Reaction mixture was diluted with EtOAc (50 mL), ice cold water (50 mL) and stirred for 15 min. The aqueous layer was extracted with EtOAc (2 * 25 mL), combined organic layer was washed with ice cold water (2 * 25 mL). The organic phase separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperi din-4-yl)methyl)isoindoline-2-carboxylate 8 (3.5 g, yield: 51%) as awhile solid. MS (ESI) m / z: 1094.89 (M+H)+.Step-7: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-4-((S)-l-((4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (9)110#583961Attorney Docket No. 00020.026.1801fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((2-(naphthalen-2- yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperi din-4- yl)methyl)isoindoline-2-carboxylate 8 (3.5 g, 3.19 mmol) in DCM (35 mL, 10 Vol) at 0°C. The resultant reaction mixture was stirred at room for 3 h. Reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (30 mL), to afford (9H-fluoren-9-yl)methyl ((S)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-4-((S)-l-((4- ((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l- oxobutan-2-yl)carbamate (9) (2.8 g, yield: 80%) as a brown solid. MS (ESI) m / z: 995.8 (M+H)+.Step-8: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4- difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolm-4-yl)methyl)piperidin- l-yl)-4-( (S)-l-( ( 4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2- carboxamido)-l-oxobutan-2-yl)carbamate (10):[000232] To a solution of CDA-5 (1.01 g, 3.35 mmol) in DMF (28 mL, 10 Vol) at 0°C was added DIPEA (1.2 ml, 6.88 mmol) followed by addition of HATU (1.07 g,2.81 mmol). The mixture was stirred at room temperature for 30 min and added (9H-fluoren-9-yl)methyl ((S)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-4-((S)-l-((4-((2-(naphthalen-2- yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (9) (2.8 g, 6.28 mmol). The resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted with 10% MeOH in DCM (25 mL) and ice water (20 mL), organic layer was separated and the aqueous layer was 111#583961Attorney Docket No. 00020.026.1801extracted with 10% MeOH in DCM (2 *25 mL). Organic layer separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (10) (2.5 g, yield: 56%) as a brown colour solid. MS (ESI) m / z:1279 (M+H)+.Step-9: Synthesis of (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobiityl)-l-((4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide (11)[000233] Diethyl amine (7.5 mL, 4 Vol) was added to a solution of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (10) (2.5 g, 1.95 mmol) in acetonitrile (10 mL, 10 Vol) at room temperature. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude. Crude material was triturated with cold EtOAc (10 mL) followed by diethyl ether (10 mL) to afford (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)-l-((4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide (11) (1.6 g, yield: 77%) as a white solid. MS (ESI) m / z: 1055.8 (M+H)+.Step-10: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-! -oxobutan-2-yl)amino)-2-oxoethyl)- 1.4.7.10-tetraazacyclododecane-l.4.7-triy I) triacetic acid (compound 7)112#583961Attorney Docket No. 00020.026.1801[000234] To a solution of (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin- 1 -y l)-4-oxobuty 1)- 1 -((4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide(ll) (1.6 g, 1.51 mmol) in acetonitrile (16 mL, 10 Vol), at room temperature, was added DIPEA (2.3 mL, 13.19 mmol) followed by DOTANHS Ester (1.14 g, 2.275 mmol). The resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude material was triturated with EtOAc (20 mL) and further purified by prep HPLC using the below conditionPrep conditionColumn GEMINI C18 (250*21.2mm)5uBuffer 0.1%TFA in WATER; B: ACN: H2O (80:20)Gradient (T / %B) 0 / 35,15 / 45.21 / 45,21.05 / 98Flow Rate lOmL / minSolubility ACN+H2O[000235] The required fractions were collected and lyophilized to afford 2,2',2"-(10-(2- (((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4.4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((2-(naphthalen-2-yl)acetamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)- 1,4,7, 10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (compound 7) (220 mg, yield: 10.1% ) as a white solid. MS (ESI) m / z: 1441.96 [M+H],1H-NMR (400 MHz, DMSO-d6): 58.72-8.71 (m. 1H), 8.62-8.58 (m. 1H), 8.4-8.2 (m, 2H), 8.1-8.0 (m, 1H), 7.9-7.7 (m, 6H), 7.5-7.4 (m, 3H), 7.4-7.31 (m, 2 H), 7.2-7.0 (m, 3H), 5.12-5.09 (m, 1H), 4.9-4.8 (m, 2H), 4.7-4.6 (m, 3H), 4.4-4.3 (m, 4H), 4.2-4.0 (m, 5H), 3.8-3.7 (m, 2H), 3.7-3.5 (m, 4H), 3.5-3.3 (m, 11 H), 3.3-3.0 (m, 10 H), 3-2.9 (m. 12 H), 2.4-2.3 (m, 2H), 2.3-#583961Attorney Docket No. 00020.026.18012.2 (m, 1H), 2.2-2.1 (m, 1H), 1.8-1.7 (m, 5H), 1.7-1.6 (m, 2H), 1.5-1.4 (m, 4H), 1.2-1 (m, 2H)Synthesis of Int#5A:Step- 1 A: Synthesis of tert-butyl 4-bromoisoindoline-2-carboxylate (2A):[000236] To a solution of 4-bromoisoindoline 1A (1 g, 0.005 mol) in 2-MeTHF (10 mL, 10 Vol) at 0°C was added EtsN (2.9 mL. 0.02 mol) and (Boc)₂O (1.47 mL, 0.0067 mol). The reaction mixture was stirred at room temperature for 2h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (30 mL), water (30 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 30 mL). Combined organic layer was washed with water (30 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 5% EtOAc in pet ether as an eluent. Required fraction were concentrated under reduced pressure to afford tert-butyl 4-bromoisoindoline-2-carboxylate 2A (1.14 g, yield: 76%) as a white solid.Step-2A: Synthesis of tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-ccirboxylcite (4A):[000237] To a solution of tert-butyl 4-bromoisoindoline-2-carboxylate 2A (500 mg, 1.67 mmol) in DMF:H₂O (9:1, 10 vol) was added 4-((4,4,5,5-tetramethyl-l,3,2-dioxaborolan- 114#583961Attorney Docket No. 00020.026.18012-yl)methylene)piperidine 3A (561.5 mg, 2.51 mmol) followed by Cs₂CO₃ (1.63 g, 5.03 mmol). The reaction mixture was degassed with argon for 15 min followed by addition of XPhos Pd G4 (80 mg, 0.67 mmol), resultant reaction mixture was stirred at 100°C for 16 h. Upon completion of the reaction, reaction mixture was filtered through celite bed washed with ethyl acetate (20 mL), filtrate was diluted with water (30 mL). The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 30 mL). Combined organic layers was washed with water (30 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 10% MeOH in DCM as an eluant. Required fractions w ere concentrated under reduced pressure to afford tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate 4A (0.415 g. yield: 79%) as a brown solid. MS (ESI) m / z: 315.1[M+H]+Step-3A: Synthesis of tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate (Int#5A[000238] To a solution of tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate 4A (680 mg, 2.162 mmol) in MeOH (10 mL) was added 10% Pd / C (680 mg, wt / wt). The resultant reaction mixture w as stirred at room temperature for 16 h under hydrogen pressure (30 psi). Upon completion of the reaction, reaction mixture was filtered through celite bed, washed with MeOH (20 mL) and filtrate w as concentrated under reduced pressure to get tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate Int#5A) (523 mg, crude) as an off white solid. MS (ESI) m / z: 317.09 [M+H]+Step-IB: Synthesis of 4-( ( 4, 4, 5, 5-tetramethyl-l, 3, 2-dioxaborolan-2-yl)methylene)piperidine (3A)#583961Attorney Docket No. 00020.026.1801[000239] To a solution of tert-butyl 4-((4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)methylene)piperidine-l -carboxylate IB (1 g, 0.003 mmol) in DCM (10 mL, 10 vol), was added 4M HC1 in 1,4-di oxane (6 mL, 6 vol) at 0°C. The reaction mixture was stirred at room temperature for Ih. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude (1.3 g). Crude was triturated with diethyl ether to afford 4-((4,4.5.5-tetramethyl-l,3,2-dioxaborolan-2-yl)methylene)piperidine 3A (635 mg, yield: 92%) as an off white solid. MS (ESI) m / z: 224 [M+H]+Example 8: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxo-4-((S)-l-((4-((4-(p-toIyI)butanamido)methyl)benzoyI)glycyl)pyrrolidine-2-carboxamido)butan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 8)Step-1: Synthesis of tert-butyl 4-((4-(p-tolyl)butanamido)methyl)benzoate (3)[000240] To a solution of 4-(p-tolyl)butanoic acid (1) (5 g, 28.08 mmol) in DMF (50 mL, 2 Vol) at 0°C was added DIPEA (7.7 mL, 42.1 mmol) followed by HATU (10 g, 28.08 mmol). The mixture was stirred at room temperature for 30 min, followed by addition of tert- 116#583961Attorney Docket No. 00020.026.1801buty l 4-(aminomethyl)benzoate (2) (5.8 g, 0.103 mol). Resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted with cold water (400 mL) and stirred for 15 min. Solid was filtered and washed with diethyl ether to afford tert-butyl 4-((4-(p-tolyl)butanamido)methyl)benzoate 3 (8 g, yield:78%) as a white solid. MS (ESI) m / z: 368.2 [M+H]+Step-2: Synthesis of 4-((4-(p-tolyl)butanamido)methyl)benzoic acid (4):[000241] TFA (40 mL, 4 vol) was added to a solution of tert-butyl 4-((4-(p-tolyl)butanamido)methyl)benzoate (3) (10 g, 27.24 mol) in DCM (100 mL, 5 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (50 mL) to afford 4-((4-(p-tolyl)butanamido)methyl)benzoic acid (4) (7.5 g, yield: 89%) as a white solid. MS (ESI) m / z: 312.1 [M+H]+Step-3: Synthesis of tert-butyl (4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycinate(6):mmol) in DMF (20 mL, 10 Vol) at 0°C, was added DIPEA (3.5 mL, 19.3 mmol ) followed by HATU (3.6 g, 9.64 mmol). The reaction mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl glycinate 5 (926 mg, 7.07 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min. Precipitated solid was filtered and wash with hexane (2* 30 mL) to afford tert-butyl (4-((4-(p-117#583961Attorney Docket No. 00020.026.1801tolyl)butanamido)methyl)benzoyl)glycinate 6 (2 g, yield: 74 %) as a white solid. MS (ESI) m / z:426.1 (M+H)+.Step-4: Synthesis of(4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycine (7):[000243] TFA (15 mL, 3 vol) was added to a solution of tert-butyl (4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycinate 6 (5 g, 11.7 mmol) in DCM (50 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (100 mL) to afford (4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycine 7 (3.2 g, yield: 80%) as a white solid. MS (ESI) m / z: 369 [ M+H|Step-5: Synthesis (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid (8):[000244] To a solution of (4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycine 7 (2 g, 5.43 mmol) in DMF (20 mL, 10 Vol) at 0°C was added DIPEA (2 mL, 10 mmol) followed by addition of HATU (3.1 g, 8.14 mmol). The reaction mixture was stirred at room temperature for 10 min followed by addition of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid Int#l (2.6 g, 5.97 mmol). The resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted with cold water (100 mL) and stirred for 15 min. Obtained white solid was triturated with diethyl ether (20 mL) to afford (S)-2-((((9H-fluoren-9-y l)methoxy )carbony l)amino)-4-((S)- 1 -((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid (8) (2 g, yield: 46%) as a pale-yellow solid. MS (ESI) m / z: 786.5 (M-H)‘.118#583961Attorney Docket No. 00020.026.1801Step-6: Synthesis of tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4- ((S)-l-((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate (9):[000245] To a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid 8 (2 g, 2.54 mmol) in DMF (20 mL, 10 Vol) at 0°C, was added DIPEA (0.9 mL, 4.57 mmol ) followed by HATU (1.47 g, 3.81 mmol). Reaction mixture was stirred at room temperature for 10 min, followed by addition of tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate Int#5A (1.4 g, 4.43 mmol). Resultant reaction was stirred at room temperature for 2h. Upon completion of the reaction, reaction was diluted with cold water (100 mL) and stirred for 15 min. Obtained white solid was triturated with diethyl ether (10 mL) to afford tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperi din-4-yl)methyl)isoindoline-2-carboxylate 9 (2 g, yield: 74 %) as a pale yellow solid. MS (ESI) m / z:1086.8 (M+H)+.Step-7: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxo-4-((S)-l-((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butan-2-yl)carbamate (10):119#583961Attorney Docket No. 00020.026.1801[000246] TFA (8 mL, 4 vol) was added to a solution of tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperi din-4-yl)methyl)isoindoline-2-carboxylate 9 (2 g, 8.3 mmol) in DCM (20 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material crude material was triturated with diethyl ether (20 mL) to afford (9H-fluoren-9-yl)methyl ((S)- 1 -(4-(isoindolin-4-y lmethyl)piperidin- 1 -yl)- 1 -oxo-4-((S)- 1 -((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butan-2-yl)carbamate 10 (1.5 g, yield: 84%) as a brown solid. MS (ESI) m / z: 986.7 [ M+H|Step-8: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxo-4-((S)-l-((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butan-2-yl)carbamate: (11)[000247] To a solution of CDA-5 (548 mg, 1.84 mmol) in DMF (5.8 mL, 10 Vol) at 0°C was added DIPEA (0.6 mL, 3.75 mmol) followed by addition of HATU (627 mg, 1.65120#583961Attorney Docket No. 00020.026.1801mmol). The reaction mixture was stirred at room temperature for 10 min followed by addition of (9H-fluoren-9-yl)methyl ((S)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxo-4-((S)-l-((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butan-2-yl)carbamate 10 (1.5 g, 1.51 mmol). Resultant reaction mixture was stirred at room temperature for 2h. Upon completion of the reaction, reaction mixture was diluted with cold water (50 mL) and stirred for 15 min. Obtained white solid was triturated with diethyl ether (300 mL) to afford (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-y l)methyl)piperidin- 1 -yl)- 1 -oxo-4-((S)- 1 -((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butan-2-yl)carbamate 11 (1.5 g, yield: 78%) as a brown solid. MS (ESI) m / z: 1271 (M+H)+.Step-9: Synthesis of (S)-N-((S)-3-amino-4-(4-((2-(2-((3S.5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobiityl)-l-((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide: (12)[000248] Diethyl amine (6 mL, 4 Vol) was added to a solution of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxo-4-((S)-l-((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butan-2-yl)carbamate 11 (1.5 g, 1.18 mmol) in acetonitrile (15 mL, 10 Vol) at room temperature. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude. Crude was triturated with cold EtOAc (10 mL) followed by diethyl ether (10 mL) to afford (S)-N-((S)-3-amino-4- 121#583961Attorney Docket No. 00020.026.1801(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)-l-((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2 -carboxamide 12 (1 g, yield: 83%) as a white solid. MS (ESI) m / z: 1048.4 (M+H)+.Step-10: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (compound 8)[000249] To a solution of (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)-l-((4-((4-(p-tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2 -carboxamide 12 (1 g, 0.95 mmol) in acetonitrile (10 mL, 10 Vol) at room temperature, was added DIPEA (1.5 mL, 8.55 mmol) followed by 2,2',2"-( 10-(2-((2.5 -dioxopy rrolidin- 1 -y l)oxy )-2-oxoethy 1)- 1,4,7, 10- 122#583961Attorney Docket No. 00020.026.1801tetraazacyclododecane-l,4,7-triyl)triacetic acid DOTA NHS ester (717 mg, 1.43 mmol). The resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. The crude material was triturated with EtOAc (20 mL) then obtained material was purified by grace reverse phase, eluted at 45-50% CH₃CN:H₂O. Required fractions were concentrated under reduced pressure and further lyophilized to afford 2,2',2"-(10-(2-(((S)-l-(4-((2-(2- ((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3- yl)acety l)isoindolin-4-yl)methy l)piperidin- 1 -y 1)- 1 -oxo-4-((S)- 1 -((4-((4-(p- tolyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butan-2-yl)amino)-2- oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 8) (225 mg, yield: 16%) as a white solid. MS (ESI) m / z: 1434.0 [M+H]+. ‘H-NMR (400 MHz, DMSO- d6): 6 8.61-8.57 (brs, 1H), 8.40-8.39 (brs, 1H), 8.31-8.21 (m, 1H), 8.16 (d, J= 5.6 Hz, 1H), 7.85-7.80 (m, 2H), 7.30 (d, J= 8.0 Hz, 2H), 7.3-7.15 (m, 2 H), 7.1-7.00 (m, 5H), 5.11-5.00 (m, 1H), 4.90-4.80 (m, 2H), 4.72-4.60 (m, 3H), 4.50-4.20 (m, 4H), 4.20-4.00 (m, 4H), 3.95- 3.80 (m, 2H), 3.70-3.50 (m, 10H), 3.40-3.10 (m, 11H), 3.10-2.90 (m, 7H), 2.85-2.75 (m, 7H), 2.45-2.40 (m, 3H). 2.30-2.25 (m, 7H). 2.24 (m, 3H), 2.2-2.01(m, 4H), 1.95-1.90 (m, 2H).1.75-1.70 (m, 6H), 1.60-1.50 (m, 2H), 1.20-0.90 (m, 2H).[000250] To a solution of 4-bromoisoindoline 1A (1 g, 0.005 mol) in 2-MeTHF (10 mL, 10 Vol) at 0°C was added Et₃N (2.9 mL. 0.02 mol) and (Boc)2O (1.47 mL, 0.0067 mol). The reaction mixture was stirred at room temperature for 2h. Upon completion of the reaction,123#583961Attorney Docket No. 00020.026.1801reaction mixture was diluted with EtOAc (30 mL), water (30 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 30 mL). Combined organic layer was washed with water (30 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 5% EtOAc in pet ether as an eluent. Required fraction were concentrated under reduced pressure to afford tert-butyl 4-bromoisoindoline-2-carboxylate 2A (1.14 g, yield: 76%) as a white solid.Step-2A: Synthesis of tert-butyl 4-(piperidin-4-ylidenemethyl)isoindolme-2-carboxylate (4A):[000251] To a solution of tert-butyl 4-bromoisoindoline-2-carboxylate 2A (500 mg, 1.67 mmol) in DMF: H2O(9:1, 10 vol) was added 4-((4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)methylene)piperidine 3A (561.5 mg, 2.51 mmol) followed by Cs₂CO₃ (1.63 g, 5.03 mmol). The reaction mixture was degassed with argon for 15 min followed by addition of XPhos Pd G4 (80 mg, 0.67 mmol), resultant reaction mixture was stirred at 100°C for 16 h. Upon completion of the reaction, reaction mixture was filtered through celite bed washed with ethyl acetate (20 mL), filtrate was diluted with water (30 mL). The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 30 mL). Combined organic layers was washed with water (30 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 10% MeOH in DCM. Required fractions were concentrated under reduced pressure to afford tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate 4A (0.415 g, yield: 79%) as a brown solid. MS (ESI) m / z: 315.1[M+H]+Step-3A: Synthesis of tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate (Int #5A124#583961Attorney Docket No. 00020.026.1801[000252] To a solution of tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate 4A (680 mg, 2.162 mmol) in MeOH (3 mL) was added 10% Pd / C (680 mg, wt / wt). The resultant reaction mixture was stirred at room temperature for 16 h under hydrogen pressure (30 psi). Upon completion of the reaction, reaction mixture was filtered through celite bed, washed with MeOH (20 mL) and filtrate was concentrated under reduced pressure to get tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate (Int#5A) (523 mg, crude) as an off white solid. MS (ESI) m / z: 317.09 [M+H]+Step-IB: Synthesis of 4-( ( 4, 4, 5, 5-tetramethyl-l, 3, 2-dioxaborolan-2-yl)methylene)piperidine (3A)[000253] To a solution of tert-butyl 4-((4, 4, 5, 5-tetramethyl-l, 3, 2-dioxaborolan-2-yl)methylene)piperidine-l -carboxylate IB (1 g, 0.003 mmol) in DCM (10 mL, 10 vol), was added 4M HC1 in 1,4-di oxane (6 mL, 6 vol) at 0°C. The reaction mixture was stirred at room temperature for Ih. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude (1.3 g). Crude was triturated with diethyl ether to afford 4-((4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methylene)piperidine 3A (635 mg, yield:92%) as an off white solid. MS (ESI) m / z: 224 [M+H]+#583961Attorney Docket No. 00020.026.1801Step-IA: 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-l-yl) (S)-pyrrolidine-l,2-dicarboxylate (3A)[000254] To a solution of (tert-butoxycarbonyl)-L-proline 1A (20 g, 93.00 mmol) in DMF (200 mL, 10 Vol) at 0°C, was added DIPEA (32 mL, 186.04 mmol) followed by HBTU (38 g, 102.32 mmol). The reaction mixture was stirred at room temperature for 30 min, followed by addition of 1 -hydroxypyrrolidine-2, 5-dione 2A (16 g, 139.50 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min. White solid was obtained then wash with Hexane (2* 100 mL) to afford 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-l-yl) (S)-pyrrolidine-l,2-dicarboxylate 3A (22 g, yield: 75 %) as a white solid.Step-2A: (S)-2-( ((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-( (S)-l-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)butanoic acid (5A)[000255] To a solution of 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-l-yl) (S)-pyrrolidine-l,2-dicarboxylate 3A (10 g, 32.05 mmol) in DCM (100 mL, 10 Vol) at 0°C, was added DIPEA (11 mL, 64.10 mmol) followed by (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-aminobutanoic acid 4A (10.8 g, 32.05 mmol). Resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was diluted with water (200 mL) and extract with 10% MeOH: DCM (2* 100 mL). Finally organic layers was separated and dried overNa₂SO₄ and concentrated to obtained (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)butanoic acid 5A (15 g, yield: 87%) as a brown gummy material.Step-3A: (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid (Int-1)[000256] TFA (30 mL, 3 vol) was added to a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)butanoic acid (10 g, 0.018 mol) in DCM (100 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (100 mL) to afford (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid (Int-1) (5 g, yield: 64%) as an off white solid. MS (ESI) m / z: 438.47 [M+H]+Example 9: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrroIidin-3-yl)acetyl)isoindoIin-4- 126#583961Attorney Docket No. 00020.026.1801yl)methyl)piperidin-l-yl)-4-((S)-l-(dodecanoylglycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 9):[000257] To a solution of dodecanoic acid 1 (10 g, 50 mmol) in DMF (100 mL, 10 Vol) at 0°C was added DIPEA (13.07 mL, 75 mmol) followed by HATU (20.9 g, 55 mmol). The mixture was stirred at room temperature for 10 min, and added tert-butyl glycinate 2 (6.55 g, 50 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (200 mL), cold water (100 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 100 mL). Combined organic layer was washed with water (200 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford crude, crude was purified by silica gel column chromatography (100-200 mesh) using 10 % Ethyl acetate in hexane as an eluant. Required fractions were concentrated under reduced pressure to afford tert-butyl dodecanoylglycinate (3) (12 g, yield: 66%) as an off white solid. MS (ESI) m / z: 314.4 [M+H]+Step-2: Synthesis of dodecanoylglycine (4):127#583961Attorney Docket No. 00020.026.1801[000258] TFA (16 mL. 4 vol) was added to a solution of tert-butyl dodecanoy Iglycinate 3 (4 g, 12.77 mmol) in DCM (20 mL, 5 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 2 h. Reaction mixture w as concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (100 mL) to afford dodecanoyl glycine 4 (2.8 g, yield: 87%) as an off white solid. MS (ESI) m / z: 258.15 [M+H]+Step-3: Synthesis of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l- (dodecanoylglycyl)pyrrolidine-2-carboxamido)butanoic acid (5):[000259] To a solution of dodecanoylglycine 4 (5 g, 19.4 mmol) in DMF (50 rnL, 10 Vol) at 0°C was added DIPEA (6.74 mL, 38.8 mmol) followed by HATU (8.8 g, 23.1 mmol). The mixture was stirred at room temperature for 30 min, and added (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid Int#6A (8.5 g, 19.4 mmol). Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (400 mL) and stirred for 15 min. Organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 100 mL). Combined organic layer was washed with water (400 rnL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-(dodecanoylglycyl)pyrrolidine-2-carboxamido)butanoic acid (5) (7.0 g, yield: 53%) as an off w hite solid. MS (ESI) m / z: 677.5 [M+H]+Step-4: Synthesis of tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-(dodecanoylglycyl)pyrrolidine-2-carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate ( 6):128#583961Attorney Docket No. 00020.026.1801[000260] To a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l- (dodecanoylglycyl)pyrrolidine-2-carboxamido)butanoic acid 5 (5 g, 7.39 mmol) in DMF (50 mL, 10 Vol) at 0°C was added DIPEA (2.5 mL, 14.79 mmol) followed by HATU (5.6 g, 14.79 mmol). The mixture was stirred at room temperature for 30 min, and added tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate Int#5A (2.8 g, 8.86 mmol). Resultant reaction mixture was stirred at room temperature for 6 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (400 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 100 mL). Combined organic layer was washed with water (400 mL). Separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4- ((S)-l-(dodecanoylglycyl)pyrrolidine-2-carboxamido)butanoyl)piperidin-4- yl)methyl)isoindoline-2-carboxylate (6) (5.5 g, yield: 76%) as a brown solid. MS (ESI) m / z: 976.20 [M+H]+Step-5: Synthesis of (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-(dodecanoylglycyl)pyrrolidine-2- carboxamido)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate (7):fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-(dodecanoylglycyl)pyrrolidine-2- carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate 6 (5.5 g, 5.64 mmol) in DCM (25 mL, 5 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 2 h. Reaction mixture was concentrated under reduced pressure to afford crude. Crude 129#583961Attorney Docket No. 00020.026.1801was triturated with diethyl ether (100 mL) to afford (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-(dodecanoylglycyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate 7 (3.5 g, yield: 71%) as a brown solid. MS (ESI) m / z: 258.15 [M+H]+Step-6: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-(dodecanoylglycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (8)carbonyl)-2-oxopyrrolidin-3-yl)acetic acid (CDA-5) (1.44 g, 4.80 mmol) in DMF (35 mL) at 0°C was added DIPEA (1.39 mL, 4.80 mmol) followed by HATU (2.28 g, 6.0 mmol). The mixture was stirred at room temperature for 15 min and added (9H-fluoren-9-yl)methyl ((S)-4-((S)- 1 -(dodecanoy Igly cyl)pyrrolidine-2-carboxamido)- 1 -(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate 7 (3.5 g, 4.0 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. Reaction mixture was diluted with EtOAc (50 mL), ice cold water (50 mL) and stirred for 15 min. The aqueous layer was extracted with EtOAc (2 * 25 mL), combined organic layer was washed with ice cold water (2 * 25 mL). The organic phase separated was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4.4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindohn-4-yl)methyl)piperidin-l-yl)-4-((S)-l-(dodecanoylglycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 8 (2.8 g, yield: 60%) as a pink solid. MS (ESI) m / z: 1158.8 (M+H)+.Step-7: Synthesis of (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin- l-yl)-4-oxobutyl)-l-(dodecanoylglycyl)pyrrolidine-2-carboxamide (9)130#583961Attorney Docket No. 00020.026.1801[000263] Diethyl amine (12 mL, 4 Vol) was added to a solution of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l- (dodecanoylglycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (8) (2.8 g, 20.7 mmol) in acetonitrile (28 mL, 10 Vol) at room temperature. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude. Crude was triturated with cold EtOAc (100 mL) followed by diethyl ether (100 mL) to afford (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)pipendin-l-yl)-4-oxobutyl)-l-(dodecanoylglycyl)pyrrolidine-2-carboxamide (9) (0.66 g, yield: 81%) as a white solid. MS (ESI) m / z: 936.6 (M+H)+.Step-9: Synthesis of 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)melhyl)piperidin-l-yl)-4-((S)-l-(dodecanoylglycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-],4.7, 10-tetraazacyclododecane-1.4.7 -triyl) triacetic acid (Compound 9)difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-y l)methy l)piperidin- 1 -y l)-4-oxobuty 1)- 1 -(dodecanoylgly cy l)pyrrolidine-2-carboxamide (9) (1.5 g, 1.607 mmol) in acetonitrile (15 mL, 10 Vol), at room temperature, was added DIPEA (2.5 mL, 14.4 mmol) followed by DOTA-NHS ESTER (1.2 g, 2.40 mmol). The resultant 131#583961Attorney Docket No. 00020.026.1801reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. The crude material was triturated with EtOAc (100 mL) and further purified by prep HPLC using the below condition.Prep conditionColumn Gemini, C18 (250*21.5mm)5pBuffer A: 0.2% FA in H2O, B:0.2 % FA in ACN: H2O (80:20)Gradient (T / %B) 0 / 40,20 / 60,22 / 60,22.05 / 98Flow Rate 12mL / minSolubility ACN+H2O[000265] The required fractions were collected and lyophilized to afford 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3 -y l)acety l)isoindolin-4-y l)methy l)piperidin- 1 -yl )-4-((S)- 1 -(dodecanoylglycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)- I,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 9) (250 mg, yield: II.8% ) as a white solid. MS (ESI) m / z: 1320.99 [M-H]⁻. ¹H-NMR (400 MHz, DMSO-d6): δ 8.26 (brs, 2H), 8.8-8.7 (brs, 1H), 8.08-8.05 (brs, 1H), 7.90-7.89 (m, 1H), 7.26-7.20 (m, 1H), 7.22-7.21 (m, 2H), 7.08-7.07 (m, 1H), 5.12-5.11 (m, 1H), 4.90-4.80 (m, 2H), 4.70-4.60 (m, 3H), 4.40-4.30 (m, 2H), 4.32-4.28 (m, 2H), 4.15-4.10 (m, 1H), 4.00-3.80 (m, 3H), 3.60-3.40 (m, 9 H), 3.20-3.10 (m, 4H), 3.05-2.95 (m, 14H), 2.80-2.60 (m, 7H), 2.45-2.40 (m, 2H), 2.30-2.25 (m, 1H), 2.15-2.10 (m, 4H), 2.09-2.00 (m, 1H), 1.90-1.80 (m, 5H), 1.65-1.55 (m, 3H), 1.45-1.40 (m, 2H), 1.35-1.20 (m, 18H), 1.10-1.05 (m, 1H), 0.84 (t, J=6.8 Hz, 3H) Synthesis of Int#5A:Step-IA: Synthesis of tert-butyl 4-bromoisoindoline-2-carboxylate (2A):132#583961Attorney Docket No. 00020.026.1801[000266] To a solution of 4-bromoisoindoline 1A (1 g, 0.005 mol) in 2-MeTHF (10 mL, 10 Vol) at 0°C was added EtsN (2.9 mL. 0.02 mol) and (Boc)₂O (1.47 mL, 0.0067 mol). The reaction mixture was stirred at room temperature for 2h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (30 mL), water (30 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 30 mL). Combined organic layer was washed with water (30 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 5% EtOAc in pet ether as an eluent. Required fraction were concentrated under reduced pressure to afford tert-butyl 4-bromoisoindoline-2-carboxylate 2A (1.14 g, yield: 76%) as a white solid.Step-2A: Synthesis of tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate (4A):[000267] To a solution of tert-butyl 4-bromoisoindoline-2-carboxylate 2A (500 mg, 1.67 mmol) in DMF: H2O(9:1, 10 vol) was added 4-((4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methylene)piperidine 3A (561.5 mg, 2.51 mmol) followed by Cs₂CO₃ (1.63 g, 5.03 mmol). The reaction mixture was degassed with argon for 15 min followed by addition of XPhos Pd G4 (80 mg, 0.67 mmol), resultant reaction mixture was stirred at 100°C for 16 h. Upon completion of the reaction, reaction mixture was filtered through celite bed washed with ethyl acetate (20 mL), filtrate was diluted with water (30 mL). The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 30 mL). Combined 133#583961Attorney Docket No. 00020.026.1801organic layers was washed with water (30 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 10% MeOH in DCM. Required fractions were concentrated under reduced pressure to afford tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate 4A (0.415 g, yield: 79%) as a brown solid. MS (ESI) m / z: 315.1[M+H]+Step-3A: Synthesis of tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate (Int#5A[000268] To a solution of tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate 4A (680 mg, 2.162 mmol) in MeOH (3 mL) was added 10% Pd / C (680 mg, wt / wt). The resultant reaction mixture was stirred at room temperature for 16 h under hydrogen pressure (30 psi). Upon completion of the reaction, reaction mixture was filtered through celite bed, washed with MeOH (20 mL) and filtrate was concentrated under reduced pressure to get tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate (Int#5A) (523 mg, crude) as an off white solid. MS (ESI) m / z: 317.09 [M+H]+Step-1 B: Synthesis of 4-( ( 4, 4, 5, 5-tetramethyl-l, 3, 2-dioxaborolan-2-yl)methylene)piperidine (3A)[000269] To a solution of tert-butyl 4-((4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)methylene)piperidine-l -carboxylate IB (1 g, 0.003 mmol) in DCM (10 mL, 10 vol), was added 4M HCl in 1,4-dioxane (6 mL, 6 vol) at 0°C. The reaction mixture was stirred at room temperature for Ih. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude (1.3 g). Crude was triturated with diethyl ether to afford134#583961Attorney Docket No. 00020.026.18014-((4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)methylene)piperidine 3A (635 mg, yield: 92%) as an off white solid. MS (ESI) m / z: 224 [M+H]+Step-IA: 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-l-yl) (S)-pyrrolidine-l,2-dicarboxylate (3A) To a solution of (tert-butoxycarbonyl)-L-proline 1A (20 g, 93.00 mmol) in DMF (200 mL, 10 Vol) at 0°C, was added DIPEA (32 mL, 186.04 mmol) followed by HBTU (38 g. 102.32 mmol). The reaction mixture was stirred at room temperature for 30 min, followed by addition of 1 -hydroxypyrrolidine-2, 5-dione 2A (16 g, 139.50 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min. White solid was obtained then wash with Hexane (2* 100 mL) to afford 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-1-yl) (S)-pyrrolidine-l,2-dicarboxylate 3A (22 g, yield: 75 %) as a white solid.Step-2A: (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)butanoic acid (5A)[000270] To a solution of 1 -(tert-butyl) 2-(2.5 -di oxo py nolidin- 1-yl) (S)-pyrrolidine-l,2-dicarboxylate 3A (10 g, 32.05 mmol) in DCM (100 mL, 10 Vol) at 0°C, was added DIPEA (11 mL, 64.10 mmol) followed by (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-aminobutanoic acid 4A (10.8 g, 32.05 mmol). Resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was diluted with water (200 mL) and extract with 10% MeOH: DCM (2* 100 mL). Finally organic layers was separated and dried over Na₂SO₄ and concentrated to obtained (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)butanoic acid 5A (15 g, yield: 87%) as a brown gummy material.Step-3A: (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid (Int-1)[000271] TFA (30 mL, 3 vol) was added to a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)butanoic acid (10 g, 0.018 mol) in DCM (100 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction,135#583961Attorney Docket No. 00020.026.1801reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (100 mL) to afford (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid (Int-1) (5 g, yield: 64%) as an off white solid. MS (ESI) m / z: 438.47 [M+H]+Example 10: Synthetic scheme for2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperazin-l-yI)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 10)Step-1: Synthesis of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate (3):[000272] To a solution of 4-(4-iodophenyl)butanoic acid 1 (30 g, 0.103 mol) in DMF (60 mL, 2 Vol) at 0°C was added DIPEA (27 mL, 0.155 mol) followed by HATU (39.3 g, 0.103 mol). The reaction mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl 4-(aminomethyl)benzoate 2 (21.4 g, 0.103 mol). Resultant reaction mixture was stirred at room temperature for 4 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (400 mL) and stirred for 15 min. The136#583961Attorney Docket No. 00020.026.1801organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 400 mL). Combined organic layer was washed with water (400 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (36.1 g, crude) as a brown liquid. MS (ESI) m / z: 480.1 [M+H]+Step-2: Synthesis of 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid (4):[000273] TFA (160 mL, 4 vol) w as added to a solution of tert-buty l 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (40 g. 0.083 mol) in DCM (200 mL. 5 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 5 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (200 mL) to afford 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid 4 (31 g, yield: 88%) as a brown solid. MS (ESI) m / z: 424.42 [M+H]+Step-3: Synthesis of tert-butyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycinateg, 23.64 mmol) in DMF (100 mL, 10 Vol) at 0°C, was added DIPEA (12.3 mL, 70.92 mmol ) followed by HATU (13.4 g, 35.46 mmol). The reaction mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl glycinate 5 (3.5 mL, 26.00 mmol). Resultant reaction mixture w as stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min.Precipitated solid w as filtered and wash with hexane (2* 100 mL) to afford tert-buty l (4-((4-#583961Attorney Docket No. 00020.026.1801(4-iodophenyl)butanamido)methyl)benzoyl)glycinate 6 (10 g, yield: 83 %) as a white solid. MS (ESI) m / z:538.28 (M+H)+.Step-4: Synthesis of (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine (7):[000275] TFA (18 mL, 3 vol) was added to a solution of tert-buty l (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycinate 6 (6 g, 0.011 mol) in DCM (60 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 5 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (100 mL) to afford (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine 7 (4.5 g, yield: 86%) as a white solid. MS (ESI) m / z: 481.13 [M+H]+Step-5: Synthesis of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-1-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid (8):[000276] To a solution of (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine 7 (2.5 g, 5.20 mmol) in DMF (25 mL, 10 Vol) at 0°C was added DIPEA (1.3 mL, 7.81 mmol) followed by addition of HATU (2.17 g, 5.72 mmol). The reaction mixture was stirred at room temperature for 10 min followed by addition of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid Int#l (2.27 g.5.20 mmol). The resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted with cold water (100 mL) and stirred for 15 min. Obtained white solid was dissolve in 10%MeOH: DCM (20 mL) then triturated with DEE (100 mL) to afford (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-138#583961Attorney Docket No. 00020.026.1801carboxamido)butanoic acid Int#8 (3.6 g, yield: 78%) as a white solid. MS (ESI) m / z: 900.47 (M+H)+.Step-6: Synthesis of tert-butyl 4-( ( 4-((S)-2-( ( ((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperazin-l-yl)methyl)isoindoline-2-carboxylate (9)[000277] To a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid Int#8 (3 g. 3.33 mmol) in DMF (30 mL, 10 Vol) at 0°C, was added DIPEA (1.1 mL, 6.67 mmol ) followed by HATU (1.26 g, 3.33 mmol). The reaction mixture was stirred at room temperature for 10 min, followed by addition of tert-butyl 4-(piperazin-l-ylmethyl)isoindoline-2-carboxylate Int#5A (1.2 g, 4.00 mmol). The resultant reaction mixture was stirred at room temperature for 2h. Upon completion of the reaction, reaction mixture was diluted with cold water (100 mL) and stirred for 15 min. Obtained white solid was dissolve in 10%MeOH: DCM (20 mL) then triturated with diethyl ether (100 mL) to afford tert-buty l 4-((4-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l -((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperazin-l-yl)methyl)isoindoline-2-carboxylate 9 (3.1 g. yield: 77 %) as a white solid. MS (ESI) m / z: 1199.8 (M+H)+.Step-7: Synthesis of (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin-4-ylmethyl)piperazin-l-yl)-l-oxobutan-2-yl)carbamate (10)139#583961Attorney Docket No. 00020.026.1801[000278] TFA (12.4 mL, 4 vol) was added to a solution of tert-butyl 4-((4-((S)-2-((((9H-fluoren-9-y l)methoxy )carbonyl)amino)-4-((S)- 1 -((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperazin-l-yl)methyl)isoindoline-2-carboxylate 9 (3.1 g, 2.58 mmol) in DCM (124 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (200 mL) to afford ((9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin-4-ylmethyl)piperazin-l-yl)-l-oxobutan-2-yl)carbamate (10) (2.8 g, yield: 92%) as an off white solid. MS (ESI) m / z: 1099.7 [M+H]+Step-8: Synthesis of (9H-fluoren-9-yl)methyl ((S)-1-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-1-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperazin-1-yl)-4-((S)-1-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-1-oxobutan-2-#583961Attorney Docket No. 00020.026.1801[000279] To a solution of CDA-5 (850 mg, 2.83 mmol) in DMF (26 ml, 10 Vol) at 0°C was added DIPEA (0.82 mL, 4.73 mmol) followed by addition of HATU (890 mg. 2.36 mmol). The reaction mixture was stirred at room temperature for 10 min followed by addition of (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin-4-ylmethyl)pi perazin- l-yl)-l -oxob utan-2-yl)carbamate (10 )(2.6 g, 2.36 mmol). Resultant reaction mixture was stirred at room temperature for 2h. Upon completion of the reaction, reaction mixture was diluted with cold water (100 mL) and stirred for 15 min. Obtained white solid was dissolve in 10%MeOH: DCM (20 mL) then triturated with DEE (100 mL) to afford (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-1-oxobutan-2-yl)carbamate (11) (2.2 g, yield: 68%) as an off white solid. MS (ESI) m / z:1383.43 (M+H)+.Step-9: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)henzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamale (12)[000280] Diethyl amine (4.8 mL, 4 Vol) was added to a solution of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3 -y l)acety l)isoindolin-4-yl)methyl)piperazin- 1 -yl)-4-((S)- 1 -((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-#583961Attorney Docket No. 00020.026.1801yl)carbamate (11) (2.2 g, 1.59 mmol) in acetonitrile (22 mL, 10 Vol) at room temperature. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude. Crude material was triturated with cold EtOAc (10 mL) followed by diethyl ether (10 mL) to afford (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate (12) (1.5 g, yield: 83%) as a white solid. MS (ESI) m / z: 1160.6 (M+H)+.Step-10: Synthesis of 2,2',2”-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4, 7, 10-tetraazacyclododecane-l,4, 7-triyl)triacetic acid (compound 10)[000281] To a solution of (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-y l)methy l)piperazin- 1 -yl)-4-oxobuty 1)- 1 -((4-((4-(4- 142#583961Attorney Docket No. 00020.026.1801iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide 12 (1 g, 0.96 mmol) in acetonitrile (10 mL, 10 Vol) at room temperature, was added DIPEA (1.3 mL. 6.97 mmol) followed by 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid DOTA NHS ester (791 mg, 1.29 mmol). The resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. The crude material was triturated with EtOAc (20 mL) then obtained material was purified by grace reverse phase, eluted at 45-50% CH₃CN:H₂O. Required fractions were concentrated under reduced pressure and further lyophilized to afford 2,2',2"-(10-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid compound 10 (210 mg, yield: 15.7%) as a white solid. MS (ESI) m / z: 1546.81 [M+H]+’H-NMR (400 MHz, DMSO-dd): 5 8.6-8.5 (brs, 2H), 8.37 (t, J= 6 Hz, 1H). 8.13 (s, 1H), 7.9-7.8 (m. 3H), 7.7-7.61 (m, 2H), 7.41-7.22 (m, 4H), 7.2-7.1 (m, 1H). 7.01-6.99 (m, 2 H), 5.11-5.00 (m, 1H), 4.90-4.80 (m, 2H), 4.72-4.60 (m, 3H), 4.40-4.30 (m, 3H), 4.20-4.00 (m, 5H), 3.82-3.50 (m, 14H), 3.30-3.00 (m, 19H), 2.91-2.75 (m, 8H), 2.65-2.50 (m, 2H), 2.40-2.35 (m, 4H), 2.25-2.10 (m, 6H), 1.95-1.90 (m, 2H), 1.75-1.70 (m, 4H), 1.60-1.50 (m, 1H)[000282] To a solution of 4-bromoisoindoline 1A (1 g, 0.005 mol) in 2-MeTHF (10 mL, 10 Vol) at 0°C was added Et₃N (2.9 mL, 0.02 mol) and (Boc)2O (1.47 mL, 0.0067 mol). The 143#583961Attorney Docket No. 00020.026.1801reaction mixture was stirred at room temperature for 2h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (30 mL), water (30 mL) and stirred for 15 min. The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 30 mL). Combined organic layer was washed with water (30 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 5% EtOAc in pet ether as an eluent. Required fraction were concentrated under reduced pressure to afford tert-butyl 4-bromoisoindoline-2-carboxylate 2A (1.14 g, yield: 76%) as a white solid.Step-2A: Synthesis of tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate (4A):[000283] To a solution of tert-butyl 4-bromoisoindoline-2-carboxylate 2A (500 mg, 1.67 mmol) in DMF: H2O(9:1, 10 vol) was added 4-((4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)methylene)piperidine 3A (561.5 mg, 2.51 mmol) followed by Cs₂CO₃ (1.63 g, 5.03 mmol). The reaction mixture was degassed with argon for 15 min followed by addition of XPhos Pd G4 (80 mg, 0.67 mmol), resultant reaction mixture was stirred at 100°C for 16 h. Upon completion of the reaction, reaction mixture was filtered through celite bed washed with ethyl acetate (20 mL), filtrate was diluted with water (30 mL). The organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 30 mL). Combined organic layers was washed with water (30 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 10% MeOH in DCM. Required fractions were concentrated under reduced pressure to afford tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate 4A (0.415 g, yield: 79%) as a brown solid. MS (ESI) m / z: 315.1[M+H]+Step-3A: Synthesis of tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate (Int#5A144#583961Attorney Docket No. 00020.026.1801[000284] To a solution of tert-butyl 4-(piperidin-4-ylidenemethyl)isoindoline-2-carboxylate 4A (680 mg, 2.162 mmol) in MeOH (3 mL) was added 10% Pd / C (680 mg, wt / wt). The resultant reaction mixture was stirred at room temperature for 16 h under hydrogen pressure (30 psi). Upon completion of the reaction, reaction mixture was filtered through celite bed, washed with MeOH (20 mL) and filtrate was concentrated under reduced pressure to get tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate (Int#5A) (523 mg, crude) as an off white solid. MS (ESI) m / z: 317.09 [M+H]+Step-IB: Synthesis of 4-( ( 4, 4, 5, 5-tetramethyl-l, 3, 2-dioxaborolan-2-yl)methylene)piperidine (3A)[000285] To a solution of tert-butyl 4-((4, 4, 5, 5-tetramethyl-l, 3, 2-dioxaborolan-2-yl)methylene)piperidine-l -carboxylate IB (1 g, 0.003 mmol) in DCM (10 mL, 10 vol), was added 4M HC1 in 1,4-di oxane (6 mL, 6 vol) at 0°C. The reaction mixture was stirred at room temperature for Ih. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude (1.3 g). Crude was triturated with diethyl ether to afford 4-((4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methylene)piperidine 3A (635 mg, yield:92%) as an off white solid. MS (ESI) m / z: 224 [M+H]+#583961Attorney Docket No. 00020.026.1801Step-IA: 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-l-yl) (S)-pyrrolidine-l,2-dicarboxylate (3A)[000286] To a solution of (tert-butoxycarbonyl)-L-proline 1A (20 g, 93.00 mmol) in DMF (200 mL, 10 Vol) at 0°C, was added DIPEA (32 mL, 186.04 mmol) followed by HBTU (38 g, 102.32 mmol). The reaction mixture was stirred at room temperature for 30 min, followed by addition of 1 -hydroxypyrrolidine-2, 5-dione 2A (16 g, 139.50 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min. White solid was obtained then wash with Hexane (2* 100 mL) to afford 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-l-yl) (S)-pyrrolidine-l,2-dicarboxylate 3A (22 g, yield: 75 %) as a white solid.Step-2A: (S)-2-( ((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-( (S)-l-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)butanoic acid (5A)[000287] To a solution of 1 -(tert-butyl) 2-(2,5-dioxopyrrolidin-l-yl) (S)-pyrrolidine-l,2-dicarboxylate 3A (10 g, 32.05 mmol) in DCM (100 mL, 10 Vol) at 0 °C, was added DIPEA (11 mL, 64.10 mmol) followed by (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-aminobutanoic acid 4A (10.8 g, 32.05 mmol). Resultant reaction mixture was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was diluted with water (200 mL) and extract with 10% MeOH: DCM (2* 100 mL). Finally organic layers was separated and dried overNa₂SO₄ and concentrated to obtained (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)butanoic acid 5A (15 g, yield: 87%) as a brown gummy material.Step-3A: (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid (Int-1)[000288] TFA (30 mL, 3 vol) was added to a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-(tert-butoxycarbonyl)pyrrolidine-2-carboxamido)butanoic acid (10 g, 0.018 mol) in DCM (100 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude material, crude material was triturated with diethyl ether (100 mL) to afford (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid (Int-1) (5 g, yield: 64%) as an off white solid. MS (ESI) m / z: 438.47 [M+H]+Example 11: Synthesis scheme for 6-((16-((6-carboxypyridin-2-yl)methyl)-l, 4, 10,13-tetraoxa-7,16-diazacyclooctadecan-7-yI)methyl)-4-(4-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2- 146#583961Attorney Docket No. 00020.026.1801cyan o-4, 4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl) butanamido) methyl) benzoyl) glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-4-oxobutanamido)picolinic acid (compound 11)Step-1: Synthesis of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate (3):[000289] To a solution of 4-(4-iodophenyl)butanoic acid 1 (30 g, 0.103 mol) in DMF (60 mL, 2 Vol) at 0°C was added DIPEA (27 mL, 0.155 mol) followed by HATU (39.3 g, 0.103 mol). The reaction mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl 4-(aminomethyl)benzoate 2 (21.4 g. 0.103 mol). Resultant reaction mixture was stirred at room temperature for 4 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (200 mL) and stirred for 15 min.Organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 400 mL). Combined organic layer was washed with water (400 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (36.1 g, crude) as a brown liquid. MS (ESI) m / z: 480.1 [M+H]+Step-2: Synthesis of 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid (4):#583961Attorney Docket No. 00020.026.1801[000290] TFA (160 mL, 4 vol) was added to a solution of tert-buty l 4-((4-(4- iodophenyl)butanamido)methyl)benzoate 3 (40 g. 0.083 mol) in DCM (200 mL. 5 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 5 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (200 mL) to afford 4-((4-(4- iodophenyl)butanamido)methyl)benzoic acid 4 (31 g, yield: 88%) as a brown solid. MS (ESI) m / z: 424.42 [M+H]+Step-3: Synthesis of tert-butyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycinateg, 23.64 mmol) in DMF (100 mL, 10 Vol) at 0 °C, was added DIPEA (12.3 mL, 70.92 mmol ) followed by HATU (13.4 g, 35.46 mmol). The mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl glycinate 5 (3.5 mL, 26.00 mmol). Resultant reaction was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min, precipitated solid was filtered and wash with hexane (2* 100 mL) to afford tert-buty l (4-((4-(4- iodophenyl)butanamido)methyl)benzoyl)glycinate 6 (10 g, yield: 83 %) as awhile solid. MS (ESI) m / z:538.28 [M+H]+.Step-4: Synthesis of (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine (7):[000292] TFA (18 mL, 3 vol) was added to a solution of tert-butyl (4-((4-(4- iodophenyl)butanamido)methyl)benzoyl)glycinate 6 (6 g, 0.011 mol) in DCM (60 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to 148#583961Attorney Docket No. 00020.026.1801afford crude. Crude was triturated with diethyl ether (100 mL) to afford (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine 7 (4.5 g. yield: 86%) as a white solid. MS (ESI)m / z: 481.13 [M+H]+Step-5: Synthesis of (S)-2-( ( f9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-( ( 4-( 4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid (8):(2.5 g, 5.20 mmol) in DMF (25 mL, 10 Vol) at 0°C was added DIPEA (1.3 mL, 7.81 mmol) followed by addition of HATU (2.17 g, 5.72 mmol). Reaction mixture was stirred at room temperature for 10 min followed by addition of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid Int#l (2.27 g, 5.20 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted with cold water (100 mL) and stirred for 15 min. obtained solid was dissolve in 10%MeOH: DCM (20 mL) then triturated with diethyl ether (100 mL) to afford (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl) butanamido) methyl) benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid 8 (3.6 g, yield: 78%) as a white solid. MS (ESI) m / z: 900.47 [M+H]+.Step-6: Synthesis of tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)gfycyl)pyrrolidine-2-carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate: (9)149#583961Attorney Docket No. 00020.026.1801[000294] To a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid 8 (9.2 g, 10.5 mmol) in DMF (90 mL, 10 Vol) at 0°C, was added DIPEA (3.66 mL, 23.00 mmol ) followed by HATU (4.38 g, 11.5 mmol), the reaction mixture was stirred for 10 min and added tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate Int#5A (3.64 g, 11.51 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (500 mL) and stirred for 15 min. Obtained white solid was dissolve in 10%MeOH: DCM (50 mL) then triturated with DEE (250 mL) to afford tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate 9 (10.2 g, yield: 83%) as a white solid. MS (ESI) m / z: 1198.69 [M+H]+.Step-7: Synthesis of (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate (10)[000295] TFA (40 mL, 4 vol) was added to a solution of tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate 9 (10 g, 8.34 mmol) in DCM (100 mL, 10 Vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (400 mL) to afford (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin- 150#583961Attorney Docket No. 00020.026.18014-ylmethyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate 10 (8 g, yield: 87%) as an off white solid. MS (ESI) m / z: 1098.94 [M+H]+Step-8: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-1-oxobutan-2-yl)carbamate (11)[000296] To a solution of CDA-5 (3.32 g, 11.05 mmol) in DMF (20 mL, 10 Vol) at 0°C was added DIPEA (2.96 mL, 17.0 mmol) followed by addition of HATU (3.55 g, 9.35 mmol). Reaction mixture was stirred at room temperature for 10 min followed by addition of (9H-fluoren-9-yl)methy 1 ((S)-4-((S)- 1 -((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate 10 (9.34 g, 8.504 mmol). Resultant reaction was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min. Obtained white solid was triturated with diethyl ether (200 mL) to afford (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-y I (acety I )i somdol i n-4-y l)methyl)piperidin- 1 -yl)-4-((S)- 1 -((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrohdine-2-carboxamido)-l-oxobutan-2-yl)carbamate 11 (8 g, yield: 62%) as an off white solid. MS (ESI) m / z: 1382.19 [M+H]+. Step-9: Synthesis of (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidme-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide (12)151#583961Attorney Docket No. 00020.026.1801[000297] Diethyl amine (0.44 mL, 4.2 mmol) was added to a solution of (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 11 (7.3 g, 5.28 mmol) in DMF (73 mL, 10 Vol) at room temperature. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with MTBE ( 730 mL) and obtained solid was filtered and wash with EtOAc (30 mL) to afford (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)pipendin-l-yl)-4-oxobutyl)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide 12 (4.9 g, yield: 79%) as a white solid. MS (ESI) m / z: 1159.78 [M+H]+.Step-10: Synthesis of 6-((16-((6-carboxypyridin-2-yl)methyl)-l,4,10,13-tetraoxa-7,16-diazacyclooctadecan- 7-yl)methyl)-4-(4-( ( (S)-l-(4-((2-(2-( ( 3S.5S)-5-( (S)-2-cyano-4.4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-4-oxobutanamido)picolinic acid (compound 11)152#583961Attorney Docket No. 00020.026.1801[000298] To a solution of 4-(3-carboxypropanamido)-6-((16-((6-carboxypyridin-2-yl)methy 1)- 1,4, 10, 13-tetraoxa-7, 16-diazacy clooctadecan-7-yl)methyl)picolinic acid Int#7A (178 mg, 0.15 mmol) in DMF (2.5 mL, 10 Vol) at 0°C, was added DIPEA (0.1 mL, 0.578 mmol) followed by addition of HATU (161 mg. 0.42 mmol). Reaction mixture was stirred at room temperature for 10 min followed by addition of (S)-N-((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide 12 (250 mg, 0.38 mmol). Resultant reaction mixture was stirred at 0 °C for 15 min. Upon completion of the reaction, reaction mixture was diluted with cold water (20 mL) and stirred for 15 min.Obtained white solid was dissolve in 10% MeOH: DCM (50 mL) then added with diethyl ether (200 mL) to afford crude. Crude was purified by prep-HPLC using below condition. Required fractions were concentrated under reduced pressure and further lyophilized to afford 6-(( 16-((6-carboxy py ri din-2 -yl)methyl)- 1,4, 10, 13 -tetraoxa-7, 16-diazacyclooctadecan-7-yl)methyl)-4-(4-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-4-oxobutanamido)picolinic acid (21 mg) as an off white solid (compound 11). MS (ESI) m / z: 1790.5 [M+H]+. ¹H-NMR (400 MHz, DMSO-d6): 5 13.2- 153#583961Attorney Docket No. 00020.026.180113.0 (brs, 1H), 10 -9.8 (brs, 1H), 8.4-8.6 (m, 1H), 8.4-8.3 (m, 1H), 8.3-8.2 (m, 1H), 8.2-8.1 (m. 2H), 7.99-7.97 (m. 1H), 7.8-7.70 (m, 3H). 7.6 (d, J= 8 Hz, 2H), 7.31 (d, J= 7.6 Hz, 2H), 7.25-7.15 (m, 2H), 7.05-6.9 (m, 3H), 5.13-5.11 (m, 1H), 4.7-4.5 (m, 9H), 4.4-4.0 (m, 10H), 3.70-3.50 (m, 12H), 3.50-3.30 (m, 17H), 3.20-3.1 (m, 2H), 2.9-2.70 (m, 6H), 2.7 -2.6 (m, 2H), 2.5-2.4 (m, 4H), 2.30-2.1 (tn, 6H), 2.1-2.0 (m, 1H), 1.9-1.70 (m, 7H), 1.6-1.4 (m, 3H), 1.01-0.9 (m, 3 H)Synthesis ofInt#7A:Step-1 A: Synthesis of methyl 6-((l,4, 10, 13-tetraoxa-7, 16-diazacyclooctadecan-7-yl)methyl)picolinate (3A)[000299] To a solution of l,4,10,13-tetraoxa-7,16-diazacyclooctadecane 1A (1 g, 39.7 mmol), in acetonitrile (20 mL, 10 vol), at 75° C was added DIPEA (5.8 g, 47.6 mmol) followed by methyl 6-(chloromethyl)picolinate 2A (18.2 mL. 79.36 mmol). Resultant reaction mixture was stirred at 75° C for 48 h. Progress of reaction was monitored by TLC. Upon completion of reaction, the reaction mixture was concentrated under reduced pressure to afford crude. Crude was purified by silica gel column chromatography (100-200 mesh), eluted with 10% MeOH in DCM to afford methyl 6-((l,4.10,13-tetraoxa-7,16-diazacyclooctadecan-7-yl)methyl)picolinate 3A (700 mg, yield:46%) as a brown liquid. MS (ESI) m / z: 412.3 [M+H]+Step-2A: Synthesis of methyl 4-amino-6-((16-((6-(methoxycarbonyl)pyridin-2-yl)methyl)-1.4,10,13-tetraoxa-7.16-diazacyclooctadecan-7-yl)methyl)picolinate ( 4A)154#583961Attorney Docket No. 00020.026.1801[000300] To a solution of methyl 6-((l,4,10,13-tetraoxa-7,16-diazacyclooctadecan-7-yl)methyl)picolinate (3A) (1 g, 2.42 mmol) in acetonitrile (20 mL, 20 Vol), at room temperature was added methyl 4-amino-6-(chloromethyl)picolinate Int#5A (0.97 g, 4.85 mmol), DIPEA (6.3 mL, 36.3 mmol). Resultant reaction mixture was stirred at 75°C for 16 h. Progress of reaction was monitored by TLC. Upon completion of reaction, the reaction mixture was concentrated under reduced pressure to afford crude. Crude was purified by silica gel column chromatography (100-200 mesh) eluted at 8% MeOH in DCM and compound contained fractions were concentrated under reduced pressure to afford methyl 4-amino-6-(( 16-((6-(methoxy carbonyl)py ridin-2-y l)methyl)- 1,4, 10, 13 -tetraoxa-7, 16-diazacyclooctadecan-7-yl)methyl)picolinate (4A) (380 mg, yield: 27%) as a brown liquid. MS (ESI) m / z: 576.4 [M+H]+.Step-3A: Synthesis of 4-((2-(methoxycarbonyl)-6-( ( 16-( ( 6-(methoxycarbonyl)pyridin-2-yl)methyl)-l, 4, 10, 13-tetraoxa- 7, 16-diazacyclooctadecan- 7-yl)methyl)pyridin-4-yl)amino)-4-[000301] To a solution of methyl 4-amino-6-((16-((6-(methoxycarbonyl)pyridin-2-yl)methyl)-1.4.10,13-tetraoxa-7.16-diazacyclooctadecan-7-yl)methyl)picolinate 4A (1 g. 1.88 mmol) in DCM (10 mL, 10 vol) at 0°C was added DIPEA (0.33 mL, 2.32 mmol) followed by succinyl dichloride 5A (0.294 g, 1.9 mmol). The reaction mixture was stirred at room temperature for 4h. Progress of reaction was monitored by TLC. Upon completion of reaction, the reaction mixture was quenched with water and neutralized with Sat. NaHCOs (20 mL) solution. Aqueous layer was separated and concentrated under reduced pressure to 155#583961Attorney Docket No. 00020.026.1801afford crude. Crude was purified by grace reverse phase column chromatography eluted with 10% CHsCN in water to afford 4-((2-(methoxycarbonyl)-6-((16-((6- (methoxy carbonyl)pyridin-2-y l)methy 1)-1,4, 10, 13-tetraoxa-7, 16-diazacyclooctadecan-7 -yl)methyl)pyridin-4-yl)amino)-4-oxobutanoic acid 6A (350 mg, yield: 31%) as a brown solid. MS (ESI) m / z: 676.55 [M+H]+.Step-4A: Synthesis of 4-(3-carboxypropanamido)-6-((16-((6-carboxypyridin-2-yl)methyl)-1.4.10,13-tetraoxa-7.16-diazacyclooctadecan- 7-yl)methyl)picolmic acid (Int- 7 A)[000302] To a solution of 4-((2-(methoxycarbonyl)-6-((16-((6- (methoxy carbonyl)pyridin-2-yl)methyl)-l,4, 10, 13-tetraoxa-7, 16-diazacyclooctadecan-7 -yl)methyl)pyridin-4-yl)amino)-4-oxobutanoic acid (6A) (300 mg, 0.44 mmol) in MeOH:THF:H2O (1:2:1, 3 mL, 10 Vol), at 0° C was added LiOH.2H₂O (74 mg, 1.77 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. Progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was poured into ice water (50 mL), and washed with EtOAc (2 x 20 mL). Organic layer was separated and aqueous layer was acidified with IN HC1 followed by lyophilization to afford 4-(3-carboxypropanamido)-6-(( 16-((6-carboxy pyridin-2-yl)methy 1)- 1,4,10,13 -tetraoxa-7, 16-diazacyclooctadecan-7-yl)methyl)picolinic acid Int-7A (0.4 g, yield: 57%) as a brown solid compound. MS (ESI) m / z: 648.54 [M+H]TSynthesis of Int#5AClStep-IB: Synthesis of tert-butyl dimethyl 4-azidopyridine-2,6-dicarboxylate (2B)#583961Attorney Docket No. 00020.026.1801[000303] To a solution of dimethyl 4-chloropyridine-2,6-dicarboxylate IB (25 g, 0.108 mmol) in DMF (250 mL, 10 Vol) at 0°C was added NaN3(68.5 g, 1.088 mmol). Resultant reaction mixture was stirred at 70 °C for 16 h. Progress of the reaction was monitored by TLC. Reaction mixture was diluted with ice water (50 mL) and precipitated solid was filter and dried under vacuum to afford dimethyl 4-azidopyridine-2,6-dicarboxylate 2B (7.2 g, 76%) as a pale-yellow solid. MS (ESI) m / z: 237 [M+H]+.Step-2B: Synthesis of methyl 4-amino-6-(hydroxymethyl)picolinate (3B):[000304] To a solution of tert-butyl dimethyl 4-azidopyridine-2,6-dicarboxylate 2B (4 g, 0.016 mol) in MeOH (40 mL, 10 vol) at 0°C, was added NaBH4(1.28 g, 0.033). Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was filtered and filtrate was concentrated under reduced pressure to get crude. Crude was purified by silica gel (100-200 mesh) eluted at 4% methanol in dichloromethane to afford methyl 4-amino-6-(hydroxymethyl)picolinate 3B (1.6 g, yield: 29%) as a pale yellow solid. MS (ESI) m / z: 183.2 [M+H]+.Step-3B: Synthesis of methyl 4-amino-6-(chloromethyl)picolinate (Int#5A)[000305] Methyl 4-amino-6-(hydroxymethyl)picolinate 4B (1 g, 5.49 mmol) was added SOCl2(10 mL, 10 vol) at 0°C. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get residue. Residue was dissolved in ethyl acetate and washed with Saturated NaHCO3solution. Organic layer was washed with water (40 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford Crude. Crude was purified by column chromatography silica gel (100-200157#583961Attorney Docket No. 00020.026.1801mesh) eluted at 5% MeOH in dichloromethane to afford methyl 4-amino-6-(chloromethyl)picolinate (Int#5A) (800 mg. yield: 73%) as a brown gummy.Example 12: Synthesis of 2,2'-(7-(2-(((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7-triazonane-l,4-diyl)diacetic acid (compound 12)Step-1: Synthesis of tert-butyl 4-((4-(4-fluorophenyl)butanamido)methyl)benzoate[000306] To a solution 4-(4-fluorophenyl)butanoic acid 1 (10 g, 54.94 mmol) in DMF (100 mL. 10 Vol) at 0°C, were added DIPEA (20 mL, 109.88 mmol) followed by HATU (22 g, 60.43 mmol). The mixture was stirred at 0°C for 10 min, and added tert-butyl 4-(aminomethyl)benzoate (11.27 g, 54.94 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. The progress of reaction was monitored by TLC. Upon completion of the reaction, reaction was quenched with ice cold water, solid obtained was dissolved in 10% MeOH in DCM, dried over anhydrous sodium sulphate, filtered and concentrated under 158#583961Attorney Docket No. 00020.026.1801reduced pressure to get crude. Crude was triturated with diethyl ether (2*20 mL) compound was dried under vacuum to afford tert-butyl 4-((4-(4-fluorophenyl) butanamido)methyl)benzoate 3 (15 g, yield: 75%) as a white solid. MS (ESI) m / z: 372.3 [M+H]+.Step-2: Synthesis of 4-((4-(4-fluorophenyl)butanamido)methyl)benzoic acid 4[000307] TFA (4 vol, 40mL) was added to a solution of tert-butyl 4-((4-(4-fluorophenyl)butanamido)methyl)benzoate 3 (10 g, 26.95 mmol) in DCM (50 mL, 5 Vol) at 0°C, Resultant reaction mixture was stirred at room temperature for 2 h. Progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (2*20 mL). to afford 4-((4-(4-fluorophenyl)butanamido)methyl)benzoic acid 4 (8 g, yield: 95%) as a white solid. MS (ESI) m / z: 316.16 [M+H]+.Step-3: Synthesis of tert-butyl(4-((4-(4-fluorophenyl)butanamido) methyl) benzoyl)15.8 mmol) in DMF (50 mL, 10 Vol) at 0 °C, were added DIPEA (5.8 mL, 31.74 mmol) followed by HATU (6.6 g, 17.45 mmol). The mixture was stirred at room temperature for 10 min, followed by addition of tert-butyl glycinate 5 (1.8 g, 14.28 mmol). Resultant reaction mixture was stirred at room temperature for 2h. Progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was quenched with ice cold water, solid obtained was dissolved in 10% MeOH in DCM, dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by triturated with diethyl ether (3* 20 mL) to afford tert-butyl (4-((4-(4- 159#583961Attorney Docket No. 00020.026.1801fluorophenyl)butanamido)methyl)benzoyl)glycinate 6 (3.75 g, yield: 50%) as a white solid. MS (ESI) m / z: 428 [M+H]+.Step-4: Synthesis of(4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycine 7[000309] TFA (30 mL, 4vol) was added to solution of tert-butyl (4-((4-(4- fluorophenyl)butanamido)methyl)benzoyl)glycinate 6 (8 g, 18.6 mmol) in DCM (80 mL, 10 vol), at 0 °C. Resultant reaction mixture was stirred at room temperature for 2 h. Progress of the reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to get crude. Crude was triturated with diethyl ether (50 mL) to get (4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycine 7 (5.2 g, Yield: 75 %) as a white solid. MS (ESI) m / z: 372 [M+H]+.Step-y. Synthesis of (S)-2-( ( ((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-( (4-((4-( 4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-ccirboxcimido)butanoic acid 8(1 g, 2.66 mmol) in DMF (10 mL, 10 Vol) at 0 °C was added DIPEA (0.7 mL, 4.02 mmol) followed by addition of HATU (1.1 g. 2.95 mmol). The mixture was stirred 10 min, and added (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2- carboxamido)butanoic acid Int#l (1.1 g, 2.68 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. The progress of reaction was monitored by TLC. Upon completion of the reaction, reaction was quench with ice cold water (20 mL). and stirred for 15 min. Solid obtained was dissolved in 10% MeOH in DCM dried over anhydrous sodium 160#583961Attorney Docket No. 00020.026.1801sulphate, filtered and concentrated under reduced pressure to get crude. Crude was triturated with diethyl ether (2*20 mL) to afford (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)- 4-((S)-l-((4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2- carboxamido)butanoic acid 8 (800 mg, yield: 38%) as a white solid. MS (ESI) m / z: 792.59 [M+H]+.Step-6: Synthesis of tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4- ((S)-l-((4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2- carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate 9((4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2- carboxamido)butanoic acid 8 (4 g, 5.05 mmol) in DMF (40 mL, 8 Vol) at 0°C was added tert-butyl 4-(piperidin-4-ylmethyl)isoindoline-2-carboxylate Int#5A (1.9 g, 6.06 mmol). Resultant reaction mixture was stirred at room temperature for 2 h. Progress of reaction was monitored by TLC. Upon completion of the reaction, reaction was filter through celite. concentrated under reduced pressure to afford crude. Crude was tnturated with diethyl ether (2*20 mL) to afford tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4- ((S)-l-((4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2- carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate 9 (3.2 g, crude) as a brown solid compound. MS (ESI) m / z: 1091.06 [M+H]+.Step-7: Synthesis of (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4- (isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate 10161#583961Attorney Docket No. 00020.026.1801[000312] TFA (2.8 mL, 4 vol) at 0 °C was added to a solution of tert-butyl 4-((l-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperidin-4-yl)methyl)isoindoline-2-carboxylate 9 (250 mg, 22.93 mmol) in DCM (2.5 mL, 10 vol). Resultant reaction mixture was stirred at room temperature for 2 h. The progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (2*20 mL) to afford (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4-(isoindolin-4-ylmethyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate 10 (200 mg. yield: 88%) as a brown solid. MS (ESI) m / z: 991.02[M+H]+.Step-8: Synthesis of (9H-f!uoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S.5S)-5-((S)-2-cyano-4.4-difluoropyrrolidme-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate162#583961Attorney Docket No. 00020.026.1801[000313] To a solution of 2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l- carbonyl)-2-oxopyrrolidin-3-yl)acetic acid (CDA-5) (592 mg, 1.96 mmol) in DMF (5 mL, 10 vol) at 0°C was added DIPEA (0.5 mL, 3.03 mmol) followed by addition of HATU (633 g, 1.66 mmol). The mixture was stirred 10 min and added (9H-fluoren-9-yl)methyl ((S)-4- ((S)-l-((4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2- carboxamido)- 1 -(4-(isoindolin-4-ylmethy l)piperidin- 1 -yl)- 1 -oxobutan-2-yl)carbamate 10 (1.5 g, 1.51 mmol) Resultant reaction mixture was stirred at room temperature for 2h. Progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was quench with ice cold water (20 mL) and stirred for 15 min. Solid obtained was dissolved in 10% MeOH in DCM, dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to get crude. Crude was purified by triturated with diethyl ether (2*20 mL ) to afford (9H-fluoren-9-yl)methyl (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)- 2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-y l)methy l)piperidin- 1 -yl)-4-((S)- 1 -((4-((4-(4- fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 11 (1.2 g, yield: 63 %) as a brown solid. MS (ESI) m / z: 1275.02 [M+H]+. Step-9: Synthesis ofN-(2-(( 2R)-2-( (S)-3-amino-4-( 4-(( 2-( 2-( ( 3S, 5S)-5-( (S)-2-cyano-4, 4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin- l-yl)-4-oxobutyl)carbamoyl)cyclopentyl)-2-oxoethyl)-4-((4-(4-fluorophenyl)butanamido)methyl)benzamide 12[000314] Diethyl amine (6.8 mL. 4 vol) was added to a solution of (9H-fluoren-9-yl)methyl (9H-fluoren-9-yl)methyl ((S)-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4- difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4- 163#583961Attorney Docket No. 00020.026.1801fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 11 (1.7 g, 1.33 mmol) in acetonitrile (17 mL, 20 vol) at room temperature. Resultant reaction mixture was stirred at room temperature for 2h. Progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (2*20 mL) to afford jV-(2-((2R)-2-(((S)-3-amino-4-(4-((2-(2-((3S, 5S)-5-((S)-2-cyano-4, 4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)carbamoyl)cyclopentyl)-2-oxoethyl)-4-((4-(4-fluorophenyl)butanamido)methyl)benzamide 12 (300 mg, yield: 57%) as a brown solid. MS (ESI) m / z: 1051.71 [M+H]+.Step-10: Synthesis of 2,2'-(7-(2-(((S)-l-(4-((2-(2-((3S.5S)-5-((S)-2-cyano-4.4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-yl)methyl)piperidin-l-yl)-4-((S)-l-((4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l, 4, 7-triazonane-l, 4-diyl)diacetic acid (compound 12)164#583961Attorney Docket No. 00020.026.1801[000315] To a solution of A-(2-((2R)-2-(((S)-3-amino-4-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-4-oxobutyl)carbamoyl)cyclopentyl)-2-oxoethyl)-4-((4-(4-fluorophenyl)butanamido)methyl)benzamide 12 (50 mg, 0.165 mmol) in acetonitrile (3 mL, 10 Vol) was added DIPEA (0.06 mL, 0.33 mmol) followed by addition of 2,2',2"-(1,4,7-triazonane-1,4,7-triyl)triacetic acid (155 mg, 0.165 mmol). Resultant reaction was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with EtOAc (20 mL), obtained material was purified by prep-HPLC, required fractions were concentrated under reduced pressure followed by lyophilization to afford 2,2'-(7-(2-(((S)-I-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-yl)methyl)piperidin- 1 -y 1) -4- (( S ) - 1 -((4-((4-(4-fluorophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4,7-triazonane-l,4-diyl)diacetic acid compound 12 (5.5 mg, yield: 7.8%) as a light brown solid. MS (ESI) m / z: 1338.02 [M+H]+'H-NMR (400 MHz, DMSO- 165#583961Attorney Docket No. 00020.026.1801d6): 5 8.6-8.5 (brs, 1H), 8.4-8.3 (brs, 1H), 79-7.7 (m, 2H), 7.31 (d, J= 8Hz, 2H), 7.3-7.1 (m, 3H), 7.1-7.0 (m, 2H), 5.12-5.09 (m, 1H), 4.90-4.70 (m, 3H), 4.72-4.60 (m, 2H), 4.50-4.40 (m, 1H), 4.4-4.3 (m, 3H), 4.2-4.1 (m, 4H), 4.0-3.70 (m, 5H), 3.7-3.5 (m, 18 H), 3.20-2.70 (m, 15H), 2.6-2.5 (m, 2H), 2.30-2.25 (m, 1H), 2.2-2.10 (m, 2H), 1.95-1.80 (m, 4H), 1.80-1.70 (m, 4H), 1.6-1.5 (m, 2H), 1.2-0.9 (m, 2H).Synthesis of Intermediate:Step- 1 A: Synthesis ofhexahydro-2alH-2a,4a,6a-triazacyclopenta[cd]pentalene 2[000316] To a solution of 1,4,7-triazonane 1 (1.8 g, 13.95 mmol) in 1,4-dioxane (18 mL, 10 Vol), followed by addition of DMF-DMF (1.9 g, 16.74 mmol). Resultant reaction mixture was stirred at 100°C for 3h. The progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford hexahydro-2alH-2a,4a.6a-triazacyclopenta[cd]pentalene 2 (1.8 g crude) pale yellow liquid#583961Attorney Docket No. 00020.026.1801Step-2A: Synthesis of l-benzyl-1,4, 7-triazonane 4[000317] To a stirred solution of hexahydro-2alH-2a,4a,6a-triazacyclopenta[cd]pentalene 2 (900 mg, 16.47 mmol) in 1,4 Dioxane (9 mL, 10 vol) at room temperature was added benzyl bromide (0.9 mL, 7.76 mmol). Resulting reaction mixture was stirred at room temperature for 16 h. After formation of intermediate, to that added KOH (2.2 g dissolved in EtOH: H2O, 20 mL). The reaction mixture was stirred at 90°C for 48 h. Progress of the reaction was monitored by TLC, upon completion of the reaction, precipitated solid was fdtered and solid was re dissolved in EtOAC and H2O (100 mL). Organic layer was separated, dried over sodium sulphate, concentrated under vacuum to afford crude. Crude was treated with 4N HC1 in 1,4- Dioxane solution and concentrated under vacuum to afford solid, the solid was triturated with n Pentane (20 mL) to afford 1 -benzvl-1,4, 7-triazonane 4 (700 mg) as a pale-yellow solid. MS (ESI) m / z: 220.21 [M+H]+StepSA: Synthesis of di-tert-butyl 2,2'-(7-benzyl-l,4, 7-triazonane -1,4 -diyl) diacetate (6):[000318] To a stirred solution of 1 -benzyl- 1,4, 7-triazonane 4 (300 mg, 1.36 mmol) in acetonitrile (3 mL, 10 Vol) at 0 °C was added K2CO3 (944 mg, 6.84 mmol) followed by tertbutyl 2-bromoacetate 5 (0.5 mL, 3.42 mmol). Resultant reaction mixture was stirred at room temperature for 2h. The progress of reaction was monitored by TLC. Upon completion of the reaction, K2CO3 was filtered and filtrated was concentrated under reduced pressure to afford di-tert-butyl 2,2'-(7-benzyl-l,4,7-triazonane-l,4-diyl)diacetate 6 (200 mg, crude) brown gummy. MS (ESI) m / z: 448.56 [M+H]+Step-4A: Synthesis of (di-tert-butyl 2,2'-(1.4. 7-triazonane- 1.4-diy I) diacetate (7):167#583961Attorney Docket No. 00020.026.1801[000319] To a solution of di-tert-butyl 2,2'-(7-benzyl-l,4,7-triazonane-l,4-diyl)diacetate 6 (1 g, 2.23 mmol) in methanol (10 mL, 10 Vol), followed by addition of 10% Pd / C (100 mg), Resultant reaction mixture was stirred under H2 pressure (30 psi) at room temperature for 2h. Progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford (di-tert-butyl 2,2'-(l,4,7-triazonane-l,4-diyl)diacetate 7 (900 mg. crude) as a brown gummy MS (ESI) m / z: 358.47 [M+H]+Step-5A: Synthesis of 2-(4, 7-bis(2-(tert-butoxy)-2-oxoethyl)-1.4, 7-triazonan-l-yl)acetic acid 9:[000320] To a solution of (di-tert-butyl 2,2'-(l,4,7-triazonane-l,4-diyl)diacetate 7 (750 mg, 2.1 mmol) in acetonitrile (3 mL, 10 Vol) at 0 °C was added Et₃N (0.6 mL, 4.2 mmol) followed by tert-butyl 2-bromoacetate 8 (0.6 mL, 4.2 mmol). Resultant reaction mixture was stirred at room temperature for Ih. The progress of reaction was monitored by TLC. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford 2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-l,4,7-triazonan-l-yl)acetic acid 9 (600 mg, crude) brown gummy. MS (ESI) m / z: 416 [M+H]+Step-6A: Synthesis of 2,2',2"-(l,4, 7-triazonane-l,4, 7 -triyl) triacetic acid (NOTA):[000321] To a solution of 2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-l,4,7-triazonan-l-yl)acetic acid 9A (500 mg, 1.204 mmol) in DCM (5 mL, 10 vol), followed by addition of TFA (2 mL, 4 vol). Resultant reaction mixture was stirred at room temperature for 2h.168#583961Attorney Docket No. 00020.026.1801Progress of reaction was monitored by TLC, upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether to afford 2,2',2"-(l,4,7-triazonane-l,4,7-triyl)triacetic acid (NOTA) (350 mg) as a white solidMS (ESI) m / z: 302.2 [M+H]+Example 13: Synthesis scheme for 2,2', 2"-(10-(2-(((S)- l-(4-(2-(2-(2-((2-(2-((3S, 5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methoxy)ethoxy)ethoxy)ethyl)piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl) butanamido) methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-1-oxobutan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (compound 13)oStep-1: Synthesis of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate (3):[000322] To a solution of 4-(4-iodophenyl)butanoic acid 1 (30 g, 0.103 mol) in DMF (60 mL, 2 Vol) at 0 °C was added DIPEA (27 mL, 0.155 mol) followed by HATU (39.3 g. 0.103 mol). The mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl 4-(aminomethyl)benzoate 2 (21.4 g, 0.103 mol). Resultant reaction mixture was stirred at room temperature for 4 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (200 mL) and stirred for 15 min. Organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 400 mL). Combined 169#583961Attorney Docket No. 00020.026.1801organic layer was washed with water (400 mL), separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford tertbutyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (36.1 g, crude) as a brown liquid. MS (ESI) m / z: 480.1 [M+H]+Step-2: Synthesis of 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid (4):[000323] TFA (160 mL, 4 vol) was added to a solution of tert-butyl 4-((4-(4-iodophenyl)butanamido)methyl)benzoate 3 (40 g, 0.083 mol) in DCM (200 mL, 5 Vol) at 0 °C. Resultant reaction mixture was stirred at room temperature for 5 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (200 mL) to afford 4-((4-(4-iodophenyl)butanamido)methyl)benzoic acid 4 (31 g, yield: 88%) as a brown solid. MS (ESI) m / z: 424.42 [M+H]+Step-3: Synthesis of tert-butyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycinate23.64 mmol) in DMF (100 mL, 10 Vol) at 0 °C, was added DIPEA (12.3 mL, 70.92 mmol ) followed by HATU (13.4 g, 35.46 mmol). The mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl glycinate 5 (3.5 mL, 26.00 mmol). Resultant reaction was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min, precipitated solid was filtered and wash with hexane (2* 100 mL) to afford tert-butyl (4-((4-(4-170#583961Attorney Docket No. 00020.026.1801iodophenyl)butanamido)methyl)benzoyl)glycinate 6 (10 g, yield: 83 %) as a white solid. MS (ESI) m / z:538.28 [M+H]+.Step-4: Synthesis of(4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine (7):[000325] TFA (18 mL, 3 vol) was added to a solution of tert-butyl (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycinate 6 (6 g, 0.011 mol) in DCM (60 mL, 10 Vol) at 0 °C. Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (100 mL) to afford (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine 7 (4.5 g, yield: 86%) as a white solid. MS (ESI)m / z: 481.13 [M+H]+Step-5: Synthesis of (S)-2-( ( f9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-( (4-((4-( 4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid (8):[000326] To a solution of (4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine 7 (2.5 g, 5.20 mmol) in DMF (25 mL. 10 Vol) at 0°C was added DIPEA (1.3 mL, 7.81 mmol) followed by addition of HATU (2.17 g, 5.72 mmol). The mixture was stirred at room temperature for 10 min followed by addition of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-pyrrolidine-2-carboxamido)butanoic acid Int#l (2.27 g, 5.20 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted with cold water (100 mL) and stirred for 15 min, obtained solid was triturated diethyl ether (10 mL) to get (S)-2-((((9H-fluoren-9-y l)methoxy )carbonyl)amino)-4-((S)- 1 -((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid 8 (3.6 g, yield: 78%) as a white solid. MS (ESI) m / z: 900.47 [M+H]+.171#583961Attorney Docket No. 00020.026.1801Step-6: Synthesis of tert-butyl 4-((2-(2-(2-(4-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-( (S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoyl)piperazin-l-yl)ethoxy)ethoxy)ethoxy)methyl)isoindoline-2-carboxylate: (9)[000327] To a solution of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)butanoic acid 8 (1.96 g, 2.18 mmol) in DMF (10.96 mL, 10 Vol) at 0°C, was added DIPEA (0.76 mL, 4.36 mmol) followed by HATU (0.91 g, 1.1 mmol), the reaction mixture was stirred for 10 min and added tert-butyl 4-((2-(2-(2-(piperazin-l-yl)ethoxy)ethoxy)ethoxy)methyl)isoindoline-2-carboxylate Int#l (0.97 g, 2.18 mmol).Resultant reaction was stirred at room temperature for 2h. Upon completion of the reaction, reaction mixture was diluted with cold water (50 mL) and stirred for 15 min. Obtained white solid was triturated with diethyl ether (250 mL) to afford tert-butyl 4-((2-(2-(2-(4-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido) butanoyl) piperazin- l-yl)ethoxy)ethoxy)ethoxy)methyl)isoindoline-2-carboxy late 9 (1.3 g, yield: 40%) as a brown solid. MS (ESI) m / z:1333.11 [M+H]+.Step-7: Synthesis of (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4-(2-(2-(2-(isoindolin-4-ylmethoxy)ethoxy)ethoxy)ethyl)piperazin-1-yl)-1-oxobutan-2-yl)carbamate (10)((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((S)-l-((4-((4-(4-iodophenyl) butanamido) methyl) benzoyl) glycyl)pyrrolidine-2-carboxamido)butanoyl)piperazin-l-yl) ethoxy) ethoxy)172#583961Attorney Docket No. 00020.026.1801ethoxy) methyl) isoindoline-2-carboxylate 9 (1 g, 7.51 mol) in DCM (10 mL, 10 Vol) at 0°C. Resultant reaction was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (10 mL) to afford (9H-fluoren-9-yl)methyl (9H-fluoren-9-yl)methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4-(2-(2-(2-(isoindolin-4-y lmethoxy)ethoxy)ethoxy)ethyl)piperazin- 1 -yl)- 1 -oxobutan-2-yl)carbamate 10 (500 mg, yield: 43%) as an off white solid. MS (ESI) m / z: 1231.97 [M+H]+Step-8: Synthesis of (9H-fluoren-9-yl)methyl ((S)-l-(4-(2-(2-(2-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methoxy)ethoxy)ethoxy)ethyl)piperazin-l-yl)-4-( (S)-l-( ( 4-((4-( 4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-1-oxobutan-2-yl)carbamate (11)[000329] To a solution of CDA-5 (350 mg, 1.15 mmol) in DMF (13 mL, 10 Vol) at 0°C was added DIPEA (0.46 mL, 2.64 mmol) followed by addition of HATU (410 mg, 1.05 mmol). Reaction mixture was stirred at room temperature for 10 min followed by addition of (9H-fluoren-9-yl) methyl ((S)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-(4-(2-(2-(2-(isoindolin-4-ylmethoxy)ethoxy)ethoxy)ethyl)piperazin- 1 -yl)- 1 -oxobutan-2-yl)carbamate 10 (1.3 g, 1.05 mmol). Resultant reaction was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was diluted with cold water (200 mL) and stirred for 15 min. Obtained white solid was triturated with diethyl ether (20 mL) to afford (9H-fluoren-9-yl)methyl ((S)-l-(4-(2-(2-(2-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-1-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl) methoxy) ethoxy) ethoxy)ethyl)piperazin-1-yl)-4-((S)-1-((4-((4-(4-iodophenyl) butanamido) methyl) benzoyl) glycyl) pyrrolidine-2-carboxamido)-1-oxobutan-2-yl)carbamate 11 (770 mg, yield: 35%) as an off white solid. MS (ESI) m / z: 1515.16 [M+H]+.173#583961Attorney Docket No. 00020.026.1801Step-9: Synthesis of (S)-N-((S)-3-amino-4-(4-(2-(2-(2-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methoxy)ethoxy)ethoxy)ethyl)piperazin-1-yl)-4-oxobutyl)-1-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide (12)[000330] Diethyl amine (3.2 mL, 4 vol) was added to a solution of (9H-fluoren-9-yl)methyl ((S)-l-(4-(2-(2-(2-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l- carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-y l)methoxy )ethoxy )ethoxy)ethy l)piperazin- 1 -yl )-4 -(( S )- 1 -((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)carbamate 11 (800 mg, 0.52 mmol) in acetonitrile (8 mL, 10 Vol) at room temperature. Resultant reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated to afford crude. Crude was triturated with diethyl ether and wash with EtOAc (30 mL) to afford (S)-N-((S)-3-amino-4-(4-(2-(2-(2-((2- (2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methoxy)ethoxy)ethoxy)ethyl)piperazin-l-yl)-4-oxobutyl)-l-((4-((4- (4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide 12 (450 mg, yield: 65%) as a white solid. MS (ESI) m / z: 1294.02 [M+H]+.Step-10: Synthesis of2,2',2"-(10-(2-(((S)-l-(4-(2-(2-(2-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methoxy)ethoxy)ethoxy)ethyl)piperazin-l-yl)-4-( (S)-l-( ( 4-( 4-( 4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-l-oxobutan-2-yl)amino)-2-oxoethyl)-l,4, 7,10-tetraazacyclododecane-l,4, 7 -triyl) triacetic acid (compound 13)174#583961Attorney Docket No. 00020.026.1801[000331] To a suspension of (S)-N-((S)-3-amino-4-(4-(2-(2-(2-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methoxy)ethoxy)ethoxy)ethyl)piperazin-l-yl)-4-oxobutyl)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide 12 (450 mg, 0.34 mmol) in acetonitrile (4.5 mL, 10 Vol) at room temperature, was added DIPEA (0.54 mL, 3.13 mmol) followed by 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid DOTA NHS ester (261 mg, 0.52 mmol). Resultant reaction was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with EtOAc (20 mL), obtained material was purified by prep-HPLC. required fractions were concentrated under reduced pressure followed by lyophilization to afford 2,2',2"-(10-(2-(((S)-l-(4-(2-(2-(2-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl) methoxy) ethoxy) ethoxy) ethyl) piperazin-l-yl)-4-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamido)-1-oxobutan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (compound 13) (70 mg, yield: 12%) as a white solid. MS (ESI) m / z: 1679.17 [M+H]+. 'H-NMR (400 MHz, DMSO-d6): 58.97(brs, 1H), 8.7-8.6 (brs, 1H), 8.45-8.44 (brs, 1H), 8.3-8.2 (brs, 2H),175#583961Attorney Docket No. 00020.026.18018.1-8.0 (m, 1H), 7.83 (d, J= 8.4 Hz, 2H), 7.61 (d, J= 8 Hz, 2H), 7.31-7.25 (m, 5H), 7.0 (d, J = 8.4 Hz. 2H), 5.12-5.08 (m. 1H), 4.90-4.80 (m. 2H), 4.72-4.60 (m. 3H), 4.5 (s, 2H), 4.4-4.3 (m, 3H), 4.3-4.0 (m, 5H) 3.85-3.60 (m, 2H), 3.60-3.40 (m, 20H), 3.40-3.30 (m, 2H), 3.2-3.1 (m, 2H), 3.1-2.9 (m, 13H), 2.9-2.61 (m, 8H), 2.6-2.50 (m, 2H), 2.5-2.4 (m, 2H), 2.4-2.2 (m, 6H), 2.20-2.1 (m, 3H), 2.2-2.18 (m, 2H), 1.9-1.60 (m, 5H), 1.5-1.4 (m, 1H).Synthesis of intfl:Step-IA: Synthesis of tert-butyl 4-(hydroxymethyl)isoindoline-2-carboxylate (2A)[000332] To a solution of 2-(tert-butyl) 4-methyl isoindoline-2, 4-dicarboxylate 1 A (60 g, 0.216 mol) in toluene (480 mL, 8 vol) at 0 °C was added 20% Red Al in toluene (102 mL, 0.367 mol). Reaction mixture was stirred at 0°C for 30 min. Resultant reaction mixture was stirred at room temperature for 2 h. After completion of reaction, reaction mixture was quenched with 20% aq. KOH solution (100 mL) diluted with water (300 mL), and extracted with ethyl acetate (2*500 mL), separated organic layer was dried over Sat. Na2SO4, concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) 10% ethyl acetate in hexane as an eluent. Compound contained fractions were concentrated under reduced pressure to afford tert-butyl 4-(hydroxymethyl)isoindoline-2-carboxylate 2A (50g, yield: 95%) as an off white solid. MS (ESI) m / z: 250.08 [M+H]+.Step-2A: Synthesis of tert-butyl 4-(bromomethyl)isoindoline-2-carboxylate (3A)#583961Attorney Docket No. 00020.026.1801[000333] To a solution of tert-butyl 4-(hydroxymethyl)isoindoline-2-carboxylate 2A (52 g, 0.208 mol) in DCM (520 mL, 10 vol) at 0 °C. was added PPh3(56 g, 0.312 mol), the reaction mixture was stirred for 30 min and added NBS (55 g, 0.208 mol). Resultant reaction was stirred at room temperature for 3 h. After completion of the reaction, reaction mixture was poured into water (200 mL), and extracted with DCM (2*500 mL) and organic layer was separated and dried over Na₂SO₄, filtered, concentrated under reduced pressure to get crude. The obtained crude was purified by silica gel column chromatography (100-200 mesh) 1% ethyl acetate in hexane as an eluent. Compound contained fractions were concentrated under reduced pressure to afford tert-butyl 4-(bromomethyl)isoindoline-2-carboxylate 3A (34 g, yield: 52%) as an light yellow solid. MS (ESI) m / z: 213.9 [M+H]+.Step-3A: Synthesis of tert-butyl 4-((2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)methyl)isoindoline-[000334] To a solution of 2,2'-(ethane-l,2-diylbis(oxy))bis(ethan-l-ol) 4A (100 mg, 0.32 mmol) in DMF (1 mL, 10 vol) at 0°C was added 60% NaH (15.3 mg, 0.64 mmol). The mixture was stirred at 0 °C for 30 min and added tert-butyl 4-(bromomethyl)isoindoline-2-carboxylate 3A (100 mg, 0.64 mmol) at 0 °C. Resultant reaction mixture was stirred at room temperature for 3 h. Upon completion of reaction, reaction mixture was poured into water (50 mL), and extracted with ethyl acetate (2*500 mL), the organic layer was separated, dried over Na₂SO₄, concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) eluted in 10% ethyl acetate in hexane as an eluent. Compound contained fractions were concentrated under reduced pressure to afford tert-butyl 4-((2-(2-(2-hydroxy ethoxy )ethoxy)ethoxy)methyl)isoindoline-2-carboxylate 5A (80 mg, crude) as a pale yellow gummy. MS (ESI) m / z: 785.6 [M+H]+.Step-4A: Synthesis of tert-butyl 4-((2-(2-(2- ((methylsulfonyl)oxy)ethoxy)ethoxy)ethoxy)methyl)isoindoline-2-carboxylate:177#583961Attorney Docket No. 00020.026.1801[000335] To a solution of tert-butyl 4-((2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)methyl)isoindoline-2-carboxylate 5A (500 mg, 1.31 mmol) in DCM (5 mL, 10 vol) at 0 °C was added triethyl amine (0.3 mL, 2.6 mmol), followed by mesyl chloride (0.15 mL, 1.96 mmol). Resultant reaction mixture was stirred at room temperature for 4 h. Upon completion of reaction, the reaction mixture was diluted with water (100 mL), and compound was extracted with EtOAc (2*50 mL) and organic layer was concentrated under reduced pressure to get tert-butyl 4-((2-(2-(2-((methylsulfonyl)oxy)ethoxy)ethoxy)ethoxy)methyl)isoindoline-2-carboxylate 6A (510 mg, crude) as a brown liquid. Crude was forwarded to the next step without purification Step-5A: Synthesis of tert-butyl 4-((2-(2-(2-(piperazin-1-yl)ethoxy)ethoxy)ethoxy)methyl)isoindoline-2-carboxylate (int#1):[000336] To a solution of tert-butyl 4-((2-(2-(2-((methylsulfonyl)oxy)ethoxy)ethoxy)ethoxy)methyl)isoindoline-2-carboxylate 6A (1 g, 2.17 mmol) in DCM (10 mL, 10 Vol) at room temperature was added piperazine (0.37 g, dissolved in DCM, 4.35 mol) followed by addition of DIPEA (0.45 mL, 0.795 mmol).Resultant reaction mixture was stirred at room temperature for 16 h. Reaction mixture was cool to room temperature and diluted with ice-cold water (300 mL) and extracted with EtOAc (2 * 50 mL). Combined organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 10% methanol in dichloromethane as an eluent. Compound contained fractions were concentrated under reduced pressure to afford tert-butyl 4-((2-(2-(2-(piperazin-l-yl)ethoxy)ethoxy)ethoxy)methyl)isoindoline-2-carboxylate Int#l (400 mg, Yield: 36%) as a pale yellow gummy liquid. MS (ESI) m / z: 450.52 [M+H]+Example 14: Synthesis of 2,2',2"-(10-((S)-18-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4- 178#583961Attorney Docket No. 00020.026.1801yl)methyl)piperidine-l-carbonyl)-l-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidin-2-yl)-l,14,20-trioxo-5,8,ll-trioxa-2,15,19-triazahenicosan-21-yl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 14):Step-1: Synthesis of (9H-fluoren-9-yl)methyl (S)-2-((14,14-dimethyl-12-oxo-3, 6.9.13-[000337] To a solution of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-proline 1 (2 g, 5.97 mmol) in DMF (20 mL, 10 Vol) at 0°C was added DIPEA (3 mL, 17.7 mmol) followed by HATU (4.4 g, 11.8 mmol). The reaction mixture was stirred at room temperature for 30 min, followed by addition of tert-butyl 3-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)propanoate 2 (1.8 g, 6.57 mmol). Resultant reaction mixture was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted with EtOAc (500 mL), cold water (200 mL) and stirred for 15 min. Organic layer was separated, and aqueous layer was further extracted with EtOAc (2 * 50 mL). Combined organic layer was washed with water (400 mL). The separated organic layer was dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford crude. Crude was purified by silica gel column chromatography (100-200 mesh) eluted at 100% ethyl acetate and compound fractions were 179#583961Attorney Docket No. 00020.026.1801concentrated under reduced pressure to afford (9H-fluoren-9-yl)methyl (S)-2-((14, 14-dimethyl- 12-oxo-3, 6, 9, 13-tetraoxapentadecyl)carbamoyl)pyrrolidine-l -carboxylate 3 (2.5 g, crude) as a brown liquid. MS (ESI) m / z: 597.38 [M+H]+Step-2: Synthesis of tert-butyl (S)-l-oxo-l-(pyrrolidin-2-yl)-5,8,ll-trioxa-2-azatetradecan-[000338] Diethyl amine (4 mL, 4 vol) was added to a solution of (9H-fluoren-9-yl)methyl (S)-2-((14,14-dimethyl-12-oxo-3,6,9,13-tetraoxapentadecyl)carbamoyl)pyrrolidine-l -carboxylate 3 (1 g, 1.67 mmol) in DCM (10 mL, 5 Vol) at 0°C. Resultant reaction was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (10 mL) to afford tert-butyl (S)-l-oxo-l-(pyrrolidin-2-yl)-5,8,11-trioxa-2-azatetradecan-14-oate 4 (0.5 g, crude) as a pale-yellow gummy. MS (ESI) m / z: 375.51 [M+H]+Step-3: Synthesis of tert-butyl (S)-l-(l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidin-2-yl)-l-oxo-5,8,ll-trioxa-2-azatetradecan- 14-oate (5):[000339] To a solution of ((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycine Int#2 (1 g, 2.08 mmol) in DMF (10 mL, 10 Vol) at 0°C was added DIPEA (0.9 mL, 5.2 mmol) followed by HATU (1.18 g, 3.12 mmol). The mixture was stirred at room temperature for 1 min, followed by addition of tert-butyl (S)-l-oxo-l-(pyrrolidin-2-yl)-5,8,11-trioxa-2-azatetradecan-14-oate 4 (0.78 g, 2.08 mmol). Resultant reaction was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mixture was diluted with cold water (100 mL) and stirred for 15 min. Filter the precipitated solid and washed with pentane (20 mL) to afford tert-butyl (S)-l-(l-((4-((4-(4- 180#583961Attorney Docket No. 00020.026.1801iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidin-2-yl)-l-oxo-5,8,l l-trioxa-2-azatetradecan- 14-oate 5 (0.38 g, yield:26%) as a pale yellow gummy. MS (ESI) m / z: 837.46 [M+H]+Step-4: Synthesis of (S)-l-(l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidin-2-yl)-l-oxo-5,8,l l-trioxa-2-azatetradecan- 14-oicacidiodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidin-2-yl)-l-oxo-5, 8,1 l-trioxa-2-azatetradecan- 14-oate 5 (1 g, 1.19 mmol) in DCM (2 mL, 2 Vol) at 0°C. Resultant reaction was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with diethyl ether (20 mL) to afford ((S)-l-(l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidin-2-yl)- 1 -oxo-5,8, 11 -trioxa-2-azatetradecan- 14-oic acid 6 (600 mg, Yield:66% ). MS (ESI) m / z: 781.39 [M+H]+Step-5: Synthesis of (9H-fluoren-9-yl)methyl ((S)-19-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4.4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-((S)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidin-2-yl)-1, 14, 19-trioxo-5, 8,1 l-trioxa-2, 15-diazanonadecan-18-yl)carbamate (7):[000341] To a solution of (S)-l-(l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidin-2-yl)-l-oxo-5, 8,1 l-trioxa-2-azatetradecan- 14-oic acid 6 (1 g. 1.21 mmol) in DMF (10 mL, 10 Vol) at 0°C was added DIPEA (0.42 mL, 2.42 mmol) followed by addition of HATU (509 mg, 1.33 mmol). The 181#583961Attorney Docket No. 00020.026.1801mixture was stirred at room temperature for 10 min and (9H-fluoren-9-yl)methyl ((S)-4-amino-l-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-oxobutan-2-yl)carbamate (Int#5) (950 mg, 1.21 mmol). Resultant reaction was stirred at room temperature for 1 h. Upon completion of the reaction, reaction mass was diluted with 10% MeOH in DCM (25 mL) and ice water (20 mL), organic layer was separated and the aqueous layer was extracted with 10% MeOH in DCM (2 *25 mL). Organic layer separated was dried over anhydrous sodium sulphate, filtered, concentrated under reduced pressure to afford (9H-fluoren-9-yl)methyl ((S)-19-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3 -y l)acety l)isoindolin-4-yl)methyl)piperidin- 1 -y 1)- 1 -((S)- 1-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidin-2-yl)-l,14,19-trioxo-5,8,l 1-trioxa-2,15-diazanonadecan-18-yl)carbamate 7 (1.3 g, crude) as a white solid. MS (ESI) m / z:1586.22 [M+H]+.Step-6: Synthesis of (S)-N-((S)-16-amino-17-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolm-4-yl)methyl)piperidin-l-yl)-12,l 7-dioxo-3, 6, 9-trioxa-l 3 -azaheptadecyl)-! -((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide (8)[000342] Diethyl amine (1.5 mL, 1 Vol) was added to a solution of (9H-fluoren-9-yl)methyl ((S)-19-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrohdin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-l-((S)-l-((4-((4-(4-iodophenyl) butanamido)methyl)benzoyl)glycyl)pyrrolidin-2-yl)-l,14,19-trioxo-5,8,l 1-trioxa-2,15-diazanonadecan-18-yl)carbamate 7 (500 mg, 0.31 mmol) in DMF (5 mL, 10 Vol) at room temperature. Resultant reaction was stirred at room temperature for 2 h. Upon completion of the reaction, reaction mass was concentrated under reduced pressure to get crude. Crude was triturated with cold EtOAc (10 mL) followed by diethyl ether (30 mL) to afford (S)-N-((S)-16-amino- 17 -(4-((2-(2-((3S,5 S)-5-((S)-2-cy ano-4,4-difluoropyrrolidine- 1 - 182#583961Attorney Docket No. 00020.026.1801carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-yl)methyl)piperidin-l-yl)-12,17-dioxo-3,6,9-trioxa-13-azaheptadecyl)-l-((4-((4-(4-iodophenyl)butanamido)methyl)benzoyl)glycyl)pyrrolidine-2-carboxamide 8 (300 mg, Yield: 71%) as a white solid. MS (ESI) m / z: 1363.99 [M+H]+.Step-7: Synthesis of 2,2',2"-(10-((S)-18-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidme-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolm-4-yl) methyl) piperidine-l-carbonyl)-l-((S)-l-((4-((4-(4-iodophenyl) butanamido) methyl) benzoyl)glycyl)pyrrolidin-2-yl)-l, 14,20-trioxo-5.8.1 l-trioxa-2,15, 19-triazahenicosan-21-yl)-1,4, 7,10-tetraazacyclododecane-l,4, 7 -triyl) triacetic acid (compound 14)4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3-yl)acetyl)isoindolin-4-y l)methyl)piperidin- 1 -y 1)- 12, 17 -dioxo-3,6,9-trioxa- 13 -azaheptadecyl)- 1 -((4-((4-(4-iodophenyl)butanamido) methyl) benzoyl)glycyl)pyrrolidine-2-carboxamide (8) (300 mg, 0.22 mmol) in acetonitrile (3 mL, 10 Vol) at room temperature, was added DIPEA (0.34 mL, 1.98 mmol) followed by 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid DOTA NHS ester (220 mg, 0.44 mmol). Resultant reaction was stirred at room temperature for 16 h. Upon completion of the reaction, reaction mixture was concentrated under reduced pressure to afford crude. Crude was triturated with EtOAc (20 mL), obtained matenal was purified by prep-HPLC, required fractions were concentrated under reduced pressure followed by lyophilization to afford 2,2',2"-(10-((S)-18-(4-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-l-carbonyl)-2-oxopyrrolidin-3 -yl)acetyl)isoindolin-4-yl)methyl)piperi dine- 1 -carbonyl)- 1-((S)-1 -((4-((4-(4-iodophenyl) butanamido) methyl) benzoyl) glycyl)pyrrolidin-2-yl)-1.14,20-trioxo-5,8,l 1-trioxa-2,15,19-triazahenicosan-21-yl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (compound 14) (22 mg, yield: 5.1%) as a light brown solid. MS (ESI) m / z: 1750.57183#583961Attorney Docket No. 00020.026.1801[M+H]+1H-NMR (400 MHz, DMSO-d6): δ 14.0-12.0 (brs, 2H), 8.8-8.7 (brs, 1H), 8.6-8.5 (brs, 1H), 8.37 (t, J= 6 Hz, 1H), 8.3-8.2 (m, 1H), 8.1-8.0 (brs, 1H), 7.9 -7.8 (m, 4H), 7.62 (d, J= 7.6 Hz, 2H), 7.31 (d, J= 8 Hz, 2H), 7.3-7.1 (m, 2H), 7.1-7.01 (m, 1H), 7.01 (d, J= 8 Hz, 2H), 5.12-5.08 (m, 1H), 4.80-4.70 (m, 2H), 4.72-4.60 (m, 3H), 4.50-4.20 (m, 5H), 4.2-4.0 (m, 6H), 3.95-3.80 (m, 3H), 3.6-3.4(m, 18H), 3.40-3.30 (m, 4H), 3.30-3.20 (m, 6H), 3.2-2.7 (m, 19H), 2.6-2.5 (m, 4H), 2.4-2.3 (m, 2H), 2.20-2.1 (m, 2H), 2.1-1.9 (m, 3H), 1.85-1.70 (m, 5H), 1.70-1.50 (m, 3H), 1.3-1.2 (m, 3H).Intermediate (Int#l) synthesis:Step-1: Synthesis tert-butyl 4-(isoindolin-4-ylmethylene)piperidine-l-carboxylate (3)[000344] To a solution of 4-bromoisoindoline 1 (10 g, 42.7 mmol) in DMF:H₂O (9:2, 10 vol) at room temperature was added tert-butyl 4-((4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)methylene)piperidine-l -carboxylate 2 (16.5 g, 51.28 mmol) followed by CS2CO3 (41.6 g, 128.2 mmol). The reaction mixture was degassed with argon for 15 min followed by addition of XPhos Pd G4 (3.67 g, 4.27 mmol), resultant reaction mixture was stirred at 100°C for 5 h. Upon completion of the reaction, reaction mixture was fdtered through celite bed washed with ethyl acetate (20 mL), fdtrate was concentrated under reduced pressure to get crude. Crude was purified by silica gel column chromatography (100-200 mesh) using 5% MeOH in DCM as an eluant. Required fractions were concentrated under reduced pressure to afford tert-butyl 4-(isoindolin-4-ylmethylene)piperidine-l -carboxylate 3 (8 g, yield: 59%) as a brown solid. MS (ESI) m / z: 315.1[M+H]+Step-2: Synthesis tert-butyl 4-(isoindolin-4-ylmethyl)piperidine-l -carboxylate (4):184#583961Attorney Docket No. 00020.026.1801[000345] To a solution of tert-butyl 4-(isoindolin-4-ylmethylene)piperidine-l-carboxylate 3 (8 g, 25.47 mmol) in MeOH (80 mL) was added 10% Pd / C (5 g, wt / wt).Resultant reaction mixture was stirred at room temperature for 16 h und...

Claims

Attorney Docket No. 00020.026.1801CLAIMS1. A complex represented by Formula (I):D-L-RAIL'EFormula (I)or a pharmaceutically acceptable salt thereof, wherein:D is a ligand that targets fibroblast activation protein (FAP);L is a linker;LEis a peptide linker comprising at least one proline;E is a secondary effector moiety; andRAis a ligand chelated or covalently linked to a therapeutic radionuclide or a diagnostic radionuclide.

2. The complex of claim 1, wherein D is represented by Formula X-B:wherein:224#583961Attorney Docket No. 00020.026.1801T is substituted or unsubstituted methylene (-CH2-), substituted or unsubstituted amino (-NH-), -O-, or -S- (e.g., wherein the substitution of T is C1-C3 alkyl, haloalkyl, or halo);J is C(RJ)0-3, wherein each RJis independently H or alkyl, or two or more RJare taken together to form oxo;R1and R2are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=C-C(O)aryl, -C=C-S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, -CONH2, and 5-tetrazolyl;R3and R4are independently selected from the group consisting of -H, -OH, F, Cl, Br, I, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl;R5, R6, R7, and R8are independently selected from the group consisting of H, alkyl, and halo; andR9, R10, and R11are independently selected from the group consisting of H,-Ci-ealkyl, -Ci-6haloalkyl, -O-Ci-ealkyl, -S-C1-6 alkyl, F, CL Br, and I.

3. The complex of claim 1, wherein D is represented by Formula X-C:(X-C)wherein:R21is -CN oru0JIOR22and R23are independently selected from the group consisting of H, C1-6 alkyl, and halo; andR24is C1-6 alkyl;225#583961Attorney Docket No. 00020.026.1801Q is -N(R25)(C(R26)(R27))t- or -O(C(R28)(R29))UC(O)-;R25is selected from H and C1-6 alkyl; andeach R26, R27, R28, and R29is independently selected from H and C1-6 alkyl; each r and s is independently 1 or 2; andeach t and u is independently 0 or 1.

4. The complex of claim 1, wherein D is represented by:R41is selected from the group consisting of -H, -CN, -B(0H)2, -C(O)alkyl, -C(O)aryl, - C=C-C(O)aryl. -C=C-S(O)2aryl. -CO2H, -SO3H, -SO2NH2, -PO3H2, and 5-tetrazolyl,R42, R43, R44, and R44are independently selected from the group consisting of -H, -OH, halogen, -Ci-6alkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl,R45is selected from the group consisting of -H, -CH2OH, and -CH(CH3)2,R46, R47, and R48are each independently selected from the group consisting of -H, -OH, oxo, halogen, CF3, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl, -NR49R50, -OR51, - Het2, and -An; each of -Ci-ealkyl being optionally substituted with from 1 to 3 substituents selected from - OH and halogen; R49, R50, and R51are each independently selected from the group consisting of -H, -OH, oxo, halogen, CF3, -Ci-ealkyl, -O-Ci-ealkyl, -S-Ci-ealkyl, and -An.An and An are each independently a 5- or 6-membered aromatic monocycle optionally comprising 1 or 2 heteroatoms selected from O, N, and S; each of An and An being 226#583961Attorney Docket No. 00020.026.1801optionally and independently substituted with from 1 to 3 substituents selected from -NR52R53, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl,R52and R53are each independently selected from the group consisting of -H, -OH, CF3, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl,Het2 is a 5- or 6-membered non-aromatic monocycle optionally comprising 1 or 2 heteroatoms selected from O, N and S; Het2 being optionally substituted with from 1 to 3 substituents selected from -NR54R55, -Ci-6alkyl, -O-Ci-6alkyl, and -S-Ci-6alkyl,R54and R55are each independently selected from the group consisting of -H, -OH, halogen, CF3, -Ci-ealkyl, -O-Ci-6alkyl, and -S-Ci-6alkyl,the fragment:represents a 5- to 10-membered N-containing aromatic or non-aromatic mono- or bicyclic heterocycle, said heterocycle optionally further comprising 1 to 3 heteroatoms selected from O, N, and S; andJ is selected from the group consisting of a bond, -Ci-3alkyl, -Ci-3alkyl-NH-, C=O, and -O-.

5. The complex of claim 1, wherein D is represented by:FAR31oy^ N^Ar’R30H Hwherein,R30is -H, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=C-C(O)aryl, -C=C-S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, or 5-tetrazolyl;R31is H or CH3; andAr1is substituted phenyl, pyridyl, chloropyridyl, or quinolinyl.227#583961Attorney Docket No. 00020.026.1801 6. The complex of claim 1, wherein D is represented by:228#583961Attorney Docket No. 00020.026.1801wherein:each m is independently 0 to 3.

7. The complex of any one of claims 1-6. wherein L is represented by:Ywherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl, 6-membered heteroaryl, and C3-6 cycloalkyl;L1is selected from a covalent bond and a linker; andL2is a linker,whereina is the point of attachment to D;P is the point of attachment to RL; andy is the point of attachment to LE.

8. The complex of any one of claims 1-7. wherein L is represented by:wherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl. 6-membered heteroaryl, and C3-6 cycloalkyl;229#583961Attorney Docket No. 00020.026.1801n is an integer from 1 to 6,q is an integer from 1 to 20; andp is 0 or 1;wherein:a is the point of attachment to D;P is the point of attachment to RL; andy is the point of attachment to LE.

9. The complex of claim 7 or 8, wherein ring A is 3 to 6-membered heterocyclyl.

10. The complex of any one of claims 7-9. wherein ring A is selected from the group consisting of:wherein \| / represents the point of attachment to L1, and s represents the point of attachment to L2.

11. The complex of any one of claims 1-10, wherein LEis represented by:e-cx ^-pwherein:each X1is an amino acid;Lcis absent or a cleavable linker; andm is an integer from 1 to 3;provided that at least one X1is proline; and230#583961Attorney Docket No. 00020.026.1801wherein 0 represents the point of attachment to L and p represents the point of attachment to E.

12. The complex of claim 11, wherein each X1is independently selected from the group consisting of absent, proline, glycine, alanine, threonine, and serine, provided that at least one X1is proline.

13. The complex of claim 11 or 12, wherein each X1is independently selected from the group consisting of proline and glycine, provided that at least one X1is proline, and m is 2.

14. The complex of any one of claims 11-13, wherein Lcis absent.

15. The complex of any one of claims 11-13, wherein Lcis a cleavable linker represented by:N O NI16. The complex of any one of claims 1-15, wherein the ligand in RAis represented by:231#583961Attorney Docket No. 00020.026.1801or232#583961Attorney Docket No. 00020.026.180117. The complex of any one of claims 1-16, wherein RAis represented by:, wherein M is a therapeutic radionuclide or a diagnostic radionuclide.

18. The complex of any one of claims 1-17, wherein E is an albumin binding ligand, 19. The complex of any one of claims 1-18, wherein E comprises a cleavable linker, 20. The complex of any one of claims 1-18, wherein E is represented by:233#583961Attorney Docket No. 00020.026.1801wherein each of X and Y is independently selected from the group consisting of halogen and C1-6 alkyl, and n is an integer from 1 to 20.

21. The complex of claim 20, wherein X is I.

22. A complex chelated or covalently linked to a radionuclide, comprising a linker represented by:owherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl, 6-membered heteroaryl, and C3-6 cycloalkyl;n is an integer from 1 to 6,q is an integer from 1 to 20;p is 0 or 1; andLEis a peptide linker comprising at least one proline.234#583961Attorney Docket No. 00020.026.180123. The complex of claim 22, wherein LEis represented by:0-(X’)m-Lc-pwherein:each X1is an amino acid;Lcis absent or a cleavable linker; andm is an integer from 1 to 3;provided that at least one X1is proline; andwherein 0 represents the point of attachment to L and p represents the point of attachment to E.

24. The complex of claim 23, wherein each X1is independently selected from the group consisting of absent, proline, glycine, alanine, threonine, and serine, provided that at least one X1is proline.

25. The complex of claim 23 or 24, wherein each X1is independently selected from the group consisting of proline and glycine, provided that at least one X1is proline, and m is 2.

26. The complex of any one of claims 23-25, wherein Lcis absent.

27. The complex of any one of claims 23-26, wherein Lcis a cleavable linker represented by:N O NI28. A complex of a compound selected from the group consisting of:235#583961Attorney Docket No. 00020.026.1801236#583961Attorney Docket No. 00020.026.1801HOOC^x / — \ COOHHOOC^x / — \ COOH237#583961Attorney Docket No. 00020.026.1801238#583961Attorney Docket No. 00020.026.1801HOOC^ / - \ COOHSO3Hor a pharmaceutically acceptable salt thereof, anda therapeutic radionuclide or a diagnostic radionuclide.

29. The complex of any one of claims 1-28, wherein the therapeutic radionuclide is selected from the group consisting of177Lu,90Y.155Tb,161Tb,225Ac,67Cu,223Ra,186Re,188Re,1311,212Pb,212Bi,213Bi. and211At.

30. The complex of any one of claims 1-29, wherein the therapeutic radionuclide is177Lu.

31. The complex of any one of claims 1-28, wherein the diagnostic radionuclide is selected from the group consisting of99mTc,111In,18F,66Ga,67Ga,68Ga,124I,123I,44Sc,47Sc,64Cu,52Mn,55Co,86Y,89Zr,149Tb,152Tb, and203Pb.239#583961Attorney Docket No. 00020.026.180132. A pharmaceutical composition comprising the complex of any one of claims 1-31, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

33. A complex of any one of claims 1-31, or a pharmaceutically acceptable salt thereof, for use in treating cancer in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of the complex, or a pharmaceutically acceptable salt thereof.

34. The complex of claim 33, wherein the cancer is selected from the group consisting of breast cancer, pancreatic cancer, mesothelioma, esophageal cancer, cholangiocellular carcinoma, head and neck cancer, carcinoma of unknown primary origin, brain metastases, sarcoma, salivary gland carcinoma, lung cancer, prostate cancer, thymus cancer, ovarian cancer, desmoid, chordoma, colorectal cancer, anal carcinoma, neuroendocrine tumors, small intestine cancer, medullary thyroid cancer, cervical cancer, hepatocellular carcinoma, gastric cancer, adenoid cystic cancer, pheochromocytoma, differentiated thyroid cancer, insulinoma, renal cell cancer, gastric cancer, renal cancer, bone metastases, lymph node metastases, lung metastases, pleura, bladder cancer, myeloma, lymphoma, brain cancer, endometrial cancer, testicular cancer, neuroblastoma, astrocytoma, meningioma, rectal cancer, cervical cancer, stomach cancer, skin cancer, thyroid cancer, and / or liver cancer.

35. A complex of any one of claims 1-31, or a pharmaceutically acceptable salt thereof, for use in treating a disease mediated by cancer-associated fibroblasts in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of the complex, or a pharmaceutically acceptable salt thereof.

36. The complex of claim 35, wherein the disease is cancer.

37. The complex of claim 36, wherein the cancer is selected from the group consisting of breast cancer, pancreatic cancer, mesothelioma, esophageal cancer, cholangiocellular carcinoma, head and neck cancer, carcinoma of unknown primary' origin, brain metastases, sarcoma, salivary' gland carcinoma, lung cancer, prostate cancer, thymus cancer, ovarian cancer, desmoid, chordoma, colorectal cancer, anal carcinoma, neuroendocrine tumors, small intestine cancer, medullary’ thyroid cancer, cervical cancer, hepatocellular carcinoma, gastric cancer, adenoid cystic cancer, pheochromocytoma, differentiated thyroid cancer, insulinoma, renal cell cancer, gastric cancer, renal cancer, bone metastases, lymph node metastases, lung metastases, pleura, bladder cancer, myeloma, lymphoma, brain cancer, endometrial cancer.240#583961Attorney Docket No. 00020.026.1801testicular cancer, neuroblastoma, astrocytoma, meningioma, rectal cancer, cervical cancer, stomach cancer, skin cancer, thyroid cancer, and / or liver cancer.

38. A complex of any one of claims 1-31, or a pharmaceutically acceptable salt thereof, for use in imaging a cancer-associated fibroblast in a patient in need thereof, comprising administering to the patient an effective amount of the complex, or a pharmaceutically acceptable salt thereof, and identifying the location of the cancer-associated fibroblast in the patient.

39. A complex of any one of claims 1-31, or a pharmaceutically acceptable salt thereof, for use in imaging activated fibroblasts in a patient with idiopathic pulmonary fibrosis, cirrhosis or liver fibrosis, Crohn’s disease, rheumatoid arthritis, ulcerative colitis, sclerosis, cardiac fibrosis, and / or kidney fibrosis in need thereof, comprising administering to the patient an effective amount of the complex of any one of claims 1-31, or a pharmaceutically acceptable salt thereof, and identifying the location of activated fibroblasts in the patient for noninvasive diagnosis.

40. A compound represented by Formula (II):D-L-RLIL'EFormula (II)or a pharmaceutically acceptable salt thereof, wherein:D is a ligand that targets fibroblast activation protein (FAP);L is a linker;LEis a peptide linker comprising at least one proline;E is a secondary effector moiety: andRLis a ligand.241#583961Attorney Docket No. 00020.026.180141. The compound of claim 40, wherein D is represented by Formula X-B:wherein:T is substituted or unsubstituted methylene (-CH2-), substituted or unsubstituted amino (-NH-), -O-, or -S- (e.g.. wherein the substitution of T is C1-C3 alkyl, haloalkyl, or halo);J is C(RJ)0-3, wherein each RJis independently H or alkyl, or two or more RJare taken together to form oxo;R1and R2are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-_ -C=C-C(O)aryl, -C=C-S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, -CONH2, and 5-tetrazolyl;R3and R4are independently selected from the group consisting of -H, -OH, F, Cl, Br, I, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl;R3, R6, R7, and R8are independently selected from the group consisting of H, alkyl, and halo; andR9, R10, andR11are independently selected from the group consisting of H,-Ci-ealkyl, -Ci ehaloalkyl, -O-Ci-ealkyl, -S-C1-6 alkyd, F, Cl, Br, and I.

42. The compound of claim 40, wherein D is represented by Formula X-C:242#583961Attorney Docket No. 00020.026.1801wherein:R21is -CN orOeach r and s is independently 1 or 2;R22and R23are independently selected from the group consisting of H, Ci-6 alkyd, and halo; andR24is C1-6 alkyl;Q is -N(R25)(C(R26)(R27))t- or -O(C(R28)(R29))UC(O)-;R25is selected from H and C1-6 alkyl; andeach R26, R27, R28, and R29is independently selected from H and Ci-6 alkyd; and each t and u is independently 0 or 1.

43. The compound of claim 40, wherein D is represented by:wherein,R30is -H, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=C-C(O)ai 4, -C=C-S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, or 5-tetrazolyl;R31is H or CH3; andAr1is substituted phenyl, pyridyl, chloropyridyl, or quinolinyl.

44. The compound of claim 40, wherein D is represented by:243#583961Attorney Docket No. 00020.026.1801R41is selected from the group consisting of -H, -CN, -B(0H)2, -C(O)alkyl, -C(O)aryl, -C=C-C(O)aryl. -C=C-S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, and 5-tetrazolyl, R42, R43a, R43band R44are each independently selected from the group consisting of -H, -OH, halogen, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl,R45is selected from the group consisting of -H, -CH2OH, and -CH(CH3)2,R46, R47, and R48are each independently selected from the group consisting of -H, -OH, oxo, halogen, CFs, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl, -NR49R50, -OR51, - Het2, and -Arc; each of -Ci-ealkyl being optionally substituted with from 1 to 3 substituents selected from -OH and halogen; R49, R50, and R51are each independently selected from the group consisting of -H, -OH, oxo, halogen, CFs, -Ci-ealkyl, -O-Ci-ealkyl, -S-Ci-ealkyl, and -An,Arc and An are each independently a 5- or 6-membered aromatic monocycle optionally comprising 1 or 2 heteroatoms selected from O, N, and S; each of Arc and An being optionally and independently substituted with from 1 to 3 substituents selected from -NR52R53, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl,R52and R53are each independently selected from the group consisting of -H, -OH, CF3, -Ci-ealkyl, -O-Ci-ealkyl, and -S-Ci-ealkyl,Het2is a 5- or 6-membered non-aromatic monocycle optionally comprising 1 or 2 heteroatoms selected from O, N and S; Het2being optionally substituted with from 1 to 3 substituents selected from -NR54R55, -Ci-6alkyl, -O-Ci-6alkyl, and -S-Ci-6alkyl,244#583961Attorney Docket No. 00020.026.1801 R54and R55are each independently selected from the group consisting of -H, -OH, halogen, CF3, -Ci-ealkyl, -O-Ci-salkyl, and -S-Ci-6alkyl,the fragment:represents a 5- to 10-membered N-containing aromatic or non-aromatic mono- or bicyclic heterocycle, said heterocycle optionally further comprising 1 to 3 heteroatoms selected from O, N, and S; andJ is selected from the group consisting of a bond, -Ci-3alkyl, -Ci-3alk l-NH-, C=O, and -O-.

45. The compound of claim 40, wherein D is represented by:245#583961Attorney Docket No. 00020.026.1801wherein:each m is independently 0 to 6.

46. The compound of any one of claims 40-45, wherein L is represented by:wherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl, 6-membered heteroaryl, and C3-6 cycloalkyl;L1is selected from a covalent bond and a linker; andL2is a linker.whereina is the point of attachment to D;246#583961Attorney Docket No. 00020.026.1801P is the point of attachment to RL; andy is the point of attachment to LE.

47. The compound of any one of claims 40-46, wherein L is represented by:wherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl, 6-membered heteroaryl, and C3-6 cycloalkyl;n is an integer from 1 to 6,q is an integer from 1 to 20; andp is 0 or 1;wherein:a is the point of attachment to D;is the point of attachment to RL; andy is the point of attachment to LE.

48. The compound of claim 46 or 47, wherein ring A is 3 to 6-membered heterocyclyl.

49. The compound of any one of claims 46-48, wherein ring A is selected from the group consisting of:247#583961Attorney Docket No. 00020.026.1801and, wherein \| / represents the point of attachment to L1, and s represents the point of attachment to L2.

50. The compound of any one of claims 40-49, wherein LEis represented by:wherein each X1is an amino acid;Lcis absent or a cleavable linker; andm is an integer from 1 to 3;provided that at least one X1is proline; andwherein 0 represents the point of attachment to L and p represents the point of attachment to E.

51. The compound of claim 50, wherein each X1is independently selected from the group consisting of absent, proline, glycine, alanine, threonine, and serine, provided that at least one X1is proline.

52. The compound of claim 50 or 51, wherein each X1is independently selected from the group consisting of proline and glycine, provided that at least one X1is proline, and m is 2.

53. The compound of any one of claims 50-52, wherein Lcis absent.

54. The compound of any one of claims 50-52, wherein Lcis a cleavable linker represented by:N O NI248#583961Attorney Docket No. 00020.026.180155. The compound of any one of claims 40-54, wherein RLis capable of chelating or covalently binding to a therapeutic radionuclide or a diagnostic radionuclide.

56. The compound of any one of claims 40-55, wherein RLis represented by:249#583961Attorney Docket No. 00020.026.1801or250#583961Attorney Docket No. 00020.026.180157. The compound of any one of claims 40-56, wherein RLis represented by:

58. The compound of any one of claims 40-57, wherein E is an albumin binding ligand.

59. The compound of any one of claims 40-58, wherein E comprises a cleavable linker.

60. The compound of any one of claims 40-58, wherein E is represented by:251#583961Attorney Docket No. 00020.026.1801wherein each of X and Y is independently selected from the group consisting of halogen and C1-6 alkyl, and n is an integer from 1 to 20.

61. The compound of claim 60, wherein X is I.

62. A compound, comprising a linker represented by:owherein:ring A is selected from the group consisting of 3 to 6-membered heterocyclyl. 6-membered heteroaryl, and C3-6 cycloalkyl;n is an integer from 1 to 6,q is an integer from 1 to 20;p is 0 or 1; andLEis a peptide linker comprising at least one proline.252#583961Attorney Docket No. 00020.026.180163. The compound of claim 62, wherein LEis represented by:e-Cx ic-gwherein:each X1is an amino acid;Lcis absent or a cleavable linker; andm is an integer from 1 to 3;provided that at least one X1is proline; andwherein 0 represents the point of attachment to L and p represents the point of attachment to E.

64. The compound of claim 63, wherein each X1is independently selected from the group consisting of absent, proline, glycine, alanine, threonine, and serine provided that at least one X1is proline.

65. The compound of claim 63 or 64, wherein each X1is independently selected from the group consisting of proline and glycine, provided that at least one X1is proline, and m is 2.

66. The compound of any one of claims 63-65, wherein Lcis absent.

67. The compound of any one of claims 63-65, wherein Lcis a cleavable linker represented by:

68. A compound of a compound selected from the group consisting of:253#583961Attorney Docket No. 00020.026.1801HOOC^ / — \ COOHHOOC^x / — \ COOH254#583961Attorney Docket No. 00020.026.1801255#583961Attorney Docket No. 00020.026.1801256#583961Attorney Docket No. 00020.026.1801HOOC^ / — \ COOHand HOOC^x / — \ COOHSO3H or a pharmaceutically acceptable salt thereof.257#583961

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