Macrocyclic peptides useful as immunomodulators

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 / 563,683 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 signalling 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 signalling (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 withMHC 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 signalling 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 ak, Nat. Tmmunok 2009; 10:29-37 and Grosso JF, et ak, 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 ak, 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 macrocylic 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 are capable of inhibiting the interaction of LAG-3 with MHC class II. These compounds have demonstrated highly efficacious binding to LAG-3, blockade of the interaction of LAG-3 with MHC Class II, and arecapable of promoting 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 aR1, R2, R3, R5, R6, R7, R8, R9, R10, R11, R12, and R13are independently selected from a natural amino acid side chain and an unnatural amino acid side chain, wherein one and only one of R5, R6, R7, is selected from the following or the natural amino acid side chain and an unnatural amino acid side chain on R5, or R6, or R7is substituted by the following: R17, -(C(R17a)2)1-3-O-X’-R30, -(C(R17a)2)1-2-phenyl-O-R17, -(C(R17a)2)1-3CONH-X’-R30,-(C(R17a)2)1-4 NHCO -X’-R30, -(C(R17a)2)1-3-O-(C(R17a)(R17))1-3-H, -(C(R17a)2)1-3CONH-(C(R17a)(R17))1-3-H, -(C(R17a)2)1-4 NHCO -(C(R17a)(R17))1-3-H, -(C(R17a)2)1-6-phenyl-(C(R17a)2)0-2-O-X’-R30, -(C(R17a)2)1-6-phenyl-(C(R17a)2)0-3-CONH-X’-R30, -(C(R17a)2)1-6-phenyl-(C(R17a)2)0-3-NHCO-X’-R30, -(C(R17a)2)1-6-phenyl-(C(R17a)2)0-2-O-(C(R17a)(R17))1-3-H, -(C(R17a)2)1-6-phenyl-(C(R17a)2)0-3-CONH-(C(R17a)(R17))1-3-H, -(C(R17a)2)1-6-phenyl-(C(R17a)2)0-3-NHCO-(C(R17a)(R17))1-3-H, -(C(R17aR17))0-2-X’-R30, -(C(R17aR17))0-4C(O)N(R16a)C(R17a)2-X’-R31, -C(R17aR17)0-2[C(O)N(R16a)C(R17aR17)1-2]w’ –X’-R31, -(C(R17aR17)1-2C(O)NR16aPEGn)m’-X’- R30; and -(C(R17aR17)1-2C(O)NR16a)m’-C(R17a)(R17)-CO2H; -(C(R17aR17)1-2C(O)NR16a-PEGn’)m’-C(R17a)(R17)-CO2H; wherein: w’ is 1-5; m’ is 0-6; n’ is 0-20; 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, provided that X’ is other than unsubstituted PEG;R30is selected from –CO2H, -C(O)NH2, and -CH3, tetrazole, and lithocolic acid and analogs of lithocolic acid; R31is -CO2H, -C(O)NH2, -CH3, tetrazole, and lithocolic acid and analogs of lithocolic acid; each R16ais independently selected from hydrogen, C1-C6alkyl, -CH2OH, -(CH2)1-4NH2, -(CH2)1-4NH-C(=NH)NH2, -CH2CO2H, -(CH2)2CO2H; each R17ais independently selected from hydrogen, C1-C6alkyl, -CH2OH, -(CH2)1-4NH2, - (CH2)1-4NH-C(=NH)NH2, -CH2CO2H, -(CH2)2CO2H; each R17is independently selected from hydrogen, -CH3, -(CH2)zNH2, – (C(R17a)2)0-4-X’-R31, -(CH2)zCO2H, –CH2OH, CH2C=CH, and -(CH2)z-triazolyl-X’-R35, wherein z is 1-6 and R35is selected from -CO2H, -C(O)NH2, CH3, -C(O)-(CH2)2–C(O)O- vitamin E,–C(O)O-vitamin E, tetrazole, and lithocolic acid and analogs; R15is selected from OH, 0-20 PEG units, NR18R19, wherein R18and R19can be independently selected from, hydrogen, 1-3 alkyl, (1-3)alkyl-5-6-carbocycle or heterocycle wherein each ring is optionally substituted with one to four groups independently selected from amino, cyano, methyl, halo, and hydroxy, -(CHR20)1-2C(O)NR18R19, -(CHR20)1-2C(O)OH, -(CHR20)1-2C(O)NH(CHR21)1-2C(O)NR18R19, -(CHR20)1-2C(O)NH(CHR21)1-2C(O)OH, -(CHR20)1-2C(O)NH(CHR21)1-2C(O)NH(CHR22)1-2NR18R19, -(CHR20)1-2C(O)NH(CHR21)1-2C(O)NH(CHR22)1-2C(O)OH; -(CHR20)1-2C(O)NH(CHR21)1-2C(O)NH(CHR22)1-2C(O)NH(CHR23)1-2C(O)NR18R19, -(CHR20)1-2C(O)NH(CHR21)1-2C(O)NH(CHR22)1-2C(O)NH(CHR23)1-2C(O)OH; wherein R20is selected from hydrogen and sidechain of a natural or unnatural amino acid and wherein R21is selected from hydrogen and sidechain of a natural or unnatural amino acid and wherein R22is selected from hydrogen and sidechain of a natural or unnatural amino acid and wherein R23is selected from hydrogen and sidechain of a natural or unnatural amino acid; and 0-2 (2-20 PEG units as spacers in between any one of the amino acids;In the second embodiment the present disclosure provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, When R16ais selected from hydrogen, R16is selected from -(C(R17aR17))0-2-X’-R30, -(C(R17aR17))1-2C(O)N(R16a)C(R17a)2-X’-R31, -C(R17aR17)1-2[C(O)N(R16a)C(R17aR17)1-2]w’–X’-R31, -(C(R17aR17)1-2C(O)NR16a)m’-X’- R30; -(C(R17aR17)1-2C(O)NR16a)m’-C(R17a)(R17)-CO2H; -(C(R17aR17)1-2C(O)NR16a)m’-C(R17a)(R17)-CO2H, and -(C(R17aR17))1-2-Phenyl-(C(R17aR17))1-2-X’-R30; and wherein a PEGn’spacer can be inserted in any part of the R16(n’ is the number of –(CH2CH2O)- unit in a PEG spacer); Alternatively, R16and R16atogether with the N atom attached formed the sidechain of either D- or L-proline, wherein the ring can be is substituted with -(C(R17aR17))0-2-X’-R30, -(C(R17aR17))1-2C(O)N(R16a)C(R17a)2-X’-R31, -C(R17aR17)1-2[C(O)N(R16a)C(R17aR17)1-2PEGn’]w’ –X’-R31, -(C(R17aR17)1-2C(O)NR16aPEGn’)m’-X’- R30; and -(C(R17aR17)1-2C(O)NR16aPEGn’)m’-C(R17a)(R17)-CO2H; -(C(R17aR17)1-2C(O)NR16a-PEGn’)m’-C(R17a)(R17)-CO2H; and wherein a PEGn’ spacer can be inserted in any part of the R16(n’ is the number of –(CH2CH2O)- unit in a PEG spacer); and wherein w’ is 1-5; m’ is 0-5; n’ is 1-12;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, provided that X’ is other than unsubstituted PEG; R30is selected from –CO2H, -C(O)NH2, and -CH3, tetrazole, and lithocolic acid and analogs of lithocolic acid; R31is -CO2H, -C(O)NH2, -CH3, tetrazole, and lithocolic acid and analogs of lithocolic acid, and biotin; each R17ais independently selected from hydrogen, C1-C2alkyl, -CH2OH, -(CH2)1-4NH2, -(CH2)1-4NH-C(=NH)NH2, -CH2CO2H, -(CH2)2CO2H, each R17is independently selected from hydrogen, -CH3, -(CH2)zNH2, – (C(R17a)2)0-4-X’-R31, -(CH2)zCO2H, –CH2OH, CH2C=CH, and -(CH2)1-3-triazolyl-X’-R35, and R35is selected from -CO2H, -C(O)NH2, CH3, tetrazole, and lithocolic acid and analogs; provided that at least one R17is other than hydrogen, -CH3, or –CH2OH; 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, squamouscell 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 compounds of formula (I), R1group is selected from sidechain of the following amino acids: phenylalanine, tyrosine, tryptophan, leucine, 2-fluorophenylalanine, 3- fluorophenylalanine, 4-fluorophenylalanine, 3,4-difluorophenylalanine, 3,5- difluorophenylalanine, 3,4-dichlorophenyl, 3,4,5-difluorophenylalanine, pentafluorophenylalanine, 3-trifluoromethylphenylalanine, 4-methylphenylalanine, 4- chlorophenylalanine, 3-methoxyphenylalananie, 4-methoxyphenylalanine, 1- naphthylalanine (1-Nal), 2-naphthylalanine (2-Nal), 3-cyanophenylalanine, 4- cyanophenylalanine, 4-difluoromethylphenylalanine, biphenylalanine, 3-(2-thienyl)- alanine, 3-(3-thienyl)-alanine, 3-benzothienylalanine (Bzt), 4-bromophenylalanine, 3- pyridinylalanine, 4-trifluoromethylphenylalanine, 4-benzoylphenylalanine (Bpa), 3- chlorophenylalanine, 4-aminophenylalanine, 4-aminomethylphenylalanine, 4- carbamoylphenylalanine, 3-carbamoylphenylalanine, 4-carboxyphenylalaine, 3-(4- thiazolyl)-alanine, 4-acetamide-phenylalanine, homo-cyclohexylalanine, 2-fuanylalanine, homo-tyrosine, 4-phenoxyphenylalanine, 4-cyclohexyloxyphenylalanine, 4- propargyloxyphenylalanine, tyrosine(O-acetic acid), 4-benoxyphenylalanine, trypophane(1-acetic acid), and 4-tert-butylphenylalanine;In compounds of formula (I), R2group is selected from sidechain of the following amino acids: alanine, 2-aminobutyric acid (Abu), cyclopropylglycine, threonine, and valine; In compounds of formula (I), R3group is selected from sidechain of the following amino acids: alanine, 3-(2-thienyl)-alanine, 3-(3-thienyl)-alanine, 3-benzothienylalanine (Bzt), 4-benzoylphenylalanine (Bpa), allothreonine, aspartic acid, citrulline (Cit), asparagine, glutamic acid, homo-glutamic acid, 2,3-diaminopropionic acid (Dap), phenylalanine, 2-fluorophenylalanine, 3-fluorophenylalanine, 4-fluorophenylalanine, 3- methylphenylalanine, 4-methylphenylalanine, 3-methoxyphenylalanine, 4- aminophenylalanine, 7-quinolinylalanine, homophenylalanine, homoglutamide, homoserine, leucine, 4-carboxyphenylalamine, tyrosine, glutamine, serine, 2-amino-4- aminobutyric acid (Dab), threonine, 4-carboxymethoxyphenyl-alanine, tryptophan, tryptophan (1-acetic acid), and tryptophane (5-benzoxy), tryptophan (1-methyl), tyrosine, and tyrosine(O-acetic acid). In compounds of formula (I), R5group is selected from sidechain of the following amino acids: alanine, citrulline, 1-aminocyclopentane-1-carboxylic acid (Cle), 3-hydroxy 1-aminocyclopentane-1-carboxylic acid (Cle(3-OH)), aspartic acid, D-alanine, 2-amino- 4-aminobutyric acid (Dab), Dab(COCH2NH2), Dab(COCH2NHBoc), Dab(COCH2CH2NH2), Dab(COCH2CH2NHBoc), (S)-2-(^2-azaneyl)-4- guanidinobutanoic acid (Dab(-CH(=NH)NH2)), Dab(CO-ethyl), Dab(CH2-4- (CH2NH2)phenyl), Dab(CH2-4-(CH2NHBoc)phenyl), Dab(CONH-isopropyl), 2,3- diaminopropionic acid (Dap), Dap(CH(=NH)NH2), D-glutamic acid, D-histidine, D- homoarginine, D-lyscine, D-lysine(COCH3), D-asparagine, D-glutamine, D-arginine, D- serine, D-valine, glutaminic acid, phenylalanine, glycine, histidine, homoarginine, lysine, leucine, asparagine, ornithine (Orn), phenylglycine, propargylalanine, glutamine, arginine, valine, tryptophan (1-acetic acid), tyrosine (O-acetic acid), tyrosine (O- propargyl), and ^-alanine; In compounds of formula (I), R6group is selected from sidechain of the following amino acids: 1-naphthylalanine (1-Nal), 2-naphthylalanine (2-Nal), alanine, biphenylalanine (Bip), 3’-carboxylic acid-biphenylalanine, 4’-carboxylic acid- biphenylalanine, 3-benzothienylalanine (Bzt), 4-benzoylphenylalanine (Bpa),phenylalanine, 3,4,5-trifluorophenylalanine, 3,4-dichlorophenylalanine, 3,5- difluorophenylalanine, 3-chlorophenylalanine, 4-chlorophenylalanine, 4- cyanophenylalanine, 4-aminomethylphenylalanine, 4-carbamoylphenylalanine, 4- carboxyphenylalaine, 4-aminophenylalanine, alpha-methylphenylalanine, beta-hydroxy- phenylalanine, isoleucine, histidine, leucine, phenyglycine (Phg), valine, tryptophan, tryptophan (1-acetic acid), tryptophan (5-benzoxy), tyrosine, tyrosine (O-acetic acid), tyrosine (O-propargyl), tyrosine (O-phenyl), and 2S)‐2‐amino‐3‐({4‐[(2S)‐2‐amino‐2‐ carboxyethyl]phenyl}formamido)propanoic acid; In compounds of formula (I), R7group is selected from sidechain of the following amino acids: alanine, aspartic acid, citrulline, 3-diaminopropionic acid (Dap), 2-amino-4- aminobutyric acid (Dab), Dap(COEt), Dap(CH(=NH)NH2), glutamic acid, glycine, histidine, 4-carboxyphenylalanine, homoarginine, lysine, asparagine, ornithine (Orn), arginine, serine, threonine, tryptophan, tryptophan (1-acetic acid), tyrosine, tyrosine (O- acetic acid), and tyrosine (O-propargyl); In compounds of formula (I), R8group is selected from sidechain of the following amino acids: 4-cyclohexyloxyphenylalanine, 1-naphthylalanine (1-Nal), 2- naphthylalanine (2-Nal), (2S)‐2‐amino‐3‐[6‐(2‐methylphenyl)pyridin‐3‐yl]propanoic acid, biphenylalanine, 2'-methoxycarbonyl-biphenylalanine, 2'-ethyl-biphenylalanine, 2'- ethyl, 4'-hydroxy-biphenylalanine, 2'-ethyl, 4'-methyoxy-biphenylalanine, 2'-fluoro- biphenylalanine, 2'-methyl-biphenylalanine, 2'-n-butoxyl-biphenylalanine, 2'-methyoxy, 4'-fluoro-biphenylalanine, 3',5'-difluoro-biphenylalanine, 3'-cyanobiphenylalanine, 3'- carboxybiphenylalanine, 3'-methyl biphenylalanine, 4'-chlorobiphenylalanine, N- cyclopropyl-4'-carboxamide-biphenylalanine, 4'-dimethylcarbamoyl-biphenylalanine, 4'- carboxybiphenylalanine, 4'-acetamido-biphenylalanine, 4'-phenoxy-biphenylalanine, (3S)‐3‐amino‐4‐[5‐(2‐methylphenyl)pyrimidin‐2‐yl]‐2‐oxobutanoic acid, 4- benzoylphenylalanine (Bpa), (2S)‐2‐amino‐3‐[4‐(1‐benzofuran‐2‐yl)phenyl]propanoic acid, (2S)‐2‐amino‐3‐[4‐(3‐cyanothiophen‐2‐yl)phenyl]propanoic acid, (2S)‐2‐amino‐3‐ {4‐[(E)‐2-phenylethenyl]phenyl}propanoic acid (Phe(4-CH=CH-phenyl)), 4- iodophenylalanine, 4-isopropylphenylalanine, 4-tert-butylphenylalanine, (2S)‐2‐amino‐3‐ [4‐(6‐methoxypyridin‐3‐yl)phenyl]propanoic acid, (2S)‐2‐amino‐3‐[4‐(pyrrolidine‐1‐ carbonyl)phenyl]propanoic acid, homoserine(O-benzyl), tryptophan, tryptophan(5- benzoxy), tyrosine(O-propargyl), and tyrosine(O-phenyl);In compounds of formula (I), R9group is selected from sidechain of the following amino acids: 1-naphthylalanine (1-Nal), 6-quinolinylalanine, 8-isoquinolinylalanine, 4- isoquinolinylalanine, alanine (3-methoxy-isoquinolin-7-yl), 6-isoquinolinylalanine, 2- naphthylalanine (2-Nal), biphenylalanine, 4-benzoylphenylalanine (Bpa), 3- benzothienylalanine (Bzt), phenylalanine, 2-chlorophenylalanine, 3,4- dichlorophenylalanine, 3,4-difluorophenylalanine, 3,5-difluorophenylalanine, 3- trifluoromethylphenylalanine, 3-chlorophenylalanine, 3-methoxyphenylalanine, 4- chlorophenylalanine, 4-fluorophenylalanine, 4-aminophenylalanine, 4- carbamoylphenylalanine, 4-tert-butylphenylalanine, homohomophenylalanine, homoserine (O-2-methyl-4-chlorophenyl), homotyrosine, tryptophan, and tryptophan(1- acetic acid); In compounds of formula (I), R10group is selected from sidechain of the following amino acids: aspartic acid, 4-carboxyphenylalanine, tyrosine(O-acetic acid), and tryptophan(1-acetic acid); In compounds of formula (I), R11group is selected from sidechain of the following amino acids: 1-naphthylalanine (1-Nal), 2-naphthylalanine (2-Nal), 2‐amino‐3‐ (1H‐indol‐4‐yl)propanoic acid, phenylalanine, 3,4-dimethoxyphenylalanine, 3- methoxyphenylalanine, 4-aminophenylalanine, 1-methyltryptophane, asparagine, tryptophan, tyrosine, 5-hydroxytryptophane, and tyrosine(O-acetic acid); In compounds of formula (I), R12group is selected from sidechain of the following amino acids: alanine, alpha, alpha-dimethylglycine (Aib), aspartic acid, D- alanine, D-aspartic acid, D-glutamic acid, D-phenylalanine, D-histidine, D-homo- arginine, D-lysine, D-leucine, D-asparagine, D-glutamine, D-arginine, D-serine, D- threonine, D-valine, D-tryptophan, D-tyrosine, glutamic acid, glycine, serine, tryptophan(1-scetivc acid), tyrosine(O-acetic acid), and tyrosine(O-propargyl); In another embodiment, compounds of formula (I), wherein, R1group is selected from sidechain of the following amino acids: L- Ala(^-3-pyridinyl) (L-3-Pya), L-Phe(4-COPh) (L-Bpa), D-Tyr(O-benzyl), L-Phe, L- Phe(2-F), L-Phe(2-Cl), L-Phe(3,4,5-tri-F), L- L-Phe(3,5-di-F), L-Phe(3-Cl), L-Phe(3-CF3), L-Phe(3-CN), L-Phe(4-CN), L-Phe(4-CONH2), L-Phe(4-COOH), L-Phe(3-CONH2), L-Phe(penta-F), L-Tyr, L-Tyr(benzyl), L-Tyr(CH2COOH), L- Tyr(propargyl), and L-Tyr(phenyl); R2group is selected from sidechain of the following amino acids: L-Ala, L-Abu, L-Gly(^-cyclopropyl), and L-Val; R3group is selected from sidechain of the following amino acids: L-alloT, L-Asp, L-Phe(4-COOH), L-homo-Glu, L-Homo-Phe, L-homo-Gln, L-homo-Ser, L-Asn, L-Gln, L-Trp, L-Trp(1-CH2COOH), L-Trp(5-OBn), L-Tyr, and L-Tyr(CH2COOH); R5group is selected from sidechain of the following amino acids: L-Dap(- C(=NH)NH2), L-Cit, L-Asp, L-Dab, L-Dab(CH2Phenyl-4-CH2NH2), L- Dab(COCH2CH2NH2), L-Dab(COCH2NH2), L-Dab(COEt), Dab(G), L-Dap, D-Glu, D- His, D-Asn, D-Gln, D-Arg, D-Val, L-Glu, L-Phe, Gly, L-His, L-Hyp, L- Dab(COCH2NHBoc), L-Dab(COCH2CH2NHBoc), L-Asn, L-Orn, L-Pro, L-Pro(NH- C(=NH)NH2), L-Pro(4-NH2), L-Phg, L-Pra, L-Gln, L-Arg, L-Val, L-Trp(1-CH2COOH), L-Tyr(CH2COOH), and L-Tyr(Pra), R6group is selected from sidechain of the following amino acids: L-1-Nal, L-2- Nal, L-Bip, L-Bip(3'-COOH), L-Bip(4'-COOH), L-Bpa, L-Bzt, L-Phe(3,4,5-tri-F), L- Phe(3,4-di-Cl), L-Phe(3,5-di-F), L-Phe(3-Cl), L-Phe(4-Cl), L-Phe(4-CN), L-Phe(4- CH2NH2), L-Phe(4-CONH2), L-Phe(4-COOH), L- L- Trp, L-Trp(1-CH2COOH), L-Trp(5-O-benzyl), L-, and L-Tyr(propargyl); R7group is selected from sidechain of the following amino acids: L- Dap(C(=NH)NH2), L-Dab, L-Dap, L-Phe(4-COOH), Gly, L-His, L-homoArg, L-Lys, L- Asn, L-Orn, L-Arg, L-Ser, L-Thr, L-Trp, L-Tyr, L-Tyr(CH2COOH), L-Tyr(propargyl), and L-Trp(CH2COOH); R8group is selected from sidechain of the following amino acids: L- Tyr(cyclohexyl), L-2-Nal, L-Bip, L-Bip(2'-COOMe), L-Bip(2'-Et), L-Bip(2'-Et,4'-OH), L-Bip(2'-Et, ,4'-OMe), L-Bip(2'-F), L-Bip(2'-Me), L-Bip(2-O-nBu), L-Bip(2'-OMe,4'-F),L-Bip(3',5'-di-F), L-Bip(3-CN), L-Bip(3'-COOH), L-Bip(3'-Me), L-Bip(4'-Cl), L-Bip(4'- CONH-cyclopropyl), L-Bip(4'-CONMe2), L-Bip(4'-COOH), L-Bip(4'-NHCOMe), L- Bip(4'-OPh), L-Bpa, L-Phe(4-(2-benzofuranyl)), L-Phe(4-(2-(3-CN-thienyl)), L-Phe(4- CH=CH-Ph), L-Phe(4-iodo), (2S)‐2‐amino‐3‐[4‐(6‐methoxypyridin‐3‐ yl)phenyl]propanoic acid, and L-Tyr(Ph); R9group is selected from sidechain of the following amino acids: L-1-Nal, L- Ala(^-quinolin-6-yl), L-Ala(^-isoquinolin-8-yl), L-Ala(^-isoquinolin-4-yl), L-Ala(^- quinolin-3-yl), L-Ala(^-3-methoxy-isoquinolin-7-yl), L-Bzt, L-Phe, L-Phe(2-Cl), L- Phe(3,4-di-F), L-Phe(3,5-di-F), L-Phe(3-CF3), L-Phe(3-OMe), L-Phe(4-Cl), L-Phe(4- CONH2), L-homohomoPhe, L-homoSer(2-Me,4-Cl-phenyl), L-homoTyr, L-Trp, ,Phe(4-COOH), and L-Tyr(CH2COOH); R11group is selected from sidechain of the following amino acids: L-1-Nal, L-2- Nal, L-Ala(^-indol-4-yl), L-Phe(3-OMe), L-Phe(4-NH2), L-Trp, L-Trp(5-OH), L- Trp(Me), and L-Tyr; R12group is selected from sidechain of the following amino , Aib, L-Asp, D-Ala, D-Asp, D-Glu, D-Phe, D-His, D-Lys, D-Leu, D-Asn,Arg, D- Ser, D-Thr, D-Trp, D-Tyr, L-Glu, Gly, L-Ser, L-Trp(1-CH2COOH), and L- Tyr(CH2COOH); R17ais each independently selected from hydrogen, CH3, -CH2OH, -(CH2)1-4NH2, -(CH2)1-4NH-C(=NH)NH2, -CH2CO2H, -(CH2)2CO2H, R17is each independently selected from hydrogen, -CH3, -(CH2)zNH2, – (C(R17a)2)0-4-X’-R31, -(CH2)zCO2H, –CH2OH, CH2C=CH, and -(CH2)1-3-triazolyl-X’-R35, and R35is selected from -CO2H, -C(O)NH2, CH3, tetrazole, and lithocolic acid and analogs; provided that at least one R17is other than hydrogen, -CH3, or –CH2OH;And one and only one of R5, R6, R7groups is selected from the following or the natural amino acid side chain and an unnatural amino acid side chain on R5, or R6, or R7groups is substituted by the following:R17, -(C(R17a)2)1-2-O-X’-R30, -(C(R17a)2)1-2-phenyl-O-R17, -(C(R17a)2)1-2CONH-X’-R30, -(C(R17a)2)1-4 NHCO -X’-R30, -(C(R17a)2)1-2-O-(C(R17a)(R17))1-3-H, -(C(R17a)2)1-3CONH-(C(R17a)(R17))1-3-H, -(C(R17a)2)1-4 NHCO -(C(R17a)(R17))1-3-H, -(C(R17a)2)1-3-phenyl-(C(R17a)2)0-2-O-X’-R30, -(C(R17a)2)1-3-phenyl-(C(R17a)2)0-3-CONH-X’-R30, -(C(R17a)2)1-3-phenyl-(C(R17a)2)0-3-NHCO-X’-R30, -(C(R17a)2)1-3-phenyl-(C(R17a)2)0-2-O-(C(R17a)(R17))1-3-H, -(C(R17a)2)1-3-phenyl-(C(R17a)2)0-3-CONH-(C(R17a)(R17))1-3-H, -(C(R17a)2)1-3-phenyl-(C(R17a)2)0-3-NHCO-(C(R17a)(R17))1-3-H, -(C(R17aR17))0-2-X’-R30, R18, R19or R20group is each independantly selected from the H, (CH2)1-4, sidechains of the following amino acid: Gly, L-Ala, L-Glu, L-Asp, L-Lys, D-Arg, L-Dap, L-Dab, L-Orn, L-Asn, D-Ser, L-Pra, D-Dap, D-Dab, D-Orn, L-mLeu, L-Pro, D-Pro, L- Trp, L-mAla, L-Tic, D-Asp, L-mPhe, D-Arg, D-Lys, L-Phe, D-Asn, D-Orn, L-Tic, and D- Ala; when NR18R19is at the C-terminal, R18and R19is each independantly selected from H, C1-6 alkyl, (CH2)1-4cycloalkyl, (CH2)1-45-6 carbocycles and heterocycles; R21, R22or R23group is each independantly selected from H, sidechains of the following amino acid: L-^-Glu, L-Dab, L-Dap, D-Arg, L-Glu, Gly, L-Ser L-Orn, L-Pra; Preferred PEG spacer: -(CH2CH2O)1-11CH2CH2OCH2CO-; In another embodiment, compounds of formula (I),wherein, R1group is selected from sidechain of the following amino acids: L-Phe(4-COPh) (L-Bpa), L-Phe, L-Phe(3,4,5-tri-F), L- L-Phe(3,5-di-F), L-Phe(3-CN), L-Phe(4-CN), L- Phe(4-CONH2), L-Phe(4-COOH), L-Phe(3-CONH2), L-Tyr(benzyl), and L- Tyr(CH2COOH); R2group is L-Val; R3group is selected from sidechain of the following amino acids: L-alloT, L-Asp, L-Trp, L-Trp(1-CH2COOH), L-Trp(5-OBn), and L-Tyr; R5group is selected from sidechain of the following amino acids: L-Asp, L-Dab, L-Dab(COCH2NH2), D-His, D-Arg, Gly, L-Hyp, and L-Tyr(CH2COOH); R6group is selected from sidechain of the following amino acids: L-2-Nal, L- Bip(3'-COOH), L-Bip(4'-COOH), L-Phe(4-Cl), L-Phe(4-CH2NH2), L-Phe(4-CONH2), L- L-Trp(1-CH2COOH), L-Trp(5-O-benzyl), L-Tyr, L- L-Tyr(propargyl);R7group is selected from sidechain of the following amino acids: L-Dap, L- Phe(4-COOH), L-Asn, L-Ser, L-Thr, L-Tyr, and L-Tyr(propargyl); R8group is selected from sidechain of the following amino acids: L-Bip, L-Bip(2'- COOMe), L-Bip(2'-Et,4'-OH), L-Bip(2'-F), L-Bip(2'-Me), L-Bip(2-O-nBu), L-Bip(2'- OMe,4'-F), L-Bip(3',5'-di-F), L-Bip(3-CN), L-Bip(3'-COOH), L-Bip(3'-Me), L-Bip(4'- Cl), L-Bip(4'-CONMe2), L-Bip(4'-COOH), L-Bip(4'-NHCOMe), L-Bip(4'-OPh), L- Phe(4-(2-benzofuranyl)), L-Phe(4-(2-(3-CN-thienyl)), L-Phe(4-CH=CH-Ph), and L- Tyr(Ph); R9group is selected from sidechain of the following amino acids: L-1-Nal, L- Ala(^-quinolin-6-yl), L-Ala(^-isoquinolin-8-yl), L-Ala(^-isoquinolin-4-yl), L-Ala(^- quinolin-3-yl), L-Ala(^-3-methoxy-isoquinolin-7-yl), L-Bzt, L-Phe, L-Phe(2-Cl), L- Phe(3,4-di-F), L-Phe(3,5-di-F), L-Phe(3-CF3), L-Phe(3-OMe), L-Phe(4-Cl), L-Phe(4-CONH2), L-homohomoPhe, L-homoSer(2-Me,4-Cl-phenyl), L-homoTyr, L-Trp, , L-Trp(5-OH), L-Trp (6-OMe), L-Trp(7-F), L-Trp(7-Me), Trp(Me), L-Tyr, L-Tyr(Bn), and L-Tyr(Me);R11group is selected from sidechain of the following amino acids: L-Ala(^-indol- 4-yl), L-Trp, L-Trp(5-OH), and L-Tyr; R12group is selected from sidechain of the following amino acids: , D-Ser, D-Trp, Gly, and L-Ser,;R17is selected from hydrogen, -(C(R17a)2)0-4-X’-R31and -(CH2)z-triazolyl-X’-R35, wherein z is 1-3 and R35is selected from -CO2H, CH3, tetrazole, -C(O)NH2, and lithocolic acid and analogs; In another embodiment, compounds of formula (I), (<100 nM) wherein, R1group is selected from sidechain of the following amino acids: L-Phe, L- Phe(3,5-di-F), L-Phe(4-CONH2), L-Phe(4-COOH), and L-Phe(3-CONH2); R3group is selected from sidechain of the following amino acids: L-Asp, L-Trp, L-Trp(1-CH2COOH), and L-Tyr; R5group is selected from sidechain of the following amino acids: L-Asp, D-Arg, and Gly; R6group is selected from sidechain of the following amino acids: L-Phe(4- COOH), L-Trp, and L-Tyr; R7group is L-Tyr; R8group is selected from sidechain of the following amino acids: L-Bip, L-Bip(2'- Me), L-Bip(3'-COOH), L-Bip(4'-CONMe2), L-Bip(4'-COOH), and L-Bip(4'-NHCOMe);R9group is selected from sidechain of the following amino acids: L-1-Nal, L- Phe(3-CF3), L-Trp, and L-Tyr; R10group is L-Asp; R11group is selected from sidechain of the following amino acids: L-Trp and L- Tyr; R12group is selected from sidechain of the following amino acids: D-Ser, and Gly; In another aspect of the above embodiment the present disclosure provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, R6groups is selected from R17, -(C(R17a)2)1-2-O-X’-R30, -(C(R17a)2)-phenyl-O-X’-R30, - (C(R17a)2)-phenyl-O-R17, As shown below in Table 1, show the compounds of the invention which activity is less than 50 nM, the data is included later in the application.Example # R3R12X H 1013H 1019 H 1020H 1021H 1022 H 1023OH 1034103510361044 1045Definitions 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: COOH COOH H2N R R NH2acidwhere R and R′ are as discussed herein. 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: 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 IC50to 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 IC50is 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 LAG-3 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 recitedrange 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 lessthan 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 molecules; (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 sidechains 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 The disclosure further relates to the polypeptides described herein wherein the sequence comprises one or more amino acid deletions from either the C-terminus and / or the N-terminus. In preferred embodiments, the following N-terminal Compound No. 99 deletion polypeptides are encompassed by the present disclosure: X1-X13, X2-X13, X3-X13, X4- X13, X5-X13, X6-X13, X7-X13, X8-X13, X9-X13, X10-X13, X11-X13, and / or X12- X13 of Compound No. 99, wherein each X is representative of an amino acid at the indicated position for each peptide as outlined herein. The present disclosure also encompasses cyclic forms of these deletion mutants using the linking chemistries described elsewhere herein. In preferred embodiments, the following C-terminal Compound No. 99 deletion polypeptides are encompassed by the present disclosure: X1-X13, X1-X12, X1-X11, X1- X10, X1-X9, X1-X8, X1-X7, X1-X6, X1-X5, X1-X4 and / or X1-X3 of Compound No. 99, wherein each X is representative of an amino acid at the indicated position for each peptide as outlined herein. The present disclosure also encompasses cyclic forms of these deletion mutants using the linking chemistries described elsewhere herein.In preferred embodiments, the following N-terminal Compound No. 1 deletion polypeptides are encompassed by the present disclosure: X1-X15, X2-X15, X3-X15, X4- X15, X5-X15, X6-X15, X7-X15, X8-X15, X9-X15, X10-X15, X11-X15, and / or X12- X15 of Compound No. 1, wherein each 1 is representative of an amino acid at the indicated position for each peptide as outlined herein. The present disclosure also encompasses cyclic forms of these deletion mutants using the linking chemistries described elsewhere herein. In preferred embodiments, the following C-terminal Compound No. 1 deletion polypeptides are encompassed by the present disclosure: X1-X15, X1-X14, X1-X13, X1- X12, X1-X11, X1-X10, X1-X9, X1-X8, X1-X7, X1-X6, X1-X5, X1-X4 and / or X1-X3 of Compound No. 1, wherein each X is representative of an amino acid at the indicated position for each peptide as outlined herein. The present disclosure also encompasses cyclic forms of these deletion mutants using the linking chemistries described elsewhere herein. In preferred embodiments, the following N-terminal Compound No. 71 deletion polypeptides are encompassed by the present disclosure: X1-X14, X2-X14, X3-X14, X4- X14, X5-X14, X6-X14, X7-X14, X8-X14, X9-X14, X10-X14, X11-X14, and / or X12- X14 of Compound No. 71, wherein each X is representative of an amino acid at the indicated position for each peptide as outlined herein. The present disclosure also encompasses cyclic forms of these deletion mutants using the linking chemistries described elsewhere herein. In preferred embodiments, the following C-terminal Compound No. 71 deletion polypeptides are encompassed by the present disclosure: X1-X14, X1-X13, X1-X12, X1- X11, X1-X10, X1-X9, X1-X8, X1-X7, X1-X6, X1-X5, X1-X4 and / or X1-X3 of Compound No. 71, wherein each X is representative of an amino acid at the indicated position for each peptide as outlined herein. The present disclosure also encompasses cyclic forms of these deletion mutants using the linking chemistries described elsewhere herein. 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 apharmaceutically 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 fromnontoxic 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 anti-oxidant. 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 compositionsof 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, mostpreferably 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 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. In another aspect, the instant disclosure provides a pharmaceutical kit of parts comprising a macrocyclic peptide and an anti-CTLA-4 antibody, as described herein. The kit may also further comprise instructions for use in the treatment of a hyperproliferative disease (such as cancer as described herein) and / or anti-viral disease. 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 description of the present disclosure herein should be construed in congruity with the laws and principals of chemical bonding. It should be understood that the compounds encompassed by the present disclosure are those that are suitably stable for use as pharmaceutical agent. One of skill in the art will know what compounds would and would not be stable based on the general principles of chemical bonding and stability. 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 ThePeptides: 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 OH Fmoc OH OHThe 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 / water / TIS / DTT (94:3:3:1), v:v:v:w; 1 mL / 100 mg of peptidyl resin) for 1.5-3 hrs at room temperature. The spent resin is then filtered off and the TFA solution was cooled and Et2O 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 a 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 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: 50oC; 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: 35oC; Gradient: 5-95% B over 4 minutes; Flow: 4.0 mL / min; Detection: UV at 220 nm.

[0001] 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 = triethylaminemin. = 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 spectrometryNMR = 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 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 acidchain coupling procedure A, Fatty acid chain coupling procedure B, General Purification Procedures. Unnatural Fmoc-Amino Acid Synthesis amd fatty acid tails were prepared according to the published methods (e.g., WO 2023 / 225661). General Purification Procedures: The crude material was purified via preparative LC / MS with the following conditions: Column: XBridge C18, 200 mm x 19 mm, 5-μm particles; Mobile Phase A: 5:95 acetonitrile: water with 0.05% trifluoroacetic acid; Mobile Phase B: 95:5 acetonitrile: water with 0.05% trifluoroacetic acid; Gradient: a 0-minute hold at certain percentage of B, then a linear increase from this percentage to a higher percentage of B over 20-30 minutes, then a 0- minute hold at 100% B; Flow Rate: 20-40 mL / min; Column Temperature: 25 C. Fraction collection was triggered by MS and UV signals. Fractions containing the desired product were combined and dried via centrifugal evaporation. If the material was not pure based on the orthogonal analytical data, it was further purified via preparative LC / MS with the following conditions: Column: XBridge C18, 200 mm x 30 mm, 5-μm particles; Mobile Phase A: 5:95 acetonitrile: water with ammonium acetate; Mobile Phase B: 95:5 acetonitrile: water with ammonium acetate; Gradient: a 0-minute hold at certain percentage of B, then a linear increase from the starting percentage of B over 20-30 minutes, then a 0-minute hold at 100% B; Flow Rate: 20-40 mL / min; Column Temperature: 25 C. Fraction collection was triggered by MS signals. Fractions containing the desired product were combined and dried via centrifugal evaporation. The yield and the purity of the product were determined. Alternatively, based on the initial analytical data, the crude material was purified via preparative LC / MS with the following conditions: Column: XBridge C18, 200 mm x 30 mm, 5-μm particles; Mobile Phase A: 5:95 acetonitrile: water with ammonium acetate; Mobile Phase B: 95:5 acetonitrile: water with ammonium acetate; Gradient: a 0-minute hold at certain percentage of B, then a linear increase from the starting percentage of B over 20 minutes, then a 0-minute hold at 100% B; Flow Rate: 40 mL / min; Column Temperature: 25 C. Fraction collection was triggered by MS signals. Fractions containingthe desired product were combined and dried via centrifugal evaporation. If the material is not pure based on the orthogonal analytical data, it was further purified via preparative LC / MS with the following conditions: Column: XBridge C18, 200 mm x 19 mm, 5-μm particles; Mobile Phase A: 5:95 acetonitrile: water with 0.05% trifluoroacetic acid; Mobile Phase B: 95:5 acetonitrile: water with 0.05% trifluoroacetic acid; Gradient: a 0- minute hold at certain percentage of B, then a linear increase from this percentage to a higher percentage of B over 20 minutes, then a 0-minute hold at 100% B; Flow Rate: 20 mL / min; Column Temperature: 25 C. Fraction collection was triggered by MS and UV signals. Fractions containing the desired product were combined and dried via centrifugal evaporation. The yield and the purity of the product were determined. Preparation of perfluorophenyl stearate, A0934-762-01 (C18) Scheme:2,2,2- trifluoroacetate (0.268 mL, 1.566 mmol), and pyridine (0.186 mL, 2.297 mmol). The vial was capped and shaken for 2 h at room temperature. The reaction mixture was concentrated and the crude product was used in the next step as such. Preparation of (S)-1-tert-butyl 5-(perfluorophenyl) 2-palmitamidopentanedioate, A067E- 136-10Scheme:- In a glass reactor equipped with a frit was added the 2-Cl-chlorotrityl resin (2.5 g, 3.85 mmol) and DCM (25 mL). To the suspension was added Fmoc-Glu-OtBu (2.048 g, 4.81 mmol) followed by DIPEA (2.062 mL, 11.55 mmol), and the reaction was allowed to shake for 3 h at room temperature. The solution was filtered through the frit and the resin was rinsed with DCM (4 x 25 mL). An aliquot of the resin was cleaved with TFA and LCMS analysis of the cleavage solution indicates clean desired product. Analysis condition E: Retention time = 1.57 min; ESI-MS(+) m / z [M+H]+: 369.9. Step 2: Preparation of (S)-5-(tert-butoxy)-5-oxo-4-palmitamidopentanoic acid (A067E- 135) The resin (3.08 g, 3.85 mmol, 1.0 eq) was swollen in DMF (20 mL) for 10 min. It was treated with 20% piperidine / DMF (10 mL x 3 min shaking) and the solution was drained through the frit before a second treatment of 20% piperidine / DMF (10 mL x 10 min shaking). The solution was drained through the frit and the resin was washed with DMF (8 x 10 mL). A solution of palmitic acid (2.96 g, 11.55 mmol, 3.0 eq) and 3H- [1,2,3]triazolo[4,5-b]pyridin-3-ol HOAt (1.572 g, 11.55 mmol, 3.0 eq) in DMF (20 mL) was added to the resin followed by the addition of N,N'-diisopropylcarbodiimide (1.127 ml, 7.36 mmol, 3.0 eq). The reaction was shaken for 16 h at rt. The reaction solution was drained through the frit and the resin was washed with DMF (4 x 20 mL) and then DCM (4 x 20 mL). The protected residue was cleaved off the resin with (20% v / v) 1,1,1,3,3,3- Hexafluoro-2-propanol (HFIP) in DCM (40 mL) for 2 h. The cleavage solution containing the crude product was obtained by filtration through the frit. The resin wasrinsed with DCM (4 x 20 mL). The combined filtrates were concentrated to dryness to give (1.21 g, 71.2%) of (S)-5-(tert-butoxy)-5-oxo-4-palmitamidopentanoic acid. Analysis condition E: Retention time = 2.26 min; ESI-MS(+) m / z [M+H]+: 442.1. Step 3: Preparation of (S)-1-tert-butyl 5-(perfluorophenyl) 2-palmitamidopentanedioate (A067E-136-10) To a pressure seal vial was added (S)-5-(tert-butoxy)-5-oxo-4-palmitamidopentanoic acid (1.21 g, 2.74 mmol, 1.0 eq), DMF (5 mL), perfluorophenyl 2,2,2-trifluoroacetate (0.937 mL, 5.48 mmol, 2.0 eq) , and pyridine (0.487 mL, 6.03 mmol, 2.2 eq). The reaction mixture was kept under a blanket of nitrogen and stirred at room temperature for 16 h. The reaction was diluted with saturated citric acid aqueous solution and then extracted with EtOAc. It was concentrated to give 1.50 g of (S)-1-tert-butyl 5-(perfluorophenyl) 2- palmitamidopentanedioate. Analysis condition E: Retention time = 2.54 min; ESI-MS(+) m / z [M+H]+: 608.1. Preparation of (S)-perfluorophenyl 5-amino-5-oxo-4-palmitamidopentanoate, A067E-180 Scheme:eq) with 20% Piperidine / DMF solution (v / v, 3 mL). The reaction was shaken for 5 min. The solution was drained through the frit and another 20% Piperidine / DMF solution (v / v, 3 mL) was added to the resin. The mixture was shaken for 5 min and the solution was drained through the frit. The resin was washed with DMF (5 x 5.0 mL) and then dilutedwith DMF (5.0 mL). To this suspension was then added a solution of (S)-2-((((9H- fluoren-9-yl)methoxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanoic acid (2.71 g, 6.36 mmol, 3.5 eq) and HATU (2.419 g, 6.36 mmol, 3.5 eq) in DMF (8.0 mL) followed by N- methylmorpholine (0.699 mL, 6.36 mmol, 3.5). The reaction was allowed to shake for 5 h. The reaction solution was drained through the frit and the resin was washed with DMF (5 x 5.0 mL) and DCM (5 x 5.0 mL). An aliquot of the resin was cleaved with TFA and LCMS analysis of the cleavage solution indicates clean desired product. Analysis condition E: Retention time = 1.13 min; ESI-MS(+) m / z [M+H]+: 368.9. Step 2: Preparation of (S)-5-amino-5-oxo-4-palmitamidopentanoic acid (A067E-179) The resin was treated with a solution of 20% piperidine / DMF (v / v, 10 mL) shaking for 3 min at room temperature. The solution was then filtered through the frit and a second treatment of 20% piperidine / DMF (v / v, 10 mL) was performed shaking for 10 min. The solution was filtered through the frit and the resin was washed with DMF (8 x 10 mL) and DCM (5 x 10 mL). A solution palmitic acid (899 mg, 3.50 mmol, 4.0 eq) and 3H- [1,2,3]triazolo[4,5-b]pyridin-3-ol (477 mg, 3.50 mmol, 4.0 eq) in DMF (8 mL) was added to the pre-loaded resin (1200 mg, 0.876 mmol, 1.0 eq) in DMF (8.0 mL) followed by the addition of N,N'-diisopropylcarbodiimide (1127 µl, 7.36 mmol, 4.0 eq). The reaction was shaken at room temperature for 3 h. The solution was drained through the frit and the resin was washed with DMF (4 x 20 mL), and DCM (4 x 20 mL). The protected Pal-Glu-amide (0.876 mmol, 1.0 eq) was cleaved off the resin with TFA cocktail (TFA / Water / TIS / DTT, 93 / 3 / 3 / 1, v / v / v) and shaken for 1 h. The cleavage solution containing the crude product was obtained by filtration. The resin was rinsed with DCM (4 x 20 mL). The combined filtrates were evaporated to give ((S)-5-amino-5-oxo-4- palmitamidopentanoic acid (323 mg, 0.840 mmol, 96%). Step 3: Preparation of (S)-perfluorophenyl 5-amino-5-oxo-4-palmitamidopentanoate (A067E-180) To (S)-5-amino-5-oxo-4-palmitamidopentanoic acid (160 mg, 0.416 mmol, 1.0 eq) was added DMF (5 mL), perfluorophenyl 2,2,2-trifluoroacetate (142 µL, 0.832 mmol, 2.0 eq), and pyridine (74.0 µL, 0.915 mmol, 2.2 eq). The reaction mixture was kept under a blanket of nitrogen and stirred at room temperature for 1 h. The crude reaction mixture was concentrated using a V-10 apparatus to give (S)-perfluorophenyl 5-amino-5-oxo-4-palmitamidopentanoate (229 mg, 0.416 mmol, 100%). Analysis condition E: Retention time = 1.34 min; ESI-MS(+) m / z [M+H]+: 551.4. Preparation of (S)-tert-butyl 1-azido-22-(tert-butoxycarbonyl)-19,24-dioxo-3,6,9,12,15- pentaoxa-18,23-diazahentetracontan-41-oate Scheme:g, was the mixture was heated to reflux.1,1-di-tert-butoxy-N,N-dimethylmethanamine (10.36 ml, 43.2 mmol, 2.68 eq) was added drop-wise over 30 min. The mixture was refluxed for 16 hours. The reaction mixture was cooled to room temperature and the solvent was removed in vacuo at 50oC. The crude material was suspended in CH2Cl2 / EtOAc (75 mL, 1:1 v / v) and stirred for 15 min. The solids were removed by filtration and washed with CH2Cl2 (40 mL). The filtrate was evaporated in vacuo to give 18-(tert-butoxy)-18-oxooctadecanoic acid (2.70 g, 7.29 mmol, 45%). 1H NMR (400MHz, METHANOL-d4) ^ 8.00 (s, 1H), 2.34 - 2.19 (m, 4H), 1.68 - 1.53 (m, 4H), 1.47 (s, 9H), 1.32 (s, 24H) Step 2: 1-(tert-butyl) 18-(2,5-dioxopyrrolidin-1-yl) octadecanedioate, A067E-200 DCC (390 mg, 1.888 mmol) was added to a solution of 18-(tert-butoxy)-18- oxooctadecanoic acid (636 mg, 1.716 mmol, 1.0 eq) and 1-hydroxypyrrolidine-2,5-dione (217 mg, 1.888 mmol, 1.1 eq) in DMF (3.4 mL). The reaction mixture was allowed to stir at room temperature overnight. Solid in suspension were removed by filtration. The obtained filtrated was concentrated using a V-10 instrument to obtain 1-tert-butyl 18-(2,5- dioxopyrrolidin-1-yl) octadecanedioate (680 mg, 1.454 mmol, 85%). Analysis condition E: Retention time = 2.33 min; ESI-MS(+) m / z [M+Na]+: 491.2.Step 3: (S)-5-(tert-butoxy)-4-(18-(tert-butoxy)-18-oxooctadecanamido)-5-oxopentanoic acid, A067E-201 Water (3.80 ml) was added to a mixture of (S)-4-amino-5-(tert-butoxy)-5-oxopentanoic acid (325 mg, 1.600 mmol, 1.1 eq), 1-tert-butyl 18-(2,5-dioxopyrrolidin-1-yl) octadecanedioate (680 mg, 1.454 mmol, 1.0 eq), sodium bicarbonate (147 mg, 1.745 mmol, 1.2 eq) in THF (11.4 ml). The resulting clear solution was stirred at room temperature for 4 h. All THF was removed by rotovap, 1M HCl (1.890 ml, 1.890 mmol, 1.3 mmol) was added and the pH was adjusted to 2-3 while the solution temperature was kept at 0oC using an ice bath. The resulting suspension was extracted with CH2Cl2. The organic fractions were combined and dried over anhydrous sodium sulfate. After filtrate and concentration the desired product was obtained. Isolated (S)-5-(tert-butoxy)-4-(18- (tert-butoxy)-18-oxooctadecanamido)-5-oxopentanoic acid (900 mg, 1.62 mmol, 100%). LCMS is good. Analysis condition E: Retention time = 2.26 min; ESI-MS(+) m / z [M+H]+: 556.1. Step 4: Preparation of (S)-tert-butyl 1-azido-22-(tert- -19,24-dioxo-3,6,9,12,15-pentaoxa-18,23-diazahentetracontan-41-oate, A067E-202 To a solution of (S)-5-(tert-butoxy)-4-(18-(tert-butoxy)-18-oxooctadecanamido)-5- oxopentanoic acid (254 mg, 0.457 mmol, 1.0 eq) in DMF (4 ml) was added DIPEA (0.239 ml, 1.371 mmol, 3.0 eq) and HATU (348 mg, 0.914 mmol, 2.0 eq).17-azido- 3,6,9,12,15-pentaoxaheptadecan-1-amine (140 mg, 0.457 mmol, 1.0 eq) was then added and the solution was allowed to stir at room temperature. The crude product was concentrated to dryness on V10 to obtain (S)-tert-butyl 1-azido-22-(tert-butoxycarbonyl)- 19,24-dioxo-3,6,9,12,15-pentaoxa-18,23-diazahentetracontan-41-oate (330 mg, 0.391 mmol, 86%). Analysis condition E: Retention time = 2.30 min; ESI-MS(+) m / z [M+H]+: 844.4. Preparation of 1-(tert-butyl) 12-(perfluorophenyl) dodecanedioateScheme: Step 1: 12-N,N-Dimethylformamide di-tert-butyl acetal (3.12 mL, 13.03 mmol) was added drop wise to a solution of dodecanedioic acid (1 g, 4.34 mmol) in toluene (20 mL) which was kept at 95 °C for 16 h. The reaction was cooled to room temperature and the solvent was removed in vacuo. The residue was dissolved in dichloromethane (15 mL). The solvent was removed in vacuum twice. The crude product was purified using CombiFlash (24 g silica gel column; Petroleum Ether : Ethylacetate as eluant). The desired fraction was eluted at 10-40 % Ethylacetate in Petroleum Ether. The desired fractions were evaporated under reduced pressure to afford the product as white solid. Analysis condition E: Retention time = 3.01 min; ESI-MS(-) m / z [M-H]+: 285.2. Step 2: 1-(tert-butyl) 12-(perfluorophenyl) dodecanedioate, A0D20-316 To a solution of 12-(tert-butoxy)-12-oxododecanoic acid (350 mg, 1.222 mmol) in DMF (1 mL) was added perfluorophenyl 2,2,2-trifluoroacetate (411 mg, 1.466 mmol), followed by pyridine (297 µL, 3.67 mmol). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was washed with citric acid and extracted into DCM. The organic fractions were dried over anhydrous sodium sulfate, filtered, and concentrated to give 1-tert-butyl 12-(perfluorophenyl) dodecanedioate (400 mg, 0.884 mmol, 72.3 % yield). Preparation of (S)-perfluorophenyl 6-(5-(tert-butoxy)-5-oxo-4- palmitamidopentanamido)hexanoate, A067E-175Scheme:In a glass reactor equipped with a frit was added the 2-chloro-chlorotrityl resin (2.5 g, 3.85 mmol) in DCM (25 mL). To this was added Fmoc-6-aminohexanoic acid (4.08 g, 11.55 mmol, 3.0 eq) followed by DIPEA (2.062 mL, 11.55 mmol, 3.0 eq). The reaction was allowed to shake at room temperature for 3 h. The reaction solution was filtered through the frit and the resin was rinsed with DCM (4 x 25 mL). An aliquot of the resin was cleaved with TFA and LCMS analysis of the cleavage solution indicated clean desired product. Analysis condition E: Retention time = 1.43 min; ESI-MS(+) m / z [M+H]+: 353.9. Step 2: Preparation of resin bound (S)-6-(4-amino-5-(tert-butoxy)-5- oxopentanamido)hexanoic acid In a glass reactor equipped with a frit, the resin (1.357 g, 3.85 mmol) was swollen in DMF (20 mL) for 10 min. The solvent was filtered through the frit and the resin was treated with 20% piperidine / DMF (v / v, 10 mL x 3 min shaking). The solution was filtered through the frit followed by a second treatment of 20% piperidine / DMF (v / v, 10 mL x 10 min shaking). The solution was filtered through the frit and the resin was washed with DMF (8 x 10 mL). To a solution of Fmoc-Glu-OtBu (3.28 g, 7.70 mmol, 2.0 eq) and 3H- [1,2,3]triazolo[4,5-b]pyridin-3-ol (1.048 g, 7.70 mmol, 2.0 eq) in DMF (10mL) was added to the resin in DMF (10 mL), followed by the addition of N,N’- diisopropylcarbodiimide (1.127 mL, 7.36 mmol, 2.0 eq). The reaction was shaken at room temperature for 16 h. The reaction solution was drained through the frit and the resin waswashed with DMF (4 x 20 mL), and DCM (4 x 20 mL). An aliquot of the resin was cleaved with TFA and LCMS analysis of the cleavage solution indicates clean desired product. Analysis condition E: Retention time = 1.22 min; ESI-MS(+) m / z [M+H]+: 482.9. Step 3: Preparation of (S)-6-(5-(tert-butoxy)-5-oxo-4-palmitamidopentanamido)hexanoic acid ( The resin was treated with 20% piperidine / DMF (10mL x 3 min shaking) followed by a second treatment of 20% piperidine / DMF (10mL x 10 min shaking). The resin was washed with DMF (8 x 10 mL) and DCM (5 x 10 mL). A solution palmitic acid (1.655 g, 6.46 mmol, 4.0 eq) and 3H-[1,2,3]triazolo[4,5-b]pyridin-3-ol (0.879 g, 6.46 mmol, 4.0 eq) in DMF (8 mL) was added to the resin (1.5 g, 1.614 mmol, 1.0 eq) in DMF (8.0 mL) followed by the addition of N,N’-diisopropylcarbodiimide (1.127 mL, 7.36 mmol, 4.0 eq). The reaction was shaken at room temperature for 16 h. The reaction solution was filtered through the frit and the resin was washed with DMF (4 x 20 mL), and DCM (4 x 20 mL). An aliquot of the resin was cleaved with TFA and LCMS analysis of the cleavage solution indicates clean desired product. Analysis condition E: Retention time = 1.94 min; ESI-MS(+) m / z [M+H]+: 499.1. The protected substrate was cleaved off the resin with 20 mL (20%, v / v) HFIP in DCM for 2 h. The cleavage solution containing the crude product was obtained by filtration through the frit. The resin was rinsed with DCM (4 x 20 mL). The combined filtrates were evaporated to give (S)-6-(5-(tert-butoxy)-5-oxo- 4-palmitamidopentanamido)hexanoic acid (630 mg, 1.136 mmol, 70.4%). Analysis condition E: Retention time = 2.20 min; ESI-MS(+) m / z [M+H]+: 555.3. Step 4: Preparation of (S)-perfluorophenyl 6-(5-(tert-butoxy)-5-oxo-4- palmitamidopentanamido)hexanoate To (S)-6-(5-(tert-Butoxy)-5-oxo-4-palmitamidopentanamido)hexanoic acid (630 mg, 1.136 mmol, 1.0 eq) was added DMF (5 mL), perfluorophenyl 2,2,2-trifluoroacetate (389 µL, 2.271 mmol, 2.0 eq) , and pyridine (202 µL, 2.498 mmol, 2.2 eq). The reaction mixture was kept under a blanket of nitrogen and stirred at room temperature for 1 h. It was concentrated to give (S)-perfluorophenyl 6-(5-(tert-butoxy)-5-oxo-4- palmitamidopentanamido)hexanoate (800 mg, 1.11 mmol, 97%). Analysis condition E: Retention time = 2.50 min; ESI-MS(+) m / z [M+H]+: 721.0.Preparation of (S)-1-tert-butyl 5-(perfluorophenyl) 2-(18-(tert-butoxy)-18- oxooctadecanamido)pentanedioate Scheme:- 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 min. A solution of Fmoc-Glu-OtBu (1.276 g, 3.00 mmol, 1.0 eq) in DCM (5 mL) was0 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 min. DIEA (0.5 mL) and methanol (3 mL) were added, and shaken for an additional 15 min. 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), ether (4 x 5mL), and dried using a flow of nitrogen. A sample of resin (13.1 mg) was treated 5 with 20% piperidine / DMF (v / v, 2.0 mL) for 10 min 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 / 200 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 shake5 for 5 min. The solution was filtered off and the resin was treated again with a solution of20% piperidine / DMF (v / v, 5 mL) for another 5 min. The reaction solution was filtered through the frit and the resin was washed with DMF (6 x 5 mL x 1 min 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 h. The resin was washed with DMF (4 x 5 mL x 1 min shaking). The protected peptide was cleaved off the resin with 20% HFIP / DCM (v / v, 30 mL) at room temperature for 2 h. 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) DMF (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 at room temperature for 16 h. 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 (R)-18-((3-azido-1-carboxypropyl)amino)-18-oxooctadecanoic acidScheme: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 min. The mixture was put under refluxing conditions overnight. The solvent was removed in vacuo at 50oC 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 (0-25% 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) ^ 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-bottomed 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 at room temperature overnight. The reaction was poured into a saturated citric acid solution and extracted with DCM (3 x). 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=7.5 Hz, 2H), 1.78 (quin, J=7.5 Hz, 2H), 1.63 - 1.52 (m, 2H), 1.48 - 1.39 (m, 12H), 1.37 - 1.20 (m, 21H). Step 3: Preparation of (R)-18-((3-azido-1-carboxypropyl)amino)-18-oxooctadecanoic acidTo a solution of 1-tert-butyl 18-(perfluorophenyl) octadecanedioate (110 mg, 0.205 mmol) and (R)-2-amino-4-azidobutanoic acid (29.5 mg, 0.205 mmol) in DMF (7 mL) was added N-ethyl-N-isopropylpropan-2-amine (26.5 mg, 0.205 mmol). The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated in vacuo and purified via ISCO to give the desired (R)-18-((3-azido-1- carboxypropyl)amino)-18-oxooctadecanoic acid (80 mg, 89%). Preparation of (S)-1-azido-16-carboxy-13,18-dioxo-3,6,9-trioxa-12,17- diazapentatriacontan-35-oic acid Scheme Step3,6,9- trioxa-12,17-diazapentatriacontan-35-oate To a solution of 2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethanamine (113 mg, 0.517 mmol, 1.1 eq) in DMF (4498 µL) was added Hunig's Base (314 µL, 1.799 mmol, 4.0 eq), then (S)-5-(tert-butoxy)-4-(18-(tert-butoxy)-18-oxooctadecanamido)-5-oxopentanoic acid (250 mg, 0.450 mmol). HATU (342 mg, 0.900 mmol, 2.0 eq) was then added, and the resulting solution was stirred at rt. DMF was removed on high vacuum, and then the residue was applied to silica gel (40 g) and eluted with DCM (100 mL), then a gradient to 75% DCM / acetone over 540 mL and finally a hold at 75% DCM / acetone for 150 mL. The desired fractions were combined. The material was taken into the next step as-is. Step 2: Preparation of (S)-1-azido-16-carboxy-13,18-dioxo-3,6,9-trioxa-12,17- diazapentatriacontan-35-oic acid To a solution of (S)-tert-butyl 1-azido-16-(tert-butoxycarbonyl)-13,18-dioxo-3,6,9-trioxa- 12,17-diazapentatriacontan-35-oate (414.0 mg, 0.548 mmol) in DCM (5476 µL) was added TFA (1266 µL, 16.43 mmol, 30.0 eq). LC / MS indicated a slow reaction, so another14 eq TFA was added and the mixture was stirred further. After another 6 h, LC / MS indicated a nearly complete reaction. Solvents were removed in vacuo. The mixture was taken up in Hunig's base / MeOH (~1%). The reaction mixture was purified by PREP HPLC in 5 injections: (30 X 100 mm HPLC Luna Axia C1850 to 100% A:B over 10 min, 5 min at 100%B (A is 90:10:0.1 water:MeOH:TFA; B is 90:10:0.1 MeOH:water:TFA)). The desired fractions were combined and concentrated to afford (S)- 1-azido-16-carboxy-13,18-dioxo-3,6,9-trioxa-12,17-diazapentatriacontan-35-oic acid (112.4 mg, 0.124 mmol, 22.64 % yield). LC / MS: (M+H)+= 644.45. Preparation of (S)-1-azido-22-carboxy-19,24-dioxo-3,6,9,12,15-pentaoxa-18,23- diazahentetracontan-41-oic acid Scheme Step 1:Octadecanedioic 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, 2.7 eq) was added drop-wise over 30 min. The mixture was reflux overnight. The solvent was removed in vacuo at 50oC 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 (0-25% acetone / CH2Cl2) to get 18- (tert-butoxy)-18-oxooctadecanoic acid (3.95 g, 10.66 mmol, 44.7 % yield). Analysiscondition 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, 1.1 eq), 1-tert-butyl 18-(2,5-dioxopyrrolidin-1-yl) octadecanedioate (2.171 g, 4.64 mmol), 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 solvents were removed. HCl (6.04 mL, 6.04 mmol, 1.3 eq) was added and the pH was adjusted to 2-3 at 0oC. The resulting suspension was extracted with CH2Cl2 (3 x). The organic layer was concentrated. 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-22-(tert-butoxycarbonyl)-19,24-dioxo- 3,6,9,12,15-pentaoxa-18,23-diazahentetracontan-41-oate To a solution of (S)-5-(tert-butoxy)-4-(18-(tert-butoxy)-18-oxooctadecanamido)-5- oxopentanoic acid (225 mg, 0.405 mmol) in DMF (4048 µL) was added Hunig's Base (212 µL, 1.214 mmol, 3.0 eq) and HATU (308 mg, 0.810 mmol, 2.0 eq).17-Azido- 3,6,9,12,15-pentaoxaheptadecan-1-amine, HCl (139 mg, 0.405 mmol, 1.0 eq) was thenadded, and the solution was stirred at rt. The crude product was purified by flash chromatography (220 g, silica gel, 10-60% acetone / CH2Cl2) to get (S)-tert-butyl 1-azido- 22-(tert-butoxycarbonyl)-19,24-dioxo-3,6,9,12,15-pentaoxa-18,23-diazahentetracontan- 41-oate (330 mg, 0.391 mmol, 97 % yield). Analysis condition D: Retention time = 2.88 min; ESI-MS(+) m / z 844.7 (M+H)+.1H NMR (500MHz, METHANOL-d4) ^ 4.30 - 4.23 (m, 1H), 3.74 - 3.60 (m, 18H), 3.60 - 3.52 (m, 2H), 3.43 - 3.35 (m, 4H), 2.34 - 2.28 (m, 2H), 2.28 - 2.19 (m, 4H), 2.15 - 2.08 (m, 1H), 1.98 - 1.87 (m, 1H), 1.69 - 1.53 (m, 4H), 1.52 - 1.44 (m, 18H), 1.41 - 1.27 (m, 24H). Step 5: Preparation of (S)-1-azido-22-carboxy-19,24-dioxo-3,6,9,12,15-pentaoxa-18,23- diazahentetracontan-41-oic acid The mixture of (S)-tert-butyl 1-azido-22-(tert-butoxycarbonyl)-19,24-dioxo-3,6,9,12,15- pentaoxa-18,23-diazahentetracontan-41-oate (330 mg, 0.391 mmol) and TFA (0.422 mL, 5.47 mmol, 14 eq) in DCM (3.0 mL) was stirred at rt for 2 h. The resulting crude product was purified by Prep-HPLC (Solvent A = 0% MeOH - 90% H2O - 0.1% TFA, Solvent B = 90% MeOH - 10% H2O - 0.1% TFA. Column: phenomenex luna 30 x 150mm, S10, Flow rate: 40 ml / min, 50 - 100% B, 10 min and stop at 13 min) to obtain (S)-1-azido-22- carboxy-19,24-dioxo-3,6,9,12,15-pentaoxa-18,23-diazahentetracontan-41-oic acid (101 mg, 0.138 mmol, 35.3 % yield). Analysis condition D: Retention time = 2.42 min; ESI- MS(+) m / z 732.5 (M+H)+. Preparation of (S)-1-azido-28-carboxy-25,30-dioxo-3,6,9,12,15,18,21-heptaoxa-24,29- diazaheptatetracontan-47-oic acid, Scheme Step3,6,9,12,15,18,21-heptaoxa-24,29-diazaheptatetracontan-47-oateTo a solution of 23-azido-3,6,9,12,15,18,21-heptaoxatricosan-1-amine (204 mg, 0.517 mmol, 1.15 eq) in DMF (4498 µL) was added Hunig's Base (314 µL, 1.799 mmol, 4.0 eq), then (S)-5-(tert-butoxy)-4-(18-(tert-butoxy)-18-oxooctadecanamido)-5-oxopentanoic acid (250 mg, 0.450 mmol). HATU (342 mg, 0.900 mmol, 2.0 eq) was then added, and the resulting solution was stirred at rt. DMF was removed on high vacuum, then the residue was applied to silica gel (40 g) and eluted with DCM (90 mL), then a gradient to 75% DCM / acetone over 540 mL and finally a hold at 75% DCM / acetone for 150 mL. The desired fractions were combined to obtain (S)-tert-butyl 1-azido-28-(tert- butoxycarbonyl)-25,30-dioxo-3,6,9,12,15,18,21-heptaoxa-24,29-diazaheptatetracontan- 47-oate (394.2 mg, 0.423 mmol, 94 % yield). Step 2: Preparation of (S)-1-azido-28-carboxy-25,30-dioxo-3,6,9,12,15,18,21-heptaoxa- 24,29-diazaheptatetracontan-47-oic acid To a solution of (S)-tert-butyl 1-azido-28-(tert-butoxycarbonyl)-25,30-dioxo- 3,6,9,12,15,18,21-heptaoxa-24,29-diazaheptatetracontan-47-oate (394.2 mg, 0.423 mmol) in DCM (4229 µL) was added TFA (456 µL, 5.92 mmol, 14 eq). Another 14 eq of TFA was added and the mixture was stirred further. After another 6 h, LC / MS indicated a nearly complete reaction. Solvents were removed in vacuo. The mixture was taken up in MeOH. The reaction mixture was purified by PREP HPLC in 7 injections: (30 X 100 mm HPLC Luna Axia C1850 to 100% A:B over 10 min, 5 min at 100%B (A is 90:10:0.1 water:MeOH:TFA; B is 90:10:0.1 MeOH:water:TFA)) to give (S)-1-azido-28-carboxy- 25,30-dioxo-3,6,9,12,15,18,21-heptaoxa-24,29-diazaheptatetracontan-47-oic acid (121.0 mg, 0.148 mmol, 34.9 % yield) after concentration. LC / MS: (M+H)+= 820.60.1H NMR (500MHz, CHLOROFORM-d) ^ 7.30 (d, J=6.4 Hz, 1H), 7.04 (t, J=5.2 Hz, 1H), 4.51 (q, J=6.2 Hz, 1H), 3.72 - 3.64 (m, 27H), 3.63 - 3.59 (m, 2H), 3.56 - 3.49 (m, 1H), 3.47 - 3.39 (m, 3H), 2.62 - 2.54 (m, 1H), 2.48 - 2.40 (m, 1H), 2.36 (t, J=7.4 Hz, 2H), 2.26 (t, J=7.6 Hz, 2H), 2.19 - 2.08 (m, 2H), 1.65 (quin, J=7.4 Hz, 4H), 1.39 - 1.24 (m, 25H). 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 acidScheme: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 50oC 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 getoxooctadecanoic 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 acidWater (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 0oC. 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's Base (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 (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 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. Theresulting 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 3-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)-2-fluoropyridine, Scheme: Step 1:bis(4- methylbenzenesulfonate) Sodium hydroxide (950 mg, 23.75 mmol) was dissolved in water (5 mL). Tetrahydrofuran (5 mL) was added to the mixture followed by tetraethylene glycol (28 mL, 162 mmol). The mixture was stirred at room temperature for 10 min. It was cooled to 0 °C. A solution of p-toluenesulfonyl chloride (3.0 g, 15.74 mmol) in tetrahydrofuran (20 mL) was added to the mixture drop-wise over 1.5 h. It was stirred at 0 °C for an additional 1 h. The mixture was placed in a refrigerator (4 °C) and held for 16 h. It was poured into ice water (150 mL). The material was extracted with dichloromethane (3 x 40 mL). The combined organics were washed with water (2 x) and brine. The organics were dried MgSO4 and it was filtered. The filtrate was concentrated to dryness. The residue (5.12 g of clear yellow oil) was purified via Biotage medium pressure liquid chromatography (Silica; Isco RediSep 120 g; 100% DCM to 2% MeOH-DCM over 1350 mL then 2% MeOH-DCM isocratic over 900 mL). The major peak was isolated. The fractions were concentrated to dryness to give ((oxybis(ethane-2,1-diyl))bis(oxy))bis(ethane-2,1-diyl) bis(4-methylbenzenesulfonate) (5.10 g, 64%) as a clear colorless oil.1H NMR (400 MHz, CDCl3) δ 7.82 (d, J=7.8 Hz, 4H), 7.36 (d, J=8.1 Hz, 4H), 4.21 - 4.16 (m, 4H), 3.75 - 3.58 (m, 12H), 2.47 (s, 6H). Step 2: Preparation of 2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl 4- methylbenzenesulfonate Tetraethylene glycol bis(p-toluenesulfonate (5.0 g, 9.95 mmol) and sodium azide (0.647 g, 9.95 mmol) were combined in ethanol (99 mL) and the mixture was heated under reflux. The reaction was stopped after 20 h. The solvent was removed in vacuo and the resulting crude material was taken up in dichloromethane (10 mL) and vacuum filtered to remove solids. The filtrate was applied to a 40 g Thomson silica gel cartridge. The product was eluted with a 0-80% ethyl acetate / hexanes gradient to give 0.93 g of pure desired product along with 1.53 g of product with 15% impurities.2-(2-(2-(2- azidoethoxy)ethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (2.49 g, 67%).1H NMR (400 MHz, CHLOROFORM-d) δ 7.78 (d, J=7.0 Hz, 2H), 7.33 (d, J=7.8 Hz, 2H), 4.20 - 4.12 (m, 2H), 3.76 - 3.48 (m,11H), 3.38 (t, J=5.1 Hz, 3H), 2.49 - 2.40 (m, 3H). Step 3: Preparation of 3-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)-2-fluoropyridine To the suspension of sodium hydride (0.120 g, 2.99 mmol) in DMF (20 mL) at 0 °C was dropwise added the solution of 2-fluoro-3-hydroxypyridine (0.338 g, 2.99 mmol) in DMF (5 mL) then followed by the dropwise addition of the solution of 2-(2-(2-(2- azidoethoxy)ethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (0.93 g, 2.490 mmol) in DMF (5 mL). The suspension was held at 0 °C for 10 min, then room temperature for 1 h and heated to 70 °C for 5 h. The solvent was removed in vacuo and the content was worked up between brine and EtOAc. The organic layer was dried over MgSO4, filtered, and concentrated in vacuo and purified using flash chromatography (IscoCombiFlash - eluted with 10-50% EtOAc in Hexanes) to give 3-(2-(2-(2-(2- azidoethoxy)ethoxy)ethoxy)ethoxy)-2-fluoropyridine (0.423 g, 1.346 mmol, 54.0 % yield) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 7.77 (dt, J=4.7, 1.5 Hz, 1H), 7.42 - 7.28 (m, 1H), 7.12 (dd, J=7.5, 4.8 Hz, 1H), 4.29 - 4.13 (m, 4H), 4.00 - 3.85 (m, 4H), 3.80 - 3.73 (m, 4H), 3.41 (t, J=5.0 Hz, 4H).Preparation of INT-1000 INT-1000 was scale as follows; to a 25-mL polypropyleneresin (70 mg, 0.05 mmol ), and the reaction vessel was placed on the Symphony peptide synthesizer. The following procedures were then performed sequentially: “Symphony Method A: Resin-swelling procedure” was followed; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Gly-OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Cys(Trt)- OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Gly-OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Tyr(tBu)- OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Asp(OtBu)- OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Trp(Boc)- OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Bip-OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Tyr(tBu)- OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Tyr(tBu)- OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Lys(Boc)- OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Asp(OtBu)- OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Tyr(tBu)-OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Val-OH; “Symphony Method A: Single-coupling procedure” was followed with Fmoc-Phe-OH followed by “Manual removal of Fmoc group procedure”; “Chloroacetic acid coupling procedure A” was followed; “Deprotection Method A” was followed; “Cyclization Method A” was followed. The crude material was purified via preparative LC / MS with the following conditions: Column: Waters XBridge C18, 19 x 200 mm, 5-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; Gradient: 10-50% B over 25 minutes, then a 4- minute hold at 50% B; Flow: 20 mL / min. Fractions containing the desired product were combined and dried via centrifugal evaporation. The yield of the product was 3.5 mg, and its estimated purity by LCMS analysis was 92%. Analysis condition B: Retention time = 1.29 min; ESI-MS(+) m / z [M+2H]2+: 971.3. Following similar procedures as described for INT-1000 and aforementioned general synthetic and purification procedures, INT-1001-INT-1011 were prepared. Preparation of Example INT-1001 The yield of the product was 4.8 mg, and its estimated purity by LCMS analysis was 94%. Analysis condition A: Retention time = 1.53 min; ESI- MS(+) m / z [M+2H]2+: 943.1.Preparation of Example INT-1002 The yield of the product was 2.3 mg, and its estimated purity by LCMS analysis was 97.8%. Analysis condition A: Retention time = 1.19 min; ESI-MS(+) m / z [M+2H]2+: 943.3. Preparation of Example INT-1003 NH The yield of the product was 4.8 mg, O HN O HN O OH O O H O and its estimated purity by LCMS S N N N N NH H HOH O H HN O NH analysis was 90.8%. Analysis NHHO H O H HN condition A: Retention time = 2.26, N NNNNO H2+O OOHO2.32 min; ESI-MS(+) m / z [M+2H] : OH NH 978.06, 977.2. Preparation of Example INT-1004 The yield of the product was 10.1 mg, and its estimated purity by LCMS analysis was 98.2%. Analysis condition A: Retention time = 2.51 min; ESI-MS(+) m / z [M+2H]2+: 978Preparation of Example INT-1005 BMT-234334-01-001 The yield of the product was 3.2 mg, and its estimated purity by LCMS analysis was 95.4%. Analysis condition A: Retention time = 1.4 min; ESI-MS(+) m / z [M+2H]2+: 943.1. Preparation of Example INT-1006 The yield of the product was 5.5 mg, and its estimated purity by LCMS analysis was 88.4%. Analysis condition B: Retention time = 1.72 min; ESI-MS(+) m / z [M+2H]2+: 942.9. Preparation of Example INT-1007 The yield of the product was 3.3 mg, and its estimated purity by LCMS analysis was 97.3%. Analysis condition B: Retention time = 1.63 min; ESI-MS(+) m / z [M+2H]2+: 927.Preparation of Example INT-1008 The yield of the product was 1.3 mg, and its estimated purity by LCMS analysis was 89.5%. Analysis condition B: Retention time = 1.45 min; ESI-MS(+) m / z [M+2H]2+: 977. Preparation of Example INT-1009 The yield of the product was 4.9 mg, and its estimated purity by LCMS analysis was 98.8%. Analysis condition A: Retention time = 1.13 min; ESI- MS(+) m / z [M+2H]2+: 1048.2. Preparation of Example INT-1010 The yield of the product was 7.1 mg, and its estimated purity by LCMS analysis was 96.6%. Analysis condition B: Retention time = 1.39 min; ESI-MS(+) m / z [M+2H]2+: 977.Preparation of Example 1001 Example 1001 procedureC” with perfluorophenyl palmitate using on a 21 ^mol scale. The crude material was purified via preparative LC / MS with the following conditions: Column: Waters X-bridge C18, 30 x 150 mm, 5-um particles: Mobile Phase A: 5:95 acetonitrile: water with 0.1% TFA; Mobile Phase B: 95:5 acetonitrile: water with 0.1% TFA. Gradient: 10-60% B over 20 minutes, then a 5 minute hold at 100% B: Flow: 45 mL / min. Fractions containing the desired product were combined and dried via centrifugal evaporation. The yield of the product was 3.1 mg, and its estimated purity by LCMS analysis was 97%. Analysis condition A: Retention time = 2.19 min. Analysis condition B: Retention time = 2.49 min; ESI-MS(+) m / z [M+2H]2+: 1090.9. The following examples were following the general synthetic sequence described for the preparation of Example 1001.Preparation of Example 1002 The yield of the product was 1.7 mg, and its estimated purity by LCMS analysis was 90.6%. Analysis condition B: Retention time = 2.45 min; ESI-MS(+) m / z [M+2H]2+: 1016.4. Preparation of Example 1003 The yield of the product was 1 mg, and its estimated purity by LCMS analysis was 92%. Analysis condition B: Retention time = 2.51 min; ESI-MS(+) m / z [M+2H]2+: 1075.1.Preparation of Example 1004 The yield of the product was 1.5 mg, and its estimated purity by LCMS analysis was 88.3%. Analysis condition A: Retention time = 1.94, 1.97 min; ESI-MS(+) m / z [M+2H]2+: 1198.18, 1198.18. Analysis condition B: Retention time = 2.31 min; ESI- MS(+) m / z [M+2H]2+: 1198.6. Preparation of Example 1005 The yield of the product was 3 mg, and its estimated purity by LCMS analysis was 95.8%. Analysis condition A: Retention time = 2.42 min. ESI-MS(+) m / z [M+2H]2+: 1198.7. Analysis condition B: Retention time = 2.74 min; ESI- MS(+) m / z [M+2H]2+: 1198.4.Preparation of Example 1006 The yield of the product was 2.2 mg, and its estimated purity by LCMS analysis was 90.9%. Analysis condition A: Retention time = 2.44 min; ESI-MS(+) m / z [M+2H]2+: 1169.3. Preparation of Example 1007 The yield of the product was 4.8 mg, and its estimated purity by LCMS analysis was 93%. Analysis condition A: Retention time = 1.92 min; ESI-MS(+) m / z [M+2H]2+: 1141.0. Analysis condition B: Retention time = 2.46 min; ESI- MS(+) m / z [M+2H]2+: 1141.1. Preparation of Example 1008 The yield of the product was 8 mg, and its estimated purity by LCMS analysis was 91.4%. Analysis condition A: Retention time = 1.64 min; ESI-MS(+) m / z [M+2H]2+: 1170.0. Analysis condition B: Retention time = 2.18 min; ESI- MS(+) m / z [M+2H]2+: 1170.0.Preparation of Example 1009- - ([1,1'-biphenyl]-4-ylmethyl)-3-((2-amino-2-oxoethyl)carbamoyl)-39-(4-carboxybenzyl)- 9,24,33-tris(4-hydroxybenzyl)-36-isopropyl-5,8,11,14,17,20,23,26,29,32,35,38,41- tridecaoxo-21-(4-(prop-2-yn-1-yloxy)benzyl)-1-thia- 4,7,10,13,16,19,22,25,28,31,34,37,40-tridecaazacyclodotetracontane-12,30-diyl)diacetic acid (20 mg, 10.25 µmol), (S)-tert-butyl 1-azido-22-(tert-butoxycarbonyl)-19,24-dioxo- 3,6,9,12,15-pentaoxa-18,23-diazahentetracontan-41-oate (12.11 mg, 0.014 mmol) was shaken in DMF (0.8n mL). In a separated 1-drum vial was added sodium (R)-2-((S)-1,2- dihydroxyethyl)-4-hydroxy-5-oxo-2,5-dihydrofuran-3-olate 28 mg and copper(II) sulfate pentahydrate 10 mg in 0.4 mL of water. The solution was shaken for 5 seconds and 0.2 mL of the solution was added to the above DMF solution. The resulting mixture wasshaken at rt for 1h to overnight. The crude was concentrated and the residue was dissolved in TFA / TIS / H2O (95:2.5:2.5) (2 mL) and stirred at rt for 1 h. The crude was concentrated, filtered, and submitted to purification. The crude material was purified via preparative LC / MS with the following conditions: Column: XBridge C18, 30 x 150 mm, 5-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; Gradient: 20- 70% B over 20 minutes, then a 2-minute hold at 100% B; Flow: 40 mL / min. Fractions containing the desired product were combined and dried via centrifugal evaporation. The yield of the product was 2.4 mg, and its estimated purity by LCMS analysis was 91.7%. Analysis condition A: Retention time = 1.56 min; ESI-MS(+) m / z [M+2H]2+: 1320.2. The following examples were prepared following the general synthetic sequence described for the preparation of Example 1009 Preparation of Example 1010 The yield of the product was 2.1 mg, and its estimated purity by LCMS analysis was 89.2%. Analysis condition A: Retention time = 1.45 min; ESI- MS(+) m / z [M+2H]2+: 1328.1. Analysis condition B: Retention time = 1.86 min; ESI- MS(+) m / z [M+2H]2+: 1329.Preparation of Example 1011 The yield of the product was 3.9 mg, and its estimated purity by LCMS analysis was 93.7%. Analysis condition A: Retention time = 1.38 min; ESI-MS(+) m / z [M+2H]2+: 1342.2.Analysis condition B: Retention time = 1.89 min; ESI-MS(+) m / z [M+2H]2+: 1342.0. Preparation of Example 1012 The yield of the product was 13.1 mg, and its estimated purity by LCMS analysis was 98%. Analysis condition A: Retention time = 1.46 min; ESI- MS(+) m / z [M+2H]2+: 1343.0. Analysis condition B: Retention time = 2.03 min; ESI- MS(+) m / z [M+2H]2+: 1342.2.Preparation of Example 1013 The yield of the product was 7.4 mg, and its estimated purity by LCMS analysis was 91%. Analysis condition A: Retention time = 1.48 min; ESI-MS(+) m / z [M+2H]2+: 1474.2. Analysis condition B: Retention time = 2.00 min; ESI-MS(+) m / z [M+2H]2+: 1474.2. Preparation of Example 1014 OH O The yield of the NH OOHO NH product was 8.4 mg, O H O O N S O N N N and its estimated purity HN O NH H O H H HNO OHby LCMS analysis was NH H O H O H HN O O N N N N N 79.5%. Analysis CH O H O H N O OH N O condition A: Retention N HOOH time = 1.58, 1.65 min; O OOHO O O O O O O O O ESI-MS(+) m / z NH O [M+2H]2+: 969.04, O N O 968.98. Analysis OH H condition B: Retention time = 2.10 min; ESI- MS(+) m / z [M+2H]2+: 1452.1.Preparation of Example 1015 The yield of the product was 3.9 mg, and its estimated purity by LCMS analysis was 93.6%. Analysis condition A: Retention time = 1.41 min; ESI-MS(+) m / z [M+2H]2+: 1369.6. Analysis condition B: Retention time = 1.83 min; ESI- MS(+) m / z [M+3H]3+: 912.4. Preparation of Example 1016 The yield of the product was 5.3 mg, and its estimated purity by LCMS analysis was 86.4%. Analysis condition B: Retention time = 1.89 min; ESI-MS(+) m / z [M+2H]2+: 1452.Preparation of Example 1017 The yield of the product was 3.6 mg, and its estimated purity by LCMS analysis was 87.1%. Analysis condition A: Retention time = 1.33 min; ESI-MS(+) m / z [M+2H]2+: 1319.3. Analysis condition B: Retention time = 1.80 min; ESI- MS(+) m / z [M+2H]2+: 1319.9. Preparation of Example 1018 The yield of the product was 1.8 mg, and its estimated purity by LCMS analysis was 83%. Analysis condition A: Retention time = 1.27 min; ESI- MS(+) m / z [M+3H]3+: 749.5.Preparation of Example 1019 The yield of the product was 11.1 mg, and its estimated purity by LCMS analysis was 97.2%. Analysis condition A: Retention time = 1.36 min; ESI-MS(+) m / z [M+2H]2+: 1568.2. Analysis condition B: Retention time = 1.95 min; ESI- MS(+) m / z [M+3H]2+: 1046. Preparation of Example 1020 The yield of the product was 22 mg, and its estimated purity by LCMS analysis was 95.2%. Analysis condition A: Retention time = 1.33 min; ESI-MS(+) m / z [M+2H]2+: 1515.1. Analysis condition B: Retention time = 1.93, 1.97 min; ESI-MS(+) m / z [M+2H]2+: 1516.1.Preparation of Example 1021 The yield of the product was 17.4 mg, and its estimated purity by LCMS analysis was 95.3%. Analysis condition A: Retention time = 1.38 min; ESI-MS(+) m / z [M+2H]2+: 1451.1. Analysis condition B: Retention time = 1.94 min; ESI-MS(+) m / z [M+2H]2+: 1451.0. Preparation of Example 1022 The yield of the product was 15.8 mg, and its estimated purity by LCMS analysis was 97.7%. Analysis condition A: Retention time = 1.39 min; ESI-MS(+) m / z [M+2H]2+: 1528.1. Analysis condition B: Retention time = 1.93 min; ESI-MS(+) m / z [M+2H]2+: 1527.4.Preparation of Example 1023 The yield of the product was 7.1 mg, and its estimated purity by LCMS analysis was 95.5%. Analysis condition A: Retention time = 1.36 min; ESI-MS(+) m / z [M+2H]2+: 1047.2. Analysis condition B: Retention time = 1.90 min; ESI- MS(+) m / z [M+2H]2+: 1569.1. Preparation of Example 1024 The yield of the product was 11 mg, and its estimated purity by LCMS analysis was 98%. Analysis condition A: Retention time = 0.98 min; ESI-MS(+) m / z [M+2H]2+: 1299.3Preparation of Example 1025 The yield of the product was 20.1 mg, and its estimated purity by LCMS analysis was 98%. Analysis condition A: Retention time = 0.94 min; ESI-MS(+) m / z [M+2H]2+: 1386.9. Preparation of Example 1027 The yield of the product was 6.7 mg, and its estimated purity by LCMS analysis was 99%. ESI- MS(+) m / z [M+2H]2+: 1196.6.Preparation of Example 10281597.9. Preparation of Example 1032 The yield of the product was 1.9 mg, and its estimated purity by LCMS analysis was 70.4%. Analysis condition B: Retention time = 1.45 min; ESI-MS(+) m / z [M+2H]2+: 1477.3.Preparation of Example 1033 The yield of the product was 9.2 mg, and its estimated purity by LCMS analysis was 95.3%. Analysis condition B: Retention time = 2 min; ESI-MS(+) m / z [M+2H]2+: 1449. Preparation of Example 1034 OH O The yield of the product was 12.2 OH OOHO OH O H O O mg, and its estimated purity by N S O N N N H H HN O NH O H HN OH LCMS analysis was 95.7%. Analysis NH H O H O H HN O N N N O N condition B: Retention time = 1.93 O CH N H O H O OHO OHOH O mi2+O n; ESI-MS(+) m / z [M+2H] : HO O 1437.4. N N N O O O O O O O O O NH O O N O OH HPreparation of Example 1035 The yield of the product was 7.1 mg, and its estimated purity by LCMS analysis was 95.4%. Analysis condition A: Retention time = 1.37 min; ESI-MS(+) m / z [M+2H]2+: 1466. Preparation of Example 1036 OH O O The yield of the product was 5.4 mg, NH OH OOHO NH O H O O and its estimated purity by LCMS N S O N N N H H H HN O NH O HN analysis was 92.4%. Analysis OH NH H O H O H HN O condition A: Retention time = 1.41 N N N O N N CH O H O H OO OHmin; ESI-2+OH MS(+) m / z [M+2H] : O N HO O O 1530.2. HO N N N O O O O O O O O O NH O O N O OH HPreparation of Example 1037 The yield of the product was 1.8 mg, and its estimated purity by LCMS analysis was 90%. Analysis condition A: Retention time = 1.41 min; ESI-MS(+) m / z [M+2H]2+: 1491.3. Preparation of Example 1038 The yield of the product was 18.7 mg, and its estimated purity by LCMS analysis was 95.4%. Analysis condition A: Retention time = 1.44 min; ESI-MS(+) m / z [M+2H]2+: 1523.Preparation of Example 1039 The yield of the product was 15.9 mg, and its estimated purity by LCMS analysis was 95.3%. Analysis condition A: Retention time = 1.47 min; ESI-MS(+) m / z [M+2H]2+: 1490. Preparation of Example 1040 The yield of the product was 50.8 mg, and its estimated purity by LCMS analysis was 87.9%. Analysis condition B: Retention time = 1.99 min; ESI-MS(+) m / z [M+2H]2+: 1437.Preparation of Example 1044 The yield of the product was 18.7 mg, and its estimated purity by LCMS analysis was 98.3%. Analysis condition A: Retention time = 1.4, 1.44 min; ESI-MS(+) m / z [M+2H]2+: 1001.16, 1000.9. Preparation of Example 1045 The yield of the product was 4.6 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition A: Retention time = 1.38, 1.51 min; ESI-MS(+) m / z [M+2H]2+: 1489.9.Preparation of Example 1046 The yield of the product was 27.4 mg, and its estimated purity by LCMS analysis was 92.7%. Analysis condition A: Retention time = 1.37 min; ESI-MS(+) m / z [M+2H]2+: 1451.1. The following examples were prepared following the general synthetic sequence described for the preparation of Example 1001. Preparation of Example 1029 The yield of the product was 5 mg, and its estimated purity by LCMS analysis was 95.6%. Analysis condition A: Retention time = 1.33 min; ESI- MS(+) m / z [M+2H]2+: 1372.5.Preparation of Example 1030 The yield of the product was 7.4 mg, and its estimated purity by LCMS analysis was 96.4%. Analysis condition A: Retention time = 1.45 min; ESI-MS(+) m / z [M+2H]2+: 1337.5. Preparation of Example 1031 The yield of the product was 0 mg, and its estimated purity by LCMS analysis was 83.7%. Analysis condition A: Retention time = 1.45 min; ESI- MS(+) m / z [M+2H]2+: 1337.5. Analysis condition B: Retention time = 2.04 min; ESI- MS(+) m / z [M+2H]2+: 892.4.Preparation of Example 1041 The yield of the product was 6.2 mg, and its estimated purity by LCMS analysis was 100%. Analysis condition A: Retention time = 2.09 min; ESI-MS(+) m / z [M+2H]2+: 1116.2. Preparation of Example 1047 The yield of the product was 6.8 mg, and its estimated purity by LCMS analysis was 92.8%. Analysis condition B: Retention time = 2.2 min; ESI-MS(+) m / z [M+2H]2+:1174.0. Preparation of Example 1048 The yield of the product was 15 mg, and its estimated purity by LCMS analysis was 95%. Analysis condition A: Retention time = 1.62 min; ESI- MS(+) m / z [M+2H]2+: 1099.2.Preparation of Example 1049 The yield of the product was 5.9 mg, and its estimated purity by LCMS analysis was 98.2%. Analysis condition B: Retention time = 2.52 min; ESI- MS(+) m / z [M+2H]2+: 1148.4. Preparation of Example 1050Preparation of Example 1051 The yield of the product was 6.6 mg, and its estimated purity by LCMS analysis was 98%. Analysis condition B: Retention time = 2.53 min; ESI- MS(+) m / z [M+2H]2+: 1125. Preparation of Example 1052 The yield of the product was 4.4 mg, and its estimated purity by LCMS analysis was 94.1%. Analysis condition A: Retention time = 2.38 min; ESI- MS(+) m / z [M+2H]2+: 1084.1. Method LAG-3 cell binding assay: Human Raji cells expressing endogenous MHCII molecules were used for binding to either human LAG-3-mFc, mouse LAG-3, or cyno LAG-3-hFc proteins. Either Flow Cytometry (FC) or High Content (HC) was employed to assess the binding events. Briefly, Raji cells were plated in a 384-well plate (Thermo 4315 for FC; Corning 354663 for HC) at a density of 24,000 cells / well for the FC readout or 8000 cells / well for the HC readout, respectively. The assay plate was incubated for 2 hour at a 37 ^C and 5% CO2incubator prior to addition of binding antigens if the assay used HC readout. Then, 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. After 30 minutes’ incubation, a corresponding detection antibody (R-Phycoerythrin conjugated anti-Mouse IgG, or anti- human IgG), Jackson Immuno Research Lab, PA) was added at 1:200 ratio, followed by another incubation for 30 minutes. For the FC readout, the assay plate was centrifuged and cell pellets were then resuspended in RPMI media. If the readout is HC, the cells were washed and then fixed by 8% formaldehyde. The binding affinity of the LAG-3 antigen was quantified by reading the plate on an iQue FC reader (Intellicyt) or a NXT HC 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 LAG-3 antigen. A: IC50< 0.01 ^M, B: 0.01 ^M <= IC50< 0.1 ^M; C: 0.1 ^M <= IC50< 1 ^M. Table 1 Biological Data Example hLAG3 / MHCII Raji Number CBA IC50 Example 1001 C Example 1002 C Example 1003 C Example 1004 C Example 1005 C Example 1006 C Example 1007 C Example 1008 C Example 1009 C Example 1010 C Example 1011 C Example 1012 B Example 1013 B Example 1014 C Example 1015 C Example 1016 B Example 1017 C Example 1018 C Example 1019 BExample hLAG3 / MHCII Raji Number CBA IC50 Example 1020 B Example 1021 B Example 1022 B Example 1023 B Example 1024 B Example 1025 B Example 1027 C Example 1028 B Example 1029 C Example 1030 C Example 1031 C Example 1032 C Example 1033 C Example 1034 B Example 1035 B Example 1036 B Example 1037 C Example 1038 B Example 1039 C Example 1040 B Example 1041 C Example 1044 B Example 1045 B Example 1046 B Example 1047 C Example 1048 C Example 1049 C Example 1050 C Example 1051 C Example 1052 C INT-1000 B INT-1001 C INT-1002 A INT-1003 B INT-1004 C INT-1005 B INT-1006 C INT-1007 A INT-1008 A INT-1009 A INT-1010 A

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, 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, 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:Table 1 Example # R3R12X H 1013H 1019 H 1020H 1021H 1022 H 1023OH 1034103510361044 N 1045HN N4. 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.

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