WO2025193583A1

WO2025193583A1 - Macrocyclic peptides useful as immunomodulators

Info

Publication number: WO2025193583A1
Application number: PCT/US2025/019126
Authority: WO
Inventors: Jennifer X. Qiao; Michael A. Poss
Original assignee: Bristol Myers Squibb Co
Current assignee: Bristol Myers Squibb Co
Priority date: 2024-03-11
Filing date: 2025-03-10
Publication date: 2025-09-18
Anticipated expiration: 2026-09-11

Abstract

In accordance with the present disclosure, macrocyclic compounds have been discovered which inhibit the LAG-3/MHC Class II protein/protein interaction, and may be useful for the amelioration of various diseases, including cancer and infectious diseases.

Description

MACROCYCLIC PEPTIDES USEFUL AS IMMUNOMODULATORS CROSS REFERENCE This application claims the benefit of U.S. Provisional Application No. 63/563,688 filed March 11, 2024 which is incorporated herein in its entirety. BACKGROUND The present disclosure provides novel macrocyclic peptides which inhibit the LAG-3/MHC Class II protein/protein interaction and are thus useful for the amelioration of various diseases, including cancer and infectious diseases. Lymphocyte activation gene-3 (LAG-3; LAG3; CD223) is a type I transmembrane protein that is expressed on the cell surface of activated CD4+ T cells, CD8+ T cells, T regulatory cells, B cells, and subsets of natural killer (NK) and dendritic cells (Triebel F, et al., J. Exp. Med.1990; 171:1393-1405; Huard, Eur. J. Immunol.1994; 24:3216-21; Grosso, J. Clin. Invest.2007; 117:3383-92; Huang, Immunity.2004; 21:503-13; Kieslow, Eur. J. Immunol.2005; 35:2081- 88; Workman CJ, et al., J. Immunol.2009; 182(4):1885- 91; Castelli, Oncoimmunology 2014; 3:11). LAG-3 is closely related to CD4, which is a co-receptor for T helper cell activation. Both molecules have four extracellular Ig-Iike domains and require binding to their ligand, major histocompatibility complex (MHC) class II, for their functional activity. In contrast to CD4, LAG-3 is only expressed on the cell surface of activated T cells and its cleavage from the cell surface terminates LAG-3 signaling. LAG-3 can also be found as a soluble protein but it does not bind to MHC class II and its function is unknown. LAG-3 is composed of the intracellular signaling domain, a transmembrane domain and 4 extracellular domains, designated Dl to D4 (Huard 1997 Proc. Natl. Acad. Sci.94:5744-9). Domain 1-2 associates with MHC class Il ligand and it has been shown that the tip of domain 1 (extra loop) forms the binding site (Huard 1997 Proc. Natl. Acad. Sci.94:5744-9). LAG-3 can also associate with alternative ligands, Galectin-3 and LSECtin, which induce its inhibitory signaling (Kouo 2015 Cancer Immunol Res.3(4):412-23; Xu 2014 Cancer Res 74(13):3418-28). Association with Galectin-3 on cells or within the extracellular matrix could downregulate T cells that would not normally engage with MHC class II, such as CD8+ T cells. Therefore, blockade of this ligand could serve as a mechanism for enhancing broad T cell function. A role of LAG-3 on T cells is to regulate T cell activation (Huard 1994 Eur. J. Immunol.24:3216- 21). LAG-3 engages with MHC class Il and this leads to down regulation of CD4+ T cells (Huard 1996 Eur. J. Immunol.26:1180-6). Upon T cell activation, LAG-3 surface expression increases. The engagement of LAG-3 dimer with ligand induces signaling through an intracellular KIEELE domain (Workman 2002 J. Immunol 169:5392-5) leading to downregulation of the T cell activity. Therefore, LAG-3 serves to modulate responses to antigens, preventing over-stimulation and maintaining immune homeostasis. It has been reported that LAG-3 plays an important role in promoting regulatory T cell (Treg) activity and in negatively regulating T cell activation and proliferation (Workman CJ, et al., J. Tmmunok 2005; 174:688-695). Both natural and induced Treg express increased LAG-3, which is required for their maximal suppressive function (Camisaschi C, et al., J. Tmmunok 2010; 184:6545-6551 and Huang CT, et al, Immunity. 2004; 21:503-513). Furthermore, ectopic expression of LAG-3 on CD4+ effector T cells reduced their proliferative capacity and conferred on them regulatory potential against third party T cells (Huang CT, et al, Immunity.2004; 21:503-513). Recent studies have also shown that high LAG-3 expression on exhausted lymphocytic choriomeningitis virus (LCMV)-specific CD8+ T cells contributes to their unresponsive state and limits CD8+ T cell antitumor responses (Blackburn SD, et al, Nat. Tmmunok 2009; 10:29-37 and Grosso JF, et al, J. Clin. Invest.2007; 117:3383-3392). In fact, LAG-3 maintained tolerance to self and tumor antigens via direct effects on CD8+T cells in 2 murine models (Grosso JF, et al, J. Clin. Invest.2007; 117:3383-3392). Epstein-Barr virus infection is yet another factor to consider in the potential induction of T cell exhaustion in hematological malignancies. It is known that EBVassociated CLL, Richter’s syndrome, and lymphoma cases are usually more aggressive than their EBV(-) counterpart (Tsimberidou AM, et al., Leuk Lymphoma 2006;47:827; Ansell SM, et al., Am J Hematol 1999;60:99.; Dolcetti R, et al., Infectious Agents and Cancer 2010;5:22; Kanakry JA, et al., Blood 2013;121:3547). Interestingly, the expression of checkpoint inhibitors like PD-Ll and LAG-3 has also been documented in EBV-associated malignancies (Green MR, et al., Clin Cancer Res 2012; 18:1611; Monti S, et al., Blood 2005;105:1851). High expression of LAG-3 has in fact been documented in chronic viral infections and its blockade with anti-LAG-3 antibodies has been able to reduce viral titers and the expression of checkpoint inhibitors in murine models (Blackburn SD, et al., Nat. Immunol.2009;10:29-37). Furthermore, LAG-3 expression, alone or in combination with other markers, has been evaluated as a prognostic or predictive marker in CLL and Hodgkin lymphoma (Zhang J, et al., BMC Bioinformatics 2010;1 l(Suppl 9):S5; Kotaskova J, et al., J Mol Diagn 2010;12(3):328— 334). LAG-3 expression on tumor-infiltrating lymphocytes (TILs) and peripheral blood also mediates T cell exhaustion in hematological malignancies (Dickinson JD, et al., Leuk Lymphoma 2006;47(2):231-44). Moreover, LAG-3 blockade with specific antibodies has shown antitumor activity in leukemia (Berrien-Elliott, M, et al., Cancer Research 2013; 73(2):605-616) and solid tumor models (Woo, S-R, et al., Cancer Research 2011; 72(4):917-927; Coding, S. R., et al., Journal of Immunology, Baltimore, Md.1950; 190(9):4899-909). Therefore, LAG-3 is a potential therapeutic target in hematological malignancies. Recent preclinical studies have documented a role for LAG-3 in CD8 T cell exhaustion, and blockade of the LAG-3/ MHC Class II interaction using LAG-3 blocking antibodies or LAG-3-Ig fusion proteins is being evaluated in a number of clinical trials in cancer patients. Additional background information can be found in WO2015/042246 A1, WP2015/116539 A1, and WO2014/008218 A1. LAG-3 blockade with macrocyclic peptide inhibitors, alone and in combination with standard of care (e.g., nivolumab, imatinib, lenalidomide) or with other checkpoint inhibitors deserves further exploration. The molecules described herein demonstrate the ability to block the interaction of LAG-3 with MHC Class II, in both biochemical and cell-based experimental systems. These results are consistent with a potential for therapeutic administration to enhance immunity in cancer or chronic infection, including therapeutic vaccine. The macrocyclic peptides described herein can inhibit the interaction of Lag-3 with MHC class II. These compounds demonstrated highly efficacious binding to LAG-3, blockade of the interaction of LAG-3 with MHC Class II and can promote enhanced T cell functional activity, thus making them candidates for parenteral, oral, pulmonary, nasal, buccal and sustained release formulations. Additionally, or alternatively, the macrocyclic peptides can possess one or more of the following functional properties described above, such as high affinity binding to human LAG-3, relatively good binding affinity to cyno LAG-3, and lack of binding to mouse LAG-3, the ability to inhibit binding of LAG-3 to MHC Class II molecules and/or the ability to stimulate antigen-specific T cell responses. In its first embodiment, the present disclosure provides a compound of formula (I) (), or a pharmaceutically acceptable salt thereof, wherein: A is selected from denotes the point of attachment to the carbonyl group and denotes the point of attachment to the nitrogen atom; n is 0 or 1; X is bond, S, O, CHR²³, NR²³ wherein R²³ is hydrogen or C1-C3 alkyl; R¹⁴ and R¹⁵ are independently selected from hydrogen and methyl; R^16a is selected from hydrogen, C1-C6 alkyl, -CH2OH, -(CH2)1-4NH2, -(CH2)1-4NH- C(=NH)NH2, -CH2CO2H and -(CH2)2CO2H; R¹⁶ is selected from -(C(R^17aR¹⁷))0-2-X’-R³⁰, -(C(R^17aR¹⁷))1-2C(O)N(R^16a)C(R^17a)2-X’-R³¹, -C(R^17aR¹⁷)1-2[C(O)N(R^16a)C(R^17aR¹⁷)1-2]w’ –X’-R³¹, -(C(R^17aR¹⁷)1-2C(O)NR^16a)m’-X’- R³⁰; and -(C(R^17aR¹⁷)_1-2C(O)NR^16a)_m’-C(R^17a)(R¹⁷)-CO₂H; -(C(R^17aR¹⁷)1-2C(O)NR^16a-PEGn’)m’-C(R^17a)(R¹⁷)-CO2H; Alternatively, R¹⁶ and R^16a together with the atoms attached may form a 4-6 membered cyclic rings, wherein the ring can be fused with another aromatic or heteroaromatic ring; the cyclic ring is subsituted with -(C(R^17aR¹⁷))0-2-X’-R³⁰, -(C(R^17aR¹⁷))1-2C(O)N(R^16a)C(R^17a)2-X’-R³¹, -C(R^17aR¹⁷)1-2[C(O)N(R^16a)C(R^17aR¹⁷)1-2PEGn’]w’ –X’-R³¹, -(C(R^17aR¹⁷)_1-2C(O)NR^16aPEG_n’)_m’-X’- R³⁰; and -(C(R^17aR¹⁷)1-2C(O)NR^16aPEGn’)m’-C(R^17a)(R¹⁷)-CO2H; -(C(R^17aR¹⁷)_1-2C(O)NR^16a-PEG_n’)_m’-C(R^17a)(R¹⁷)-CO₂H; wherein: w’ is 1-5; m’ is 0-6; n’ is 0-20; X’ is a chain of between 1 and 172 atoms wherein the atoms are selected from carbon and oxygen and wherein the chain may contain one, two, three, or four groups selected from -NHC(O)-, and -C(O)NH-, -NHC(O)NH-, embedded therein; and wherein the chain is optionally substituted with one to six groups independently selected from – CO2H, -C(O)NH2, and –(CH2)1-2CO2H, provided that X’ is other than unsubstituted PEG; R³⁰ is selected from –CO2H, -C(O)NR^wR^x, and -CH3 wherein R^w and R^x are independently selected from hydrogen and C₁-C₆alkyl, tetrazole, and lithocholic acid and analogs of lithocholic acid; R³¹ is selected from -CO2H, -C(O)NR^wR^x, -CH3, alexa-5-SDP, tetrazole, and lithocholic acid and analogs of lithocholic acid, and biotin; each R^17a is independently selected from hydrogen, C₁-C₆alkyl, C₁-C₆OH, C₁- C6NH2, C1-C6NH-C(=NH)NH2, COOH, CONH2, C1-C6CO2H, C1-C6CONH2, C1-C6aryl, C₁-C₆heteroaryl; each R¹⁷ is independently selected from hydrogen, -CH₃, -(CH₂)_zNH₂, – (C(R^17a)2)0-4-X’-R³¹, -(CH2)zCO2H, –CH2OH, CH2C=CH, and -(CH2)z-triazolyl-X’-R³⁵, wherein z is 1-6 and R³⁵ is selected from -CO₂H, -C(O)NR^wR^x, CH₃, biotin, -C(O)- (CH2)2–C(O)O-vitamin E,–C(O)O-vitamin E, tetrazole, and lithocholic acid and analogs; provided that at least one R¹⁷ is other than hydrogen, -CH3, or –CH2OH; R^b, R^d, R^e, and R^k are each independently selected from hydrogen or methyl; R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², and R¹³ are independently selected from a natural amino acid side chain and an unnatural amino acid side chain or form a ring with the corresponding vicinal R group as described below; R^f is hydrogen or methyl, or R^f and R⁶ together with the atoms to which they are attached may form a ring selected from either the D enantiomer or the L enantiomer of azetidine, pyrrolidine, morpholine, piperidine, piperazine, and tetrahydrothiazole; wherein each ring is optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, and hydroxy; R^g is hydrogen or methyl, or R^g and R⁷ together with the atoms to which they are attached may form a ring selected from either the D enantiomer or the L enantiomer of azetidine, pyrrolidine, morpholine, piperidine, piperazine, and tetrahydrothiazole; wherein each ring is optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, and hydroxy; Rⁱ is hydrogen or methyl, or Rⁱ and R⁹, together with the atoms to which they are attached may form a ring selected from azetidine, pyrrolidine, morpholine, piperidine, piperazine, and tetrahydrothiazole; wherein each ring is optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, hydroxy, and phenyl or fused with another aromatic ring optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, hydroxy, and phenyl; and wherein the pyrrolidine and the piperidine ring are optionally fused to a cyclohexyl, phenyl, or indole group; R^m is hydrogen or C_1-6 alkyl, or R^m and R¹³, together with the atoms to which they are attached may form a ring selected from azetidine, pyrrolidine, morpholine, piperidine, piperazine, and tetrahydrothiazole; wherein each ring is optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, hydroxy, and phenyl or fused with another aromatic ring optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, hydroxy, and phenyl; and wherein the pyrrolidine and the piperidine ring are optionally fused to a cyclohexyl, phenyl, or indole group; In one aspect of the first embodiment: the present disclosure provides a compound of formula (Ia), or a pharmaceutically acceptable salt thereof

naturally occurring amino acid consisting of D-Ala and D-Phe; Alternatively when R^e is methyl, R⁵ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of Phe, Phe(2-F), Phe(4-F), Trp, Tyr, Tyr(CH₂phenyl), and D-Phe; When R^f is hydrogen, R⁶ is selected from the sidechain of D-Ala, D-Asp, D-Asn, D-Glu, D-Gln, D-Leu, D-Trp, D-Tyr or Lys. Alternatively when R^f is methyl, R⁶ is selected from the sidechain of Gly, D-Ala, D-Leu, Ala or Leu, or alternatively, R^f and R⁶ together with the atoms to which they are attached can form a ring selected from the D enantiomer of azetidine, pyrrolidine, morpholine, and piperidine; wherein each ring is optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, and hydroxy, and wherein each ring can be fused with a six-membered aromatic or heteroaromatic ring; R^g is hydrogen or methyl, or R^g and R⁷ together with the atoms to which they are attached can form a pyrrolidine ring which is optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, and hydroxy; Rⁱ is hydrogen or methyl, or Rⁱ and R⁹, together with the atoms to which they are attached, can form a ring selected from pyrrolidine, morpholine, piperidine; wherein each ring is optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, hydroxy, and phenyl or fused with another aromatic ring optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, hydroxy, and phenyl; and wherein the pyrrolidine and the piperidine ring are optionally fused to a cyclohexyl, phenyl, or indole group; R^m is hydrogen or methyl, or R^m and R¹³, together with the atoms to which they are attached, can form a pyrrolidine, morpholine, piperidine, piperazine, and tetrahydrothiazole; wherein each ring is optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, hydroxy, and phenyl or fused with another aromatic ring optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, hydroxy, and phenyl; and wherein the pyrrolidine and the piperidine ring are optionally fused to a cyclohexyl, phenyl, or indole group; R¹⁰ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of Bzt, Trp, Trp(7-F), Trp(7-F), and Trp (1-Me); R¹² is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of Bzt, Phe, Trp, and Tyr. m’ is 0-5; n’ is 0-11; In another embodiment the present disclosure provides a method of enhancing, stimulating, and/or increasing the immune response in a subject in need thereof, said method comprising administering to the subject a therapeutically effective amount of at least one macrocyclic peptide described herein. In another embodiment the method further comprises administering an additional agent prior to, after, or simultaneously with the macrocyclic, peptide or peptides described herein. In another embodiment the additional agent is an antimicrobial agent, an antiviral agent, a cytotoxic agent, and/or an immune response modifier. In another embodiment the present disclosure provides a method of inhibiting growth, proliferation, or metastasis of cancer cells in a subject in need thereof, said method comprising administering to the subject a therapeutically effective amount of one or more macrocyclic peptides described herein. In another embodiment the cancer is selected from melanoma, renal cell carcinoma, squamous non-small cell lung cancer (NSCLC), non-squamous NSCLC, colorectal cancer, castration-resistant prostate cancer, ovarian cancer, gastric cancer, hepatocellular carcinoma, pancreatic carcinoma, squamous cell carcinoma of the head and neck, carcinomas of the esophagus, gastrointestinal tract and breast, and a hematological malignancy. In another embodiment the present disclosure provides a method of treating an infectious disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of at least one macrocyclic peptide described herein. In another embodiment the infectious disease is caused by a virus. In another embodiment, the virus is selected from HIV, Hepatitis A, Hepatitis B, Hepatitis C, herpes virus, and influenza. In another embodiment the present disclosure provides a method of treating septic shock in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of one or more macrocyclic peptides described herein. In another embodiment the present disclosure provides a method blocking the interaction of LAG-3 with MHC Class II molecule in a subject, said method comprising administering to the subject a therapeutically effective amount of at least one macrocyclic peptide described herein. In one aspect of the embodiments, compounds of formula (Ia), R¹ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of 1- naphthylalanine (1-Nal), 2-naphthylalanine (2 Nal), 3-(2-thienyl)-alanine, 3-(3-thienyl)- alanine, 3-benzothienylalanine (Bzt), 4-pyridinylalanine (4-Pya), biphenylalanine (Bip), 4-bromophenylalanine (Bpa), phenylalanine, tyrosine, tryptophan, 2-fluorophenylalanine, 3-fluorophenylalanine, 4-fluorophenylalanine, 3,4-difluorophenylalanine, 3,5- difluorophenylalanine, 3,4,5-difluorophenylalanine, pentafluorophenylalanine, 3- methylphenylalanine, 4-methylphenylalanine, 4-chlorophenylalanine, 3- methoxyphenylalananie, 4-methoxyphenylalanine, 4-cyanophenylalanine, 4- difluoromethylphenylalanine, 4-trifluoromethylphenylalanine, 3-chlorophenylalanine, 4- aminophenylalanine, 4-aminomethylphenylalanine, 4-carbamoylphenylalanine, 4- carboxyphenylalaine, 3-(4-thiazolyl)-alanine, ^-phenyl-phenylalanine, 4- phenoxyphenylalanine, and 4-cyclohexyloxyphenylalanine;. In compounds of formula (Ia) when R^b is hydrogen, R² is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of 2- naphthylalanine, 2-pyridinylalanine, 3-pyridinylalanine, 4-pyridinylalanine, 3-(6-(o- tolyl)pyridinylalanine, tert-butylglycine, 3-benzothienylalanine, phenylalanine, 2- fluorophenylalanine, 3-fluorophenylalanine, 4-fluorophenylalanine, 2- methylphenylalanine, 3-methylphenylalanine, 4-methylphenylalanine, 3- chlorophenylalanine, 4-chlorophenylalanine, 3-methoxyphenylalananie, 4- methoxyphenylalanine, 3-cyanophenylalanine, 4-cyanophenylalanine, 4- difluoromethylphenylalanine, 3-aminomethylphenylalanine, 4- aminomethylphenylalanine, 4-carbamoylphenylalanine, 4-carboxyphenylalanine, 3- carboxyphenylalanine, 3-hydroxylphenylalanine, tyrosine, valine, 4-(prop-2-yn-1- yloxy)phenylalanine, 4-benzoxyphenylalanine, 4-carboxymethoxyphenylalanine, lysine, and 4-allyloxyphenylalanine;. Alternatively, when R^b is methyl, R² is selected from the sidechain of alanine, glycine, phenylalanine, tyrosine. In compounds of formula (Ia), preferred R³ is the sidechain of aspartic acid. In compounds of formula (Ia) when R^d is hydrogen, R⁴ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of 4- pyridinylalanine, alanine, aspartic acid, histidine, asparagine, glutamic acid, homo- glutamic acid, 2,3-diaminopropionic acid (Dap), phenylalanine, 3-carboxyphenylalamine, 4-carboxyphenylalamine, tyrosine, glutamine, arginine, 2-amino-4-aminobutyric acid (Dab), ornithine (Orn), threonine, lysine, lysine (COCH₃), propargylglycine, 4-(prop-2- yn-1-yloxy)phenylalanine, 4-carboxymethoxyphenylalanine, tryptophan, tryptophan (1- acetic acid), and 4-pyridinylalanine; Alternatively, when R^d is methyl, R⁴ is selected from the sidechain of serine, alanine, glycine, tyrosine, aspartic acid, and threonine; In compounds of formula (Ia), when R^e is hydrogen, R⁵ is selected from the sidechain of a non naturally occurring amino acid consisting of D-alanine and D-Phe; Alternatively, when R^e is methyl, R⁵ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of tryptophan, tyrosine, phenylalanine, 2-F-phenylalanine, 3-F-phenylalanine, 4-F-phenylalanine, and 4-benzyltyrosine. In compounds of formula (Ia), when R^f is hydrogen, R⁶ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of glycine, D- alanine, D-leucine, D-aspartic acid, D-asparagine, D-glutamic acid, D-glutamine, serine, D-lysine, D-tryptophan, and D-tyrosine; Alternatively, when R^f is methyl, R⁶ is selected from the sidechain of L-alanine, D-alanine, L-leucine, D-leucine, and glycine; alternatively, R^f and R⁶ together with the atoms to which they are attached can form a ring selected from the D enantiomer of azetidine, pyrrolidine, morpholine, and piperidine; wherein each ring is optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, and hydroxy, and wherein each ring can be fused with a six-membered aromatic or heteroaromatic ring; In compounds of formula (Ia) when R^g is hydrogen, R⁷ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of glycine, t- butylglycine, 4-pyridinylphenylalanine, aspartic acid, asparagine, glutamic acid, 2,3- diaminopropionic acid (Dap), 2-amino-4-aminobutyric acid (Dab), ornithine, threonine, lysine, lysine (COCH3), serine, homo-serine, arginine, ornithine, histidine, glutamine, alanine, propargylglycine, 4-(prop-2-yn-1-yloxy)phenylalanine, tryptophan (1-acetic acid), 4-trifluoromethylphenylalanine, and 3-carboxyphenylalanine. Alternatively, when R^g is methyl, R⁷ is selected from the sidechain of glycine, alanine, aspartic acid, serine, threonine, and tyrosine; In compounds of formula (Ia) R⁸ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of alanine, glycine, L-aspartic acid, D- aspartic acid, asparagine, glutamic acid, 2-amino-4-aminobutyric acid (Dab), threonine, lysine, serine, homo-serine, methyl-homo-serine, tryptophan (1-acetic acid), and 3- carboxyphenylalanine; In compounds of formula (Ia) when Rⁱ is hydrogen, R⁹ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of glutamic acid, glycine, serine, homo-serine, methyl-homo-serine, propargylglycine, glutamine, lysine, arginine, tyrosine, and valine; When Rⁱ is methyl, preferred R⁹ side chains are: alanine and serine; Alternatively, when Rⁱ is n-butyl or –CH2COOH, R⁹ is the sidechain of glycine; Alternatively Rⁱ and R⁹, together with the atoms to which they are attached, can form a ring selected from pyrrolidine, morpholine, piperidine; wherein each ring is optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, hydroxy, and phenyl or fused with another aromatic ring optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, hydroxy, and phenyl; and wherein the pyrrolidine and the piperidine ring are optionally fused to a cyclohexyl, phenyl, or indole group; In compounds of formula (Ia) R¹⁰ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of tryptophan, 7-methyltryptophan, benzothienylalanine, 7-fluorotryptophan, 7-methyltryptophan, 3-methylphenylalanine, tryptophan(1-methyl). In compounds of formula (Ia) when R^k hydrogen, R¹¹ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of tryptophan, tyrosine, 4-(prop-2-yn-1-yloxy)phenylalanine, propargylglycine, 3- hydroxyphenylalanine, N-methyltryptophan, 7-methyltryptophan, 3-pyridinylalanine, phenylalanine, 4-carboxyphenylalanine, 4-benzoxyphenylalanine, 1-napththylalanine, 3- carboxyphenylalanine, 4-aminomethylphenylalanine, 4-methoxyphenylalanine, 5- cyanotryptophan, 2-mehylphenylalanine, 2-methyltryptophan, 2-napththylalanine, 4- fluorophenylalanine, glutamine, arginine, valine, tert-butylglycine, glycine, lysine, 1- acetic acid-tryptophan, 3-methylphenylalanine, biphenylalanine, biphenylalanine, 3- benzothienylalanine, 4-(4-pyridinyl)phenylalanine, 4’-carboxy-4-biphenylalanine, 3’- carboxy-4-biphenylalanine, 3-quinolinylalanine, 6-quinolinylalanine, 6- isoquinolinylalanine and isotryptophan; Alternatively when R^k is methyl, R¹¹ is selected from the sidechain of glycine, tyrosine, and tryptophan; In compounds of formula (Ia) R¹² is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of tryptophan, tyrosine, phenylalanine, and 3-benzothienylalanine (Bzt). In compounds of formula (Ia) where R^m is hydrogen, R¹³ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of glycine, propargylglycine, alanine, aspartic acid, asparigine, arginine, glutamic acid, glutamine, norvaline, serine, lysine, 4-carboxyphenylalanine, 1-acetic acid-tryptophan, 3- carboxyphenylalanine, 4-carbamoylphenylalanine, tetrazolylalanine, methyl-homo-serine, 4-carboxymethoxyphenylalanine, 4-pyridinylalanine, 3-pyridinylalanine, benzoxyphenylalanine, 4-(prop-2-yn-1-yloxy)phenylalanine, 2,3-diaminopropionic acid (Dap), 2-amino-4-aminobutyric acid (Dab), cyclopropylalanine; and ethyl; Alternatively, when R^m is methyl, R¹³ is selected from the sidechain of glycine, alanine, aspartic acid, glutamic acid, and tyrosine; Alternatively, when R^m is n-butyl or –CH2COOH, R⁹ is the sidechain of glycine; In another aspect of the embodiments, (binding less than 1000 nM) In compounds of formula (Ia), R¹ is selected from the sidechain of a naturally or non naturally occurring amino acid consisting of L-Ala(^-4-pyridinyl) (L-4-Pya), L-Phe, L-Phe(3-F), L-Phe(4-F), L-Phe(3,5-di-F), L-Phe(3,4,5-tri-F), L-Phe(3-OMe), L-Phe(4- COOH), L-^-phenyl-phenylalanine, and L-Ala(^-3-pyridinyl (L-3-Pya),; R² is the sidechain of L-Tyr; R³ is the sidechain of L-Asp; R⁴ is selected from the sidechain of L-4-Pya, L-Asp, L-Dap, L-Dab, L-Glu, L-Lys, L-Asn, L-Thr, and L-Tyr; When R^e is methyl, R⁵ is selected from the sidechain of L-Phe and L-Tyr; When R^f is hydrogen, R⁶ is the sidechain of D-Glu; Alternatively, when R^f is methyl, R⁶ is selected from the sidechain of Gly, L-Glu, L-Tyr, and dedrydraalanine; Alternatively, R^f and R⁶ together with the atoms to which they are attached can form a ring selected from the sidechain of D-Azt and D-Pro; R⁷ is selected from the sidechain of L-Asp, L-Dap, L-Dab, L-Lys, L-Asn, L-4- Pya, L-Ser, L-Phe(4-CF3), L-Phe(4-COOH), L-Tyr, and L-Trp(CH2COOH); R⁸ is selected from the sidechain of L-Ala, L-Asp, and L-Dab; When Rⁱ is hydrogen, R⁹ is the sidechain of L-Ser; Alternatively, when Rⁱ is methyl, R⁹ is the sidechain of L-Ser; Alternatively, Rⁱ and R⁹ together with the atoms to which they are attached can form a ring selected from the sidechain of D-Pro, L-Pro, D-Hyp, L-Hyp, and L-Morph; R¹⁰ is the sidechain of L-Trp; R¹¹ is selected from the sidechain of L-Trp, L-Tyr, L-Phe(4-CN), and L-alanine(^- 2-thienyl); R¹² is the sidechain of L-Trp; R¹³ is selected from the sidechain of L-4-Pya, L-3-Pya, L-Ala, L- L-Asp, L-Dab, L-Glu, and L-Ser; R^16a is selected from hydrogen, methyl; Alternatively, R¹⁶ and R^16a together with the atoms attached may form 5- membered cyclic rings consistent of Pro or Tic; Each R^17a is independently selected from hydrogen, the sidechain of the following natural or nonnatural occurring amino acids: L-Ala, D-Ala, L-Asp, D-Asp, L-Dap, L- Dab, D-Dap, D-Dab, L-Lys, D-Lys, L-Asn, D-Asn, L-Orn, D-Orn, L-Glu, Gly, L-Arg, D- Arg, L-Ser, D-Ser, L-Phe, D-Phe, L-tetrazolyl-alanine, L-Trp, D-Pro, D-Phe, L-Pra, L- Trp, L-^-Glu, and L-^-Asp; n’ = 0-11 X’ is a chain of between 8 and 60 atoms wherein the atoms are selected from carbon and oxygen and wherein the chain may contain one, two, three, or four groups selected from -NHC(O)-, and -C(O)NH-, -NHC(O)NH-, embedded therein; and wherein the chain is optionally substituted with one to six groups independently selected from – CO₂H, -C(O)NH₂, and –(CH₂)_1-2CO₂H and ; R³⁰ is selected from -CO₂H, -C(O)NH₂, -CH₃, tetrazole, and lithocholic acid; R³¹ is selected from -CO₂H, -C(O)NH₂, -CH₃, tetrazole, and lithocholic acid; R³⁵ is selected from -CO₂H, - C(O)NH₂, -CH₃, tetrazole, and lithocholic acid; In another aspect of the embodiment (binding less than 20 nM), In compounds of formula (Ia), R⁴ is selected from the sidechain of L-Asp, L-Glu, L-Lys, L-Asn, L-Thr, and L-Tyr; R⁸ is selected from the sidechain of L-Ala, and L-Asp; R¹¹ is selected from the sidechain of L-Trp, L-Tyr, and L-Phe(4-CN); R¹² is the sidechain of L-Trp; In another aspect of the embodiment (binding less than or equal to 2.5 nM), In compounds of formula (Ia), R⁴ is selected from the sidechain of L-Asp, L-Glu, L-Lys, L-Asn, and L-Thr; R⁸ is the sidechain of L-Asp; When Rⁱ is hydrogen, R⁹ is the sidechain of L-Ser; Alternatively, when Rⁱ is methyl, R⁹ is the sidechain of L-Ser; Alternatively, Rⁱ and R⁹ together with the atoms to which they are attached can form a ring selected from the sidechain of D-Pro, L-Pro, L-Hyp, and L-Morph; R¹¹ is selected from the sidechain of L-Trp and L-Tyr; As shown below in Tables 1-14, the following compounds of the invention show activity at less than or equal to 0.5 nM in the assay included later in the application.

Example # n m R 1253 2 1 COOH

Table 2 n m R X

Example R^f R⁶ Rⁱ R⁹ m

Table 4 Example X¹ Rⁱ R⁹ X Example X¹ Rⁱ R⁹ X # Example X¹ Rⁱ R⁹ X # Example X¹ Rⁱ R⁹ X # Example X¹ Rⁱ R⁹ X # Example X¹ Rⁱ R⁹ X

Example R⁴ X R⁷ R¹³ n #

Example R¹³ R¹⁵ R¹⁶ n #

Example # R¹³ R¹⁵ R¹⁶ R¹⁷ n

abe 8 Example # L X

Table 9 Exam R¹⁵ L₁ L₂ L₃ L₄ R l # Exam R¹⁵ L₁ L₂ L₃ L₄ R ple # Table 10 Example L R¹⁹ #2

Table 11 Example R¹ R⁴ Rⁱ R⁹ R¹¹ L #

Table 12 Ex. # R¹³ R¹⁵ L n Ex. # R¹³ R¹⁵ L n 1318 -NHCH2CO- 1 Table 13 Ex. # R⁴ R¹⁵ L n

Table 14 Ex. # L n Definitions The definitions provided herein apply, without limitation, to the terms as used throughout this specification, unless otherwise limited in specific instances. Those of ordinary skill in the art of amino acid and peptide chemistry are aware that an amino acid includes a compound represented by the general structure: w Unless otherwise indicated, the term "amino acid" as employed herein, alone or as part of another group, includes, without limitation, an amino group and a carboxyl group linked to the same carbon, referred to as "α" carbon, where R and/or R′ can be a natural or an un-natural side chain, including hydrogen. The absolute "S" configuration at the "α" carbon is commonly referred to as the "L" or "natural" configuration. In the case where both the "R" and the "R′"(prime) substituents equal hydrogen, the amino acid is glycine and is not chiral. The term “naturally occurring amino acid side chain,” as used herein, refers to side chain of any of the naturally occurring amino acids (i.e., alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine,-histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine) usually in the S-configuration (i.e., the L-amino acid). The term “non-naturally occurring amino acid side chain,” as used herein, refers to a side chain of any naturally occurring amino acid usually in the R-configuration (i.e., the D-amino acid) or to a group other than a naturally occurring amino acid side chain in R- or S-configuration (i.e., the D- or L-amino acid, respectively) selected from: C₂-C₇alkenyl, C₁-C₃alkoxyC₁-C₃alkyl, C₁-C₆alkoxycarbonylC₁-C₃alkyl, C₁- C7alkyl, C1-C3alkylsulfanylC1-C3alkyl, amidoC1-C3alkyl, aminoC1-C3alkyl, azaindolylC1- C₃alkyl, benzothiazolylC₁-C₃alkyl, benzothienylC₁-C₃alkyl, benzyloxyC₁-C₃alkyl, carboxyC1-C3alkyl, C3-C6cycloalkylC1-C3alkyl, diphenylmethyl, furanylC1-C3alkyl, imidazolylC₁-C₃alkyl, naphthylC₁-C₃alkyl, pyridinylC₁-C₃alkyl, thiazolylC₁-C₃alkyl, thienylC1-C3alkyl; biphenylC1-C3alkyl wherein the biphenyl is optionally substituted with a methyl group; indolylC1-C3alkyl, wherein the indolyl part is optionally substituted with one group selected from C₁-C₃alkyl, carboxyC₁-C₃alkyl, halo, hydroxy, and phenyl, wherein the phenyl is further optionally substituted by one, two, or three groups independently selected from C₁-C₃alkoxy, C₁-C₃alkyl, and halo; NR^aR^b(C₁-C₇alkyl), wherein R^a and R^b are independently selected from hydrogen, C2-C4alkenyloxycarbonyl, C1-C3alkyl, C1-C3alkylcarbonyl, C3-C6cycloalkylcarbonyl, furanylcarbonyl, and phenylcarbonyl. When the alkyl linker contains more than one carbon an additional NR^aR^b group can be on the chain. NR^cR^dcarbonylC1-C3alkyl, wherein R^c and R^d are independently selected from hydrogen, C₁-C₃alkyl, and triphenylmethyl; phenylC₁-C₃alkyl wherein the phenyl part is optionally substituted with one, two, three, four, or five groups independently selected from C1-C4alkoxy, C1-C4alkyl, C1- C₃alkylsulfonylamino, amido, amino, aminoC₁-C₃alkyl, aminosulfonyl, carboxy, cyano, halo, haloC1-C3alkyl, hydroxy, -NC(NH2)2, nitro, and –OP(O)(OH)2; and phenoxyC1-C3alkyl wherein the phenyl is optionally substituted with a C1-C3alkyl group. The term “C₂-C₄alkenyl,” as used herein, refers to a straight or branched chain group of two to four carbon atoms containing at least one carbon-carbon double bond. The term “C2-C7alkenyl,” as used herein, refers to a straight or branched chain group of two to seven carbon atoms containing at least one carbon-carbon double bond. The term “C₂-C₄alkenyloxy,” as used herein, refers to a C₂-C₄alkenyl group attached to the parent molecular moiety through an oxygen atom. The term “C1-C3alkoxy,” as used herein, refers to a C1-C3alkyl group attached to the parent molecular moiety through an oxygen atom. The term “C1-C4alkoxy,” as used herein, refers to a C1-C4alkyl group attached to the parent molecular moiety through an oxygen atom. The term “C1-C6alkoxy,” as used herein, refers to a C1-C6alkyl group attached to the parent molecular moiety through an oxygen atom. The term “C1-C3alkoxyC1-C3alkyl,” as used herein, refers to a C1-C3alkoxy group attached to the parent molecular moiety through a C₁-C₃alkyl group. The term “C₁-C₆alkoxycarbonyl,” as used herein, refers to a C₁-C₆alkoxy group attached to the parent molecular moiety through a carbonyl group. The term “C1-C6alkoxycarbonylC1-C3alkyl,” as used herein, refers to a C1- C₆alkoxycarbonyl group attached to the parent molecular moiety through a C₁-C₃alkyl group. The term “C1-C3alkyl,” as used herein, refers to a group derived from a straight or branched chain saturated hydrocarbon containing from one to three carbon atoms. The term “C₁-C₄alkyl,” as used herein, refers to a group derived from a straight or branched chain saturated hydrocarbon containing from one to four carbon atoms. The term “C1-C6alkyl,” as used herein, refers to a group derived from a straight or branched chain saturated hydrocarbon containing from one to six carbon atoms. The term “C₁-C₃alkylcarbonyl,” as used herein, refers to a C1-C3alkyl group attached to the parent molecular moiety through a carbonyl group. The term “C1-C3alkylsulfanyl,” as used herein, refers to a C1-C3alkyl group attached to the parent molecular moiety through a sulfur atom. The term “C₁-C₃alkylsulfanylC₁-C₃alkyl,” as used herein, refers to a C₁- C3alkylsulfanyl group attached to the parent molecular moiety through a C1-C3alkyl group. The term “C₁-C₃alkylsulfonyl,” as used herein, refers to a C₁-C₃alkyl group attached to the parent molecular moiety through a sulfonyl group. The term “C1-C3alkylsulfonylamino,” as used herein, refers to a C1- C₃alkylsulfonyl group attached to the parent molecular moiety through an amino group. The term “amido,” as used herein, refers to –C(O)NH₂. The term “amidoC₁-C₃alkyl,” as used herein, refers to an amido group attached to the parent molecular moiety through a C1-C3alkyl group. The term “amino,” as used herein, refers to –NH2. The term “aminoC1-C3alkyl,” as used herein, refers to an amino group attached to the parent molecular moiety through a C₁-C₃alkyl group. The term “aminosulfonyl,” as used herein, refers to an amino group attached to the parent molecular moiety through a sulfonyl group. The term “azaindolylC1-C3alkyl,” as used herein, refers to an azaindolyl group attached to the parent molecular through a C₁-C₃alkyl group. The azaindolyl group can be attached to the alkyl moiety through any substitutable atom in the group. The term “benzothiazolylC1-C3alkyl,” as used herein, refers to an benzothiazolyl group attached to the parent molecular through a C₁-C₃alkyl group. The benzothiazolyl group can be attached to the alkyl moiety through any substitutable atom in the group. The term “benzothienylC1-C3alkyl,” as used herein, refers to a benzothienyl group attached to the parent molecular through a C₁-C₃alkyl group. The benzothienyl group can be attached to the alkyl moiety through any substitutable atom in the group. The term “benzyloxy,” as used herein, refers to a benzyl group attached to the parent molecular moiety through an oxygen atom. The term “benzyloxyC₁-C₃alkyl,” as used herein, refers to a benzyloxy group attached to the parent molecular moiety through a C1-C3alkyl group. The term “biphenylC1-C3alkyl,” as used herein, refers to a biphenyl group attached to the parent molecular moiety through a C₁-C₃alkyl group. The biphenyl group can be attached to the alkyl moiety through any substitutable atom in the group. The term “carbonyl,” as used herein, refers to –C(O)-. The term “carboxy,” as used herein, refers to –CO2H. The term “carboxyC1-C3alkyl,” as used herein, refers to a carboxy group attached to the parent molecular moiety through a C₁-C₃alkyl group. The term “cyano,” as used herein, refers to –CN. The term “C3-C6cycloalkyl,” as used herein, refers to a saturated monocyclic, hydrocarbon ring system having three to six carbon atoms and zero heteroatoms. The term “C3-C6cycloalkylC1-C3alkyl,” as used herein, refers to a C3-C6cycloalkyl group attached to the parent molecular moiety through a C₁-C₃alkyl group. The term “C3-C6cycloalkylcarbonyl,” as used herein, refers to a C3-C6 cycloalkyl group attached to the parent molecular moiety through a carbonyl group. The term “furanylC1-C3alkyl,” as used herein, refers to a furanyl group attached to the parent molecular moiety through a C₁-C₃alkyl group. The furanyl group can be attached to the alkyl moiety through any substitutable atom in the group. The term “furanylcarbonyl,” as used herein, refers to a furanyl group attached to the parent molecular moiety through a carbonyl group. The terms “halo” and “halogen,” as used herein, refer to F, Cl, Br, or I. The term “haloC1-C3alkyl,” as used herein, refers to a C1-C3alkyl group substituted with one, two, or three halogen atoms. The term “halomethyl,” as used herein, refers to a methyl group substituted with one, two, or three halogen atoms. The term “hydroxy,” as used herein, refers to –OH. The term “imidazolylC1-C3alkyl,” as used herein, refers to an imidazolyl group attached to the parent molecular moiety through a C1-C3alkyl group. The imidazolyl group can be attached to the alkyl moiety through any substitutable atom in the group. The term “indolylC1-C3alkyl,” as used herein, refers to an indolyl group attached to the parent molecular moiety through a C₁-C₃alkyl group. The indolyl group can be attached to the alkyl moiety through any substitutable atom in the group. The term “naphthylC1-C3alkyl,” as used herein, refers to a naphthyl group attached to the parent molecular moiety through a C₁-C₃alkyl group. The naphthyl group can be attached to the alkyl moiety through any substitutable atom in the group. The term “nitro,” as used herein, refers to –NO2. The term “NR^aR^b,” as used herein, refers to two groups, R^a and R^b, which are attached to the parent molecular moiety through a nitrogen atom. R^a and R^b are independently selected from hydrogen, C2-C4alkenyloxycarbonyl, C1-C3alkylcarbonyl, C₃-C₆cycloalkylcarbonyl, furanylcarbonyl, and phenylcarbonyl. The term “NR^aR^b(C₁-C₃)alkyl,” as used herein, refers to an NR^aR^b group attached to the parent molecular moiety through a C1-C3alkyl group. The term “NR^cR^d,” as used herein, refers to two groups, R^c and R^d, which are attached to the parent molecular moiety through a nitrogen atom. R^c and R^d are independently selected from hydrogen, C1-C3alkyl, and triphenylmethyl. The term “NR^cR^dcarbonyl,” as used herein, refers to an NR^cR^d group attached to the parent molecular moiety through a carbonyl group. The term “NR^cR^dcarbonylC₁-C₃alkyl,” as used herein, refers to an NR^cR^dcarbonyl group attached to the parent molecular moiety through a C1-C3alkyl group. The term “phenoxy,” as used herein, refers to a phenyl group attached to the parent molecular moiety through an oxygen atom. The term “phenoxyC₁-C₃alkyl,” as used herein, refers to a phenoxy group attached to the parent molecular moiety through a C1-C3alkyl group. The term “phenylC1-C3alkyl,” as used herein, refers to a phenyl group attached to the parent molecular moiety through a C₁-C₃alkyl group. The term “phenylcarbonyl,” as used herein, refers to a phenyl group attached to the parent molecular moiety through a carbonyl group. The term “pyridinylC1-C3alkyl,” as used herein, refers to a pyridinyl group attached to the parent molecular moiety through a C₁-C₃alkyl group. The pyridinyl group can be attached to the alkyl moiety through any substitutable atom in the group. The term “sulfanyl,” as used herein, refers to –S-. The term “sulfonyl,” as used herein, refers to –SO2-. The term “thiazolylC1-C3alkyl,” as used herein, refers to a thiazolyl group attached to the parent molecular moiety through a C₁-C₃alkyl group. The thiazolyl group can be attached to the alkyl moiety through any substitutable atom in the group. The term “thienylC1-C3alkyl,” as used herein, refers to a thienyl group attached to the parent molecular moiety through a C₁-C₃alkyl group. The thienyl group can be attached to the alkyl moiety through any substitutable atom in the group. The term "treating" refers to: (i) preventing a disease, disorder, or condition from occurring in a patient that may be predisposed to the disease, disorder, and/or condition but has not yet been diagnosed as having it; (ii) inhibiting the disease, disorder, or condition, i.e., arresting its development; and (iii) relieving the disease, disorder, or condition, i.e., causing regression of the disease, disorder, and/or condition and/or symptoms associated with the disease, disorder, and/or condition. Unless otherwise indicated, the term "alkyl" as employed herein alone or as part of another group includes, without limitation, both straight and branched chain hydrocarbons, containing 1 to 40 carbons, preferably 1 to 20 carbons, more preferably 1 to 8 carbons, in the normal chain, such as methyl, ethyl, propyl, isopropyl, butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, the various branched chain isomers thereof, and the like. Further, alkyl groups, as defined herein, may optionally be substituted on any available carbon atom with one or more functional groups commonly attached to such chains, such as, but not limited to alkyl, aryl, alkenyl, alkynyl, hydroxy, arylalkyl, cycloalkyl, cycloalkylalkyl, alkoxy, arylalkyloxy, heteroaryloxy, heteroarylalkyloxy, alkanoyl, halo, O _{hydroxyl, thio, nitro, cyano, carboxyl, carbonyl ( ), carboxamido, amino, alkylamino,} dialkylamino, amido, alkylamino, arylamido, heteroarylamido, azido, guanidino, amidino, phosphonic, phosphinic, sulfonic, sulfonamido, haloaryl, CF3, OCF2, OCF3, aryloxy, heteroaryl, cycloalkylalkoxyalkyl, cycloheteroalkyl and the like to form alkyl groups such as trifluoro methyl, 3-hydroxyhexyl, 2-carboxypropyl, 2-fluoroethyl, carboxymethyl, cyanobutyl and the like. Unless otherwise indicated, the term "cycloalkyl" as employed herein alone or as part of another group includes, without limitation, saturated or partially unsaturated (containing 1 or 2 double bonds) cyclic hydrocarbon groups containing 1 to 3 rings, appended or fused, including monocyclic alkyl, bicyclic alkyl and tricyclic alkyl, containing a total of 3 to 20 carbons forming the rings, preferably 4 to 7 carbons, forming each ring; which may be fused to 1 aromatic ring as described for aryl, which include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, cyclododecyl, cyclohexenyl, an n atoms with 1 or more groups selected from hydrogen, halo, haloalkyl, alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, trifluoromethyl, trifluoromethoxy, alkynyl, cycloalkylalkyl, fluorenyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, heteroaryl, arylalkyl, aryloxy, aryloxyalkyl, arylalkoxy, arylthio, arylazo, heteroarylalkyl, heteroarylalkenyl, O _{heteroarylheteroaryl, heteroaryloxy, hydroxy, nitro, oxo, cyano, carboxyl, carbonyl ( ),} carboxamido, amino, substituted amino wherein the amino includes 1 or 2 substituents (which are alkyl, aryl or any of the other aryl compounds mentioned in the definitions), amido, azido, guanidino, amidino, phosphonic, phosphinic, sulfonic, sulfonamido, thiol, alkylthio, arylthio, heteroarylthio, arylthioalkyl, alkoxyarylthio, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, arylaminocarbonyl, alkoxycarbonyl, aminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylamino, arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl, arylsulfonylamino or arylsulfonaminocarbonyl, or any of alkyl substituents as set out above. The term "aryl" as employed herein alone or as part of another group refers, without limitation, to monocyclic and bicyclic aromatic groups containing 6 to 10 carbons in the ring portion (such as phenyl or naphthyl) and may optionally include one to three additional rings fused to "aryl" (such as aryl, cycloalkyl, heteroaryl or heterocycloalkyl rings) and may be optionally substituted through any available carbon atoms with 1 or more groups selected from hydrogen, alkyl, halo, haloalkyl, alkoxy, haloalkoxy, alkenyl, trifluoromethyl, trifluoromethoxy, alkynyl, cycloalkylalkyl, fluorenyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, heteroaryl, arylalkyl, aryloxy, aryloxyalkyl, arylalkoxy, arylthio, arylazo, heteroarylalkyl, heteroarylalkenyl, heteroaryloxy, heteroarylalkyloxy, heteroarylalkyloxyalkyl, hydroxy, nitro, oxo, cyano, amino, substituted amino wherein the amino includes 1 or 2 substituents (which are alkyl, cycloalkyl, heterocycloalkyl, heteroaryl, or aryl or any of the other aryl compounds mentioned in the definitions), thiol, alkylthio, arylthio, heteroarylthio, arylthioalkyl, alkoxyarylthio, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, cycloalkylaminocarbonyl, arylaminocarbonyl, heteroarylaminocarbonyl, heteroarylalkylaminocarbonyl alkoxycarbonyl, aminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylamino, arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl, arylsulfonylamino or arylsulfonaminocarbonyl, or any of alkyl substituents as set out above. The term "arylalkyl" as used herein alone or as part of another group refers, without limitation, to alkyl groups as defined above having an aryl substituent, such as benzyl, phenethyl or naphthylpropyl, wherein said aryl and/or alkyl groups may optionally be substituted as defined above. The term "alkoxy", "aryloxy", "heteroaryloxy", "arylalkyloxy", or "heteroarylalkyloxy" as employed herein alone or as part of another group includes, without limitation, an alkyl or aryl group as defined above linked through an oxygen atom. The term "heterocyclo", "heterocycle", "heterocyclyl" or "heterocyclic", as used herein, represents, without limitation, an unsubstituted or substituted stable 4-, 5-, 6-, or 7-membered monocyclic ring system which may be saturated or unsaturated, and which consists of carbon atoms and from one to four heteroatoms selected from nitrogen, sulfur, oxygen and/or a SO or SO2 group, wherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. The heterocyclic ring may be attached at any heteroatom or carbon atom which results in the creation of a stable structure. Examples of such heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl, piperidinyl, piperazinyl, oxopyrrolidinyl, oxopiperazinyl, oxopiperidinyl and oxadiazolyl. Optionally a heterocyclo group may be substituted with one or more functional groups, such as those described for "alkyl" or "aryl". The term "heterocycloalkyl" as used herein alone or as part of another group refers, without limitation, to alkyl groups as defined above having a heterocycloalkyl substituent, wherein said "heterocyclo" and/or alkyl groups may optionally be substituted as defined above. The term "heteroaryl" as used herein refers, without limitation, to a 5-, 6- or 7- membered aromatic heterocyclic ring which contains one or more heteroatoms selected from nitrogen, sulfur, oxygen and/or a SO or SO2 group. Such rings may be fused to another aryl or heteroaryl ring and include possible N-oxides; examples of such heteroaryl groups include, but are not limited to, furan, pyrrole, thiophene, pyridine, pyrimidine, pyrazine, pyridazine, isoxazole, oxazole, imidazole and the like. Optionally a heteroaryl group may be substituted with one or more functional groups commonly attached to such chains, such as those described for "alkyl" or "aryl". The term "heteroarylalkyl" as used herein alone or as part of another group refers, without limitation, to alkyl groups as defined above having a heteroaryl substituent, wherein said heteroaryl and/or alkyl groups may optionally be substituted as defined above. The "inhibitory concentration" of LAG-3 inhibitor is intended to mean the concentration at which a compound screened in an assay of the disclosure inhibits a measurable percentage of the interaction of LAG-3 with MHC Class II molecules. Examples of "inhibitory concentration" values range from IC₅₀ to IC₉₀, and are preferably, IC50, IC60, IC70, IC80, or IC90, which represent 50%, 60%, 70%, 80% or 90% reduction in LAG-3/MHC Class II molecules binding activity, respectively. More preferably, the "inhibitory concentration" is measured as the IC50 value. It is understood that another designation for IC50 is the half-maximal inhibitory concentration. Binding of the macrocyclic peptides to LAG-3 can be measured, for example, by methods such as homogeneous time-resolved fluorescence (HTRF), Surface Plasmon Resonance (SPR), isothermal titration calorimetry (ITC), nuclear magnetic resonance spectroscopy (NMR), and the like. Further, binding of the macrocyclic peptides to LAG- 3 expressed on the surface of cells can be measured as described herein in cellular binding assays. Administration of a therapeutic agent described herein includes, without limitation, administration of a therapeutically effective amount of therapeutic agent. The term "therapeutically effective amount" as used herein refers, without limitation, to an amount of a therapeutic agent to treat or prevent a condition treatable by administration of a composition of the LAG-3/MHC Class II molecules binding inhibitors described herein. That amount is the amount sufficient to exhibit a detectable therapeutic or preventative or ameliorative effect. The effect may include, for example and without limitation, treatment or prevention of the conditions listed herein. The precise effective amount for a subject will depend upon the subject's size and health, the nature and extent of the condition being treated, recommendations of the treating physician, and therapeutics or combination of therapeutics selected for administration. Thus, it is not useful to specify an exact effective amount in advance. In another aspect, the disclosure pertains to methods of inhibiting growth of tumor cells in a subject using the macrocyclic peptides of the present disclosure. As demonstrated herein, the macrocyclic peptides of the present disclosure are capable of binding to LAG-3, disrupting the interaction between LAG-3 and MHC class II molecules. As a result, the macrocyclic peptides of the present disclosure are potentially useful for modifying an immune response, treating diseases such as cancer or infectious disease, stimulating a protective autoimmune response or to stimulate antigen-specific immune responses (e.g., by coadministration of lag03 blocking peptides with an antigen of interest). In order that the present disclosure may be more readily understood, certain terms are first defined. Additional definitions are set forth throughout the detailed description. The terms "Programmed Death Ligand 1", "Programmed Cell Death Ligand 1", "Protein PD-L1", "PD-L1", "PDL1", "PDCDL1", "hPD-L1", "hPD-LI", "CD274" and "B7-H1" are used interchangeably, and include variants, isoforms, species homologs of human PD-L1, and analogs having at least one common epitope with PD-L1. The complete PD-L1 sequence can be found under GENBANK® Accession No. NP_054862. The terms "Programmed Death 1", "Programmed Cell Death 1", "Protein PD-1", "PD-1", "PD1", "PDCD1", "hPD-1" and "hPD-I" are used interchangeably, and include variants, isoforms, species homologs of human PD-1, and analogs having at least one common epitope with PD-1. The complete PD-1 sequence can be found under GENBANK® Accession No. U64863. The terms "cytotoxic T lymphocyte-associated antigen-4", "CTLA-4", "CTLA4", "CTLA-4 antigen" and "CD152" (see, e.g., Murata, Am. J. Pathol., 155:453-460 (1999)) are used interchangeably, and include variants, isoforms, species homologs of human CTLA-4, and analogs having at least one common epitope with CTLA-4 (see, e.g., Balzano, Int. J. Cancer Suppl., 7:28-32 (1992)). The complete CTLA-4 nucleic acid sequence can be found under GENBANK® Accession No. L15006. The term "immune response" refers to the action of, for example, lymphocytes, antigen presenting cells, phagocytic cells, granulocytes, and soluble macromolecules produced by the above cells or the liver (including macrocyclic peptides, cytokines, and complement) that results in selective damage to, destruction of, or elimination from the human body of invading pathogens, cells or tissues infected with pathogens, cancerous cells, or, in cases of autoimmunity or pathological inflammation, normal human cells or tissues. A "signal transduction pathway" refers to the biochemical relationship between a variety of signal transduction molecules that play a role in the transmission of a signal from one portion of a cell to another portion of a cell. As used herein, the phrase "cell surface receptor" includes, for example, molecules and complexes of molecules capable of receiving a signal and the transmission of such a signal across the plasma membrane of a cell. An example of a "cell surface receptor" of the present disclosure is the PD-1 receptor. The term "macrocyclic peptide derivatives" refers to any modified form of the macrocyclic peptides disclosed herein, e.g., mutations, isoforms, peptides with altered linker backbones, conjugates with an antibody and/or another agent, etc.. As used herein, a (preferred?) macrocyclic peptide of the present disclosure that "specifically binds to human LAG-3" is intended to refer to a macrocyclic peptide that binds to human LAG-3 with an IC₅₀ of less than about 1000 nM, less than about 300 nM, less than about less than about 100 nM, less than about 80 nM, less than about 60 nM, less than about 40 nM, less than about 20 nM, less than about 15 nM, less than about 10 nM, less than about 5 nM, less than about 1 nM, or less. In this context, the term "about" shall be construed to mean anywhere between ± 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nM more or less than the cited amount. The term "treatment" or "therapy" refers to administering an active agent with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve, or affect a condition (e.g., a disease), the symptoms of the condition, or to prevent or delay the onset of the symptoms, complications, biochemical indicia of a disease, or otherwise arrest or inhibit further development of the disease, condition, or disorder in a statistically significant manner. As used herein, "about" or "comprising essentially of" mean within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, "about" or "comprising essentially of" can mean within one or more than one standard deviation per the practice in the art. Alternatively, "about" or "comprising essentially of" can mean a range of up to 20%. Furthermore, particularly with respect to biological systems or processes, the terms can mean up to an order of magnitude or up to 5-fold of a value. When particular values are provided in the application and claims, unless otherwise stated, the meaning of "about" or "comprising essentially of" should be assumed to be within an acceptable error range for that particular value. As described herein, any concentration range, percentage range, ratio range or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated. Variant Macrocyclic Peptides In yet another embodiment, a macrocyclic peptide of the disclosure comprises amino acid sequences that are homologous to the amino acid sequences of the macrocyclic peptides described herein, and wherein the macrocyclic peptides retain the desired functional and/or biological properties of the macrocyclic peptide of the disclosure. For example, the disclosure provides a macrocyclic peptide, or antigen-binding portion thereof, comprising: an amino acid sequence that is at least 80% homologous to an amino acid sequence selected from the compounds described herein; and the macrocyclic peptide exhibits one or more of the following properties: (a) the macrocyclic peptide binds to human LAG-3 with an IC₅₀ of 200 nM or less; (b) the macrocyclic peptide does not substantially bind to human CD4; (c) the macrocyclic peptide binds to human LAG-3 and one or more of the following: cynomolgus monkey LAG-3, and/or mouse LAG-3; (d) the macrocyclic peptide inhibits the binding of LAG-3 to MHC Class II moleucules; (e) the macrocyclic peptide inhibits tumor cell growth in a cellular assay and/or in vivo assay; In other embodiments, the macrocyclic peptide amino acid sequences may be about 80%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the sequences set forth above. In this context, the term "about" shall be construed to mean anywhere between 1, 2, 3, 4, or 5 percent more or less than the cited amount. A macrocyclic peptide of the present disclosure having sequences with high identity (i.e., 80% or greater) to the sequences set forth above, can be obtained by mutating the sequences during chemical synthesis, for example, followed by testing of the altered macrocyclic peptide for retained function (i.e., the functions set forth in (a) through (i) above) using the functional assays described herein. The biological and/or functional activity of the variant macrocyclic peptide amino acid sequences may be at least about 1x, 2x, 3x, 4x, 5x, 6x,7x, 8x, 9x, or 10x more than the reference macrocyclic peptide on which the variant is based. In this context, the term "about" shall be construed to mean anywhere between 0.1x, 0.2x, 0.3x, 0.4x, 0.5x, 0.6x, 0.7x, 0.8x, or 0.9x more or less than the cited amount. As used herein, the percent homology between two amino acid sequences is equivalent to the percent identity between the two sequences. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences (i.e., % homology = # of identical positions / total # of positions.times.100), taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm, as described in the non-limiting examples below. The percent identity between two amino acid sequences can be determined using the algorithm of Meyers E. et al., (Comput. Appl. Biosci., 4:11-17 (1988)) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. In addition, the percent identity between two amino acid sequences can be determined using the Needleman et al. (J. Mol. Biol., 48:444-453 (1970)) algorithm which has been incorporated into the GAP program in the GCG® software package (available at www.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. Macrocyclic Peptides with Conservative Modifications In yet another embodiment, a macrocyclic peptide of the disclosure comprises amino acid sequences that are homologous to the amino acid sequences of the macrocyclic peptides described herein, and wherein the macrocyclic peptides retain the desired functional and/or biological properties of the macrocyclic peptide of the disclosure. For example, the disclosure provides a macrocyclic peptide, or antigen-binding portion thereof, comprising: an amino acid sequence that is at least 80% homologous to an amino acid sequence selected from the macrocyclic peptides described herein, wherein one or more amino acids have been substituted with a conservative amino acid; and the macrocyclic peptide exhibits one or more of the following properties: (a) the macrocyclic peptide binds to human LAG-3 with an IC50 of 200 nM or less; (b) the macrocyclic peptide does not substantially bind to human CD4; (c) the macrocyclic peptide binds to human LAG-3 and one or more of the following: cynomolgus monkey LAG-3, and/or mouse LAG-3; (d) the macrocyclic peptide inhibits the binding of LAG-3 to MHC Class II moleucules; (e) the macrocyclic peptide inhibits tumor cell growth in a cellular assay and/or in vivo assay; As used herein, the term "conservative sequence modifications" is intended to refer to amino acid modifications that do not significantly affect or alter the binding characteristics of the macrocyclic peptide containing the amino acid sequence. Such conservative modifications include amino acid substitutions, additions and deletions. Modifications can be introduced into an antibody of the disclosure by standard techniques known in the art, such as substitution of peptide amidites during chemical synthesis, site- directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions are ones in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, one or more amino acid residues within the antigen binding regions of macrocyclic peptides of the disclosure can be replaced with other amino acid residues from the same side chain family and the altered antibody can be tested for retained function (i.e., the functions set forth in (a) thru (i) above) using the functional assays described herein. Conservative amino acid substitutions may also be selected from one or more non-naturally occurring amino acids disclosed herein. Pharmaceutical Compositions In another aspect, the present disclosure provides a composition, e.g., a pharmaceutical composition, containing one or a combination of macrocyclic peptides, or antigen-binding portion(s) thereof, of the present disclosure, formulated together with a pharmaceutically acceptable carrier. Such compositions may include one or a combination of (e.g., two or more different) macrocyclic peptides, or immunoconjugates or bispecific molecules of the disclosure. For example, a pharmaceutical composition of the disclosure can comprise a combination of macrocyclic peptides (or immunoconjugates or bispecifics) that bind to different epitopes on the target antigen or that have complementary activities. Pharmaceutical compositions of the disclosure also can be administered in combination therapy, i.e., combined with other agents. For example, the combination therapy can include a macrocyclic peptide combined with at least one other anti- inflammatory or immunosuppressant agent. Examples of therapeutic agents that can be used in combination therapy are described in greater detail below in the section on uses of the macrocyclic peptides of the disclosure. As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. Preferably, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion). Depending on the route of administration, the active compound, i.e., a macrocyclic peptide, immunoconjugate, or bispecific molecule, may be coated in a material to protect the compound from the action of acids and other natural conditions that may inactivate the compound. The pharmaceutical compounds of the disclosure may include one or more pharmaceutically acceptable salts. A "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the parent compound and does not impart any undesired toxicological effects (see e.g., Berge, S.M. et al., J. Pharm. Sci., 66:1-19 (1977)). Examples of such salts include acid addition salts and base addition salts. Acid addition salts include those derived from nontoxic inorganic acids, such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, phosphorous and the like, as well as from nontoxic organic acids such as aliphatic mono- and dicarboxylic acids, phenyl- substituted alkanoic acids, hydroxy alkanoic acids, aromatic acids, aliphatic and aromatic sulfonic acids and the like. Base addition salts include those derived from alkaline earth metals, such as sodium, potassium, magnesium, calcium and the like, as well as from nontoxic organic amines, such as N,N′-dibenzylethylenediamine, N-methylglucamine, chloroprocaine, choline, diethanolamine, ethylenediamine, procaine and the like. A pharmaceutical composition of the disclosure also may include a pharmaceutically acceptable antioxidant. Examples of pharmaceutically acceptable antioxidants include: (1) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil- soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like. Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions of the disclosure include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of presence of microorganisms may be ensured both by sterilization procedures, supra, and by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin. Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. The use of such media and agents for pharmaceutically active substances is known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the pharmaceutical compositions of the disclosure is contemplated. Supplementary active compounds can also be incorporated into the compositions. Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. 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. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, monostearate salts and gelatin. Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by sterilization microfiltration. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a 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 (lyophilization) that yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the subject being treated, and the particular mode of administration. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will generally be that amount of the composition which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 0.01 percent to about ninety-nine percent of active ingredient, preferably from about 0.1 percent to about 70 percent, most preferably from about 1 percent to about 30 percent of active ingredient in combination with a pharmaceutically acceptable carrier. Dosage regimens are adjusted to provide the optimum desired response (e.g., a therapeutic response). For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of therapeutic situation. It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the disclosure are dictated by and directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals. For administration of the macrocyclic peptide, the dosage ranges from about 0.0001 to 100 mg/kg, and more usually 0.01 to 5 mg/kg, of the host body weight. For example dosages can be 0.3 mg/kg body weight, 1 mg/kg body weight, 3 mg/kg body weight, 5 mg/kg body weight or 10 mg/kg body weight or within the range of 1-10 mg/kg. An exemplary treatment regime entails administration once per day, bi-weekly, tri-weekly, weekly, once every two weeks, once every three weeks, once every four weeks, once a month, once every 3 months or once every three to 6 months. Preferred dosage regimens for a macrocyclic peptide of the disclosure include 1 mg/kg body weight or 3 mg/kg body weight via intravenous administration, with the antibody being given using one of the following dosing schedules: (i) every four weeks for six dosages, then every three months; (ii) every three weeks; (iii) 3 mg/kg body weight once followed by 1 mg/kg body weight every three weeks. In some methods, two or more macrocyclic peptides with different binding specificities are administered simultaneously, in which case the dosage of each compound administered falls within the ranges indicated. The compounds are usually administered on multiple occasions. Intervals between single dosages can be, for example, weekly, monthly, every three months or yearly. Intervals can also be irregular as indicated by measuring blood levels of macrocyclic peptide to the target antigen in the patient. In some methods, dosage is adjusted to achieve a plasma antibody concentration of about 1- 1000.mu.g/ml and in some methods about 25-300.mu.g/ml. Alternatively, the macrocyclic peptide can be administered as a sustained release formulation, in which case less frequent administration is required. Dosage and frequency vary depending on the half-life of the macrocyclic peptide in the patient. The dosage and frequency of administration can vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, a relatively low dosage is administered at relatively infrequent intervals over a long period of time. Some patients continue to receive treatment for the rest of their lives. In therapeutic applications, a relatively high dosage at relatively short intervals is sometimes required until progression of the disease is reduced or terminated, and preferably until the patient shows partial or complete amelioration of symptoms of disease. Thereafter, the patient can be administered a prophylactic regime. Actual dosage levels of the active ingredients in the pharmaceutical compositions of the present disclosure may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. The selected dosage level will depend upon a variety of pharmacokinetic factors including the activity of the particular compositions of the present disclosure employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compositions employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts. A "therapeutically effective dosage" of a macrocyclic peptide of the disclosure preferably results in a decrease in severity of disease symptoms, an increase in frequency and duration of disease symptom-free periods, or a prevention of impairment or disability due to the disease affliction. For example, for the treatment of tumors, a "therapeutically effective dosage" preferably inhibits cell growth or tumor growth by at least about 20%, more preferably by at least about 40%, even more preferably by at least about 60%, and still more preferably by at least about 80% relative to untreated subjects. The ability of a compound to inhibit tumor growth and/or HIV can be evaluated in an animal model system predictive of efficacy in human tumors or viral efficacy. Alternatively, this property of a composition can be evaluated by examining the ability of the compound to inhibit, such inhibition in vitro by assays known to the skilled practitioner. A therapeutically effective amount of a therapeutic compound can decrease tumor size, decrease viral load, or otherwise ameliorate symptoms in a subject. One of ordinary skill in the art would be able to determine such amounts based on such factors as the subject's size, the severity of the subject's symptoms, and the particular composition or route of administration selected. A composition of the present disclosure can be administered via one or more routes of administration using one or more of a variety of methods known in the art. As will be appreciated by the skilled artisan, the route and/or mode of administration will vary depending upon the desired results. Preferred routes of administration for macrocyclic peptides of the disclosure include intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, spinal or other parenteral routes of administration, for example by injection or infusion. The phrase "parenteral administration" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion. Alternatively, a macrocyclic peptide of the disclosure can be administered via a non-parenteral route, such as a topical, epidermal or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually or topically. The active compounds can be prepared with carriers that will protect the compound against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for the preparation of such formulations are patented or generally known to those skilled in the art. See, e.g., Robinson, J.R., ed., Sustained and Controlled Release Drug Delivery Systems, Marcel Dekker, Inc., New York (1978). Therapeutic compositions can be administered with medical devices known in the art. For example, in a preferred embodiment, a therapeutic composition of the disclosure can be administered with a needleless hypodermic injection device, such as the devices disclosed in U.S. Patent Nos. 5,399,163, 5,383,851, 5,312,335, 5,064,413, 4,941,880, 4,790,824, or 4,596,556. Examples of well-known implants and modules useful in the present disclosure include: U.S. Patent No. 4,487,603, which discloses an implantable micro-infusion pump for dispensing medication at a controlled rate; U.S. Patent No. 4,486,194, which discloses a therapeutic device for administering medication through the skin; U.S. Patent No. 4,447,233, which discloses a medication infusion pump for delivering medication at a precise infusion rate; U.S. Patent No. 4,447,224, which discloses a variable flow implantable infusion apparatus for continuous drug delivery; U.S. Patent No. 4,439,196, which discloses an osmotic drug delivery system having multi-chamber compartments; and U.S. Patent No. 4,475,196, which discloses an osmotic drug delivery system. These patents are incorporated herein by reference. Many other such implants, delivery systems, and modules are known to those skilled in the art. In certain embodiments, the macrocyclic peptides of the disclosure can be formulated to ensure proper distribution in vivo. For example, the blood-brain barrier (BBB) excludes many highly hydrophilic compounds. To ensure that therapeutic compounds of the disclosure cross the BBB (if desired), they can be formulated, for example, in liposomes. For methods of manufacturing liposomes, see, e.g., U.S. Patent Nos. 4,522,811, 5,374,548, and 5,399,331. The liposomes may comprise one or more moieties which are selectively transported into specific cells or organs, thus enhance targeted drug delivery (see, e.g., Ranade, V.V., J. Clin. Pharmacol., 29:685 (1989)). Exemplary targeting moieties include folate or biotin (see, e.g., U.S. Patent No. 5,416,016 to Low et al.); mannosides (Umezawa et al., Biochem. Biophys. Res. Commun., 153:1038 (1988)); macrocyclic peptides (Bloeman, P.G. et al., FEBS Lett., 357:140 (1995); Owais, M. et al., Antimicrob. Agents Chemother., 39:180 (1995)); surfactant protein A receptor (Briscoe et al., Am. J. Physiol., 1233:134 (1995)); p120 (Schreier et al., J. Biol. Chem., 269:9090 (1994)); see also Keinanen, K. et al., FEBS Lett., 346:123 (1994); Killion, J.J. et al., Immunomethods 4:273 (1994). Peptide Synthesis The macrocyclic peptides of the present disclosure can be produced by methods known in the art, such as they can be synthesized chemically, recombinantly in a cell free system, recombinantly within a cell or can be isolated from a biological source. Chemical synthesis of a macrocyclic peptide of the present disclosure can be carried out using a variety of art recognized methods, including stepwise solid phase synthesis, semi- synthesis through the conformationally-assisted re-ligation of peptide fragments, enzymatic ligation of cloned or synthetic peptide segments, and chemical ligation. A preferred method to synthesize the macrocyclic peptides and analogs thereof described herein is chemical synthesis using various solid-phase techniques such as those described in Chan, W.C. et al, eds., Fmoc Solid Phase Synthesis, Oxford University Press, Oxford (2000); Barany, G. et al, The Peptides: Analysis, Synthesis, Biology, Vol.2 : "Special Methods in Peptide Synthesis, Part A", pp.3-284, Gross, E. et al, eds., Academic Press, New York (1980); in Atherton, E., Sheppard, R. C. Solid Phase Peptide Synthesis: A Practical Approach, IRL Press, Oxford, England (1989); and in Stewart, J. M. Young, J. D. Solid-Phase Peptide Synthesis, 2nd Edition, Pierce Chemical Co., Rockford, IL (1984). The preferred strategy is based on the (9-fluorenylmethyloxycarbonyl) group (Fmoc) for temporary protection of the ^-amino group, in combination with the tert-butyl group (tBu) for temporary protection of the amino acid side chains (see for example Atherton, E. et al, "The Fluorenylmethoxycarbonyl Amino Protecting Group", in The Peptides: Analysis, Synthesis, Biology, Vol.9 : "Special Methods in Peptide Synthesis, Part C", pp.1-38, Undenfriend, S. et al, eds., Academic Press, San Diego (1987). The peptides can be synthesized in a stepwise manner on an insoluble polymer support (also referred to as "resin") starting from the C-terminus of the peptide. A synthesis is begun by appending the C-terminal amino acid of the peptide to the resin through formation of an amide or ester linkage. This allows the eventual release of the resulting peptide as a C-terminal amide or carboxylic acid, respectively. The C-terminal amino acid and all other amino acids used in the synthesis are required to have their ^-amino groups and side chain functionalities (if present) differentially protected such that the ^-amino protecting group may be selectively removed during the synthesis. The coupling of an amino acid is performed by activation of its carboxyl group as an active ester and reaction thereof with the unblocked ^-amino group of the N-terminal amino acid appended to the resin. The sequence of ^-amino group deprotection and coupling is repeated until the entire peptide sequence is assembled. The peptide is then released from the resin with concomitant deprotection of the side chain functionalities, usually in the presence of appropriate scavengers to limit side reactions. The resulting peptide is finally purified by reverse phase HPLC. The synthesis of the peptidyl-resins required as precursors to the final peptides utilizes commercially available cross-linked polystyrene polymer resins (Novabiochem, San Diego, CA; Applied Biosystems, Foster City, CA). Preferred solid supports are: 4- (2',4'-dimethoxyphenyl-Fmoc-aminomethyl)-phenoxyacetyl-p-methyl benzhydrylamine resin (Rink amide MBHA resin); 9-Fmoc-amino-xanthen-3-yloxy-Merrifield resin (Sieber amide resin); 4-(9-Fmoc)aminomethyl-3,5- dimethoxyphenoxy)valerylaminomethyl-Merrifield resin (PAL resin), for C-terminal carboxamides. Coupling of first and subsequent amino acids can be accomplished using HOBt, 6-Cl-HOBt or HOAt active esters produced from DIC/HOBt, HBTU/HOBt, BOP, PyBOP, or from DIC/6-C1-HOBt, HCTU, DIC/HOAt or HATU, respectively. Preferred solid supports are: 2-chlorotrityl chloride resin and 9-Fmoc-amino-xanthen-3-yloxy- Merrifield resin (Sieber amide resin) for protected peptide fragments. Loading of the first amino acid onto the 2-chlorotrityl chloride resin is best achieved by reacting the Fmoc- protected amino acid with the resin in dichloromethane and DIEA. If necessary, a small amount of DMF may be added to solubilize the amino acid. The syntheses of the peptide analogs described herein can be carried out by using a single or multi-channel peptide synthesizer, such as an CEM Liberty Microwave synthesizer, or a Protein Technologies, Inc. Prelude (6 channels) or Symphony (12 channels) or Symphony X (24 channels) synthesizer. Useful Fmoc amino acids derivatives are shown below. Examples of Orthogonally Protected Amino Acids used in Solid Phase Synthesis

The peptidyl-resin precursors for their respective peptides may be cleaved and deprotected using any standard procedure (see, for example, King, D.S. et al, Int. J.

Peptide Protein Res.. 36:255-266 (1990)). A desired method is the use of TFA in the presence of water, TIS as scavenger, and DTT or TCEP as the disulfide reducing agent.

Typically, the peptidyl-resin is stirred in TFA/TIS/DTT (96:3:1), v:v:w; 1 mL/100 mg of peptidyl resin) for 1-3 hrs at room temperature. The spent resin is then filtered off, the

TFA solution was cooled, and Et20 solution was added. The precipitates were collected by centrifuging and decanting the ether layer (3 x). The resulting crude peptide is either redissolved directly into DMF or DMSO or CH3CN/H2O for purification by preparative

HPLC or used directly in the next step.

Peptides with the desired purity can be obtained by purification using preparative

HPLC, for example, on a Waters Model 4000 or a Shimadzu Model LC-8A liquid chromatography. The solution of crude peptide is injected into an YMC S5 ODS (20 x 100 mm) column and eluted with a linear gradient of MeCN in water, both buffered with 0.1% TFA, using a flow rate of 14-20 mL/min with effluent monitoring by UV absorbance at 217 or 220 nm. The structures of the purified peptides can be confirmed by electro-spray MS analysis. Analytical Data: Mass Spectrometry: “ESI-MS(+)” signifies electrospray ionization mass spectrometry performed in positive ion mode; “ESI-MS(-)” signifies electrospray ionization mass spectrometry performed in negative ion mode; “ESI-HRMS(+)” signifies high-resolution electrospray ionization mass spectrometry performed in positive ion mode; “ESI-HRMS(-)” signifies high-resolution electrospray ionization mass spectrometry performed in negative ion mode. The detected masses are reported following the “m/z” unit designation. Compounds with exact masses greater than 1000 were often detected as double-charged or triple-charged ions. The crude material was purified via preparative LC/MS. Fractions containing the desired product were combined and dried via centrifugal evaporation. Analytical LC/MS Condition A: Column: Waters Acquity UPLC BEH C18, 2.1 x 50 mm, 1.7-μm particles; Mobile Phase A: 5:95 acetonitrile:water with 10 mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile:water with 10 mM ammonium acetate; Temperature: 50 °C; Gradient: 0-100% B over 3 minutes, then a 0.75-minute hold at 100% B; Flow: 1.0 mL/min; Detection: UV at 220 nm. Analytical LC/MS Condition B: Column: Waters Acquity UPLC BEH C18, 2.1 x 50 mm, 1.7-μm particles; Mobile Phase A: 5:95 acetonitrile:water with 0.1% trifluoroacetic acid; Mobile Phase B: 95:5 acetonitrile:water with 0.1% trifluoroacetic acid; Temperature: 50 °C; Gradient: 0-100% B over 3 minutes, then a 0.75-minute hold at 100% B; Flow: 1.0 mL/min; Detection: UV at 220 nm. Analytical LC/MS Condition C: Column: Waters Acquity UPLC BEH C18, 2.1 x 50 mm, 1.7-μm particles; Mobile Phase A: 5:95 acetonitrile:water with 10 mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile:water with 10 mM ammonium acetate; Temperature: 70 °C; Gradient: 0-100% B over 3 minutes, then a 2.0-minute hold at 100% B; Flow: 0.75 mL/min; Detection: UV at 220 nm. Analytical LC/MS Condition D: Column: Waters Acquity UPLC BEH C18, 2.1 x 50 mm, 1.7-μm particles; Mobile Phase A: 5:95 acetonitrile:water with 0.1% trifluoroacetic acid; Mobile Phase B: 95:5 acetonitrile:water with 0.1% trifluoroacetic acid; Temperature: 70 °C; Gradient: 0-100% B over 3 minutes, then a 2.0-minute hold at 100% B; Flow: 0.75 mL/min; Detection: UV at 220 nm. Analytical LC/MS Condition E: Column: Kinetex XB C18, 3.0 x 75 mm, 2.6-μm particles; Mobile Phase A: 10 mM ammonium formate in water:acetonitrile (98:2); Mobile Phase B: 10 mM ammonium formate in Water:acetonitrile (02:98); Gradient: 20- 100% B over 4 minutes, then a 0.6-minute hold at 100% B; Flow: 1.0 mL/min; Detection: UV at 254 nm. Analytical LC/MS Condition F: Column: Ascentis Express C18, 2.1 x 50 mm, 2.7-μm particles; Mobile Phase A: 10 mM ammonium acetate in water:acetonitrile (95:5); Mobile Phase B: 10 mM ammonium acetate in Water:acetonitrile (05:95), Temperature: 50 ^oC; Gradient: 0-100% B over 3 minutes; Flow: 1.0 mL/min; Detection: UV at 220 nm. Analytical LC/MS Condition G: Column: X Bridge C18, 4.6 x 50 mm, 5-μm particles; Mobile Phase A: 0.1% TFA in water; Mobile Phase B: acetonitrile, Temperature: 35 ^oC; Gradient: 5-95% B over 4 minutes; Flow: 4.0 mL/min; Detection: UV at 220 nm. The following abbreviations are employed in the Examples and elsewhere herein: Ph = phenyl Bn = benzyl i-Bu = iso-butyl i-Pr = iso-propyl Me = methyl Et = ethyl Pr = n-propyl Bu = n-butyl t-Bu = tert-butyl Trt = trityl TMS = trimethylsilyl TIS =triisopropylsilane Et2O = diethyl ether HOAc or AcOH = acetic acid MeCN or AcCN = acetonitrile DMF = N,N-dimethylformamide EtOAc = ethyl acetate THF = tetrahydrofuran TFA = trifluoroacetic acid TFE = α,α,α-trifluoroethanol Et₂NH = diethylamine NMM = 4-methylmorpholine NMP = N-methylpyrrolidone DCM = dichloromethane TEA = triethylamine min. = minute(s) h or hr = hour(s) L = liter mL or ml = milliliter ^L = microliter g = gram(s) mg = milligram(s) mol = mole(s) mmol = millimole(s) meq = milliequivalent rt or RT = room temperature sat or sat'd = saturated aq. = aqueous mp = melting point BOP reagent = benzotriazol-1-yloxy-tris-dimethylamino-phosphonium hexafluorophosphate (Castro's reagent) PyBOP reagent = benzotriazol-1-yloxy-tripyrrolidino phosphonium hexafluorophosphate HBTU = 2-(1H-Benzotriazol-1-yl)-1,1,3,3-tetramethyluronim hexafluorophosphate HATU = O-(7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronim hexafluorophosphate HCTU = 2-(6-Chloro-1-H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate T3P = 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphorinane-2,4,6-trioxide DMAP = 4-(dimethylamino)pyridine DIEA = diisopropylethylamine Fmoc or FMOC = fluorenylmethyloxycarbonyl Boc or BOC = tert-butyloxycarbonyl HOBT or HOBT^H₂O = 1-hydroxybenzotriazole hydrate Cl-HOBt = 6-Chloro-benzotriazole HOAT = 1-hydroxy-7-azabenzotriazole HPLC = high performance liquid chromatography LC/MS = high performance liquid chromatography/mass spectrometry MS or Mass Spec = mass spectrometry NMR = nuclear magnetic resonance Sc or SC or SQ = sub-cutaneous IP or ip = intra-peritoneal General Procedures: All manipulations were performed under automation on a Prelude or a Symphony, or Symphony X peptide synthesizer (Protein Technologies). All procedures were performed according to the published methods (e.g., WO 2023/225661). Prelude: Resin-swelling procedure, Single-coupling procedure, Single-coupling extended time procedure, Chloroacetic Anhydride coupling, Single-Coupling Manual Addition Procedure A, Single-Coupling Manual Addition Procedure B, Manual removal of Fmoc group procedure: Symphony: Resin-swelling procedure, Single-coupling procedure, Single-coupling extended time procedure, Double-coupling extended time procedure, Chloroacetic Anhydride coupling: Symphony X: Resin-swelling procedure, Single-coupling procedure, Single-coupling 3 deprotections procedure, Single-coupling extended time procedure, Single-coupling 3 deprotections extended time procedure, Pre-activated single-coupling procedure, Single- Coupling Manual Addition Procedure A, Single-Coupling Manual Addition Procedure B: Chloroacetic Anhydride coupling, Final rinse and dry procedure. The following procedures were performed according to the published methods (e.g., WO 2023/225661). Global Deprotection Method, Cyclization Method, N-Methylation on-Resin Method A, N- Methylation On-resin Method B (Turner, R.A. et al, Org. Lett., 15(19):5012-5015 (2013)), N-Alkylation On-resin Procedure Method A, N-Alkylation On-resin Procedure Method B, N-Nosylate Formation Procedure, N-Nosylate Removal Procedure, General Procedure for Preloading amines on the PL-FMP resin, General Procedure for Preloading Fmoc-Amino Acids on Cl-trityl resin, Click Reaction On-Resin Method A, Click Reaction On-Resin Method B, Suzuki Reaction On-resin Procedure, Fatty acid chain coupling procedure A, Fatty acid chain coupling procedure B, General Purification Procedures. Unnatural Fmoc-Amino Acid Synthesis and fatty acid tail synthesis were performed according to the published methods (e.g., WO 2023/225661). Preparation of 1-(tert-butyl) 18-(perfluorophenyl) octadecanedioate, (C18) Scheme: Step 1: Preparation of 18-(tert-butoxy)-18-oxooctadecanoic acid, Octadecanedioic acid (5.0 g, 15.90 mmol) was suspended in toluene (28.4 ml) and the mixture was heated to reflux..1,1-di-tert-butoxy-N,N-dimethylmethanamine (10.22 ml, 42.6 mmol) was added dropwise over 30 minutes. The mixture was put under refluxing conditions overnight. The solvent was removed in vacuo at 50 ^oC and the crude material was suspended in DCM/EtOAc ( 75 mL.1:1 ) and stirred for 15 min. The solids were removed by filtration and washed with DCM ( 25 mL). The filtration was evaporated in vacuo. The crude product was purified by flash chromatography ( 0 to 25% Acetone / DCM ) to get 18-(tert-butoxy)-18-oxooctadecanoic acid (1.72 g, 4.64 mmol, 29.2 % yield). ¹H NMR (500 MHz, METHANOL-d4) Shift 2.32 - 2.17 (m, 4H), 1.66 - 1.54 (m, 4H), 1.50 - 1.30 (m, 32H). Step 2: Preparation of 1-(tert-butyl) 18-(perfluorophenyl) octadecanedioate, To a 50 ml round bottom flask was added 18-(tert-butoxy)-18-oxooctadecanoic acid (807 mg, 2.178 mmol), N,N-Dimethylformamide (8 mL), pyridine (379 mg, 4.79 mmol), and perfluorophenyl 2,2,2-trifluoroacetate (1220 mg, 4.36 mmol). The flask was sealed with a septum and kept under a blanket of nitrogen and stirred overnight at room temperature. The next day the reaction was poured into a saturated citric acid soution and extracted with DCM three times. The organic layers were combined and washed with brine, dried over Na2SO4 and evaporated in vacuo. The crude product 1-tert-butyl 18- (perfluorophenyl) octadecanedioate (1.1 g, 2.050 mmol, 94 % yield) was used as is without purification.¹H NMR (400 MHz, CHLOROFORM-d) δ 2.70 - 2.63 (m, 2H), 2.22 (t J=75 Hz 2H) 178 ( uin J=75 Hz 2H) 163 - 152 (m 2H) 148 - 139 (m 12H) 5-(tert-butoxy)-5-oxopentanoic acid, To a glass reaction vessel equipped with a frit was added the 2-chloro-chlorotrityl resin mesh 50-150, (1.54 meq / gram, 1.94 grams, 3.0 mmole) to be swollen in DCM (5 mL) for 5 minutes. A solution of Fmoc-Glu-OtBu (1.276 g, 3.00 mmol, 1.0 eq ) in DCM (5 mL) was added to the resin followed by DIPEA (2.61 ml, 15.00 mmol, 5.0 eq). The reaction was shaken at room temperature for 60 minutes. Add in DIEA (0.5 mL) and Methanol (3 mL), shaken for an additional 15 minutes. The reaction solution was filtered through the frit and the resin was rinsed with DCM (4 x 5 mL), DMF (4 x5 mL), DCM (4 x 5mL), diethyl ether (4 x 5mL), and dried using a flow of nitrogen. A sample of resin (13.1 mg) was treated with 20% piperidine / DMF (v/v, 2.0 mL) for 10 minutes with shaking.1 mL of this solution was transferred to a 25.0 mL volumetric flask and diluted with methanol to a total volume of 25.0 mL. A blank solution of 20% piperidine /DMF (v/v, 1.0 mL) was diluted up with methanol in a volumetric flask to 25.0 mL. The UV was set to 301nm and zero with the blank solution followed by the reading of the solution, Absorbance = 1.9411 (1.9411/20 mg)*6.94 = 0.6736. Loading of the resin was measured to be 0.6736 mmol/g. Step 2: Preparation of (S)-5-(tert-butoxy)-4-(18-(tert-butoxy)-18-oxooctadecanamido)-5- oxopentanoic acid, In a glass reaction vessel equipped with a frit was added the previous pre-loaded resin and a solution of 20% piperidine / DMF (v/v, 5 mL) and the suspension was allowed to shake for 5 mins. The solution was filtered off and the resin was treated again with a solution of 20% piperidine / DMF (v/v, 5 mL) for another 5 mins. The reaction solution was filtered through the frit and the resin was washed with DMF (6 x 5 mL x 1 minute shaking). A solution of 18-(tert-butoxy)-18-oxooctadecanoic acid (593 mg, 1.6 mmol, 1.6 eq) and 1- hydroxy-7-azabenzotriazole (218 mg, 1.600 mmol, 1.6 eq) in DMF (4 mL) was added to the resin followed by N,N’-diisopropylcarbodiimide (251 µl, 1.600 mmol, 1.6 eq). The reaction was shaken at room temperature for 16 hours. The resin was washed with DMF (4 x 5 mL x 1 minute shaking). The protected peptide was cleaved off the resin with 20% HexaFluoro IPA/ DCM (v/v, 30 mL) for 2 hours at room temperature. The cleavage solution containing the crude product was obtained by filtration. The resin was rinsed with DCM (2 x 5 mL). The combined filtrate were evaporated, chased with DCM (2 x 5 mL) to afford (S)-5-(tert-butoxy)-4- (18-(tert-butoxy)-18-oxooctadecanamido)-5-oxopentanoic acid as an oil (378 mg, 0.680 mmol, 68%). Analysis condition E: Retention time = 1.32 min; ESI-MS(+) m/z [M+H]⁺: 557.5. Step 3: Preparation of 1-(tert-butyl) 5-(perfluorophenyl) (18-(tert-butoxy)-18- oxooctadecanoyl)-L-glutamate, To a pressure seal vial was added (S)-5-(tert-butoxy)-4-(18-(tert-butoxy)-18- oxooctadecanamido)-5-oxopentanoic acid (378 mg, 0.680 mmol, 1.0 eq) d MF (5 mL), perfluorophenyl 2,2,2-trifluoroacetate (233 µl, 1.360 mmol, 2.0 eq), and pyridine (121 µl, 1.496 mmol, 2.2 eq). The reaction mixture was kept under a blanket of nitrogen and stirred for 16 hours at room temperature. The reaction was poured into a saturated citric acid solution and extracted with CH₂Cl₂ (3 x). The organic layers were combined and washed with brine, dried over Na2SO4 and evaporated in vacuo. The crude material was purified by chromatography on silica gel (40 g) and eluted with 100 hexanes to 30% ethyl acetate/hexanes. The appropriate fractions were combined to obtain (S)-1-tert-butyl 5- (perfluorophenyl) 2-(18-(tert-butoxy)-18-oxooctadecanamido)pentanedioate (249 mg, 0.345 mmol, 50.7 % yield).¹H NMR (400MHz, CHLOROFORM-d) ^ 6.08 (d, J=7.8 Hz, 1H), 4.63 (td, J=7.8, 5.0 Hz, 1H), 2.89 - 2.65 (m, 2H), 2.43 - 2.31 (m, 1H), 2.29 - 2.18 (m, 4H), 2.14 - 2.02 (m, 1H), 1.59 - 1.43 (m, 18H), 1.37 - 1.22 (m, 24H). Preparation of (S)-1-azido-40-carboxy-37,42-dioxo-3,6,9,12,15,18,21,24,27,30,33- undecaoxa-36,41-diazanonapentacontan-59-oic acid (ELN), 17‐{[(1S)‐3‐[(35‐azido‐3,6,9,12,15,18,21,24,27,30,33‐undecaoxapentatriacontan‐1‐ yl)carbamoyl]‐1‐carboxypropyl]carbamoyl}heptadecanoic acid (IUPAC) (Azide-Peg11-^Glu-FDA18)

Scheme: Step 1: Preparation of 18-(tert-butoxy)-18-oxooctadecanoic acid Octadecanoic acid (7.5 g, 23.85 mmol) was suspended in toluene (42.6 mL) and the mixture was heated to reflux.1,1-Di-tert-butoxy-N,N-dimethylmethanamine (15.33 mL, 63.9 mmol) was added drop-wise over 30 min. The mixture was reflux overnight. The solvent was removed in vacuo at 50 ^oC and the crude material was suspended in CH2Cl2/EtOAc (110 mL.1:1) and stirred for 15 min. The solids were removed by filtration and washed with CH2Cl2 (40 mL). The filtration was evaporated in vacuo. The crude product was purified by flash chromatography (SG, 0-25% acetone/CH₂Cl₂) to get 18-(tert-butoxy)-18-oxooctadecanoic acid (3.95 g, 10.66 mmol, 44.7 % yield). Analysis condition D: Retention time = 5.04 min; ESI-MS(+) m/z 297.3 [M – OC(CH₃)₃].¹H NMR (500MHz, METHANOL-d4) 2.29 (t, J=7.5 Hz, 2H), 2.22 (t, J=7.4 Hz, 2H), 1.67 - 1.53 (m, 4H), 1.50 - 1.42 (m, 9H), 1.40 - 1.25 (m, 24H). Step 2: Preparation of 1-tert-butyl 18-(2,5-dioxopyrrolidin-1-yl) octadecanedioate DCC (5.11 mL, 5.11 mmol, 1.1 eq) was added to a solution of 18-(tert-butoxy)-18- oxooctadecanoic acid (1.72 g, 4.64 mmol) and 1-hydroxypyrrolidine-2,5-dione (0.588 g, 5.11 mmol, 1.1 eq) in DMF (48 mL). The mixture was stirred at rt overnight. The mixture was filtered and concentrated to get 1-tert-butyl 18-(2,5-dioxopyrrolidin-1-yl) octadecanedioate which was used as is in the next step. Step 3: Preparation of (S)-5-(tert-butoxy)-4-(18-(tert-butoxy)-18-oxooctadecanamido)- 5-oxopentanoic acid Water (5.80 mL) was added to a mixture of (S)-4-amino-5-(tert-butoxy)-5- oxopentanoic acid (1.038 g, 5.11 mmol), 1-tert-butyl 18-(2,5-dioxopyrrolidin-1-yl) octadecanedioate (2.171 g, 4.64 mmol, 1.10 eq), sodium bicarbonate (0.468 g, 5.57 mmol, 1.2 eq) in THF (17.41 mL). The resulting clear solution was stirred at rt for 4 h. The solvent was removed in vacuo. HCl (6.04 mL, 6.04 mmol, 1.3 eq) was added and the pH was adjusted to 2-3 at 0 ^oC. The resulting suspension was extracted with CH2Cl2 (3x). The organic fractions were combined, dried over anhydrous sodium sulfate, and filtered. The organic solution was concentrated on rotovap. The resulting crude product was purified by flash chromatography (acetone/CH₂Cl₂0-25%) to afford (S)-5-(tert-butoxy)- 4-(18-(tert-butoxy)-18-oxooctadecanamido)-5-oxopentanoic acid (2.29 g, 4.12 mmol, 89 % yield) as a white solid. Analysis condition D: Retention time = 2.74 min; ESI-MS(+) m/z 555.6 (M+H)⁺.¹H NMR (500MHz, METHANOL-d₄) ^ 4.32 (dd, J=9.0, 5.3 Hz, 1H), 2.45 - 2.33 (m, 2H), 2.30 - 2.06 (m, 5H), 1.99 - 1.82 (m, 3H), 1.78 - 1.53 (m, 2H), 1.53 - 1.44 (m, 18H), 1.44 - 1.26 (m, 24H) Step 4: Preparation of (S)-tert-butyl 1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate To a solution of (S)-5-(tert-butoxy)-4-(18-(tert-butoxy)-18-oxooctadecanamido)- 5-oxopentanoic acid (438 mg, 0.789 mmol, 1.5 eq) in DMF (1593 µL) was added Hunig'sBase (275 µL, 1.577 mmol, 3.0 eq) and HATU (400 mg, 1.051 mmol, 2.0 eq).35- azido-3,6,9,12,15,18,21,24,27,30,33-undecaoxapentatriacontan-1-amine (300 mg, 0.526 mmol) was then added, and the solution stirred at rt overnight. The mixture was poured into water and extracted with CH2Cl2 (3 x). The combined organic extracts were dried over MgSO₄, filtered, and concentrated in vacuo to get (S)-tert-butyl 1-azido-40-(tert- butoxycarbonyl)-37,42-dioxo-3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41- diazanonapentacontan-59-oate, which is used as is in the next step. Analysis condition D: Retention time = 2.84 min; ESI-MS(+) m/z 1109.1 (M+H)⁺. Step 5: Preparation of 17‐{[(1S)‐3‐[(35‐azido‐3,6,9,12,15,18,21,24,27,30,33‐ undecaoxapentatriacontan‐1‐yl)carbamoyl]‐1‐carboxypropyl]carbamoyl}heptadecanoic acid The mixture of (S)-tert-butyl 1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (280 mg, 0.253 mmol) and TFA (3 mL, 38.9 mmol) in DCM (3.0 mL) was stirred at rt for 2 h. The resulting crude product was purified by Prep-HPLC (Solvent A = 10% MeOH-90% H₂O- 0.1% TFA, Solvent B = 90% MeOH-10% H2O - 0.1% TFA. Column: phenomenex luna 30 x 100mm, S10, Flow rate: 40 ml / min, 50 - 100% B, 10 min and stop at 12 min) to obtain (S)-1-azido-40-carboxy-37,42-dioxo-3,6,9,12,15,18,21,24,27,30,33-undecaoxa- 36,41-diazanonapentacontan-59-oic acid (124 mg, 0.124 mmol, 49.3 % yield). Analysis condition D: Retention time = 2.43 min; ESI-MS(+) m/z 996.9 (M+H)⁺.¹H NMR (500MHz, METHANOL-d4) ^ 4.44 - 4.35 (m, 1H), 3.84 - 3.27 (m, 48H), 2.39 - 2.11 (m, 7H), 2.04 - 1.87 (m, 1H), 1.71 - 1.55 (m, 4H), 1.44 - 1.18 (m, 24H). Preparation of Example 1000 loaded with oxy)carbonyl)amino)undecanoic acid on a 50 µmol scale, and the reaction vessel was placed on the Symphony peptide synthesizer. The following procedures were then performed sequentially: “Symphony Resin-swelling procedure” was followed; “Symphony Pre-activated Single-coupling procedure” was followed with 1-(9H-fluoren- 9-yl)-3-oxo-2,7,10-trioxa-4-azadodecan-12-oic acid; “Symphony Single-coupling procedure” was followed with Fmoc-Asp(tBu)-OH; “Symphony Single-coupling procedure” was followed with Fmoc-Cys(Trt)-OH; “Symphony Single-coupling procedure” was followed with Fmoc-Asp(tBu)-OH; “Symphony Single-coupling procedure” was followed with Fmoc-Trp(Boc)-OH; “Symphony Single-coupling procedure” was followed with Fmoc-Tyr(tBu)-OH; “Symphony Single-coupling procedure” was followed with Fmoc-Trp(Boc)-OH; “Symphony Single-coupling procedure” was followed with Fmoc-Ser(tBu)-OH; “Symphony Single-coupling procedure” was followed with Fmoc-Asp(tBu)-OH; “Symphony Single-coupling procedure” was followed with Fmoc-Asp(tBu)-OH; “Symphony Single-coupling procedure” was followed with Fmoc-D-Pro-OH; “Symphony double-coupling procedure” was followed with Fmoc-N-Me-Phe-OH; “Symphony double-coupling extended time procedure” was followed with Fmoc- Asn(Trt)-OH; “Symphony double-coupling extended time procedure” was followed with Fmoc- Asp(tBu)-OH; “Symphony Single-coupling procedure” was followed with Fmoc-Tyr(tBu)-OH; “Symphony Pre-activated Single-coupling procedure” was followed with (S)-2-((((9H- fluoren-9-yl)methoxy)carbonyl)amino)-3-(3,4,5-trifluorophenyl)propanoic acid (Fmoc- Phe(3,4,5-trifluoro)-OH; “Symphony Chloroacetic Anhydride coupling procedure” “Global Deprotection Method” was followed; “Cyclization Method” was followed. The crude material was purified via preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5-µm particles; Mobile Phase A: 5:95 acetonitrile: water with 10-mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile: water with 10- mM ammonium acetate; Gradient: 10-50% B over 20 minutes, then a 4-minutes hold at 100% B; Flow: 20 mL/min. Fractions containing the desired product were combined and dried via centrifugal evaporation. The yield of the product was 10.5 mg, and its estimated purity by LCMS analysis was 98.4%. Analysis condition B: Retention time = 1.66 min; ESI-MS(+) m/z [M+2H]²⁺: 1213.2. The following examples were prepared, using 2-chlorotrityl resin pre-loaded with 11- ((((9H-fluoren-9-yl)methoxy)carbonyl)amino)undecanoic acid on a 50 µmol scale, following the general synthetic sequence described for the preparation of Example 1000. Ex. Structure Ex. Structure No. Yield (mg); HPLC purity No. Yield (mg); HPLC purity Ex. Structure Ex. Structure No. Yield (mg); HPLC purity No. Yield (mg); HPLC purity Ex. Structure Ex. Structure Ex. Structure Ex. Structure N Yil HPL it N Yil HPL it Preparation of Example 1008 Example 1008 ((6S,9S,12S,15S,18S,24R,27S,30S,33S,36S,38aS,44S,47S,49aR)-30,33,36-tris((1H- indol-3-yl)methyl)-24-(((S)-1-((2-((2-(((S)-3-amino-1-carboxypropyl)amino)-2- oxoethyl)amino)-2-oxoethyl)amino)-3-carboxy-1-oxopropan-2-yl)carbamoyl)-6-benzyl- 15-(4-hydroxybenzyl)-7-methyl-5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo- 18-(3,4,5-trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 9,12,27,44,47-pentayl)pentaacetic acid (58.6 mg, 25 µmol) following the “Fatty acid chain coupling procedure A” general procedure with perfluorophenyl tetradecanoate. The crude material was purified via preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5-µm particles; Mobile Phase A: 5:95 acetonitrile: water with 10-mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile: water with 10- mM ammonium acetate; Gradient: 40-80% B over 20 minutes, then a 4-minutes hold at 100% B; Flow: 20 mL/min. Fractions containing the desired product were combined and dried via centrifugal evaporation. The yield of the product was 3.3 mg, and its estimated purity by LCMS analysis was 95.1%. Analysis condition B: Retention time = 2.53 min; ESI-MS(+) m/z [M+2H] ²⁺: 1291.7. The following examples were prepared using exocyclic amines such as 1008A, 1010A, following the “Fatty acid chain coupling procedure A or B” general procedure as shown in Example 1008. Ex Structure Ex Structure Ex. Structure Ex. Structure Ex. Structure Ex. Structure

Ex Structure Ex Structure Ex. Structure Ex. Structure E t t E t t Ex Str tr Ex Str tr Ex. Structure Ex. Structure No. Yield (mg); HPLC purity No Yield (mg); HPLC purity Ex. Structure Ex. Structure Ex Str t r Ex Str t r Preparation of Example 1055 Ex , chain coupling procedure B” general procedure with (S)-1-tert-butyl 5-(perfluorophenyl) 2-(18-(tert-butoxy)-18-oxooctadecanamido)pentanedioate. The crude material was purified via preparative LC/MS. The yield of the product was 17.5 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition A: Retention time = 1.18 min; ESI-MS(+) m/z [M+2H] ²⁺: 1047.3. Preparation of Example 1057 Example 1 chain coupling procedure A” general procedure with (R)-perfluorophenyl 4- ((3R,5R,8R,9S,10S,13R,14S,17R)-3-hydroxy-10,13-dimethylhexadecahydro-1H- cyclopenta[a]phenanthren-17-yl)pentanoate. The crude material was purified via preparative LC/MS. The yield of the product was 3.7 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition A: Retention time = 1.34 min; ESI-MS(+) m/z [M+2H] ²⁺: 1540.16.

Preparation of Example 1058 Example 1 chain coupling procedure B” general procedure with 11-(tert-butyl) 16-(perfluorophenyl) hexadecanedioate. The crude material was purified via preparative LC/MS. The yield of the product was 7.5 mg, and its estimated purity by LCMS analysis was 96.4%. Analysis condition B: Retention time = 1.80 min; ESI-MS(+) m/z [M+2H] ²⁺: 1495.3.

Preparation of Example 1063 Example 1063 was prepa n vessel was added 2-chlorotrityl resin pre-loaded with 11-((((9H-fluoren-9- yl)methoxy)carbonyl)amino)undecanoic acid on a 0.10 mmol scale, and the reaction vessel was placed on the Prelude peptide synthesizer. The following procedures were then performed sequentially: “Prelude Method: Resin-swelling procedure” was followed; “Prelude Method: Single-coupling procedure” was followed with (S)-2-((((9H-fluoren-9- yl)methoxy)carbonyl)amino)pent-4-ynoic acid; “Prelude Method: Single-coupling procedure” was followed with 1-(9H-fluoren-9-yl)-3- oxo-2,7,10-trioxa-4-azadodecan-12-oic acid; “Prelude Method: Single-coupling procedure” was followed with Fmoc-Glu(OtBu)-OH; “Prelude Method: Single-coupling procedure” was followed with 1-(9H-fluoren-9-yl)-3- oxo-2,7,10-trioxa-4-azadodecan-12-oic acid; “Prelude Method: Single-coupling procedure” was followed with (S)-4-((((9H-fluoren-9- yl)methoxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanoic acid; “Prelude Method: Single-coupling procedure” was followed with Fmoc-Gly-OH; “Prelude Method: Single-coupling procedure” was followed with Fmoc-Cys(Trt)-OH; “Prelude Method: Single-coupling procedure” was followed with Fmoc-Asp(OtBu)-OH; “Prelude Method: Single extended time-coupling procedure” was followed with Fmoc- Trp(Boc)-OH; “Prelude Method: Single extended time-coupling procedure” was followed with Fmoc- Trp(Boc)-OH; “Prelude Method: Single extended time-coupling procedure” was followed with Fmoc- Trp(Boc)-OH; “Prelude Method: Single extended time-coupling procedure” was followed with Fmoc- Pro-OH; “Prelude Method: Single extended time-coupling procedure” was followed with Fmoc- Asp(OtBu)-OH; “Prelude Method: Single-coupling procedure” was followed with Fmoc-Asp(OtBu)-OH; “Prelude Method: Single-coupling procedure” was followed with Fmoc-D-Pro-OH; “Prelude Method: Single-coupling procedure” was followed with Fmoc-Nme-Tyr(tBu)- OH; “Prelude Method: Single-coupling procedure” was followed with Fmoc-Asn(Trt)-OH; “Prelude Method: Single-coupling procedure” was followed with Fmoc-Asp(tBu)-OH; “Prelude Method: Single-coupling procedure” was followed with Fmoc-Tyr(tBu)-OH; “Prelude Method: Single-coupling procedure” was followed with (S)-2-((((9H-fluoren-9- yl)methoxy)carbonyl)amino)-3-(3,4,5-trifluorophenyl)propanoic acid; “Prelude Method: Chloroacetic anhydride coupling” was followed; “Deprotection Method A” was followed; “Cyclization Method A” was followed. The crude material was purified via preparative LC/MS. The yield of the product was 1.6 mg, and its estimated purity by LCMS analysis was 95.5%. Analysis condition B: Retention time = 1.64 min; ESI-MS(+) m/z [M+3H] ³⁺: 971.9. The following examples were prepared using exocyclic amines following the “Fatty acid chain coupling procedure A or B” general procedure as shown in Example 1008.

Preparation of Example 1064 The yield of the product was 2.1 mg and its p p The yield of the product was 3.8

Preparation of Example 1093 The yield of the product was 1.3 , 1010A, following the “Fatty acid chain coupling procedure A or B” general procedure as shown in Example 1008. Example number Structure Yield (m ) HPLC urit LCMS Retention time (RT) m/z (Anal tical The following examples were prepared, using 2-chlorotrityl resin pre-loaded with 11- ((((9H-fluoren-9-yl)methoxy)carbonyl)amino)undecanoic acid on a 50 µmol scale, following the general synthetic sequence described for the preparation of Example 1000. Ex. Structure Ex. Structure No. Yield (mg); HPLC purity No. Yield (mg); HPLC purity Preparation 23.5 mg, 96.6%.

Preparation 34.3 mg, 99%. P ti f E l 1117 The yield of the product was 25.6 mg, 100%. Analysis condition A: Retention time = 1.27 min; ESI-MS(+) m/z [M+2H]²⁺: 1492.1. The following examples were prepared, using 2-chlorotrityl resin pre-loaded with 11- ((((9H-fluoren-9-yl)methoxy)carbonyl)amino)undecanoic acid on a 50 µmol scale, following the general synthetic sequence described for the preparation of Example 1000. Preparation of Example 1120 Preparation of Example 1125

Preparation of Example 1137 Preparation of Example 1139

Preparation of Example 1141 Preparation of Example 1143 , correspoding fatty acid tail on a 50 or 100 µmol scale, following the general synthetic sequence described for the preparation of Example 1063.

Preparation of Example 1150 Example 11 30,33,36-tris benzyl-24-(((S)-1-carboxybut-3-yn-1-yl)carbamoyl)-15-(4-hydroxybenzyl)-7,44- dimethyl-5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,47-diyl)diacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition A: Retention time = 1.41 min; ESI-MS(+) m/z [M+2H]²⁺: 1026. Example 1150 was prepared in a 12 ^mol scale as follows: Compound from Example 1150A (24 mg, 0.012 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42- dioxo-3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (14.27 mg, 0.013 mmol) (14.27 mg, 0.013 mmol) were dissolved in DMF (1 mL). In a separated 20 ml-scintillation vial was added sodium (R)-2-((S)-1,2-dihydroxyethyl)-4- hydroxy-5-oxo-2,5-dihydrofuran-3-olate (280 mg) and copper(II) sulfate pentahydrate 100 mg in 5 mL of water. The solution was shaken for 30-60 seconds and 0.15 mL of the solution was added to the above DMF solution. The resulting mixture was shaken at RT for 3 h. The mixture was concentrated. The residue was dissolved in TFA/TIS/H₂O (96:3:1) (2 mL) and stirred at RT for 20 min. Cold Et2O was added. The mixture was centrifuged and the solvents were decanted. The crude product was dissolved in DMF, filtered and submitted to purification. The crude material was purified via preparative LC/MS. The yield of the product was 3.2 mg, and its estimated purity by LCMS analysis was 85.9%. Analysis condition B: Retention time = 2.03 min; ESI-MS(+) m/z [M+3H]³⁺: 1016.2. P ti f E l 11 1 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine-24- carboxamido)pent-4-ynoic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The crude material was purified via preparative LC/MS. Analysis condition A: Retention time = 1.41 min; ESI-MS(+) m/z [M+2H]²⁺: 1047. Example 1151 was prepared, on a 17 ^mol scale, using the product from Example 1151 A (36 mg, 0.017 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (20.98 mg, 0.019 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 2.6 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition B: Retention time = 2.05 min; ESI-MS(+) m/z [M+3H]³⁺: 1030. Preparation of Example 1152 Example 11 , , synthetic sequence described for the preparation of Example 1151. The crude material was purified via preparative LC/MS. The yield of the product was 1.6 mg, and its estimated purity by LCMS analysis was 81.2%. Analysis condition A: Retention time = 1.69 min; ESI-MS(+) m/z [M+3H]³⁺: 1029.9. Examples 1151 and 1152 are two pure fractions with different retention time but same molecular weight.

Preparation of Example 1153 OH H N O O H Example 1153 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The crude material was purified via preparative LC/MS. The yield of the product was 81.8 mg, and its estimated purity by LCMS analysis was 95.6%. Analysis condition B: Retention time = min; ESI-MS(+) m/z [M+2H]²⁺: 1050.03. Example 1153 was prepared on a 21 ^mol scale, using the product from Example 1153 A (44 mg, 0.021 mmol), tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (25.6 mg, 0.023 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 8.3 mg, and its estimated purity by LCMS analysis was 95.3%. Analysis condition B: Retention time = 2.12 min; ESI-MS(+) m/z [M+3H]³⁺: 1032.0 Preparation of Example 1154 hydroxybenzyl)-7-methyl-5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18- (3,4,5-trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,27-diyl)diacetic acid, was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The crude material was purified via preparative LC/MS. The yield of the product was 37 mg, and its estimated purity by LCMS analysis was 96.1%. Analysis condition B: Retention time = 1.46 min; ESI-MS(+) m/z [M+2H]²⁺: 1054.1. Example 1154 was prepared on a 14 ^mol scale, using the product from Example 1154A (30 mg, 0.014 mmol), tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (17.4 mg, 0.016 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 4.7 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition A: Retention time = 1.73 min; ESI-MS(+) m/z [M+3H]³⁺: 1035.0. Preparation of Example 1155 Example 1155 was pre [(3S,6S,9R,15S,18S,21 , , , , , , , , )‐ ‐{[( )‐ ‐amno‐ ‐ cyanopropyl]carbamoyl}‐15‐benzyl‐3,6,18,36‐tetrakis(carboxymethyl)‐24‐[(4‐ hydroxyphenyl)methyl]‐39,42,45‐tris[(1H‐indol‐3‐yl)methyl]‐16‐methyl‐ 2,5,8,14,17,20,23,26,29,35,38,41,44,47‐ tetradecaoxo‐27‐[(3,4,5‐trifluorophenyl)methyl]‐31‐thia‐ 1,4,7,13,16,19,22,25,28,34,37,40,43,46‐ tetradecaazatricyclo[46.3.0.0^9,13]henpentacontan‐21‐yl]acetic acid on a 50 ^mol scale, following the “Fatty acid chain coupling procedure B” general procedure with 1‐tert‐butyl 2,3,4,5,6‐pentafluorophenyl dodecanedioate. The crude material was purified via preparative LC/MS. The yield of the product was 5.0 mg, and its estimated purity by LCMS analysis was 94%. Analysis condition A: Retention time = 1.44 min; ESI-MS(+) m/z [M+2H]²⁺: 1155.2. Analysis condition B: Retention time = 1.75 min; ESI-MS(+) m/z [M+2H]²⁺: 1155.5.

Preparation of Example 1156 Example 11 30,33,36-tris benzyl-24-(((S)-1-carboxybut-3-yn-1-yl)carbamoyl)-15-(4-hydroxybenzyl)-7,44- dimethyl-5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,27-diyl)diacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The yield of the product was 49 mg, and its estimated purity by LCMS analysis was 99.3%. Analysis condition B: Retention time = 1.57 min; ESI-MS(+) m/z [M+2H]²⁺: 1026.1. Example 1156 was prepared, on a 19 ^mol scale, using the product from Example 1156A (39 mg, 0.019mmol), tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (23.2 mg, 0.021 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 5.7 mg, and its estimated purity by LCMS analysis was 92.8%. Analysis condition A: Retention time = 1.51 min; ESI-MS(+) m/z [M+3H]³⁺: 1015.9.

indol-3-yl)methyl)-9-(2-amino-2-oxoethyl)-47-(2-aminoethyl)-6-benzyl-24-(((S)-1- carboxybut-3-yn-1-yl)carbamoyl)-15-(4-hydroxybenzyl)-7,27-dimethyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,44-diyl)diacetic acid, was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The crude material was purified via preparative LC/MS. The yield of the product was 55.1 mg, and its estimated purity by LCMS analysis was 87.3%. Analysis condition A: Retention time = 1.34 min; ESI-MS(+) m/z [M+2H]²⁺: 1026.0. Analysis condition B: Retention time = 1.62 min; ESI-MS(+) m/z [M+2H]²⁺: 1026.0. Example 1157 was prepared, using on a 21 ^mol scale, using the product from Example 1157A (43 mg, 0.023 mmol), tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42- dioxo-3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (25.6 mg, 0.023 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 3 mg, and its estimated purity by LCMS analysis was 95.1%. Analysis condition B: Retention time = 2.02 min; ESI-MS(+) m/z [M+3H]³⁺: 1016.1.

30,33,36-tris((1H-indol-3-yl)methyl)-9-(2-amino-2-oxoethyl)-47-(4-aminobutyl)-6- benzyl-12-(carboxymethyl)-15-(4-hydroxybenzyl)-7,27,44-trimethyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine-24- carboxamido)pent-4-ynoic acid, was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The crude material was purified via preparative LC/MS. The yield of the product was 35.5 mg, and its estimated purity by LCMS analysis was 98.6%. Analysis condition B: Retention time = 1.58 min; ESI-MS(+) m/z [M+2H]²⁺: 1017.4. Example 1158 was prepared, on a 16 ^mol scale, using the product from Example 1158A (33 mg, 0.016 mmol), tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (19.8 mg, 0.018 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 5.7 mg, and its estimated purity by LCMS analysis was 92.9%. Analysis condition A: Retention time = 1.62 min; ESI-MS(+) m/z [M+2H]²⁺: 1516.1.

tris((1H-indol-3-yl)methyl)-47-(4-aminobutyl)-27-(2-aminoethyl)-6-benzyl-12- (carboxymethyl)-15-(4-hydroxybenzyl)-7,44-dimethyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-9-(pyridin-4-ylmethyl)-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine-24- carboxamido)pent-4-ynoic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The crude material was purified via preparative LC/MS. The yield of the product was 67.8 mg, and its estimated purity by LCMS analysis was 96.8%. Analysis condition A: Retention time = 1.49 min; ESI-MS(+) m/z [M+2H]²⁺: 1049.2. Example 1159 was prepared, on a 21 ^mol scale, using the product from Example 1159A (44 mg, 0.021 mmol), tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (25.6 mg, 0.023 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 3.6 mg, and its estimated purity by LCMS analysis was 90.5%. Analysis condition A: Retention time = 1.64, 1.69 min; ESI-MS(+) m/z [M+2H]²⁺, [M+3H]³⁺: 1547.2, 1032.0.Analysis condition B: Retention time = 1.9 min; ESI-MS(+) m/z [M+3H]³⁺: 1032.1. Preparation of Example 1160 Example 11 30,33,36-tris (((S)-1-carboxybut-3-yn-1-yl)carbamoyl)-15-(4-hydroxybenzyl)-7-methyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-27-(pyridin-3-ylmethyl)-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,44-diyl)diacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition B: Retention time = 1.38 min; ESI-MS(+) m/z [M+2H]²⁺: 1071.1. Example 1160 was prepared, on a 15 ^mol scale, using the product from Example 1160A (33 mg, 0.015 mmol), tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (18.8 mg, 0.017 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 5.4 mg, and its estimated purity by LCMS analysis was 94.7%. Analysis condition A: Retention time = 1.57 min; ESI-MS(+) m/z [M+2H]²⁺: 1568.9. Preparation of Example 1161 Exam 2,2'-( indol-3-yl)methyl)-9-(2-amino-2-oxoethyl)-47-(4-aminobutyl)-27-(2-aminoethyl)-6- benzyl-24-(((S)-1-carboxybut-3-yn-1-yl)carbamoyl)-15-(4-hydroxybenzyl)-7-methyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,44-diyl)diacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The crude material was purified via preparative LC/MS Analysis condition B: Retention time = 1.55 min; ESI-MS(+) m/z [M+2H]²⁺: 1054.2. Example 1176 was prepared, on a 11 ^mol scale, using the product from Example 1176A (24 mg, 0.011 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (13.9 mg, 0.013 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 2.1 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition A: Retention time = 1.6 min; ESI-MS(+) m/z [M+3H]³⁺: 1035.1. Preparation of Example 1162 , , , , , , , , , , , , , , , indol-3-yl)methyl)-9-(4-aminobutyl)-6-benzyl-24-(((S)-1-carboxybut-3-yn-1- yl)carbamoyl)-15-(4-hydroxybenzyl)-27-(hydroxymethyl)-7-methyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,44,47-triyl)triacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition A: Retention time = 1.27 min; ESI-MS(+) m/z [M+2H]²⁺: 1048.2. Example 1162 was prepared using the product from Example 1162A and tert-butyl (S)-1- azido-40-(tert-butoxycarbonyl)-37,42-dioxo-3,6,9,12,15,18,21,24,27,30,33-undecaoxa- 36,41-diazanonapentacontan-59-oate, following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 6.2 mg, and its estimated purity by LCMS analysis was 82.5%. Analysis condition A: Retention time = min; ESI-MS(+) m/z [M+3H]³⁺: 1032.0. Preparation of Example 1163 , , , , , , , , , , , , , , , indol-3-yl)methyl)-9-(2-amino-2-oxoethyl)-27-(2-aminoethyl)-6-benzyl-24-(((S)-1- carboxybut-3-yn-1-yl)carbamoyl)-15-(4-hydroxybenzyl)-7-methyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,44,47-triyl)triacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The yield of the product was 37.8 mg, and its estimated purity by LCMS analysis was 85.5%. Analysis condition A: Retention time = 1.31 min; ESI-MS(+) m/z [M+2H]²⁺: 1048.2. Example 1163 was prepared, on a 18 ^mol scale, using the product from Example 1163A (37 mg, 0.018 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (21.5 mg, 0.019 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 3.1 mg, and its estimated purity by LCMS analysis was 88.6%. Analysis condition A: Retention time = 1.43 min; ESI-MS(+) m/z [M+2H]²⁺: 1545.5. Preparation of Example 1164 Example 11 30,33,36-tris 3-yn-1-yl)carbamoyl)-15-(4-hydroxybenzyl)-7,44-dimethyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-9-(pyridin-4-ylmethyl)-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43] tetradecaazacyclopentatetracontine-12,27-diyl)diacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition B: Retention time = 1.52 min; ESI-MS(+) m/z [M+2H]²⁺: 1057.3. Example 1164 was prepared, on a 8 ^mol scale, using the product from Example 1164A (16.9 mg, 0.008 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (9.8 mg, 0.0088 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 2.8 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition B: Retention time = 2.05 min; ESI-MS(+) m/z [M+3H]³⁺: 1036.9. Preparation of Example 1165 Exa 30,33,36-tris((1H-indol-3-yl)methyl)-9,47-bis(2-amino-2-oxoethyl)-6-benzyl-24-(((S)-1- carboxybut-3-yn-1-yl)carbamoyl)-15-(4-hydroxybenzyl)-7,27-dimethyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,44-diyl)diacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition A: Retention time = 1.26 min; ESI-MS(+) m/z [M+2H]²⁺: 1033. Example 1165 was prepared, on a 17 ^mol scale, using the product from Example 1165A (35.9 mg, 0.017 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (21.2 mg, 0.019 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 13.7 mg, and its estimated purity by LCMS analysis was 98.6%. Analysis condition B: Retention time = 2.11 min; ESI-MS(+) m/z [M+3H]³⁺: 1020.8. Preparation of Example 1166 indol-3-yl)methyl)-9-(aminomethyl)-6-benzyl-24-(((S)-1-carboxybut-3-yn-1- yl)carbamoyl)-15-(4-hydroxybenzyl)-27-(hydroxymethyl)-7-methyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,44,47-triyl)triacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition B: Retention time = 1.54 min; ESI-MS(+) m/z [M+2H]²⁺: 1027. Example 1166 was prepared, on a 7.8 ^mol scale, using the product from Example 1166A (16 mg, 0.008 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42- dioxo-3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (9.5 mg, 0.009 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 3.5 mg, and its estimated purity by LCMS analysis was 98.5%. Analysis condition B: Retention time = 2.06 min; ESI-MS(+) m/z [M+2H]²⁺: 1525.2. Preparation of Example 1167 Example 11 , ((6S,9S,12S,15S,18S,24R,27S,30S,33S,36S,38aS,44S,47S,49aR)-30,33,36-tris((1H- indol-3-yl)methyl)-47-(4-aminobutyl)-9-(2-aminoethyl)-6-benzyl-24-(((S)-1-carboxybut- 3-yn-1-yl)carbamoyl)-15-(4-hydroxybenzyl)-7-methyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-27-(pyridin-4-ylmethyl)-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,44-diyl)diacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition A: Retention time = 1.38 min; ESI-MS(+) m/z [M+2H]²⁺: 1069. Example 1167 was prepared, on a 19 ^mol scale, using the product from Example 1167A (41 mg, 0.019 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (22.3 mg, 0.020 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 1.2 mg, and its estimated purity by LCMS analysis was 98.2%. Analysis condition A: Retention time = 1.61 min; ESI-MS(+) m/z [M+2H]²⁺: 1569.1. Preparation of Example 1168 Exampl ((6S,9S,12S,15S,18S,24R,27S,30S,33S,36S,38aS,44S,47S,49aR)-30,33,36-tris((1H- indol-3-yl)methyl)-9-(2-amino-2-oxoethyl)-47-(4-aminobutyl)-6-benzyl-24-(((S)-1- carboxybut-3-yn-1-yl)carbamoyl)-15-(4-hydroxybenzyl)-7-methyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-27-(pyridin-4-ylmethyl)-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,44-diyl)diacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition B: Retention time = 1.52 min; ESI-MS(+) m/z [M+3H]³⁺: 719.2. Example 1168 was prepared, on a 23 ^mol scale, using the product from Example 1168A (49.7 mg, 0.023 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (26.8 mg, 0.024 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 5.9 mg, and its estimated purity by LCMS analysis was 98.5%. Analysis condition B: Retention time = 1.97 min; ESI-MS(+) m/z [M+2H]²⁺: 1575.9. Preparation of Example 1169 Exampl ((6S,9S indol-3-yl)methyl)-9-(2-amino-2-oxoethyl)-47-(4-aminobutyl)-6-benzyl-12- (carboxymethyl)-15-(4-hydroxybenzyl)-7,44-dimethyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-27-(pyridin-4-ylmethyl)-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetra decaazacyclopentatetracontine-24-carboxamido)pent-4-ynoic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The yield of the product was 53.3 mg, and its estimated purity by LCMS analysis was 99.1%. Analysis condition B: Retention time = 1.48 min; ESI-MS(+) m/z [M+2H]²⁺: 1056.1. Example 1169 was prepared, on a 19 ^mol scale, using the product from Example 1169A (40.4 mg, 0.019 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (22.3 mg, 0.020 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 4.2 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition B: Retention time = 1.98 min; ESI-MS(+) m/z [M+3H]³⁺: 1036.2. Preparation of Example 1170 Example 117 ((6S,9S,12S, indol-3-yl)methyl)-9-(2-amino-2-oxoethyl)-44-(2-aminoethyl)-6-benzyl-12- (carboxymethyl)-15-(4-hydroxybenzyl)-7,27-dimethyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-47-(pyridin-4-ylmethyl)-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetra decaazacyclopentatetracontine-24-carboxamido)pent-4-ynoic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The yield of the product was 55.7 mg, and its estimated purity by LCMS analysis was 95.7%. Analysis condition A: Retention time = 1.48 min; ESI-MS(+) m/z [M+2H]²⁺: 1042.2. Example 1170 was prepared, on a 19 ^mol scale, using the product from Example 1170A (40 mg, 0.019 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (23.4 mg, 0.021 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 4.5 mg, and its estimated purity by LCMS analysis was 96%. Analysis condition B: Retention time = 1.92 min; ESI-MS(+) m/z [M+3H]³⁺: 1027.1. Preparation of Example 1171 Examp ((6S,9S,1 , , , , , , , , , , , , , indol-3-yl)methyl)-9-(2-amino-2-oxoethyl)-6-benzyl-24-(((S)-1-carboxybut-3-yn-1- yl)carbamoyl)-15-(4-hydroxybenzyl)-47-(hydroxymethyl)-7,27-dimethyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 12,44-diyl)diacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The yield of the product was 36.1 mg, and its estimated purity by LCMS analysis was 96.1%. Analysis condition 2: Retention time = 1.72 min; ESI-MS(+) m/z [M+2H]²⁺: 1019.01. Example 1171 was prepared, on a 15 ^mol scale, using the product from Example 1171A (31 mg, 0.015 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (18.6 mg, 0.017 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 7.4 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition A: Retention time = 1.56 min; ESI-MS(+) m/z [M+2H]²⁺: 1517.1. Preparation of Example 1172 Exampl ((6S,9S,12 , , , , , , , , a , , , a )- , , -rs(( - indol-3-yl)methyl)-9-(2-amino-2-oxoethyl)-47-(2-aminoethyl)-6-benzyl-12- (carboxymethyl)-15-(4-hydroxybenzyl)-7,27,44-trimethyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine-24- carboxamido)pent-4-ynoic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The yield of the product was 57.4 mg, and its estimated purity by LCMS analysis was 99.3%. Analysis condition B: Retention time = 1.56 min; ESI-MS(+) m/z [M+2H]²⁺: 1004.2. Example 1172 was prepared, on a 13 ^mol scale, using the product from Example 1172A (26.5 mg, 0.013 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (15.4 mg, 0.014 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 5.8 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition B: Retention time = 2.13 min; ESI-MS(+) m/z [M+2H]²⁺: 1502.2. Preparation of Example 1173 E xample 1173A: (5S,17S,31S)-5-(aminomethyl)-17-(2-carboxyethyl)-4,7,16,19,28- pentaoxo-31-(2-((6S,9S,12S,15S,18S,24R,27S,30S,33S,36S,38aS,44S,47S,49aR)- 30,33,36-tris((1H-indol-3-yl)methyl)-12,27,44,47-tetrakis(carboxymethyl)-9- (hydroperoxymethyl)-6,15-bis(4-hydroxybenzyl)-7-methyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine-24- carboxamido)acetamido)-9,12,21,24-tetraoxa-3,6,15,18,27-pentaazadotriacontanedioic acid was prepared, using Dap on chorotrityl resin on a 30 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition B: Retention time = 1.33 min; ESI-MS(+) m/z [M+2H]²⁺: 1354.0. Example 1173 was prepared, on a 25 ^mol scale, using the crude product from Example 1173A (67.8 mg, 25 µmol) following the “Fatty acid chain coupling procedure A” general procedure with perfluorophenyl tetradecanoate and crude perfluorophenyl icosanoate (23.93 mg, 50.0 µmol). The crude material was purified via preparative LC/MS. The yield of the product was 11.6 mg, and its estimated purity by LCMS analysis was 99.4%. Analysis condition B: Retention time = 2.72 min; ESI-MS(+) m/z [M+2H]²⁺: 1501.2. The following examples were prepared, on a 50 or 100 ^mol scale, using the crude parent macrocycle with a side chain amine, which was generated using the general synthetic sequence of Example 1000, following the “Fatty acid chain coupling procedure A or B” general procedure with the corresponding fatty acid tail. Preparation of Example 1174 The yield of the product was 9.9

Preparation of Example 1181 M M2H²⁺

Preparation of Example 1186 187 min; ESI- Preparation of Example 1193

Preparation of Example 1200

Preparation of Example 1202 Preparation of Example 1204

Preparation of Example 1210 P ti f E l 1212

Preparation of Example 1216 P ti f E l 1218 Preparation of Example 1225 The yield of the sis was 95.6%. Analysis co . ; ]²⁺: 1553.1. Preparation of Example 1226 The ield of the Preparation of Example 1227 Preparation of Example 1229 The yield of the Example 1230A: 2,2',2' 36S,38aS,44S,47S,49aR)-30, ((carboxymethyl)amino)-1-oxobutan-2-yl)carbamoyl)-6-benzyl-15-(4-hydroxybenzyl)-7- methyl-5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 9,12,27,44,47-pentayl)pentaacetic acid was prepared, using Gly on chorotrityl resin on a 100 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition A: Retention time = 1.06 min; ESI-MS(+) m/z [M+2H]²⁺: 1087. Example 1230 was prepared, on a 40 ^mol scale, using the crude product of Example 1230A (112 mg, 40 µmol), following the “Fatty acid chain coupling procedure B” general procedure with 1‐tert‐butyl 2,3,4,5,6‐pentafluorophenyl (48 mg, 100 µmol) The crude material was purified via preparative LC/MS. The yield of the product was 8.7 mg, and its estimated purity by LCMS analysis was 98.1%. Analysis condition B: Retention time = 1.71 min; ESI-MS(+) m/z [M+3H]³⁺: 848. Preparation of Example 1231 Th i ld f th d t d Preparation of Example 1233

Preparation of Example 1234 Preparation of Example 1236 Th ild f th Preparation of Example 1238 ( Preparation of Example 1240 Preparation of Example 1242 Preparation of Example 1244 Prearation of Examle 1246 Preparation of Example 1255

parent macrocycle with a side chain amine, which was generated using the general synthetic sequence of Example 1000, following the “Fatty acid chain coupling procedure A or B” general procedure with the corresponding fatty acid tail.

Preparation of Example 1258 Preparation of Example 1264 Preparation of Example 1269

Preparation of Example 1277 parent macrocycle with a side chain amine, which was generated using the general synthetic sequence of Example 1000, following the “Fatty acid chain coupling procedure A or B” general procedure with the corresponding fatty acid tail.

Preparation of Example 1285 Preparation of Example 1287

Preparation of Example 1288 Preparation of Example 1291 general synthetic sequence of Example 1000. Preparation of Example 1292

reparaon o xampe

Preparation of Example 1299 Preparation of Example 1303 Example 13 ((6S,9S,12S, 49aR)-30,33,36-tris((1H-indol-3-yl)methyl)-6-benzyl-24-(((S)-4-carboxy-1-(((S)-1- carboxybut-3-yn-1-yl)amino)-1-oxobutan-2-yl)carbamoyl)-15-(4-hydroxybenzyl)-7- methyl-5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 9,12,27,44,47-pentayl)pentaacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. The yield of the product was 21.2 mg, and its estimated purity by LCMS analysis was 92.2%. Analysis condition A: Retention time = 0.96 min; ESI-MS(+) m/z [M+2H]²⁺: 1120.0. Example 1303 was prepared, on a 30 ^mol scale, using the product from Example 1303A (67.2 mg, 0.030 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (33.3 mg, 0.030 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 4.1 mg, and its estimated purity by LCMS analysis was 97.2%. Analysis condition B: Retention time = 1.92 min; ESI-MS(+) m/z [M+3H]³⁺: 1079.3. The following examples were prepared, on a 50 or 100 ^mol scale, using the crude parent macrocycle with a side chain amine, which was generated using the general synthetic sequence of Example 1000, following the “Fatty acid chain coupling procedure A or B” general procedure with the corresponding fatty acid tail. Preparation of Example 1304

Example 1310 was prepared, on a 30 ^mol scale, using the product from Example 1317, 2,2',2'',2''',2''''-((6S,9S,12S,15S,18S,24R,27S,30S,33S,36S,38aS,44S,47S,49aR)- 30,33,36-tris((1H-indol-3-yl)methyl)-6-benzyl-24-((2-(((S)-1-((10-carboxydecyl)amino)- 1-oxopent-4-yn-2-yl)amino)-2-oxoethyl)carbamoyl)-15-(4-hydroxybenzyl)-7-methyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 9,12,27,44,47-pentayl)pentaacetic acid (70.5 mg, 0.030 mmol) and tert-butyl (S)-1-azido- 40-(tert-butoxycarbonyl)-37,42-dioxo-3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41- diazanonapentacontan-59-oate (33.3 mg, 0.030 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was mg, and its estimated purity by LCMS analysis was 100%. Analysis condition A: Retention time = 1.6 min; ESI-MS(+) m/z [M+2H]²⁺: 1673.3. Preparation of Example 1311 OH H N O O H Example 1 2,2',2'',2''',2''''-((6S,9S,12S,15S,18S,24R,27S,30S,33S,36S,38aS,44S,47S,49aR)-30,33,36- tris((1H-indol-3-yl)methyl)-6-benzyl-24-(((S)-1-((2-((10-carboxydecyl)amino)-2- oxoethyl)amino)-1-oxopent-4-yn-2-yl)carbamoyl)-15-(4-hydroxybenzyl)-7-methyl- 5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18-(3,4,5- trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 9,12,27,44,47-pentayl)pentaacetic acid (70.5 mg, 0.030 mmol) and tert-butyl (S)-1-azido- 40-(tert-butoxycarbonyl)-37,42-dioxo-3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41- diazanonapentacontan-59-oate (33.3 mg, 0.030 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 6.7 mg, and its estimated purity by LCMS analysis was 95.9%. Analysis condition B: Retention time = 2.1 min; ESI-MS(+) m/z [M+3H]³⁺: 1116.1. Preparation of Example 1312 Example 1312 w p p , , g p p , 2,2',2'',2''',2''''-((6S,9S,12S,15S,18S,24R,27S,30S,33S,36S,38aS,44S,47S,49aR)-30,33,36- tris((1H-indol-3-yl)methyl)-6-benzyl-24-(((S)-1-((2-((2-((10-carboxydecyl)amino)-2- oxoethyl)amino)-2-oxoethyl)amino)-1-oxopent-4-yn-2-yl)carbamoyl)-15-(4- hydroxybenzyl)-7-methyl-5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18- (3,4,5-trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 9,12,27,44,47-pentayl)pentaacetic acid (72.5 mg, 0.030 mmol) and tert-butyl (S)-1-azido- 40-(tert-butoxycarbonyl)-37,42-dioxo-3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41- diazanonapentacontan-59-oate (33.3 mg, 0.030 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 3.3 mg, and its estimated purity by LCMS analysis was 98.8%. Analysis condition B: Retention time = 2.07 min; ESI-MS(+) m/z [M+3H]³⁺: 1135.1. Preparation of Example 1313 Example 1313 was , , , 2,2',2'',2''',2''''-((6S,9S,12S,15S,18S,24R,27S,30S,33S,36S,38aS,44S,47S,49aR)-30,33,36- tris((1H-indol-3-yl)methyl)-6-benzyl-24-(((S)-4-carboxy-1-((2-(((S)-1-((10- carboxydecyl)amino)-1-oxopent-4-yn-2-yl)amino)-2-oxoethyl)amino)-1-oxobutan-2- yl)carbamoyl)-15-(4-hydroxybenzyl)-7-methyl-5,8,11,14,17,20,26,29,32,35,38,43,46,49- tetradecaoxo-18-(3,4,5-trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 9,12,27,44,47-pentayl)pentaacetic acid (74.4 mg, 0.030 mmol) and tert-butyl (S)-1-azido- 40-(tert-butoxycarbonyl)-37,42-dioxo-3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41- diazanonapentacontan-59-oate (33.3 mg, 0.030 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 5.9 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition A: Retention time = 1.54 min; ESI-MS(+) m/z [M+3H]³⁺: 1159.15.

((6S,9S,12S,15S,18S,24R,27S,30S,33S,36S,38aS,44S, 47S,49aR)-30,33,36-tris((1H-indol-3-yl)methyl)-6-benzyl-24-(((S)-1-carboxybut-3-yn-1- yl)carbamoyl)-15-(4-hydroxybenzyl)-7-methyl-5,8,11,14,17,20,26,29,32,35,38,43,46,49- tetradecaoxo-18-(3,4,5-trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 9,12,27,44,47-pentayl)pentaacetic acid was prepared, using Gly on chlorotrityl resin on a 100 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition B: Retention time = 1.45, 1.48 min; ESI-MS(+) m/z [M+2H]²⁺: 1149.2. Example 1314 was prepared, on a 30 ^mol scale, using the product from Example 1314A (63.3 mg, 0.0307 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (34.3 mg, 0.032 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 5.8 mg, and its estimated purity by LCMS analysis was 98.4%. Analysis condition B: Retention time = 1.95 min; ESI-MS(+) m/z [M+3H]³⁺: 1098.2. E ((6S,9S,12S,15S,18S,24R,27S,30S,33S,36S,38aS,44S, 47S,49aR)-30,33,36-tris((1H-indol-3-yl)methyl)-6-benzyl-24-((2-(((S)-1- ((carboxymethyl)amino)-1-oxopent-4-yn-2-yl)amino)-2-oxoethyl)carbamoyl)-15-(4- hydroxybenzyl)-7-methyl-5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18- (3,4,5-trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 9,12,27,44,47-pentayl)pentaacetic acid was prepared, using Gly on chlorotrityl resin on a 100 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition A: Retention time = 1.0 min; ESI-MS(+) m/z [M+2H]²⁺: 1113. Example 1315 was prepared, on a 30 ^mol scale, using the product from Example 1315A (66.7 mg, 0.0307 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (34.3 mg, 0.032 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 1.2 mg, and its estimated purity by LCMS analysis was 98.7%. Analysis condition B: Retention time = 1.99 min; ESI-MS(+) m/z [M+3H]³⁺: 1074.1. The following examples were prepared, on a 50 or 100 ^mol scale, using the general synthetic sequence of Example 1000. Preparation of Example 1316 The ield of the roduct was 203

Preparation of Example 1318

Preparation of Example 1321 Exa , , , , ((6S,9S,12S,15S,18S,24R,27S,30S,33S,36S,38aS,44S, 47S,49aR)-30,33,36-tris((1H-indol-3-yl)methyl)-6-benzyl-24-(((S)-1-carboxybut-3-yn-1- yl)carbamoyl)-15-(4-hydroxybenzyl)-7-methyl-5,8,11,14,17,20,26,29,32,35,38,43,46,49- tetradecaoxo-18-(3,4,5-trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine- 9,12,27,44,47-pentayl)pentaacetic acid was prepared, using Pra on chlorotrityl resin on a 50 ^mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition A: Retention time = 1.05 min; ESI-MS(+) m/z [M+2H]²⁺: 1056.1. Example 1321 was prepared, on a 50 µmol scale, using the product from Example 1321A (106 mg, 0.050 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (58.2 mg, 0.053 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 10.0 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition A: Retention time = 1.29 min; ESI-MS(+) m/z [M+3H]³⁺: 1036.1,. Preparation of Example 1322 carboxyethyl)-5,14,17,26,29-pentaoxo-28-(prop-2-yn-1-yl)-2-(2- ((6S,9S,12S,15S,18S,24R,27S,30S,33S,36S,38aS,44S,47S,49aR)-30,33,36-tris((1H- indol-3-yl)methyl)-6-benzyl-9,12,27,44,47-pentakis(carboxymethyl)-15-(4- hydroxybenzyl)-7-methyl-5,8,11,14,17,20,26,29,32,35,38,43,46,49-tetradecaoxo-18- (3,4,5-trifluorobenzyl)hexatetracontahydro-1H,5H-dipyrrolo[2,1-a1:2',1'- r][1]thia[4,7,10,13,16,19,22,25,28,31,34,37,40,43]tetradecaazacyclopentatetracontine-24- carboxamido)acetamido)-9,12,21,24-tetraoxa-6,15,18,27,30- pentaazahentetracontanedioic acid (87 mg, 0.030 mmol), which was synthesized following the general sequence of Example 1000, and tert-butyl (S)-1-azido-40-(tert- butoxycarbonyl)-37,42-dioxo-3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41- diazanonapentacontan-59-oate (34.9 mg, 0.032 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 7.1 mg, and its estimated purity by LCMS analysis was 92.9%. Analysis condition A: Retention time = 1.31 min; ESI-MS(+) m/z [M+3H]³⁺: 1299. Preparation of Example 1323 Example 1323A was prepared, using Gly on chlorotrityl resin on a 50 mol scale, following the general synthetic sequence described for the preparation of Example 1000. Analysis condition 2: Retention time = 1.49 min; ESI-MS(+) m/z [M+2H]²⁺: 1386.9. Example 1323 was prepared, on a 50 ^mol scale, using the crude product of Example 1323A (139 mg, 0.050 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42- dioxo-3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (58.2 mg, 0.053 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 29.7 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition A: Retention time = 1.23 min; ESI-MS(+) m/z [M+3H]³⁺: 1257.1. Preparation of Example 1324 Example 1324 w as prepare , on a mo scae, usng e cru e ( )‐ ‐[ ‐( ‐{ ‐[( )‐ 2‐[2‐(2‐{2‐[(4S)‐4‐(2‐{[(3S,6S,9R,15S,18S,21S,24S,27S,33R,36S,39S,42S,45S,48S)‐15‐ benzyl‐3,6,18,21,36‐pentakis(carboxymethyl)‐24‐[(4‐hydroxyphenyl)methyl]‐39,42,45‐ tris[(1H‐indol‐3‐ yl)methyl]‐16‐methyl‐2,5,8,14,17,20,23,26,29,35,38,41,44,47‐tetradecaoxo‐27‐[(3,4,5‐ trifluorophenyl)methyl]‐31‐thia‐1,4,7,13,16,19,22,25,28,34,37,40,43,46‐ tetradecaazatricyclo[46.3.0.0^9,13]henpentacontan‐33‐yl]formamido}acetamido)‐4‐ carboxybutanamido]ethoxy}ethoxy)acetamido]‐4‐ carboxybutanamido]ethoxy}ethoxy)acetamido]pent‐4‐ynoic acid (81 mg, 0.030 mmol) and tert-butyl (S)-1-azido-40-(tert-butoxycarbonyl)-37,42-dioxo- 3,6,9,12,15,18,21,24,27,30,33-undecaoxa-36,41-diazanonapentacontan-59-oate (34.5 mg, 0.032 mmol), following the general synthesis described for the preparation of Example 1150. The crude material was purified via preparative LC/MS. The yield of the product was 5.8 mg, and its estimated purity by LCMS analysis was 98.1%. Analysis condition B: Retention time = 1.89 min; ESI-MS(+) m/z [M+3H]³⁺: 1238.1. The following examples were prepared, on a 50 or 100 ^mol scale, using the crude parent macrocycle with a side chain amine, which was generated using the general synthetic sequence of Example 1000, following the “Fatty acid chain coupling procedure A or B” general procedure with the corresponding fatty acid tail. Preparation of Example 1325 Prearation of Examle 1329 LAG-3 cell binding assay: Human Raji cells expressing endogenous MHC Class II molecules were used for binding to either human LAG-3-mFc, mouse LAG-3, or cyno LAG-3-hFc proteins. Briefly Raji cells were plated in a 384-well plate (Corning 354663) at a density of 8000 cells/well. After 2 hour incubation at a 37 ^C and 5% CO2 incubator, LAG-3 antigen (hLAG-3 –mFc, mLAG-3-mFc, or cLAG-3-hFc) were added to all wells at a final concentration of 0.088, 0.25, or 0.072 µg/ml and incubated for 30 minutes. Then, a corresponding detection antibody (R-Phycoerythrin conjugated anti-Mouse IgG, or anti-human IgG), Jackson Immuno Research Lab, PA) was added. The binding affinity of the LAG-3 antigen was quantified by reading the plate on a NXT High Content Reader (ThermoFisher). To assess the potency of LAG-3 compounds to block the binding of LAG-3 antigen to the MHCII molecules expressed on the Raji cell surface, compounds were serially diluted and added to the Raji cells prior to the addition of an appropriate LAG3 antigen. A: IC₅₀ < 0.001 ^M, B: 0.001 ^M <= IC₅₀ < 0.01 ^M; C: 0.01 ^M <= IC50 < 0.1 ^M; D: 0.1 ^M <= IC50 < 1 ^M. Table 1 Biological Data for Examples 1001-1342 ^{Example No.} _{hLag3 Raji CBA HC IC50} 1⁰⁴² ^{Example No} _{hLa 3 Raji CBA HC IC}

Claims

CLAIMS We claim: 1. A compound selected from Example 1001, Example 1002, Example 1003, Example 1004, Example 1005, Example 1006, Example 1007, Example 1008, Example 1009, Example 1010, Example 1011, Example 1012, Example 1013, Example 1014, Example 1015, Example 1016, Example 1017, Example 1018, Example 1019, Example 1020, Example 1021, Example 1022, Example 1023, Example 1024, Example 1025, Example 1026, Example 1027, Example 1028, Example 1029, Example 1030, Example 1031, Example 1032, Example 1033, Example 1034, Example 1035, Example 1036, Example 1037, Example 1038 and Example 1039, Example 1040, Example 1041, Example 1042, Example 1043, Example 1044, Example 1045, Example 1046, Example 1047, Example 1048, Example 1049, Example 1050, Example 1051, Example 1052, Example 1053, Example 1054, Example 1055, Example 1056, Example 1057, Example 1058, Example 1059, Example 1060, Example 1061, Example 1062, Example 1063, Example 1064, Example 1065, Example 1066, Example 1067, Example 1068, Example 1069, Example 1070, Example 1071, Example 1072, Example 1073, Example 1074, Example 1075, Example 1076, Example 1077, Example 1078, Example 1079, Example 1080, Example 1081, Example 1082, Example 1083, Example 1084, Example 1085, Example 1086, Example 1087, Example 1088, Example 1089, Example 1090, Example 1091, Example 1092, Example 1093, Example 1094, Example 1095, Example 1096, Example 1097, Example 1098, Example 1099, Example 1100, Example 1101, Example 1102, Example 1103, Example 1104, Example 1105, Example 1106, Example 1107, Example 1108, Example 1109, Example 1110, Example 1111, Example 1112, Example 1113, Example 1114, Example 1115, Example 1116, Example 1117, Example 1118, Example 1119, Example 1120, Example 1121, Example 1122, Example 1123, Example 1124, Example 1125, Example 1126, Example 1127, Example 1128, Example 1129, Example 1130, Example 1131, Example 1132, Example 1133, Example 1134, Example 1135, Example 1136, Example 1137, Example 1138, Example 1139, Example 1140, Example 1141, Example 1142, Example 1143, Example 1144, Example 1145, Example 1146, Example 1147, Example 1148, Example 1149, Example 1150, Example 1151, Example 1152, Example 1153, Example 1154, Example 1155, Example 1156, Example 1157, Example 1158, Example 1159, Example 1160, Example 1161, Example 1162, Example 1163, Example 1164, Example 1165, Example 1166, Example 1167, Example 1168, Example 1169, Example 1170, Example 1171, Example 1172, Example 1173, Example 1174, Example 1175, Example 1176, Example 1177, Example 1178, Example 1179, Example 1180, Example 1181, Example 1182, Example 1183, Example 1184, Example 1185, Example 1186, Example 1187, Example 1188, Example 1189, Example 1190, Example 1191, Example 1192, Example 1193, Example 1194, Example 1195, Example 1196, Example 1197, Example 1198, Example 1199, Example 1200, Example 1201, Example 1202, Example 1203, Example 1204, Example 1205, Example 1206, Example 1207, Example 1208, Example 1209, Example 1210, Example 1211, Example 1212, Example 1213, Example 1214, Example 1215, Example 1216, Example 1217, Example 1218, Example 1219, Example 1220, Example 1221, Example 1222, Example 1223, Example 1224, Example 1225, Example 1226, Example 1227, Example 1228, Example 1229, Example 1230, Example 1231, Example 1232, Example 1233, Example 1234, Example 1235, Example 1236, Example 1237, Example 1238, Example 1239, Example 1240, Example 1241, Example 1242, Example 1243, Example 1244, Example 1245, Example 1246, Example 1247, Example 1248, Example 1249, Example 1250, Example 1251, Example 1252, Example 1253, Example 1254, Example 1255, Example 1256, Example 1257, Example 1258, Example 1259, Example 1260, Example 1261, Example 1262, Example 1263, Example 1264, Example 1265, Example 1266, Example 1267, Example 1268, Example 1269, Example 1270, Example 1271, Example 1272, Example 1273, Example 1274, Example 1275, Example 1276, Example 1277, Example 1278, Example 1279, Example 1280, Example 1281, Example 1282, Example 1283, Example 1284, Example 1285, Example 1286, Example 1287, Example 1288, Example 1289, Example 1290, Example 1291, Example 1292, Example 1293, Example 1294, Example 1295, Example 1296, Example 1297, Example 1298, Example 1299, Example 1300, Example 1301, Example 1302, Example 1303, Example 1304, Example 1305, Example 1306, Example 1307, Example 1308, Example 1309, Example 1310, Example 1311, Example 1312, Example 1313, Example 1314, Example 1315, Example 1316, Example 1317, Example 1318, Example 1319, Example 1320, Example 1321, Example 1322, Example 1323, Example 1324, Example 1325, Example 1326, Example 1327, Example 1328, Example 1329, Example 1330, Example 1331, Example 1332, Example 1333, Example 1334, Example 1335, Example 1336, Example 1337, Example 1338, Example 1339, Example 1340, Example 1341, Example 1342, or a pharmaceutically acceptable salt thereof. 2. A compound according to claim 1 wherein the compound shows activity less than or equal to 1 µM in the LAG-3 cell binding assay. 3. The compound or a pharmaceutically acceptable salt thereof according to claim 2 as shown in the following tables: O OH R O m HN H Ex. # n m R

n m R X Table 3 Ex. # R^f R⁶ Rⁱ R⁹ m 1263 H (S)- 1 O Table 4 Ex. # X¹ Rⁱ R⁹ X Ex. # X¹ Rⁱ R⁹ X Ex. # X¹ Rⁱ R⁹ X 1227 OH

E^{x. #} R⁴ X R⁷ R¹³ n

E

a e Ex. # R¹³ R¹⁵ R¹⁶ R¹⁷ n 1^{070 H 1}

Table 8 Ex. No. L X

Table 9 Ex. R¹⁵ L₁ L₂ L₃ L₄ R No Ex R¹⁵ L₁ L₂ L L₄ R

Table 11 Example R¹ R⁴ Rⁱ R⁹ R¹¹ L #

a e Ex. # R¹³ R¹⁵ L n 112

Table 13

E 1299 1 4. A pharmaceutical composition comprising one or more compounds according to claim 1 in a pharmaceutically acceptable carrier. 5. A pharmaceutical composition comprising one or more compounds according to claim 2 in a pharmaceutically acceptable carrier. 6. A pharmaceutical composition comprising one or more compounds according to claim 3 in a pharmaceutically acceptable carrier.