Methods of using cyclic peptides that bind to TNFR1

Abstract

Provided herein, in certain aspects, are methods for inhibiting a TNFR1 -mediated activity using a compound that binds to a binding pocket of TNFR1 between CRD3 and CRD4 of TNFR1.

Description

LVPHLGDREK TASENHLRHC RDSVCPQGKY LSCSKCRKEM IHPQNNSICC GQVEISSCTV TKCHKGTYLY DRDTVCGCRK NDCPGPGQDT NQYRHYWSEN DCRECESGSF LFQCFNCSLC LNGTVHLSCQ EKQNTVCTCH AGFFLRENEC VSCSNCKKSL ECTKLCLPQI ENVKGTEDSG TT (SEQ ID NO: I) As used herein. SEQ ID NO:1 refers to both the extracellular domain of membrane-bound TNFRl and sTNFR and unless otherwise indicated, is used to encompass binding of the compound to both or either TNFRl forms. [0071 [ In addition to its extracellular domain, membrane-bound TNFRl also has a transmembrane and a cytoplasmic domain. The full sequence of membrane-bound TNFRl is represented by SEQ ID NO:2 below, wherein the extracellular domain is underlined, the transmembrane domain is bolded, and the cytoplasmic domain is double underlined: MGLSTVPDLL LPLVLLELLV GIYPSGVIGL VPHLGDREKR DSVCPQGKYI HPQNNSICCT KCHKGTYLYN DCPGPGQDTD CRECESGSFT ASENHLRHCL SCSKCRKEMG QVEISSCTVD RDTVCGCRKN QYRHYWSENL FQCFNCSLCL NGTVHLSCQE KQNTVCTCHA GFFLRENECV SCSNCKKSLE CTKLCLPQIE NVKGTEDSGT TVLLPLVIFF GLCLLSLLFI GLMYRYQRWK SKLYSIVCGK STPEKEGELE GTTTKPLAPN PSFSPTPGFT PTLGFSPVPS STFTSSSTYT PGDCPNFAAP RREVAPPYQG ADPILATALA SDPTPNPLQK WEDSAHKPQS LDTDDPATLY AWENVPPLR WKEFVRRLGL SDHETDRLEL pNGRCLREAQ YSMLATWRRR TPRREATLEL LGRVLRDMDL LGCLEDIEEA LCGPAALPPA PSLLR (SEQ IDNO:2).

[0072] In SEQ ID NO:2, the initial 29 amino acids are a signal sequence and may or may not be present m the TNFRl to which the compounds disclosed herein are bound to inhibit TNFRl-mediated activity. TNFRl, without a signal sequence is represented below as SEQ ID NO:3: LVPHLGDREK RDSVCPQGKY IHPQNNSICC TKCHKGTYLY NDCPGPGQDT DCRECESGSF TASENHLRHC LSCSKCRKEM GQVEISSCTV DRDTVCGCRK NQYRHYWSEN LFQCFNCSLC LNGTVHLSCQ EKQNTVCTCH AGFFLRENEC VSCSNCKKSL ECTKLCLPQI ENVKGTEDSG TTVLLPLVIF FGLCLLSLLF IGLMYRYQRW KSKLYSIVCG KSTPEKEGEL EGTTTKPLAP NPSFtgPTPGFJTP^^ LQNGRCLREA QYSMLATWRR RTPRREATLE LLGRVLRDMD LLGCLEDTEE ALCGPAALPP APSLLR (SEQ ID NO:3).

[0073] Binding of the compounds disclosed herein to one or more residues of SEQ ID NO: 1 of TNFR1 inhibit the downstream activity of the TNFRl. As used herein, binding of the compound to the binding pocket that inhibits TNFRl activity will be made with reference to SEQ ID NO: I. However, it will be understood by one of skill in the art that the same binding pocket is also present in SEQ ID NO:2 and SEQ ID NO: 3. Thus, binding of the compounds disclosed herein will also inhibit downstream activity of a TNFRl that comprises an amino acid sequence of SEQ ID NO:2 and SEQ ID NO:3. One of skill in the art will further understand that a particular numbered residue of SEQ ID NO:1, e.g., Seri 18, also includes and refers to the same residue in SEQ ID NO:2 after accounting for the leading 29 ammo acids of the signaling sequence present in SEQ ID N0:2. That is, the amino acid in SEQ ID N0:2 that corresponds to Seri 18 in SEQ ID NO:1 is Serl47. 7.2.1. TNFRI Binding Pocket and Interaction Residues

[0074] In some aspects, provided herein are methods of inhibiting TNFRI -mediated activity by binding of a compound to a binding site of TNFRI. Other methods are contemplated and described elsewhere herein.

[0075] In some embodiments of the methods disclosed herein, TNFRI is membrane-bound TNFRI. In some embodiments, the membrane-bound TNFRI comprises an amino acid sequence according to SEQ ID NO: 1. In some embodiments, the membrane-bound TNFRI comprises an amino acid sequence according to SEQ ID NO:2. In some embodiments, the membrane-bound TNFRI comprises an amino acid sequence according to SEQ ID NO:3. In some embodiments, the membrane-bound TNFRI is present as a monomer one TNFRI species. In some embodiments, the membrane-bound TNFRI is present as a pre-assembled (e.g., not yet bound to aTNFL) dimer comprising two TNFRI species. In some embodiments, the membrane-bound TNFRI is present as a. pre-assembled (e g., not yet bound to a TNFL) trimer comprising three TNFRI species.

[0076] In some embodiments, TNFRI is sTNFRl. In some embodiments, the sTNFRl comprises an amino acid sequence according to SEQ ID NO: 1. As used herein, and unless otherwise indicated, reference to “TNFRI” includes either or both of membrane-bound TNFRI and sTNFRl.

[0077] In some embodiments of the methods disclosed herein, the binding pocket of TNFRI comprises one or more residues of SEQ ID NO: 1. In some embodiments, the TNFRI binding pocket is recognizable from the primary' sequence, secondary7 structure, or tertiary structure of the ammo acid sequence of the TNFRI. In some embodiments, the TNFRI binding pocket is between two cysteine-rich domains (CRD) of SEQ ID NO:1. In some embodiments, the TNFRI binding pocket is between CRD3 and CRD4 of SEQ ID NO: 1. In some embodiments, the TNFRI binding pocket is between CRD3 and CRD4 of SEQ ID NO:2. In some embodiments, the TNFRI binding pocket is between CRD3 and CRD4 of SEQ ID NO:3. In some embodiments, the TNFRI binding pocket is the TNFR I binding pocket as shown in FIG. 1. In some embodiments, the TNFRI binding pocket is a binding pocket for a compound according to Formula (II). SEQ ID NO:6, 7, 8, 9, Compound A, B. C, D, or a pharmaceutically acceptable salt thereof, as described herein.

[0078] In some embodiments of the methods disclosed herein, the TNFR I binding pocket comprises or consists of the amino acid residues of Asnl34, Thrl35, CysI37, and AsnI48 of SEQ ID NO: I. In some embodiments, the TNFRl binding pocket comprises or further comprises the ammo acid residues of Serll8 through Asnl22 (e.g., Serll8-Leull9-Cysl20-Leul21-Asnl22) of SEQ ID NO: I. In some embodiments, the TNFRl binding pocket comprises or further comprises the amino acid residues of Cysl37 through Cysl39 (e.g. Cysl37-Thrl38-Cysl39) of SEQ ID NO:1. In some embodiments, the TNFRl binding pocket comprises or further comprises the ammo acid residues of Leul45 and Asnl48 through Cysl50 (e.g, Asnl48-Giul49-CysI50) of SEQ ID NO:1. In some embodiments, the TNFRl binding pocket comprises or consists of the amino acid residues of Serll8-Leul 19-Cysl20-Leul21-Asnl22. Cysl37-Thrl38-Cysl39, Leul45, and Asnl48-Glul49-Cysl50 of SEQ ID NO:1. In some embodiments, when a compound as described herein binds at a TNFRl binding pocket, all of the residues recited for the TNFRl binding pocket are bound by the compound. In some embodiments, when a compound as described herein binds at a TNFRl binding pocket, one or more but not ah of the residues recited for the TNFRl binding pocket are bound by the compound. In some embodiments, the TNFRl binding pocket further comprises additional TNFRl residues, such as those additional residues of the TNFRl binding pocket as described herein.

[0079] In some embodiments of the methods disclosed herein, the binding between a residue of the TNFRl binding pocket and a compound as described herein is via a non-covalent interaction. In some embodiments, a non-covalent interaction as described herein may be donated by a residue of the binding pocket of the TNFRl and accepted by the compound. In some embodiments, a non-covalent interaction as described herein may be donated by the compound and accepted by a residue of the binding pocket of the TNFRl. For example, a pi-effect interaction may involve a cation from the binding pocket (e.g, a hydrogen from the backbone of a residue of the binding pocket) being donated to and accepted by a pi orbital of the compound, to form a cation-pi interaction. Cation-pi interactions can occur between cationic side chains of either a lysine or arginine and an aromatic. In some embodiments, a residue of the TNFRl binding pocket binds to a compound as described herein via a lipophilic interaction and / or hydrophobic interaction (e.g, minimization of non-polar surface area exposure to polar molecules). In some embodiments, a residue of the TNFRl binding pocket binds to a compound as described herein via an electrostatic interaction (e.g. Coulombic attraction interaction). In some embodiments, a residue of the TNFRl binding pocket binds to a compound as described herein via an ionic interaction (e.g., electrovalent interaction). In some embodiments, a residue of the TNFRl binding pocket binds to a compound as described herein via an H-bond interaction (e.g, backbone or side chain H-bond interaction). In some embodiments, a residue of the TNFRl binding pocket binds to a compound as described herein via a Van der Waals interaction (e g., a dipole-dipole interaction; dipole-induced dipole interaction; or London dispersion forces). In some embodiments, a residue of the TNFR1 binding pocket binds to a compound as described herein via a pi-effect interaction (e.g., a pi-pi interaction; CH-pi interaction; cation-pi interaction; anion-pi interaction; orpolar-pi interaction) to a residue of the TNFR1. In some embodiments, a pi-effect interaction is a stacking interaction (e g., a pi-pi interaction). In some embodiments, the pi-effect interaction is a non-polar pi-effect interaction. In some embodiments, a polar-pi interaction is a polar hydrogen-pi interaction. In some embodiments, a polar-pi interaction is a polar nitrogen-pi interaction. In some embodiments, a non-covalent interaction may be characterized as more than one type of non-covalent interaction as described herein.

[0080] In some embodiments of the methods disclosed herein, a residue of a TNFR1 binding pocket as described herein binds to a compound at a functional group of a compound. In some embodiments, the functional group of the compound is a functional group on the side chain or backbone moiety. In some embodiments, afunctional group of the compound is an amino group (e.g., NH group, NH2 group, or NHs* group), carbonyl group, carboxylate group, or a cyclic pi-system of the compound (e.g., an arene, aryl, biaryl. heteroaryl, or heterobiaryl). In some embodiments, the functional group of the compound is a carbonyl oxygen of the backbone. In some embodiments, the functional group of the compound is a hydrogen of a backbone amino group. [GC§1 ]    In some embodiments of the methods disclosed herein, the TNFR1 binding pocket comprises one or more of the amino acids corresponding to residues 134. 135, 137. and 148 of SEQ ID NO: I and optionally one or more additional residue of SEQ ID NO: 1. In some embodiments of the methods disclosed herein, the TNFR1 binding pocket comprises one or more of the amino acids corresponding to residues 118-122, 137-139, 145, and 148-150 of SEQ ID NO: 1 and optionally one or more additional residue of SEQ ID NO: 1. In some embodiments, the TNFR1 binding pocket is defined by one or more of ammo acid residues Seri 18, Leull9, Cysl20, Leul21, Asnl22, Cysl37, Thrl38, Cysl39, Leul45, Asnl48, Glut49, and Cysl50 of TNFR1 (SEQ ID NO: 1) and optionally one or more additional residue SEQ ID NO: 1. In some embodiments, the one or more additional residue is selected from the group consisting of Asnl 34 and Thrl35 of SEQ ID NO: 1.

[0082] In some embodiments of the methods disclosed herein, the TNFR1 binding pocket comprises or consists of one or more amino acids corresponding to residues selected from 118, 119, 120. 122, 134, 135, 137, 138, 139, 145, 148, 149. and 150 of the TNFR1 (SEQ IDNO:1). In some embodiments, the TNFR1 binding pocket comprises or consists of one or more ammo acid residues selected from residues Serll8, LeuI19, Cysl20, Leul21, Asnl22, Asnl34. Thr235, Cysl37, Thrl38, Cysl39, Leul45, Asnl48, Glul49, and Cysl50 of the TNFRI (SEQ ID NO: I ). [0083 ]    In some embodiments of the methods disclosed herein, the binding of the compound at the TNFRI binding pocket to a specified residue is mediated by the backbone or side chain of the specified residue (e.g., a backbone NH, backbone carbonyl, or side chain functional group of the specified TNFRI residue interacts with the compound). In some embodiments, one or more residues of the TNFRI binding pocket mediates a non-covalent interaction with a compound as described herein. In some embodiments, one or more residues of the TNFRI binding pocket mediates an interaction with a compound as described herein via a bridging water. In some embodiments, the one or more residues of TNFRI mediating the interaction between the binding pocket and a compound as described herein is selected from Seri 18, Leu 119. CysI20, Leu 121, Asnl22, Cysl37, Thrl38, Cysl39. Leul45, W 148. Glul49, and Cysl50 of the TNFRI (SEQ ID NO:1). [0084 [    In some embodiments of the methods disclosed herein, a residue of the TNFRI binding pocket (e.g.. Seri 18, Leul 19, Cysl20, Leul21, Asn 122, Cysl37, Thrl38, Cysl 39, Leul45. Asn 148, Glul 49, and / or Cysl50 of SEQ ID NO: 1) mediates a non-covalent interaction with a compound as described herein. In some embodiments, a residue of the TNFRI binding pocket (e.g., Seri 18, Leu 119, Cysl 20, Leul21, Asn 122, Cysl37, Thr'138, Cysl 39, Leu 145, Asnl48, Glul49, and / or Cysl50 of SEQ ID NO: 1) mediates a lipophilic interaction and / or hydrophobic interaction with a compound as described herein. In some embodiments, a. residue of the TNFRI binding pocket (e.g., Seri 18, Leul 19. Cysl 20, Leul 21, Asnl22, Cysl 37, Thrl38, Cysl39, Leul45, Asnl48, Glul49, and / or Cysl50 of SEQ ID NO:1) mediates an electrostatic interaction with a compound as described herein. In some embodiments, a residue of the TNFRI binding pocket (e.g.. Seri 18, Leul 19, Cysl 20, Leul2L Asnl22, Cysl 3 7, Thrl38, Cysl 39, Leul45, Asn 148. Glul49, and / or Cysl50 of SEQ ID NO: 1) mediates an ionic interaction with a compound as described herein. In some embodiments, a residue of the TNFRI binding pocket (e.g., Seri 18, Leul 19, Cysl20, Leul21, Asn 122, Cysl37, Thrl38, Cysl39, Leul45, Asnl48, Glu'149, and / or Cysl50 of SEQ ID NO: 1) mediates an H-bond interaction with a. compound as described herein. In some embodiments, a residue of the TNFRI binding pocket (e.g. Seri 18, Leul 19, Cysl 20, Leul21, Asn 122, Cysl37, Thrl38, Cysl39, Leul45, Asn 148, Glul49, and / or Cysl 50 of SEQ ID NO: 1) mediates a Van der Waals interaction with a compound as described herein. In some embodiments, a residue of the TNFRI binding pocket (e.g., Seri 18, Leul 19, Cysl20, Leul21, Asn 122, Cysl37, ThrI38, Cysl39. Leul45, AsnI48, Glul49, and / or Cysl50 of SEQ ID NO:1) mediates a Van der Waals interaction (eg, a dipole-dipole interaction; dipole-induced dipole interaction; or London dispersion force) with a compound as described herein. In some embodiments, a residue of the TNFRI binding pocket (e.g., Seri 18, Leul 19, Cysl 20, Leu121, Asnl22. Cysl37, Thrl38. Cysl 39, Leul45. Asnl48, Glut 49, and / or Cysl 50 of SEQ ID NO:1) mediates a pi-effect interaction (¾ a pi-pi interaction; CH-pi interaction; cation-pi interaction; anion-pi interaction; or polar-pi interaction) with a compound as described herein. In some embodiments of the methods disclosed herein, a residue of the TNFRI binding pocket (e.g., SerlHs. Leul 19, Cysl20. Leul 2L Aso 122. Cysl37. Thrl38, Cysl39. Leu Li v Asnl48. Glut49, and / or Cysl 50 of SEQ ID NO: 1) mediates an interaction with a compound as described herein via a bridging a water molecule. In some embodiments, a residue of the TNFRI binding pocket (e.g., Seri 18, Leul 19, Cysl20, Leul21, Asnl22, Cysl37, Thrl38, Cysl39, Leul45, Asnl48, Glut 49. and / or CysL50 of SEQ ID NO: 1) mediates more than one non-covalenl interaction as described herein, which may be of different types.

[0085] In some embodiments of the methods disclosed herein, residues of the TNFRI binding pocket may mediate an H-bond interaction between the TNFRI and a compound as described herein, either directly or indirectly through a bridging water.

[0086] In some embodiments of the methods disclosed herein, residue Asnl34 of the TNFRI mediates one or two H-bond interactions between the TNFRI and a compound as described herein. In certain embodiments, residue Cys 137 of SEQ ID NO: 1 mediates one or two H-bond interactions between the TNFRI and a compound as described herein. In certain embodiments, residue Asnl48 of SEQ ID NO:1 mediates one or two H-bond interactions between the TNFR I and a compound as described herein. In certain embodiments, residue Asnl48 of SEQ ID NO: 1 mediates one or two H-bond interactions between the TNFRI and a compound as described herein. In certain embodiments, one or two of the H-bond interactions between the TNFRI and a compound are via a bridging water. In certain embodiments, residue Thrl35 of SEQ ID NO: 1 mediates one or two H-bond interactions between the TNFRI and a compound as described herein.

[0087] In some embodiments of the methods disclosed herein, the binding interactions between the TNFR I and compound as described herein are as depicted in any one of figures disclosed herein. 7.3. Methods

[0088] The present disclosure provides a method of inhibiting TNFRI -mediated activity, comprising selectively contacting the TNFRI with a compound as disclosed herein, such as in Section 7.4, that binds to TNFRI at a binding pocket between CRD3 and CRD4 of the TNFRI, wherein the binding pocket is defined according to any embodiment as disclosed herein, such as in Section 7.2.1. In some embodiments, the compound is a compound of Formula (II), SEQ ID N0s;6. 7, 8, 9, Compounds A. B, C. D, or a pharmaceutically acceptable salt of any of the foregoing.

[0089] In another aspect, the present disclosure provides a method of inhibiting oligomerization of two or more TNFRl receptor species comprising contacting the TNFRl with a compound disclosed herein, such as in Section 7.4, that competes for binding to the TNFRl al a binding pocket between CRD3 and CRD4 of the TNFRl, wherein the binding pocket is defined according to any embodiment as disclosed herein, such as in Section 7.2.1. In some embodiments, the compound is a compound of Formula (II), SEQ ID NOs:6, 7, 8, 9, Compound A, B, C, D. or a pharmaceutically acceptable sail of any of the foregoing.

[0090] In another aspect, the present disclosure provides a method of inhibiting oligomerization of two or more TNFRl receptor species comprising contacting the TNFRl with a first compound that competes with a second compound for binding to the TNFRl at a binding pocket between CRD3 and CRD4 of the TNFRl. In a specific embodiment, the second compound is any compound disclosed herein, such as in Section 7.4. In a specific embodiment, the binding pocket is defined according to any embodiment as disclosed herein, such as in Section 7.2.1. In some embodiments, the second compound is a compound of Formula (II), SEQ ID NOs:6, 7, 8, 9, Compound A, B, C, D, or a pharmaceutically acceptable salt of any of the foregoing. [009 Q    In another aspect, the present disclosure provides method of inhibiting a TNFR1 - mediated activity comprising contacting a first TNFRl with a compound as disclosed herein, such as in Section 7.4, that inhibits interaction with a second TNFRl with the first TNFRl. In some embodiments, the first TNFRl is TNFL-bound TNFRl. In some embodiments, the second TNFRl is TNFL-bound TNFRl. In some embodiments, the TNFL is TNFa.

[0092] In another aspect, the present disclosure provides method of inhibiting a TNFRl -mediated activity comprising contacting a first TNFRl with a first compound that competes with a second compound for binding to the TNFRl at a binding pocket between CRD3 and CRD4 of the TNFRl. In a specific embodiment, the second compound is any compound disclosed herein, such as m Section 7.4. In a specific embodiment, the binding pocket is defined according to any embodiment as disclosed herein, such as in Section 7.2.1. In some embodiments, the second compound is a compound of Formula (II), SEQ ID NOs:6, 7, 8, 9, Compound A, B, C, D, or a pharmaceutically acceptable salt of any of the foregoing. In certain embodiments, the first compound inhibits interaction of a second TNFRl with the first TNFRl. In some embodiments, the first TNFRI is TNFL-bound TNFRI. In some embodiments, the second TNFRI is TNFL-bound TNFRI. In some embodiments, the TNFL is INFa.

[0093] In another aspect, the present disclosure provides method of inhibiting a TNFL-mediated activity comprising contacting aTNFRJ with a compound as disclosed herein, such as in Section 7.4, that inhibits interaction with a second TNFRI with the first TNFRI. In some embodiments, the first TNFRI is TNFL-bound TNFRI. In some embodiments, the second TNFRI is TNFL-bound TNFRI. In some embodiments, the TNFL is TNFa.

[0094] In another aspect, the present disclosure provides method of inhibiting a TNFL-mediated activity- comprising contacting a first TNFRI with a first compound that competes with a second compound for binding to the TNFRI at a binding pocket between CRD3 and CRD4 of the TNFRI. In a specific embodiment, the second compound is any compound disclosed herein, such as in Section 7.4. In a specific embodiment, the binding pocket is defined according to any embodiment as disclosed herein, such as in Section 7.2.1. In some embodiments, the second compound is a compound of Formula (II), SEQ ID NOs:6, 7, 8, 9, Compound A, B, C, D, or a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the TNFL is TNFa.

[0095] In another aspect, the present disclosure provides a method of inhibiting oligomerization of two or more TNFR1 receptor species comprising contacting a first TNFRI with a compound, with a compound as disclosed herein, such as in Section 7.4, that inhibits the interaction of the first TNFRI with a second TNFRI. In some embodiments, the first TNFRI is TNFL-bound TNFRI. in some embodiments, the second TNFR1 is TNFL-bound TNFRI. In some embodiments, the TNFL is TNFa.

[0096] In another aspect, the present disclosure provides a method of inhibiting oligomerization of tw o or more TNFRI receptor species comprising contacting a first TNFRI with a first compound that competes with a second compound for binding to the TNFRI at a binding pocket between CRD3 and CRD4 of the TNFRI. In a specific embodiment, the second compound is any compound disclosed herein, such as in Section 7.4. In a specific embodiment, the binding pocket is defined according to any embodiment as disclosed herein, such as in Section 7,2.1. In some embodiments, the second compound is a compound of Formula (II), SEQ ID NOs:6, 7. 8, 9, Compound A, B, C, D, or a pharmaceutically acceptable salt of any of the foregoing. In certain embodiments, the first compound inhibits interaction of a second TNFRI with the first TNFRI. In some embodiments, the first TNFRI is TNFL-bound TNFRI. In some embodiments, the second TNFRI is TNFL-bound TNFRI. In some embodiments, the TNFL. is TNFa. In some embodiments, the first TNFR1 is TNFL-bound TNFRL Tn some embodiments, the second TNFR1 is TNFL-bound TNFRL In some embodiments, the TNFL is TNFa.

[0097] As used herein, TNFRl-mediated activity refers io any downstream biological activity of TN.FR1 activation by any TNFL (including TNFa) including, but not limited to., inflammation, cytokine signaling, apoptosis, cell proliferation, cell survival, cell differentiation, MAPK signaling, NF-kB signaling, and any combination thereof. As used herein, reference to the compound binding to TNFR1 also encompasses binding to sTNFRL

[0098] In certain embodiments, the compound binds the TNFR1 at a binding pocket, wherein the binding of the compound to the binding pocket blocks the TNFRl-mediated activity. In certain embodiments, the compound binds to the extracellular domain of TNFR 1. In certain embodiments, the compound binds to sTNFRL In certain embodiments, the compound or does not bind to TNFR1 in the binding pocket specific for the ligand of TNFR1 (e g., TNFa), In certain embodiments, the compound binds to the TNFR1 at a region distinct from where the ligand of TNFR 1 (e.g., TNFa) binds. In certain embodiments, the binding of the compound to the TNFR1 is non-competitive with the binding of tire ligand of TNFR1 (eg., TNFa), In certain embodiments, the compound binds to TNFR1 at an interface of a first TNFR! that interacts with a second TNFR1 once each of the first and second TNFRI is bound by a ligand of TNFR1 (e.g., TNFa). In certain embodiments, the interface is a clustering interface. In certain embodiments, the compound blocks a second TNFL-bound TNFR1 from binding to or interacting with the first TNFL-bound TNFRL In some embodiments, the compound binds to a binding pocket located between CRD3 and CRD4 of TNFRL hi some embodiments, the binding of the compound to the TNFR] at the binding pocket inhibits oligomerization or clustering of TNFRL In certain embodiments, the compound binds to a binding pocket as disclosed in Section 7.2.1 herein.

[0099] In certain embodiments, binding of the compound as disclosed herein to TNFR1 disrupts the interaction between TNFL-bound TNFR1 receptor species. In certain embodiments, binding of the compound as disclosed herein to TNFR1 inhibits the interaction between TNFL-bound TNFR1 monomers. In certain embodiments, binding of the compound as disclosed herein to TNFR! prevents interaction between TNFL-bound TNFR1 receptor species. In some embodiments, binding of the compound as disclosed herein to TNFR1 prevents oligomerization of TNFR1 upon binding by a TNFL to the TNFRL

[00100] In some embodiments of the methods described herein, the compound is a compound as disclosed herein, such as in Section 7.4, In some embodiments, the compound is a compound of Formula (II), SEQ ID NOs:6, 7, 8, 9, Compound A, B, C, D, or a pharmaceutically acceptable salt of any of the foregoing. 7.4. Compounds that Bind to TNFRI JOOlOlj In certain aspects of the various methods provided herein, the present disclosure compounds are provided that bind to a binding pocket of the TNFRI. Exemplary compounds are described in any embodiment in Section 7.2.1. In certain embodiments, the compound does not bind to TNFRi in the binding pocket specific for ligand of TNFRI (e.g., TNFa). In certain embodiments, the compound does not bind to a binding pocket of TNFRI located between cysteine rich domain 2 (CRD2) and CRD3. In certain embodiments, the compound binds to an interface, for example, a clustering interface, of TNFR I. In certain embodiments, the compound binds to TNFRI between CRD3 and CRD4 of TNFRI. In certain embodiments, the TNFRI binding pocket is defined by ammo acid residues Asn 134, Thrl35, Cysl37, and Asnl48 of SEQ ID NO:1. In certain embodiments, the TNFR I binding pocket is defined or further defined by ammo acid residues Seri 18 through Asnl22, Cysl37 through Cysl39, and Asn 148 through Cysl50 of SEQ ID NO:1. In certain embodiments, the TNFRI binding pocket is defined or further defined by amino acid residues Cysl37 through Cysl39, Leul45, and Asnl48 through Cysl.50 of SEQ ID NO: 1. In certain embodiments, the TNFRI binding pocket is defined or further by amino acid residues Cysl37 through Cysl39 and Asnl48 through Cysl50 of SEQ ID NO: 1. In certain embodiments, the TNFRi binding pocket is defined or further defined by ammo acid residues Serll8 through Asnl22, Cysl37 through Cysl39, Leu 145, and Asn 148 through Cysl50 of SEQ ID NO:1. In certain embodiments, the compound binds to TNFRI at one or more of residues Asnl34. Thrl35, Cysl 3 7, and Asn 148 of SEQ ID NO: 1. In certain embodiments, the compound binds to TNFRI at. two or more of residues Asnl34, Thrl35, Cysl37, and Asn 148 of SEQ ID NO:1. In certain embodiments, the compound binds to TNFRI at three or more of residues Asnl34. Thrl35, Cysl37. and Asnl48 of SEQ ID NO:1. In certain embodiments, the compound binds to TNFRI at each of residues Asn 134, Thrl35, Cysl37, and Asnl48 of SEQ ID NO: I.

[00102] In some embodiments of the methods disclosed herein, a compound as described herein inhibits oligomerization of a TNFL-bound TNFRI, which inhibits one or more biological functions of the TNFRI. In some embodiments, a compound as described herein blocks the interface of a first TNFL-bound TNFR I al which a second TNFL-bound TNFRI would otherwise bind to form an oligomeric TNFRI cluster, which inhibits one or more biological functions of one or both of the first and second TNFL-bound TNFRI. In some embodiments, the blocking is a steric blocking. In some embodiments, a compound as described herein binds to a first TNFL-bound TNFRI betw een CRD3 and CRD4, which blocks an interface of the first TNFL-bound TNFRI at which a second TNFL-bound TNFRI would otherwise bind to form an cluster, and in turn, inhibits one or more biological functions of TNFRI. In some embodiments, the biological function of TNFR1 is inflammation (e g., in a subject). In some embodiments, the biological function of TNFR1 is cytokine signaling. In some embodiments, the biological function of TNFR1 is apoptosis. In some embodiments, the biological function of TNFR1 is necroptosis. In some embodiments, the biological function of TNFR1 is cell proliferation. In some embodiments, the biological function of TNFR1 is cell survival. In some embodiments, the biological function of TNFR1 is cell differentiation. In some embodiments, the biological function of TNFR1 is MAPK signaling. In some embodiments, the biological function of TNFR! is NF-kB signaling.

[00103] In certain embodiments of the methods provided herein, the compound that binds to TNFR1 as disclosed herein comprises a peptide. In certain embodiments, the compound that binds io TNFR1 as disclosed herein is a peptide. In some embodiments, the compound is cyclic. In certain embodiments, the compound is a cyclic peptide.

[00104] In certain embodiments of the methods provided herein, the cyclic peptide comprises or consists of an amino acid sequence having a length of from 9 to 28 amino acid residues (e.g., 9, 10, 11. 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26. 27, or 28 amino acids). In some embodiments, the cyclic peptide comprises or consists of an amino acid sequence having a length of from 10 to 24 ammo acid residues. In some embodiments, the cyclic peptide comprises or consists of an amino acid sequence having a length of from 11 to 20 ammo acid residues. In some embodiments, the cyclic peptide comprises or consists of an amino acid sequence having a length of from 11 to 18 amino acid residues. In some embodiments, the cyclic peptide comprises or consists of an amino acid sequence having a length of from 11 to 1 7 ammo acid residues, in some embodiments, the cyclic peptide comprises or consists of an amino acid sequence having a length of from 12 to 16 amino acid residues. In some embodiments, the cyclic peptide comprises or consists of an amino acid sequence having a length of from 13 to 15 ammo acid residues. In some embodiments, the cyclic peptide comprises or consists of an amino acid sequence having a length of 12 amino acids. In some embodiments, the cyclic peptide comprises or consists of an amino acid sequence having a length of 13 amino acids. In some embodiments, the cyclic peptide comprises or consists of an amino acid sequence having a length of 14 amino acids. In certain embodiments, the cyclic peptide comprises or consists of an amino acid sequence having a length of 15 ammo acids. In some embodiments, the cyclic peptide comprises or consists of an ammo acid sequence having a length of 16 amino acids. In certain embodiments, the compound further comprises anon-peptidic linker.

[00105] In certain embodiments of the methods provided herein, the compound is a macrocyclic peptide. In some embodiments, the macrocyclic peptide comprises or consists of an amino acid sequence having a length of from 9 io 28 amino acid residues (e.g., 9. 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 2.4, 25. 26, 27, or 28 amino acids). In some embodiments, the macrocyclic peptide comprises or consists of an amino acid sequence having a length of from 10 to 24 amino acid residues In some embodiments, the macrocyclic peptide comprises or consists of an ammo acid sequence having a length of from 11 to 20 ammo acid residues. In some embodiments, the macrocyclic peptide comprises or consists of an amino acid sequence having a length of from 12 to 16 amino acid residues. In some embodiments, the macrocyclic peptide comprises or consists of an ammo acid sequence having a length of from 13 to 15 ammo acid residues. In some embodiments, the macrocyclic peptide comprises or consists of an ammo acid sequence having a length of 12 amino acids. In some embodiments, the macrocyclic peptide comprises or consists of an amino acid sequence having a length of 13 amino acids. In some embodiments, the macrocyclic peptide comprises or consists of an ammo acid sequence having a length of 14 amino acids. In certain embodiments, the macrocyclic peptide comprises or consists of an amino acid sequence having a length of 15 amino acids. In some embodiments, the macrocyclic peptide comprises or consists of an amino acid sequence having a length of 16 amino acids. In certain embodiments, the compound further comprises a non-peptidic linker.

[00106] In some embodiments, the compound is a compound as described herein. In some embodiments, a compound as described herein that binds to TNFR1 at a binding pocket as described herein is a lipophilic compound comprising one or more lipophilic moieties. In some embodiments, a compound as described herein that binds to TNFR1 at a binding pocket as described herein is hydrophobic compound comprising one or more hydrophobic moieties.

[00107] In certain embodiments of the methods provided herein, the compound that binds to TNFRi as disclosed herein has a molecular weight of from about 1200 Da to about 3000 Da. In certain embodiments, the compound has a molecular weight of from about 1200 Da to about 2500 Da. In certain embodiments, the compound has a molecular weight of from about 1200 Da to about 2250 Da. In certain embodiments, the compound has a molecular weight of from about from about 1500 Da to 2250 Da. In certain embodiments, the compound has a molecular weight of from about from about 1500 Da to 2500 Da. In certain embodiments, the compound has a molecular weight of from about from about 1500 Dato 3000 Da. In certain embodiments, the compound has a molecular weight of from about 1750 Da to about 3000 Da. In certain embodiments, the compound has a molecular weight of from about 1750 Da to about 2500 Da. In certain embodiments, the compound has a molecular weight of from about 1750 Da to about 2250 Da.

[00108] In certain embodiments of the methods provided herein, the compound that binds to TNFRl as disclosed herein comprises one or more moieties selected from: (a) an H-bond interaction moiety capable of accepting an H-bond from the side chain of residue Asnl48 of the TNFRl; (b) an H-bond interaction moiety capable of donating an H -bond to the side chain of residue Asnl48 of the TNFRl; (c) an H-bond interaction moiety capable of accepting an H-bond from the backbone of residue Cysl37 of the TNFRl; and (d) an H-bond interaction moiety’ capable of donating an H-bond to the backbone of residue Cysl37 of the TNFRl.

[00109] In certain embodiments of the methods provided herein, the compound that binds to TNFRl as disclosed herein comprises one or more moieties selected from: (a) an H-bond interaction moiety capable of donating an H-bond to the backbone of residue Thrl35 of the TNFRl ; and (b) an H-bond interaction moiety capable of accepting an H-bond from the sidechain residue Asnl34 of the TNFRl.

[00110] In certain embodiments of the methods provided herein, the compound that binds to TNFRl as disclosed herein comprises an H-bond interaction moiety capable of accepting an H-bond from the side chain of residue Asnl34 (e.g., the side chain amine of residue Asnl34) of SEQ ID NO: 1. In certain embodiments, the compound comprises an H-bond interaction moiety capable of donating an H-bond to the side chain of residue Asnl34 (e.g., the side chain aminocarbonyl oxygen of residue Asn 134 of TNFRl) of SEQ ID NO:1.

[00111] In certain embodiments, the compound comprises an H-bond interaction moiety capable of donating an H-bond to the backbone of residue Thrl35 (e.g., the carbonyl oxygen of the backbone of residue Thrl 35 of TNFRl) of SEQ ID NO:1.

[00112] In certain embodiments, the compound comprises an H-bond interaction moiety capable of accepting an H-bond from the backbone of residue Cysl37 (e.g., the amide nitrogen of the backbone of residue Cy s 137 of TNFR l ) of SEQ ID NO:1. In certain embodiments, the compound comprises an H-bond interaction moiety capable of donating an H-bond to the backbone of residue Cysl37 (e.g.. the carbonyl oxygen of the backbone of residue Cysl37 of TNFRl) of SEQ H)NO:L

[00113] In certain embodiments of the methods provided herein, the compound that binds to TNFRl as disclosed herein comprises an H-bond interaction moiety capable of accepting an H-bond from the side chain of residue Asnl48 (e.g., the side chain amine of residue Asnl48 of TNFRl) of SEQ ID NO:1. In certain embodiments, the compound comprises an H-bond interaction moiety capable of donating an H-bond to the side chain of residue Asnl48 (e.g., the side chain aminocarbonyl oxygen of residue Asnl48 of TNFRl) of SEQ ID NO: 1.

[00114] In certain embodiments of the methods provided herein, the compound comprises two or more of the H-bond interaction moieties described above. In certain embodiments of the methods provided herein, the compound comprises three or more of the H-bond interaction moieties described above. In certain embodiments of the methods provided herein, the compound comprises four or more of the H-bond interaction moieties described above. In certain embodiments of the methods provided herein, the compound comprises five or more of the H-bond interaction moieties described above. In certain embodiments of the methods provided herein, the compound comprises six or more of the H-bond interaction moieties described above. In certain embodiments of the methods provided herein, tire compound comprises all of the H-bond interaction moieties described above.

[00115] As used herein, the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, and thirteenth amino acids of the compound are as depicted with reference to Formula (II) in FIG. 2.

[00116] In certain embodiments of the methods provided herein, the compound binds to the TNFRl binding pocket according to any embodiment disclosed herein via an aliphatic or aliphatic-ary 1 (e.g., alkyl or aryl-alkyl) side chain on the first amino acid residue of the compound. In certain embodiments, the compound binds to the TNFRl binding pocket according to any embodiment disclosed herein via a bicyclic aryl or heteroaryl moiety of an eighth amino acid of the compound. In certain embodiments, the compound binds to the TNFR1 binding pocket according to any embodiment disclosed herein via an hydroxymethyl moiety of a ninth amino acid of the compound. In certain embodiments, the compound binds to the TNFRl binding pocket according to any embodiment disclosed herein via two or three moieties selected from the group consisting of an aliphatic (e.g., alkyl) or aliphatic-aiyl (e.g.. aryl-alkyl) side chain on the first amino acid residue of the compound, a bicyclic ary l or heteroaryl moiety’ of the eighth amino acid of the compound, a hydroxymethyl moiety of the ninth amino acid of the compound, and an aryl moiety of the tenth amino acid of the compound.

[00117] In certain embodiments of the methods provided herein, the H-bond interaction moiety capable of accepting an H-bond from the amino hydrogen of the backbone of residue Cysl 37 of TNFRl is present on the backbone or side chain of the eighth amino acid of the compound. In certain embodiments, the H-bond interaction moiety capable of donating an H-bond to the carbonyl oxygen of the backbone of residue Cysl 37 of TNFRl is present on the backbone or side chain of the eighth ammo acid of the compound. In certain embodiments, the eighth amino acid of the compound is 3-(2-naphthyl)-L-alanme (2Nal), (5)-2-amino-3-(7-chloro-lH-mdoI-3-yl)propanoic add (Trp7Cl), or L-try ptophan (W).

[00118] In certain embodiments of the methods provided herein, the H-bond interaction moiety capable of accepting an H-bond from the side chain of residue Asnl34 of the TNFR1 is present on the backbone or side chain of the tenth amino acid of the compound. In certain embodiments, the H-bond interaction moiety capable of accepting an H-bond from the side chain of residue Asnl34 of the TNFRJ is a carbonyl moiety. In certain embodiments, the H-bond interaction moiety7 capable of accepting an H-bond from the side chain of residue Asnl34 of the TNFR1 is a carboxylate-substituted or hydroxy-substituted phenyl moiety. In certain embodiments, the tenth amino acid of the compound is L-tyrosine (Y) or L-phenylalanine-4-carboxylic acid (Phe4COOH).

[00119] In certain embodiments of the methods provided herein, the H-bond interaction moiety- capable of accepting an H-bond from the side chain of residue Asnl48 of the TNFR1 is present on the backbone or side chain of the third amino acid of the compound. In certain embodiments, the H-bond interaction moiety- capable of donating an H-bond to the side chain of residue Asnl48 of the TNFR1 is present on the backbone or side chain of the third amino acid of the compound. In certain embodiments, the H-bond interaction moiety capable of donating an H-bond to Asnl48 comprises a hydroxyl moiety or an amine moiety. In certain embodiments, the H-bond interaction moiety capable of donating an H-bond to Asnl48 comprises an hydroxy alkyl moiety. In certain embodiments, the third ammo acid of the compound is (5)-piperazme-2-carboxylic acid (Pip2c), (5)-piperidme-2~carboxylic acid (Pip), or L-serine (S).

[00120] In certain embodiments of the methods provided herein, an H-bond interaction moiety capable of donating an H-bond to the backbone of residue Thrl35 of SEQ ID NO: 1 is present on the backbone or side chain of the ninth amino acid of the compound. In certain embodiments, the H-bond interaction moiety capable of donating an H-bond to the carbonyl oxygen of the backbone of residue Thrl35 of SEQ ID NO:1 comprises a hydroxyl moiety7. In certain embodiments, the H-bond interaction moiety capable of donating an H-bond to the carbonyl oxygen of the backbone of residue Thrl35 of SEQ ID NO: 1 comprises an hydroxyalky7! moiety7, for example an hydroxymethyl moiety7. In certain embodiments, the ninth ammo acid of the compound is a-methyl-L-serme (aMeS) or i,-serine (S).

[00121] In certain embodiments, the thirteenth amino acid of the compound is a-methyl-L-tryptophan (aMeW), L-lryplophan (W), or (2S,3S)-2-ammo-3-(7-melhoxy-l / / -indol-3-yl)butanoic acid (SbMeW7OMe).

[00122] In certain embodiments of the methods provided herein, the first amino acid comprises an aliphatic side chain or aryl-containing side chain. In certain embodiments, the first amino acid comprises a Ci-6 alkyl side chain or a C1-3 alkylaryl side chain. In certain embodiments, the first amino acid comprises a methyl, ethyl, propyl, butyl moiety. In certain embodiments, the side chain of the first amino acid is propy l. In certain embodiments, the side chain of the first amino acid is butyl. In certain embodiments, the side chain of the first amino acid is substituted with a monocyclic aryl moiety (e.g., phenyl). In certain embodiments, the side chain of the first amino acid is phenyl. In certain embodiments, the first ammo acid of the compound is L-norleucine (Nie), L-phenylalanine (F), or (>S’)-2-(amino)heptanoic acid (Ahp).

[00123] In certain embodiments of the methods provided herein, the eighth amino acid comprises a side chain comprising an optionally substituted bicyclic aryl or heteroaryl moiety. In certain embodiments, the eighth amino acid comprises aside chain comprising an optionally substituted indolyl or naphthyl moiety. In certain embodiments, the eighth amino acid of the compound is L-tryptophan (W), 3-(2-naphthyl)-L-alanine (2Nal), or (5)-2-amino-3-(7-chloro-l / ?-indol-3-yl)propanoic acid (Trp7Cl).

[00124] In certain embodiments, the tenth amino acid comprises a side chain comprising substituted or unsubstituted benzyl. In certain embodiments, the side chain of the tenth amino acid comprises benzyl substituted with a carboxylic acid (-COOH) moiety. In certain embodiments, the side chain of the tenth amino acid comprises benzyl para-substituted with a carboxylic acid (-COOH) moiety. In certain embodiments, the side chain of the tenth amino acid comprises benzyl substituted with a hydroxyl (-OH) moiety. In certain embodiments, the side chain of the tenth amino acid comprises benzyl para-substituted with a hydroxyl (-OH) moiety. In certain embodiments, the tenth amino acid of the compound is L-tyrosine (Y). or (5)-4-(2-amino-2“Carboxyethyl)benzoic acid (Phe4COOH).

[00125] Without being bound by theory, it is believed that binding of the compounds to TNFRl leads to a conformational change of an interface of a first TNFL-bound TNFR1 trimertrimer complex at which a second TNFL-bound TNFRl tnmerrtrimer complex would otherwise bind or interact io form an oligomeric TNFL-bound TNFRl cluster. For example, such conformational changes can lead to steric inhibition of the binding of the second TNFL-bound TNFRl tnmerrtrimer complex to the first TNFL-bound TNFRl. This conformational change effectively inhibits oligomerization of the first and second TNFRl receptor species. Inhibition of oligomerization, in turn, inhibits TNFRl-mediated activity that would otherwise occur if the TNFR 1 monomers had proceeded to oligomerize. In some embodiments. TNFRl -mediated activity is inflammation, cytokine signaling, apoptosis, necroptosis, cell proliferation, cell survival, cell differentiation, MAPK signaling, NF-kB signaling, or any combination thereof. In some embodiments, TNFRl-mediated activity is inflammation. In some embodiments, TNFR1 -mediated activity is cytokine signaling. In some embodiments, TNFRl-mediated activity is apoptosis. In some embodiments, TNFR1 -mediated activity is necroptosis. In some embodiments, TNFRl-mediated activity is cell proliferation. In some embodiments. TNFRl-mediated activity' is cell survival. In some embodiments, TNFRl-mediated activity' is cell differentiation. In some embodiments, TNFRl-mediated activity is MAPK signaling. In some embodiments, TNFRl-mediated activity is NF-kB signaling, or any combination thereof.

[00126] In certain embodiments of the methods provided herein, the compound that binds to TNFR1 comprises an amino acid sequence, wherein the first amino acid in the sequence comprises an aliphatic or aryl-alkyl moiety. In certain embodiments, the third amino acid in the sequence comprises a hydroxyl moiety or an amine moiety. In certain embodiments, the eighth amino acid in the sequence comprises an optionally substituted bicyclic aryl or heteroaryl moiety. In certain embodiments, the ninth amino acid in the sequence comprises an hydroxymethyl moiety. In certain embodiments, the tenth ammo acid in the sequence comprises an optionally substituted aryl moiety-.

[00127] In certain embodiments of the methods provided herein, the compound that binds to TNFR1 comprises an amino acid sequence, wherein: the first amino acid in the sequence comprises an alkyl or aryl-alkyl moiety; the third amino acid in the sequence comprises a hydroxyl moiety or an amine moiety; the eighth ammo acid in the sequence comprises a bicyclic aryl or bicyclic heteroaryl moiety; the ninth amino acid in the sequence comprises a hydroxymethyl moiety-; and the tenth amino acid in the sequence comprises an aryl-methyl moiety.

[00128] In certain embodiments of the methods provided herein, the compound that binds to TNFR1 comprises an amino acid sequence, wherein: the first amino acid in the sequence comprises a Cue alkyl or aryl(Co--3 alkyl) moiety' (e.g., pentyl, butyl or benzyl); the third amino acid in the sequence comprises an hydroxymethyl moiety, a pipendinyl moiety’, or a piperazinyl moiety': the eighth amino acid in the sequence comprises a naphthyl, indolyl, naphthylniethyl or indolylmethyl moiety; the ninth amino acid in the sequence comprises an hydroxymethyl moiety; and the tenth amino acid in the sequence comprises a substituted benzyl moiety (e.g., phenyl substituted (e.g, para-substituted) with a hydroxyl moiety or a carboxylic acid moiety).

[00129] In certain embodiments of the methods provided herein, the compound that binds to TNFR1 comprises an amino acid sequence, wherein: the first amino acid in the sequence comprises (5)-2-aminoheptanoic acid, L-norleucine, or L-phenylalanine; the third amino acid in the sequence comprises (S)-piperazine-2-carboxy1ic acid, (S)-piperidine-2-carboxyhc acid, orL-serine; the eighth amino acid in the sequence comprises 3-(2-naphthyl)-L-alanine, (S)-2-amino-3-(7-chloro-Z / f-indo]-3-yl)propanoic acid, or L-tiy ptophan; the ninth amino acid in the sequence comprises a-melhyl-L-serine or L-serine; and the tenth ammo acid in the sequence comprises L-tyrosine or L-phenylalanine-4-carboxylic acid.

[00130] In certain embodiments of the methods provided herein, the compound that binds to TNFR1 comprises an amino acid sequence according to SEQ ID NO:6: CIAc’-Ahp-lPal-S-SbMeW-dP-N-Phe4COOH-2Mal-aMeS-Phe4COOH-H-Sar-aMeW-C*-NH2 (SEQ ID NO :6) or pharmaceutically acceptable salt thereof, wherein * indicates attachment to form a cyclic peptide, and where each moiety is described herein, e.g., in Table 1.

[00131] In certain embodiments of the methods provided herein, the compound that binds to TNFR1 comprises an amino acid sequence according to SEQ ID NO:7: Nle*-4Pal-Pip2c-W-NMeE-N-Phe4COOH-Trp7Cl-aMeS-Phe4COOH-Y-Sar-SbMeW7OMe-bhcLeu* (SEQ ID NO:7) or pharmaceutically acceptable salt thereof, wherein * indicates attachment to form a cyclic peptide, and where each moiety is described herein, eg., in Table 1.

[00132] In certain embodiments of the methods provided herein, the compound that binds to TNFR1 comprises an amino acid sequence according to SEQ ID NO:8: Nle*-4Pal-Pip-W-NMeE-N-4Pal-Trp7Cl-aMeS-Phe4COOH-Pbe3CONH2-Sar-SbMeW7OMe-bhcLeu* (SEQ ID NO: 8) or pharmaceutically acceptable salt thereof, wherein * indicates attachment to form a cy clic peptide, and where each moiety is described herein, e.g., in Table 1.

[00133] In certain embodiments of the methods provided herein, the compound that binds to TNFR1 comprises an ammo acid sequence according to SEQ ID NO:9: ClAc*-F-Y-S-W-Sar-N-Y-W-S-Y-Y-Sar-W-C*-G-E-E-NH2 (SEQ ID NO:9) or pharmaceutically acceptable salt thereof, wherein * indicates attachment to form a cyclic peptide, and where each moiety is described herein, e.g, in Table 1.

[00134] In certain embodiments of the methods provided herein, the compound is a compound of Formula (I) or a pharmaceutically acceptable salt thereof: (I) wherein: R1 is selected from hydrogen, CMo alkyl, (Cw alkyl>2 ammo(Cc,.i0 alkyl), (C^ alkyl)o-2 amino(CQ.10 alkyl)oxy(C0^ alkyl), (CM alkvd)3N+(C0.6 alkyl), atyl(C0.i0 alkyl), heteroary!(C0.w alkyl), (Ch.^.)^cloaIkyl(Cg_jQ alkyl), helerocycloalkyl(Cg_]g alkyl), fluoioalkyl, Cp-io alkenyl, (C0.6 alkyl)carbonylamino(C0^ alkyl), (C0.6 alkyl) 0-2 aminocarbonyl(C0^ alkyl), (Cw alkyl)o-2 aminocarbonylamino(C0^ alkyl), arylcarbonylammo(C0.,6 alkyl), a.rylaminocarbonyl(C0..6 alkyl), heteroarylcarbonylammo(C0.6 alkyl), heteroaiylaminocarbonyl(C0.6 alkyd), Cb6 alkyloxy, (Ci.5 aikyl)oxy(C0.6 alkyl), ((C3.]2)cycloalkyl)oxy(Co.6 alkyl), ((C3.]2)cycloalkyl C0,6 alkyI)oxy(C0.6 alkyl). (C0.6 alkyl)carboxy(Cc.,6 alkyl), N'^N' :::N-(C0.6 alkyl), and H2N-C( M!)MI |C„, alkyl), wherein is substituted by 0, 1, 2, 3. or 4 R substituents each independently selected from alkyl, amino, cyano, halo, hydroxy, (C3.12)cycloalkyloxy, and CM alkyloxy: each is independently selected from hydrogen, hydroxy, CMalkyl, fluoro, and alkyloxy: R^b is selected from aiyl, heteroaryl, cycloalkyl, and heterocycloalkyl, wherein said heteroaryl contains at least one nitrogen atom, wherein R^b js substituted by 0, 1. or 2 R2c independently selected from C-.,6 alkyl, amino(C0.6 alkyl), (C^ alkyl)o-2 amino(C0.6 alkyl), (Cj,6 alkyljjhTfCg.g alkyl), aminocarbonyl(C0. 6 alkyl), (Ci,6 alkyl)o-2 aminocarbonyl(C0.6 alkyl), hydroxy, C3.6alkyloxy, halo, (C^ alkyl)o-2 amino(C0^ alkyloxy), (Cw alkyl )3N+(CG.6 alkyloxy), (C;.6 alkyloxy^arbonylCC^ alkyl), carboxy(C0.6 alkyd), carboxy (C ,.6 alkyl)oxy(C(w. alkyl), C^haloalkyl. C^haloalkyloxy, and C,.6 alkyloxy; R^ais selected from hydrogen, hydroxy, hydroxyfC^ alkyl), amino, amino(CM alkyl), Ci.lo alkyl, (C5.!2)cyc1oalkyl(C0.i0alkyl), (Cc..6 alkyd)thio(C\..6 alkyl), and carboxy^C^ alkyl), wherein R^a may be substituted by 0, 1, or 2 R^£ substituents; R^b is selected from hydrogen, C^o alkyl, hydroxy(Cj.6 alkyl), amino(C14o alkyl), (Cb6 alkyl)o-2 aminoCCvto alkyl), (C].6 alkyl)3N'(C;.s alkyl), Cb6 haloalky 1, aryl(Co.io alkyl), heteroaiyl(C0.ie alkyd), (C3.12)cycIoalkyl(CG.10 alkyl), heterocycloalkyl(CG.1G alkyl), (Cw alkydloxytCi.g alkyd), (C3.i2)cycloalkyloxy(Ci.6alkyl). carboxy(C j..6alkyl), aminocarbonyl(Cj^ alkyd), (CM alkyl)o-2 aminocarbonyl(Cj.6 alkyd), aminocar bonylammo(CMalkyl), (Cwalkyd)o-2 aminocarbonylamino(C -,.6 alkyl), (Co^ alkyDthiolCj^ alkyl), (Cj^ alky 1)SO2(CM alkyl), and (Cf.6 alkyl)sulfinyl(C1.6 alkyl), wherein may be substituted by 0, 1, or 2 R^e substituents; each R^c is independently selected from halo, alkyl, amino, (Ci,6 alkyl)o-2 amino(C0„ 6 alkyl), (Cj.6alkyr)3N+-. (Cj.6alkyl)SO2(C0.6 alkyd), cyano, cyano(Cj.6 alkyl), hydroxy. hydroxy(C3.6alkyl), (C^galkyd)oxy(C0.5alkyl), aminocarbonyl(C0.6alkyl), and (()0^)carboxy(C0. 6 alkyl), and wherein R^a and R^b, together with the atoms to which they are attached, may form a saturated ring system, wherein said saturated ring system may be substituted by 0, 1, or 2 R^c substituents; Rda is selected from hydrogen, CM alkyl, hydroxy, CM alkyloxy, and fluoro; R^b is selected from bicyclic heteroaryd(C0.3 alkyl), and bicyclic aryl(CG_3 alkyl), wherein R^b js substituted with 0, 1, or 2 R^c substituents each independently selected from halo, hydroxy, cyano, nitro, carboxy, carboxy (Cj_6 alkyl), (Cw alkyl oxy )carbonyl(C0^ alkyl), alkyloxy. (Cj^alkyl)oxy(Cw alkyl), alkyl, and Cj.6haloalkyl; Rsa is selected from hydrogen, C14o alkyl, Ci4G fluoroalkyl, carboxy (Ci4G alkyl), hydroxy, bydroxy(C140 alkyl), cyanoCCuo alkyl), heterocycloalkyl(Co.so alkyl), aminocarbonyl(C].6 alkyl), (Cb6alky1 )o-2 aminocarbonyXQ.g alkyl). (carboxytC4_10alky1 ))oxy (CM alkyl), aiyl(CG.6 alkyl), (C3.12)cycloalkyl(CG.6 alkyl), heteroaryl(C0.6 alkyl), (Cf^ alkyd) oxy(Cw alkyl), (C,_6 alkydoxy)carbonyl(Co.6 alkyl), amino, amino(C1.6 alkyl), (C^ alkyd)o-2 amino(C0^ alkyl), amino(Ci.6 alkyl)carbonylamino(C].6 alkyl), (Cw alkyl)o-2 amino(Ci.6 alkyl)carbonyiamino(Ci.6 alkyl). (Cb6 alkvl;-:\ (C ,... alkyl), (Cj.galkyljsNXCj.g alkyl)carbonylamino(C6 alkyl), and C140fluoroalkyl, and wherein R3ais substituted by 0, 1, or 2 R^d substituents; js selected from hydrogen, Ci_10 alkyl, hydroxy(Cn0alkyl), carboxy(C^alkyl). (Cb 6 alkyDoxyfCj.c, alkyl), aminocarbonyKC^g alkyl), (CM alkyl)o-2 aminocarbonyl(Ci_6 alkyl), carboxy(Cf.io alkyl)oxy(C^ alkyl), cyano(Cl40 alkyl), amino(CM alkyl), (CM alkyl)o-2 aminojC^alkyl), aminoCC^alkyl)carbonylamino(Cj.6alkyl), (Cj_6alkyl)o-2 amino(C1.6 alkyl)carbonylamino(C1.6 alkyl), (Cb6 alkyDsN'(C2.6 alkyl)oxy(Cw alkyl), (Cw alkyl)3N+(C;.6 alkyl)carbonylamino(Cj^ alkyl), (Cj_6 alkyl)sN+(Cw alkyl), (heteroeycloalkyl(C040 alkyl), (C3. i2)cycloalkyd(C().w alkyl), and Cj.iohaloalkyl, wherein R$h is substituted by 0, 1. 2. or 3 R5® substituents, wherein RSa and R5^, together with the atoms to which they are attached, form a saturated mono- or bi-cyclic ring, wherein said mono- or bi-cyclic ring is substituted with 0, 1, 2, or 3 ant| q, I, 2, or 3 R$e substituents; each RSd js independently selected from selected from halo, hydroxy, hydroxy(C].i0 alkyl), Cbl0 alkyl, carboxy, carboxy(Cw alkyl), Cw alkyloxy, (C1.6alkyl)oxy(C1.6 alkyl), ammo, aminofCfog alkyl), (Cw alkyl)o-2 amino(C0.6 alkyl), amino(Cw alkyl)carbony1amino(C0^ alkyl), (C^alkyl)o-2 amino(Ci.6alkyl)carbonylammo(Co^alkyl), (C]_6alkyl)3N+(Co-6 alkyl), (Cb.6 alkyl)3br(C2.6 alkyl)oxy(C0.6 alkyl), (Cb6 atkyl)3N'(Cb6 alkyl)carbonylamino(C0.6 alkyl), (carboxy(Cl^alkyl))oxy(Ci.6alkyl), cyano(C<,_6 alkyl), tetrazolyl(C0.6alkyl), and Cw haloalky 1, and tw'o R^d substituents together with the atom they are attached to may join together to form a saturated ring; each R5® is independently selected from halo, hydroxy. hydroxy(C].!0 alkyl). Ci.lo alkyl, carboxy, carboxy(Cb6 alkyl). C-,.6 alky I oxy, (CMalkyl)oxy(C]^ alkyl), amino, amino(CM alkyl), (CM alkyl)o-2 amino(Ce.6 alkyl), armno(C!.6 aJkyl)carbonylamino(C0.6 alkyl), (Ct.s alkyl)o-2 amino(Ci.6alkyl)carbonylamino(C0.,6alkyl), (C;.6alkyl)3N+(C0.6 alkyl), (CMalkyI)jN"(C2.6 alkyl)oxy(C0.6 alkyl). (Cj.6alkyl)3N4(C5.6alkyl)carbonylamino(C0.6allyl), (carboxy(Cj_6 aikyOjoxyCCpg alkyl), cyano(C0^ alkyl), tetrazolyi(C0.6 alkyl), and    haloalky 1, and two R$e substituents together with the atom they are attached to may join together to form a saturated ring; R^c is hydrogen. Ci.4 alkyl, hydroxy(CM alkyl), Cb4 alkyloxy, or (CMalkyl)oxy(Cj.4 alkyd); is selected from hydrogen, hydroxy, amino, CM alkyloxy, and CM alkyl; R$b is selected from hydrogen, alkyl, hydroxy, Cw alkyloxy, and fluoro; R^c is selected from Cj.lo alkyl, alkyloxy, (C^alkyOoxyCC^ alkyl), Cpjo haloalkyl, aryl(C0^ alkyl), heteroaryl(C0.!0 alkyl), (Cw alkyl)SO2(Cc,.6 alkyl), hydroxy, hydroxy (C^ alkyl), aminofCo^ alkyl), (Cj^ alkyl)o-2 amino(C0^ alkyl), aminocarbonyl(C&.6 alkyl), (Chalky! )0-2 aminocarbonyl(C0^ alkyl), (C5.6 aikyl)carbonylamino(C0.6 alkyl), (CM alky 1)0-2 aminocarbonylamino(C0.6 alky l), and carboxy(C0.6 alkyl); R^a is selected from hydrogen, CM alkyl, hydroxy, CM alkyloxy. and fluoro: R^b is selected from aiyl(C0„6 alkyl), heteroand(C0^ alkyl), and (C3.i2)cycloalkyl(C0.6 alkyl), wherein R^b is substituted by 0, 1, 2, or 3 R^c substituents each independently selected from Ci-6 alkyl. C^ofluoroalkyl, C^o fluoroalkyloxy, (C^alkyDo^ ammo(C0.5 alkyl), (C].6 alkyl)3N (C0.5 alkyl), carboxy(CM alkyl), (Cs,6 alkylo.xy)carbonyl(C0.6 alkyl). carboxy(C1.6 alkyl)oxyr(C0^ alkyl), ammocarbonyl(C0.6 alkyl), (C w alkyl)o-2 aminocarbonyl(C0^ alkyd), hydroxy, hydroxyiC^ alkyl). halo, -(Cq-s alkyl)--(S(=:O)?OH), —(C0_5 alkyl)-(S(:=O)2NH2), amino(C0^ alkyl)oxy(C0.6 alkyl), (Cw alkyl)o-2 amino(C0^ alkyl)oxy(C0.6 alkyl), (C alkyl)3N*(Co.6 alkyl)oxy(C0.6 alkyl). aminocarbonylarnino(C0.6 alkyl). (Cj„6 alky 1)0-2 anhnocarbonylamino(C0.6 alkyl), Cvwhaloalkyl, C^johaloalkyloxy, and (Cj^ allyl)oxy(C0.6 alkyl); R^a is selected from hydrogen, alkyl, hydroxy, C1-4 alkyloxy, Cw fluoroalkyl, Cp4 fluoroalkyloxy, and halo; R^b is selected from bicyclic aty l(C ,: alkyl) and bicyclic heteroaryl(C0.3 alkyl), wherein R^b is substituted by 0, 1, or 2 R§c substituents each independently selected from CM alkyl, halo, cyano, nitro, carboxy, amino, hydroxy, fluoroalkyl, Cj^ fluoroalkyloxy, Cb6 alkyloxy, amino(Cj.3 alkyl), and hydroxy(Ci_6 alkyl); R9 is selected from hydrogen, and Cb4 alkyl; ^10a is selected from hydrogen, hydroxy, C]_6 alkyd, CM alkyloxy, and fluoro; RlOb js seiected from (C3..i2)cycloalkyl(C0.3 alkyl), aryl(C0.3 alkyl), and heteroaryl(C0.3 alkyl), wherein said heteroaryl contains 1, 2, or 3 nitrogen atoms, wherein Rl^b 1S substituted by 0, 1. or 2 R1^ wherein each Rl®c is independently selected from Cho alkyl. Cho fluoroalkyl, C3.6 haloalky 1, Ci_10fluoroalkyloxy, amino, amino(CMalkyl), (Cw alkyl)o-2 ammo(Cc.,6 alkyl), (Cw alkyljsN (C0.6 alkyl), hydroxy, hydroxy(C w alkyl), cyano, halo, aminocarbonyl, aminocarbonyi(Cj_6 alkyl), (Q.g alkyl)0-2 aminocarbonyl(C0.6 alkyl), (CM a1kyl)o-2 aminocarbonylamino(C0.6 alkyl). (CV6 alkyl)carbonylamino(C0.6 alkyl), carboxy (C0.6 alkyl), (C 1.6 alkoxy)carbonyl(C0^ alkyl), carboxy(CM alkyl)oxy(C0.6 alkyd), ~-(C0.5 alkyl)-(S(==O)2OH), -(C0.5 alkyl)-(S(::::O)2NH2X amino(C0.6 alkyl)oxy(Cc._6 alkyl), (CM alkyl)0-2 amino(C0^ alkyf)oxy(C0.6 alkyl). (Cj 4 dkyl^NXC^ aikyl)oxy(Cc,.6 alkyl), (Cw alkyl)oxy(C0.6 alkyl), Cj^ haloalkyloxy, (C3.12)cycloalkyl(C0.6 alkyl), and heterocycloalkyl(C0.6 alkyl); is selected from hydrogen, hydroxy, C-._6 alkyl, Cj^alkyloxy, and fluoro; Rllb is selected from aryl(C0.3 alkyl), heteroaryl(Co-3 alkyl), wherein said heteroaryl contains 1,2, or 3 nitrogen atoms, and H2N-€(”NH)NH-(Cj^ alkyl), wherein is substituted by 0, 1,2, 3, or 4 substituents each independently selected from Cw alkyl, amino, amino(CV6 alkyl), (C;.calkyl)o-2 amino(C0^ alkyl), (Cj^ alkj4)3N+(C0.6 alkyl), hydroxy, hydroxy(Cj_6 alkyl), cyano, halo, aminocarbonyl, aminocarbonyl(C|.6 alkyl), (C 3.6 alkyl)o-2 ammocarbonyl(C0,6 alkyl), carboxy(C0^ alkyl), carboxy(C3.6 alkyl)oxy(C0^ alkyl), ammo(C0.6 alkyl)oxy(C0.6 alkyl). (Cb6 a1kyl)o-2 amino(C0.6 alkyl)oxy(C0^ alkyd), (Cw alkylhN (Ct,.6 alkyl)oxy(C0^ alkyd). (C,^ alkyl)oxy(C0^ alkyd). C5.s haloalkyloxy, haloalkyl, (C3.]2)cy,'cloalkyl(C0^ alkyl), heterocycloalkyl(C0^ alkyl), ((C3.6alkyl)carbonyl)heterocycloalkyl (Co-10 alkyl), and ((C3.6 alkyl)carbonyloxy)heterocycloalkyl(C0.10 alkyl), R^a is selected from hydrogen, hydroxy, amino, Cho alky l, and (C3.12)cycloalkyl(Co.6 alkyl); is selected from hydrogen, CT.W alkyl, and (C3.12)cycloalkyl(C0.6 alkyl); wherein ^12a atK| R12b, together with the atoms to which they are attached, may form a saturated ring; is selected from hydrogen, hydroxy. Cj.6 alkyl, Cb4 alkyloxy, and fluoro; R13b js selected from hydrogen, and alkyl; r!3c js selected from a bicyclic nitrogen-containing heteroaryl having 1 or 2 nitrogen and bicyclic-aryl and wherein r!3c is substituted independently by 0, 1, or 2 R13d substituents each independently selected from halo, alkyl, carboxy(C0.4alkyl), Ci^haloalkyloxy, and C3. 4 alkyloxy; alkyloxy, m is selected from 1,2, 3, or 4: n is selected from 1, 2, 3. or 4; and js selected from hydrogen, Cj.s alkyl, aryl(C0.6 alkyl), and heteroaryl(C0.6 alkyl), wherein R^^ 1S substituted by 0, 1, 2, or 3 halo groups.

[00135] In a first embodiment of the invention. R^ is selected from aminomethyl, aminoethyl, aminopropyl, aminobutyl, ammopentyl. phenyl. phenylmethyL phenylethyl, phenylpropyl. styryl, biphenyl, naphthyl, pyridyl, pyridazinyl, pyrimidyl, pyrazinyl, imidazolyl, pyrazolyl, furyl, thiophenyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl. oxadiazolyl, biphenylmethyl, naphthylmethyl, pyridylmethyl, pyridazinylmethyl, pyrimidylmethyl, pyrazinylmethyl, imidazolylmethyl, pyrazolylmethyl, furylmethyl, thiophenylmethyl, oxazolylmethyl, isoxazolylmethyl. thiazolylmethyl, isothiazolylmethyl, oxadiazolylmethyl, bicyclo[1 .1.1 .]pentyl, (bicyclo[ 1.1.1. ]pentyl)methyl, phenylcarbonylaminoethyl, aminocarbonylmethyl, aminocarbonylethyl, aminocaibonylpropyl, aminocarbonylisopropyl, aminocarbonylbutyl, aminocarbonylaminomethyl, aminocarbonylaminoethyl, aminocarbonylaminopropyl, aminocarbonylammobulyl, methyl, ethyl, propyl, isopropyl, butyl, .vec-butyL isobutyl, ter / -butyl, w-pentyl, isopentyl, neopentyl, w-hexyl, isohexyl, n-heptyl, n-octyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclobutylmethyl, cyclopenty Imethyl, cyclohexylmethyl, adamantyl, carboxy, carboxymelhyl, carboxyethyl, azidomethyl, azidoelhyl, azidopropyl, phenylaminocarbonylmethyl, pyridyl aminocarbonylmethyl, (pyridylcarbonylamino)methyl, guanidine, guanidinomethyl, guanidinoethyi, guanidinopropyl, guanidinobutyl, fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, pentafluoroethyl, fluoropropyl, difluoropropyl, trifluoropropyl, pentafluoropropyl, heptafluoropropyl, trifluorobutyl, / VJV7 / -trimethylmethylammonium, TVJVJV-trimethyleth-1 -ylammonium, ,V^,AMrimethylpropan-l-ylammonium, AuVA'-trimethylbut-l-ylammonium. methylamino, methyl aminomethyl, methylaminoethyl, methylaminopropyl, methylaminobutyl, dimethylamino, dimethylaminomethyl, dimethylaminoethyl, dimethylaminopropyl, dimethylaminobutyl, ethylamino, ethylaminomethyl, ethylaminoethyl, ethylaminopropyl, ethy I aminobutyl, di ethy I amino, di ethy I aminomethy I, diethyl aminoethyl, di ethy I aminopropy 1, diethylaminobutyl, aminoetboxy, aminoethoxymethyl, isoxazolylcarbonylaminomethyl, methoxy, ethoxy, propoxy, isopropoxy, cyclopropoxy, cyclopropoxymethyl, cyclopropylmethoxy, cyclopropylniethoxymethyl, cyclopropylmethoxyethyi. methoxymethyl, ethoxymethyl, methoxyethyl, ethoxyethyl, vinyl, prop-2-enyl, but-3-enyl, and pent-4-enyl. and the other groups are as provided in the general Formula (I) above.

[00136] In a second embodiment of the invention, Ris selected from 3-aminopropyl, 4-aminobutyl. phenyl, phenylmethyl, bicyclo[l.l.l.|pentyl, phenylcarbonylaminoethyl, aminocarbonylmethyl, aminocarbonylaminoethyl, n-propyl, M-butyl, isobutyl, isopentyl, n-pentyl, w-hexyl, 1-azidoethyl, 2-azidoethyl, azidopropyl, 3-azidopropyl, pyndylaminocarbony Imethyl, 3-guanidinopropy 1, 2,2-difluoropropyl, 4,4,4-trifluorobuty I, ALA(AMrimethylpropan-l-ylammonium, methylaminopropyl, dimethylaminopropyl, isoxazolylcarbonylaminomethyl. ethoxy, methoxy-methyl, prop-2-enyl, cyclopropylmethoxy. and aminoethoxy, and the other groups are as provided in the general Formula (I) above or as in the first embodiment.

[00137] In a third embodiment of the invention.     substituents are each independently selected from C1-6 alkyl, amino, cyano, halo, and hydroxy, and the other groups are as provided in the general Formula (I) above or as in the first through second embodiments.

[00138] In a fourth embodiment of the invention, each js independently selected from hydrogen, hydroxy, methyl, ethyl, methoxy, ethoxy, fluoro, and the other groups are as provided in the general Formula (I) above, or as in the first through third embodiments.

[00139] In a fifth embodiment of the invention, each js independently selected from hydrogen, hydroxy, and methyl, and the other groups are as provided in the general Formula (I) above, or as in the first through fourth embodiments.

[00140] In a sixth embodiment of the invention, R^b is selected from phenyl, benzyl, biphenyl, naphthyl, pyridyl, pyridazinylpyrimidyl, pyrazinyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, and bicyclo[l.I.l]pentyl, and the other groups are as provided in the general Formula. (I) above, or as in the first through fifth embodiments.

[00141] In a seventh embodiment, R^b is selected from phenyl, pyridyl, pyrimidyL pyridazinyl, imidazolyl, and bicycloj 1.1. l]pentyl. and the other groups are as provided in the general Formula (I) above, or as in the first through sixth embodiments.

[00142] In an eighth embodiment, each R^c is independently selected from aminomethyl, hydroxy, methoxy, ethoxy, di fluoromethoxy, trifluoromethoxy. fluoromethyl, difluoromethyl, trifluoromethyl, trifluoroethyl, methyl, ethyl. ;?-propyl, isopropyl, cyclopropyl, w-butyl, tertbutyl, fluoro, chloro, bromo, iodo, aminoethoxy, A’-methylaminoethoxy, A-ethylaminoethoxy, A’,A-dimethylaminoethoxy,            O , carboxy, carboxymethoxy, (carboxymethoxy )methyl, aminocarbonyl, AlAMimethylammocarbonyi, and aminocarbonylmethyl, and the other groups are as provided in the general Formula (I) above, or as in the first through seventh embodiments.

[00143] In a ninth embodiment, each !S independently selected from aminomethyl, hydroxy, methoxy, fluoro, carboxy, and aminocarbonyl, and the other groups are as provided in the general Formula (I) above, or as in the first through eighth embodiments.

[00144] In a tenth embodiment of the invention, R-$a is selected from hydrogen, methyl, ethyl, n-propyl. isopropyl, w-bulyl. 2-methylpropyl, te / 7-butyl, cyclopropyl, cyclopropy Imethyl, cyclopropylethyl, cyclobutyl, cyclobutylmethyl, cyclobutylethyl, aminoethyl, aminopropyl. aminobutyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, carboxymethyl, carboxyethyl, carboxypropyl. carboxy butyl, thioethyl. and thiopropyl, wherein     may be substituted by 0. 1, or 2 substituents each independently selected from fluoro, chloro, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, sec-butyl, ferz-butyl, amino, aminomethyl, JV-melhylamino, N-methylaminomethyl. / V-ethylamino, / V-ethylaminomethyL AUV-dimethylamino, AtyV-dimethylaminomethyl, AtyV-diethylammo, AtyV-diethylammomethyl, AfAtyV-trimethy I ammonium, AtyV^-trimethylmethylammonium, hydroxy, hydroxymethyl, SO2CH5, CH2SO2CH3, CH2CH2SO2CH3, cyano, cyanomethyl, methoxy, ethoxy, methoxymethyl, methoxyethyl, carboxy, carboxymethyl, and carboxy ethyl, and the other groups are as provided in the general Formula (I) above, or as in the first through nineth embodiments.

[00145] In a eleventh embodiment of the invention, R^ajg selected from hydrogen, methyl, ethyl, propyl. 2-methylpropyl, butyl, aminoethyl, 2-aminoethyl, aminopropyl, 3-aminopropyl, hydroxy ethyl, 2-hydroxyethyl, hydroxypropyl, 2-hydroxypropyl, 3-hydroxypropyl, carboxyethyl, 2-carboxy ethyl, and thioethyl, wherein R^a may be substituted by 0. 1, or 2 R^c substituents each independently selected from methyl, isopropyl, cyclopropyl, ammo, N-methylamino, hydroxy,-SO2CH3, -CH2SO2C.H3, cyano, methoxy, and carboxy, and the other groups are as provided in the general Formula (1) above, or as in the first through tenth embodiments.

[00146] In a twelfth embodiment of the invention, R-^ is selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, .sec-butyl, tert-butyl, w-pentyl, isopentyl, neopentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclobutylmetyl, cyclopentylmethyl, cyclohexylmethyl, bicyclof 1.1.1 ]pentylmethyI, hydroxymethyl. 1-hydroxyethyl. 2-hydroxyethyl. bydroxypropyl, 3-hydroxypropyl. 1-methyl-1-hydroxyethyl, hydroxyisopropyl, hydroxybutyl, methoxymethyl, niethoxyethyl, methoxypropyl, ethoxymethyl, ethoxy ethyl, aminomethyl, 2-aminoethyl, A-methylamino methyl, T^ / V-dimethylaminomethjd, N-methylaminoethyl, A’,A-dimethylaminoethyl, / V-methylaminopropyl, AW-dimethylaminopropyl, 1-aminopropyl, 2-ammopropyl, 3-aminopropyl. 2-aminoprop-2-yl, fluoromethyl. difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, benzyl. 3-pyridinylmethyl, 4-pyridinylmethyl, imidazolylmethyl, thiazolylmethyl, oxazolylmethyl. thiophenylmethyl, furanylmethyl, pyrazolylmethyl, AApyrazolylmethyl. 1-phenylethyl, 1-(4-pyndinylfethyl, aminocarbonylmethyl, aminocarbonylethyl, aminocarbonylpropyl. (AtyV-dimethyl)aminocarbonylmethyl, (AfiV-dmiethyl)ammocarbonylethyl, thiomethyl, thioethyl, thiopropyl,-CH2CH2SO2CH3, carboxymethyl, carboxyethyl, 2-carboxyethyl, carboxypropyl, 3-carboxypropyl, carboxybutyl, 4-carboxybutyl, piperazinylmethyl. morpholinomethyl, piperidinylmethyl, azetidinylmethyl, tetrahydropyranylmethyL aminocarbonylaminomethyl, aminocarbonylaminoethyl. aminocarbonylaminopropyl. and aminocarbonylaminobutyl, wherein R^b may be substituted by 0, 1, or 2 substituents each independently selected from fluoro, chloro, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, «-butyl, sec-butyl, / er / -butyl, amino, aminomethyl, JV-methylamino. AAmethylaminomethyl, JV-ethylamino, 7V-ethylaminomethyl, AtiV-dimethylamino, A7 / -dimethylaminomethyL A'jV-diethylamino, A’JV-diethylaminomethyl, ACA’JV-tri methyl ammonium, AyAfAMrimethylmethylammonium, hydroxy, hydroxymethyl, -SO2CH3, -CH2SO2CH3, -CH2CH2SO2CH3, cyano, cyanomethyl, methoxy, ethoxy, methoxymethyl, methoxyethyl, carboxy, carboxyniethyl, and carboxy ethyl, and the other groups are as provided in the general Formula (I) above, or as in the first through eleventh embodiments.

[00147] In a thirteenth embodiment of the invention, R^b js selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, isobutyl, w-butyl, cyclopropyl, cyclobutyl, hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, hydroxypropyl, 3~hydroxy propyl, 1 -methyl-1 -hydroxyethyl, methoxyethyl, aminomethyl. 2-aminoethyl, AAmethylaminomethyl, 1-methyl-l-aminoethyl. 2-aminoprop-2-yl, 1-phenylmethyl, benzyl, imidazolylmethyl, thiazolylmethyl, aminocarbonylmethyl, aminocarbonylethyl, thiomethyl, -CH2CH2SO2CH3, carboxymethyl, 2-carboxyethyl, 3-carboxypropyl, 4-carboxybutyl, piperazinylmethyl, and aminocarbonylaminopropyl, wherein R^b may be substituted by 0, 1, or 2 R^c substituents each independently selected from methyl, isopropyl, cyclopropyl, amino, A’-methyl amino, hydroxy,

[00148] -SO2CH3, --CH2SO2CH3. cyano, methoxy, and carboxy, and the other groups are as provided in the general Formula (I) above, or as in the first through twelfth embodiments.

[00149] In a fourteenth embodiment of the invention, R$a         together with the atoms to which they are attached, form a saturated ring system substituted by 0,I, or 2, R3c substituents and the ring system is selected from: O , and the other groups are as provided in the genera] Formula (I) above, or as in the first through thirteenth embodiments.

[00150] In a fifteenth embodiment of the invention, and R-Ib together with the atoms to which they are attached, form a saturated ring system substituted by 0, 1. or 2, R^c substituents and the other groups are as provided in the general Formula (I) above, or as in the first through fourteenth embodiments.

[00151] In a sixteenth embodiment of the invention, is selected from hydrogen, methyl, ethyl, propyl, hydroxy, methoxy, and fluoro, and the other groups are as provided in the general Formula (1) above, or as in the first through fifteenth embodiments.

[00152] In a seventeenth embodiment of the invention. R^a !S hydrogen or methyl, and the other groups are as provided in the general Formula (1) above, or as in the first through sixteenth embodiments.

[00153] In ftn eighteenth embodiment of the invention, R^^ is selected from indolyl, naphthyl, quinohnyl. pyrrolo[2,3~b]pyridmyl. [l,2,4]triazoio[L5-a]pyridine, LH-pyrazolo[3.4-b]pyridme, indazolyl, benzothiazolyl, and benzothiophenyl, wherein substituted with 0, 1, or 2 R4c substituents each independently selected from fluoro, chloro, bromo, iodo, cyano, nitro, hydroxy, methyl, ethyl, propyl, isopropyl, carboxy, carboxy methyl, and carboxyethyl, and the other groups are as provided in the general Formula (J) above, or as in the first through seventeenth embodiments.

[00154] In a nineteenth embodiment of the invention, is selected from indolyl, naphthyl, quinolinyl, pyrrolo[2,3-b]pyridinyl, lH-pyrazoIo]3,4- / ?]pyridine, and indazolyl, wherein R^b substituted with 0, I, or 2 R^c substituents each independently selected from fluoro, chloro, methyl, and carboxymethyl, and the other groups are as provided in the general Formula (I) above, or as in the first through eighteenth embodiments.

[00155] In a twentieth embodiment of the invention, R^a is selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, H-butyl, isobuty1, cyclobutyl, n-pentyl, isopentyl, neopentyl, cyclopentyl, cyclobutyl, cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, cyclobutylethyl, cyclopentylmethyl, cyclopentylethyl, cyclohexylmethyl, cyclohexylethyl, phenyl, benzyl, phenylethyl, phenylpropyl, oxazolylmethyl, thiazolylmethyl, imidazolylmethyl, tnazolylmethyl, oxadiazolylniethyl, thiadiazolylmethyl, oxazolylethyl, thiazolylethyl, imidazolylethyl, triazolylethyl. oxadiazolylethyl, thiadiazolylethyl, oxazolylpropyl, thiazolylpropyl, imidazolylpropyl, triazolylpropyl, oxadiazolylpropyl, thiadiazolylpropyl, azetidinylmetbyl, azetidinylethyl, oxetanylmethyl, oxetany!methyl, pyrrolidinylmethyl. pyirolidinylethyl, tetrahydrofuranylmethyl, tetrahydrofuranylethyl, piperidinylmethyl, pipendinylethyl, piperazinylmethyl, piperazinylethyl, tetrahydropyranylmethyl, tetrahydropyranylmethyl, hydroxy ethyl, hydroxypropyl, hydroxy isopropyl, hydroxybutyl, 3-hydroxy-2,2-dimethy Ipropyl. cyclopropylmethyl, 1 -hydroxypropan-2-yl, 2-hydroxyethyl, 3-hydroxypropyl. 2-hydroxy isopropyl, methoxyethyh methoxy propyl, ethoxy ethyl, ethoxypropyl, cyanomethyl, cyanoethyl, cyanopropyl, cyanobutyl, carboxymethyl, carboxyethyl, carboxypropyl, carboxybutyl, 2-fluoroethyl, 2,2-difluoroethyl, 2,2,2-lrifluoroethyl, 2-ammoethyl. 3-aminopropyl, 3-ammo~2.2-dimethylpropyl, cyclopropylmethyl, 4-aminobutyl, aminomethylcarbonylaminoethyl, aminoethyl carbonylaminoethyl, aminomethyl carbonylaminopropyl, aminoethyl aminocarbonylmethyl, aminoethylaminocarbonylethyl, aminoethylaminocarbonylpropyl, aminohexylcarbonylaminoethyl, aminohexylcarbonylaminoethyl, (A-methylamino)ethyl, (N-methylamino)propyl, (A-ethylamino)ethyl. (A,A-diethylamino)propyl, (N.N-dimethylamino)ethyl, (A7,A-dimethylanhno)propyL (A7,AMiethylammo)ethyl, (A^V-diethylaminojpropyl, (AAAA-triniethyiammoniwniethyl, (A’y\f,AMrimethylammonium)propyl, (A',AJV-triethylammonium)ethyl, (N,A^V-triethylammonium)propyl, (A-methylamino)methylcarbonylaminoethyl, (A-me1hylamino)ethylcarbonylaminoethyl, (A7-methylamino)methylcarbonylaminopropyl, (A7-ethylamino)methylcarbonylaminoethyl, (A"-ethylamino)ethylcarbonylaminoethyl, (A’-etliylamino)methylcarbonylaminopropyl, (N-methylamino)penlylcarbonylaminoethyl, (A-metliylanuno)perUylcarbonylaminoelhyl, (A-methylamirm)pentylcarbonylammopropyl. (AJV-dimethylamino)methylcarbonylaminoethyL (A,A-dimethylamino)ethylcarbonylaminoethyl, (N,N-dnnethylamino)methy I carbonylaminopropyl, (A,A-diethylamino)methylcarbonylaminoethyl, (AA-diethylamino)ethylcarbonylaminoethyl, (AJV-diethylamino)methylcarbonylaminopropyl, (AW-dimethylamino)pentylcarbonylaminoethyl, (A',A7-dimethylamino)pentylcarbonylaminoethyl, (A,A-dimethylamino)pentylcarbonylaminopropyl, A^A-trimethyl-ethan-l-ammonium, A, A, AMrimethyl-propan-l-ammonium, (N,N,N-trimethylammonium)methylcarbonylaminoethyJ, (AAV,A7-trimethylanmionium)ethylcarbonylammoethyl, (NyN,N-trimethylammonium)methylcarbonylaminopropyl, (Ar.Ar,A7-trimethylammoniumjethylcarbonylaminopropyl, trimethylammonium)pentj'lcaibonylaminoethyL (NJ\r,N-triniethylammomum)pentylcarbonylaminopropyl, (carboxymethyl)oxyethyl, and (carboxymethyl)oxypropyl, wherein R^a js substituted by 0, 1, 2, or 3 R^d substituents each independently selected from chloro, fluoro, hydroxy, hydroxymethyl, hydroxy ethyl, methyl, ethyl, n-propyl, isopropyl, w-butyl. isobutyl, cyclobutyl, n-pentyl, isopentyl, neopentyl, carboxy, carboxymethyl, carboxyethyl, methoxy, ethoxy, methoxymethyl, methoxyethyl, methoxy propyl, ammo, aminomethyl, aminoethyl, A-methylamino, (A-methylamino)methyl, (A-methylaminojethyl, A,A-dimethylamino, (A7_AMmiethylamino)methyl, (NJ\r-dimethylamino)ethyl. A7,AMiethylamino, (A’.A-diethylamino)methyl, (A^V-diethylamino)ethyl, aminomethyl carbonylamino, aminoethylcarbonylamino, aminopentylcarbonylamino, aminomethylcarbonylaminomethyl, aminoethyl carbonylaminomethyl, (A-methylamino)methylcarbonylamino, (A-methylamino)ethylcarbonylamino, (TV- methylamino)methylcarbonylammomethyl, (A7-methyIamino)ethylcarbonylaminomethyl, (N.N-dimethylammo)methylcarbonylamino, (AW-dimethylamino)ethylcarbonylamino, (A'’,A-dimethylammolmethylcarbonylaminomethyl, (A^V-dimelhyiamino)ethylcarbonylaminomelhyl, (AyV-diethylamino)methylcarbonylamino, (A,A?-diethylamino)ethylcarbonylamino, (N,N~ diethylamino)methylcarbonylaminomethyl, (AtyV-dietliylamino)ethyicarbonylaminomethyl, Ar,A'yV-trimethylammonium, (Aty¥Ar4rimethylammomum)methyl, (N,N,N~ trimethylammonium)ethyl, (A(.tyAtyrimethylammonium)ethoxy, trimetbylammonium)ethoxymethyl, (ApV,A-trimethylammoniutn)meihylcarbonylamino, (N,N,N-triethylammonium)methylcarbonylamino, (AfA(Atyrimethylamnionium)ethylcarbonylarnmo, (AfAyA-trimethylammonium)penty Icarbonylamino, irimethy 1 ammoni urn)melhy Icarbony 1 ammomethy 1, (N,N,N-trimethylammonium)etbylcarbonylaminomethyl, cyano, cyanomethyl, cyanoethyl. tetrazoyl, tetrazoylmethyl, tetrazoylethyl, carboxymethoxy, carboxy ethoxy, carboxy methoxymethyl, and carboxy ethoxymethyl, and the other groups are as provided in the general Formula (I) above, or as in the first through nineteenth embodiments. JOO 156]   In a twenty-first embodiment of the invention, R$a is selected from hydrogen, methyl, ethyl, .u-propyl, cyciopropylmethyl, (1,3,4-oxadiazol-2-yl)ethyl, 2-hydroxyethvl, 3-hydroxypropyl, carboxymethyl, 2-carboxyeihyl, 3-carboxypropyl, 2-hydroxyethyl, 2-hydroxy-1-methylethyl, hydroxypropyl. 3-hydroxy-2,2-dimethylpropyL methoxyethyl, methoxypropyl, aminocarboxy ethyl, 2-fluoroethyl, 2,2-difluoroethyl, carboxymethoxy ethyl. 2-aminoethyL 3-ammopropyl, (A'jV-dimethylammo)ethy 1, Ay AfA’-trimethyl-ethan-1 -ammonium, (A'uV,A-trimethylammomurn)ethoxyethyl, (ApVJV-trimethylamnionium)methylcarbonylaminoethyl, and (carboxymethyl)oxyethyl, wherein R5a is substituted by 0, 1, 2. or 3 R5^ substituents each independently selected from fluoro, hydroxy, hydroxymethyl, methyl, carboxy, carboxymethyl, methoxy, amino, A^V-dimethylamino, AT,AyV-trimethylammonium, (.V,AyV-trimethylammoniumfethoxy. (Ay¥,A-trimethylammonium)methy]carbonylamino, cyano, tetrazoyl. and carboxymethoxy, and the other groups are as provided in the general Formula (1) above, or as in the first through twentieth embodiments.

[00157] In a twenty-second embodiment of the invention, R®b is selected from hydrogen, methyl, ethyl, isopropyl, n-propyl. cyclopropyl, isobutyl, n-butyl. sec-butyl, isobutyl, tert-butyl, cyclobutyl, cyciopropylmethyl, oxetanylmethyl, tetrahydrofurylmethyl, tetrahydropyranylmethyl, hydroxymethyl, hydroxyethyl, hydroxy propyl, methoxymethyl, methoxyethyl, methoxy propyl, carboxy methyl, carboxy ethyl, carboxy propyl, carboxybutyl, (carboxymethoxy)methyl, (carboxymethoxy)ethyL (carboxyethoxy)methyl, (carboxyethoxy )ethyl, aminomethyl, aminoethyl, aminopropyl, aminobutyl, (A-methylaniino)methyl, (W-methylamino)ethy L QV-methylamino)propyL (A’-methylannno)buty 1, (ACV-dimethylamino)methyl, (A?,A-dimethylamino)ethyl, (AW-dimethylamirm)propyl, (N,N-dimetbylamino)butyl, ( / VJV-diethylamino)methyl, (AyAdiethylamino)etbyl, (W,A’~ diethylaminojpropyl. (A;,A-diethyIamino)butyl, A,AA,A-trimethylmethan-1 -ylammonium, N,N,N-trimethy leth an- 1-yl ammonium, A / AV^-triethyleihan-1 -ylammonium, AyAyV-trimethylpropan-l-ylarnmonium, A7,ApV-trimetbylbutan-1-ylammonium, (N,N,N-trimetbylammonium)ethoxymethyl, ( / VpV,,¥-trimethylammonium)ethoxyethyl, (A'JVoV-triethylammonium)ethoxy ethyl, (A^A^V-trimethylammonium)ethoxypropyl, (A'.A',A-trimethylammonium)ethoxybutyl, aminocarbonylmethyl, aminocarbonylethyl, aminocarbonylpropyl, aminocarbonylbutyl, (A’-melhylamino)carbonylmethyl, (N-methylamino)carbonyleihyl. (TV-methylamino)carbonylpropyl, (A-metbylamino)carbonylbutyl, (A’,A-dimethylamino)carbonylmethyl, (^V-dimethylamino)carbonylethyl, (N.N~ dimethylamino)carbonylpropyl, (Ar,AMimethylammo)carbonyIbutyl, aminomethyl carbonylaminoethyl, aminomethyl carbonylaminopropyl, aminoethylcarboiiylaminoethyl, aminoethylcarbonylaminopropyl, (ApV-dimethylamino)methylcarbonylaminoethyl, (A^V-dimethylamino)methylcarbonylaminopropyl, (ArjV-dimethylamino)ethylcarbonylaminoethyl, (AVvMimethylamino)ethylcarbonylaminopropyl, (A / pV,A7-trimethyIammonium)methylcarbonylaminoethyl, triethylammomum)methylcarbonylaminoethyl, (ApVA-trimethylammonium)methylcarbonylammopropyl, (AAVyV-trimetbylammoniumjelhylcarbonylaminoethyl, (N,N,N~ trimethylammonium)ethylcarbonylaminopropyl, (AvVAMrimethylammonium)ethoxyethyl, (ALV,A-tmnethylammonium)ethoxypropyl, 2-fluoroethyl, 2,2-difluoroethyI, 2,2,2-trifluoroethyl, 3,3,3-trifluoropropyl, cyanomethyl, cyanoethyl, cyanopropyl, cyanoisopropyl, and cyanobutyl, wherein is substituted by 0, 1, 2, or 3 R®e substituents each independently selected from chloro, fluoro, hydroxy, hydroxymethyl, hydroxyethyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, cyclobutyl, n-pentyl, isopentyl, neopentyl, carboxy, carboxymelhyl, carboxyethyl, methoxy, ethoxy, methoxymethyl, methoxyethyl, methoxypropyl, amino, aminomethyl, aminoethyl, A7~methylamino, (Af-methylamino)methyl, (A-methylamino)ethyl, AiA-dimethylamino, (AV^-dimethylamino)methyl, (A7A'-dirnethylamino)ethy1, N.N-diethylamino, (AyA-diethylamino)methyl, (A,;V-diethylamino)ethyl, aminomethylcarbonyl amino, aminoetbylcarbonylamino. aminopenty]carbonylamino, aminomethylcarbonylaminomethyl, ammoethylcarbonylaminomethyl, (A-methylamino)methylcarbonylamino, (AA- methylamino)ethylcarbonylamino, ( / V-methy1amino)methylcarbonylaminomethyl, ( / V-methylamino)ethylcarbonylaminomethyl, (AfAMimethylamjno)methylcarboiiylammo, (N,N-dimethylamino)ethylcarbonylamino, (AAV-dimethylamino)methylcarbonylaminomethyL (N,N-dimetbylamino)ethylcarbonylaminomethyl, (AfA~diethylarnino)methylcarboi!yIammo, (N,N-die±ylamino)ethylcarbonylamino, (7VJV-diethylamino)methylcarbonylaminomethyl, (2V^V-diethylamino)ethylcarbonylaminomethyi, ?<Ar,Ar“triniethylanimonium, (N,N,N-trimethylammonium)methyl, (AAAfAr-trimethylammonium)ethyl, (N,N,N~ trimetbylammonium)ethoxy, (AAA, A-trimetbylammoni um)6thoxymethyl, (AyALA1-trimethylammoniumjmethylcarbonylammo. (AjVA-triethylanimonium)methylcarbonylamino, (AfAfA4rimethyiammonium)ethylcarbonyiamino, (N,N,N-trimethylammonium)pentylcaibonylamino, CAyAAV-trimethylammonium)methylcarbonylaminomethyl, (N,N,N-trimethylammoniumlethylcarbonylannnomethyl, cyano, cyanomethyl, cyanoethyl, tetrazoyl, tetrazoy line thy 1, tetrazoylethyl, carboxymethoxy, carboxy ethoxy, carboxymethoxymethyl, and carboxyethoxymethyl, and the other groups are as provided in the general Formula (I) above, or as in the first through twenty-first embodiments.

[00158] In a twenty-third embodiment of the invention, R^^ is selected from hydrogen, methyl, ethyl, n-propyl, cyclopropylmethyl, hydroxymethyl, 2-hydroxyethyl, 2-carboxyethyl, 3-carboxypropyl, 2-aminoethyl, (AJV-dimethylamino)ethyl,2-methoxyethyl, N,N,N~ trimethylethan-1-ammonium, (A.ArA-tnmethyl-ammonium)methylcarbonylaminoethyl, (AAyA-trimethyl-ammonium)ethoxyethyl, 2-fluoroethyl, 2,2-difluoroethyl, and (carboxymethoxy)ethyl, wherein R5^ is substituted by 0,1, 2, or 3 R5® substituents each independently selected from fluoro, hydroxy, hydroxymethyl, methyl, carboxy, carboxymethyl, methoxy, amino, A,A-dimethylamino, AiAiAMrimethylammonrum. (AyAAV-trimethylammonmm)ethoxy, (N,N,N-trimethylammonium)methylcarbonylamino, cyano, tetrazoyl, and carboxy methoxy, and the other groups are as provided in the general Formula (1) above, or as in the first through twenty-second embodiments.

[00159] In a twenty-fourth embodiment of the invention, R5C is hydrogen, methyl, or ethyl, and the other groups are as provided in the general Formula (I) above, or as in the first through twenty -third embodimen ts.

[00160] In a twenty-fifth embodiment of the invention. R$c is hydrogen, or methyl, and the other groups are as provided in the general Formula (I) above, or as in the first through twentyfourth embodiments.

[00161] In a twenty-sixth embodiment of the inventio, R$a and together with the atoms to which they are attached, form a saturated mono- or bi-cyclic ring system substituted with 0. 1. 2, or 3 R3^ and 0. 1,2, or 3 R$e substituents, wherein said mono- or bi-cyclic ring system is selected from; the general Formula (I) above, or as in the first through the sixteenth and the twenty-first through the twenty-fifth embodiments.

[00162] In a twenty-seventh embodiment of the invention, and together with the atoms to which they are attached, form a saturated mono- or bi-cyclic ring system substituted with 0, 1. 2, or 3 R^^ and 0, 1, 2, or 3 R5® substituents, wherein said mono- or bi-cyclic ring system is selected from. Formula (I) above, or as in the first through the sixteenth and the twenty-first through the twenty -sixth embodiments.

[00163] In a twenty-eighth embodiment, R$a is selected from hydrogen, hydroxy, methyl, ethyl, propyl, and methoxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the twenty-seventh embodiments.

[00164] In a twenty-nmeth embodiment, R^a is selected from hydrogen and methyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the twenty -ei ghth embodimen ts.

[00165] In a thirtieth embodiment, R$b is selected from hydrogen, hydroxy, methyl, ethyl, propyl, methoxy, and ethoxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the twenty-nmeth embodiments.

[00166] In a thirty-first embodiment, R^b js selected from hydrogen, hydroxy, and methyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the thirtieth embodiments.

[00167] In a thirty-second embodiment, R6c is selected from aminocarbonyl, (aminocarbonyl)methy 1, (aminocarbony! )ethyl, (aminocarbonyl)propyl, (7V-methylaminojcarbonyl, (A-methylamino)carbonylmethyl, (IV-methylanimo)carbonyletiiyl, (IV-methylamino)carbonylpropyi, (TVgV-dimethylamino)carbonyl, (N.N-dimethy1amino)carbonylmethyl, (.¥,>V-dimethylammo)carbonylethyl, (N,N~ dimethylamino)carbonylpropyl, (AyV-di ethylammo)carbonyl, (N,N~ diethylammo)carbonylmethyl, (A(AMiethylarmno)carbonylethyi, (N,N~ diethylamino)carbonylpropyl, aminocarbonylamino, (aminocarbonylamino)methyl, (ammocarbonyiamino)ethyl, (ammocarbonylammo)propyl, methoxy, methoxymethyl. methoxyothy1, ethoxy, ethoxy methyl, ethoxyethyl, methylsulfonyl, (methylsulfoiiyl)methyl, (methylsulfonyl)ethyl, (methylsulfonyl)propyL amino, aminomethyl, aminoethyl, aminopropyl, ammoisopropyL aminobutyl, carboxy, carboxymethyl, carboxyethyl, hydroxy, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxyisopropyl, trifluoromethyl, 2,2,2-trifluoroethyl, methyl, ethyl, isopropyl, w-propyl, isobutyl, ^-butyi, sec-butyl, isobutyl, tert-butyl, phenyl, and benzyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the thirty-first embodiments.

[00168] In a thirty-third embodiment, R^4’ is selected from aminocarbonyl, (aminocarbonyl)methyL ( / VJ' / -dimethylamino)carbonylmethyl, aminocarbonyl amino, (aminocarbonylamino)ethyl, methoxymethyl, (niethylsulfonyl)methyl, hydroxy, and methyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the thirty'-second embodiments.

[00169] In a thirty-fourth embodiment, R^a js selected from hydrogen, methyl, ethyl, hydroxy, methoxy, and ethoxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the thirty-third embodiments.

[00170] In a thirty-fifth embodiment, R / a is selected from hydrogen, methyl, and hydroxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the thirty -fourth embodiments.

[00171] In a thirty-sixth embodiment, R^b selected from phenyl, biphenyl, naphthyl, pyridyl, pyridazinyl, pyrimidyl, pyrazinyl. imidazolyl, pyrazolyl, furyl, thiophenyl, oxazolyl, isoxazolyL thiazolyl, isothiazolyl, oxadiazolyl, 3-oxoisoindolinyl, and bicyclofl. 1.1 ]pentyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the thirty -fifth embodiments.

[00172] In a thirty-seventh embodiment, R^b js selected from phenyl, pyridyl, pyrimidyl, fund, thiazolyl, and 3-oxoisoindolinyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the thirty-sixth embodiments.

[00173] In a thirty-eighth embodiment of the invention, R^c is selected from fluoro, chloro, bromo, iodo, hydroxy, methoxy, ethoxy, difluoromethoxy, tri fluoromethoxy, carboxy, carboxymethyl, methoxycarbonyl, methoxy carbonylmethyl, ethoxycarbonyl, carboxy methoxy, carboxyethoxy, carboxymethoxymethyl, aminocarbonyl, aminocarbonylmethyl, (N,N-dimethylamino)carbonyl, (AW-dimethylaminolcarbonylmethyl, ammo, aminomethyl, N.N-dimethylammo. (A'jV~dimethy4amino)methyl, ( / V,AMiethylamino)methyl, N,N,N-trimethylammonium, AdMAMrimethydmethyl ammonium, ATV^Mri ethylmethyl ammonium, methyl, ethyl, propyl, isopropyl, difluoromethyl, trifluoromethyl, -SO2OH.-CH2SO2OH, - aminoethoxy, aminoethoxymethyl, (fVJV-dimethylamino)ethoxy, jV^ / Mrimethyleth-1 -oxy-ammonium, aminocarbonylamino, and aminocarbonyl aminomethyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the thirty-seventh embodiments.

[00174] In a thirty-ninelh embodiment of the invention, is selected from hydroxy, carboxymethoxy, fluoro, chloro, methoxy, carboxy, carboxy methyl, methoxycarbonyl, aminocarbonyl, aminomethyl, -SOaOH. -SO2NH2, -CH2SO2OH. aminoethoxy, and aminocarbonylamino, and the other groups are as provided in the general Formula (I) above, or as in the first through the thirty-eighth embodiments.

[00175] hi a fortieth embodiment, R8a is selected from hydrogen, methyl, ethyl, hydroxy, methoxy, and ethoxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the thirty-nineth embodiments.

[00176] In a forty-first embodiment, R8« is selected from hydrogen, and methyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the fortieth embodiments.

[00177] In a forty-second embodiment of the invention, is selected from mdolyl. naphthyl, indolylmethyl, naphthylmethyl, quinolinyl, pyrrolo[2.3-Z»]pyridinyl, indazolyl, benzothiazolyl, and benzothiophenyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the forty-first embodiments.

[00178] In a forty-third embodiment of the invention, R^b js selected from indoly], naphthyl, and pyrrolo[2,3-Z>]pyridinyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the forty-second embodiments.

[00179] In a forty-fourth embodiment of the invention, each independently is selected from methyl, ethyl, isopropyl, fluoro, chloro, bromo, iodo, cyano, amino, aminomethyl, nitro, hydroxy, hydroxymethyl, carboxy, fluoromethyl, difluoromethyl, trifluoromethyl, trichloromethyl, methoxy, ethoxy, and trifluoromethoxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the forty-third embodiments.

[00180] In a forty-fifth embodiment of the invention, each R8« independently is selected from fluoro, chloro, bromo, and cyano, and the other groups are as provided in the general Formula (I) above, or as in the first through the forty-fourth embodiments.

[00181] In a forty-sixth embodiment of the invention, is selected from hydrogen, methyl, ethyl, and propyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the forty-fifth embodiments.

[00182] In a forty-seventh embodiment of the invention,    is hydrogen or methyl, and the other groups are as provided in the general Formula (1) above, or as in the first through the fortysixth embodiments.

[00183] In a forty-eighth embodiment of the invention. is selected from hydrogen, hydroxy, methyl, ethyl, propyl, methoxy, ethoxy, and propoxy, and the other groups are as provided in the general Formula (1) above, or as in the first through the forty-seventh embodiments.

[00184] In a forty-nmeth embodiment of the invention. is selected from hydrogen, and hydroxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the forty-eighth embodiments.

[00185] In a fiftieth embodiment of the invention. R10b is selected from phenyl, benzyl, biphenyl, biphenylmethyl, pyridyl, pyridyl methyl, pyridazinyl, pyrimidyl, pyrazmyl, imidazolyl, imidazolylmethyl, pyrazolyl, furyl, furylmethyl, oxazolyl, oxazolylmethyl, thiazolyl, indolyl, [l,2,4]iriaz.olo[l,5-a]pyndine, and bicyclofl. 1.1]pentyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the forty-nineth embodiments.

[00186] In a fifty-first embodiment of the invention, Ri^b is selected from phenyl, and pyrimidyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the fiftieth embodiments.

[00187] In a fifty-second embodiment, each Rl®c is independently selected from methyl, ethyl, w-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, cyclobutyl, u-pentyl, isopentyl, neopentyl, cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, cyclopentyl, cyclohexyl, cyclopentylmethyl, cyclohexylmethyl, fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, trifluoromethoxy, 2,2,2-trifluoroethoxy, fluoro, chloro, bromo, iodo, hydroxy, hydroxymethyl, hydroxyethyl, cyano, amino, aminomethyl, aminoethyl, A'-methylamino, (N-methylammo)methyi, fiV-methy kimmoietm1, A.A-dimeim iammo. (AAA-dimethylamino)methyl, (A^-dimethylamino)ethyl, ACN,AMrimethylammonium, AWAMrimethylmethan-ylammonium, aminocarbonyl, aminocarbonylmethyl, aminocarbonylethyl, (A’-methylamino)carbonyl, (N-methylamino)carbonylmethyl, (AlA-dimethylamino)carbonyl, dimethylamino)carbonylmethyl, aminocarbonylamino, aminocarbonylaminomethyl, methylcarbonylamino, methylcarbonylaminomethyl, carboxy, carboxymethyl, carboxyethyl, meth oxy carboxy, carboxy methoxy, carboxy ethoxy, carboxymethoxymethyl, carboxyethoxymethyl, S( (ty-OH. CH2(S(=O)2OH), S( ObXH.. CH .(S{ O} XH .). aminoethoxy, aminopropoxy, (A'-niethylammo)ethoxy, (A-ethylamino)ethoxy, (N,N-diW^^AWiV°)eihoxy, (AAAMiethylamino)ethoxy, (7V,2VpV-trimethylammonium)ethoxy, methoxy, ethoxy, methoxymethyl, ethoxymethyl, di fluoromethoxy, trifluoromethoxy, and 2,2,2-trifluoroethoxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the fifty-first embodiments.

[00188] In a fifty-third embodiment, each R^c is independently selected from fluoro, carboxy, carboxymethyl, (carboxymethyl)oxy, aniinocarbonyi, amino, aminomethyl, -SO2OH, -SO2NH2. hydroxy, and aminocarbonylamino, and the other groups are as provided in the general Formula (I) above, or as in the first through the fifty-second embodiments.

[00189] In a fifty-fourth embodiment of the invention, R^^3 is selected from hydrogen, hydroxy, methyl, ethyl, methoxy, and ethoxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the fifty-third embodiments.

[00190] In a fifty-fifth embodiment of the invention, Rlla is selected from hydrogen, and hydroxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the fifty-fourth embodiments.

[00191] In a fifty-sixth embodiment, js selected from (I-i2N-C(==NH)-NH)methyl, (H.XC( Ml)(HgX C( XH) XHipropxL U hX ( ( Xii > XHtyuld. phenyl, benzyl, pyridinyl, pyridinylmethyl, indolyl, indolylmethyl, pyridazinyl, pyridazinylmethyl, pyrimidyl, pyrimidylmethyl, pyrazinyl, pyraziny Im ethyl, imidazolyl, imidazolylmethyl, pyrazolyl, pyrazolylmethyl, [l,2,4|triazolo[l,5-a|pyridine, oxazolyl, oxazoly I methyl, thiazolyl, and thiazolylmethyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the fifty-fifth embodiments.

[00192] In a fifty-seventh embodiment, R^^ is selected from phenyl, (H X C{ M h NH)ethyl, pyridinyl, indolyl. and imidazolyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the fifty-sixth embodiments.

[00193] In a fifty-eighth embodiment of the invention, each c is independently selected from fluoro, chloro, bromo, iodo, (carboxymethyl)oxy, (carboxymethyl)oxymethyl, (carboxy ethyljoxy, hydroxy, hydroxymethyl, hydroxyethyl, methoxy, ethoxy, difluoromethoxy, trifluoromethoxy, carboxy, carboxymethyl, aminocarbonyl, aminocarbonylmethyl, (N-melhylaniino)carbonyl, (IV-methylamino)carbonylmethyl, (,V,A’-dimethylamino)carbom (N,N-dimethylamino)carbonylmethyl, amino, A-methylamino, AtyV-dimethylamino, N.N-diethylamino, aminomethyl, (AtyV-dimethylamino)methyl, (AtyX-diethylamino)methyl, N,N,N-trimethylammonium, XfAXV-trimethylmeth-1 -yl-ammonium, N,N,N-{n ethyl ammonium, N,N,N-triethylmeth-1-yl-ammonium, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, difluoromethyl, trifluoromethyl, aminoethoxy, aminopropoxy, aminoethoxymethyl, (AfAMimethylamino)ethoxy, AfAyV-trimethyleth-l-yloxy-ammonium, cyano, and methylcarbonylpiperazyl [QV-acetyl)piperazyl], and the other groups are as provided in the general Formula (I) above, or as in the first through the fifty-seventh embodiments.

[00194] In a fifty -nineth embodiment of the invention, each R^c is independently selected from (carboxymethyl)oxy, fluoro, chloro, hydroxy, methoxy, trifluoromethoxy, carboxy. carboxymethyl, aminocarbonyl, aminocarbonylmethyl, aminomethyl, aminoethoxy, (N,N~ dimethylammo)ethoxy, and methylcarbonylpiperazyl (A-acetyl)piperazyl or , and the other groups are as provided in the general Formula (I) above, or as in the first through the fifty -eighth embodiments.

[00195] In a sixtieth embodiment of the invention,      js selected from hydrogen, methyl, ethyl, propyl, cyclopropyl, and isopropyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the fifty-nmeth embodiments.

[00196] In a sixty-first embodiment of the invention, R^a js selected from hydrogen. methyl, ethyl, and «-propyl, and the other groups are as provided in the general Formula (I) above, or as m the first through the sixtieth embodiments.

[00197] In a sixty-second embodiment, R*2b JS selected from hydrogen, methyl, ethyl, and propyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the sixty-first embodiments

[00198] In a sixty-third embodiment, R^b js selected from hydrogen, and methyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the sixty-second embodiments.

[00199] In a sixty-forth embodiment of the invention, R^a and R together with the atoms to which they are attached form a saturated ring selected from1^ , / X VNx and            , and the other groups are as provided in the general Formula (I) above, or as in the first through the sixty-third embodiments.

[00200] In a sixty-fifth embodiment of the invention. R^’a anc| R12b together with the atoms to which they are attached form , and the other groups are as provided in the general Formula (J) above, or as in the first through the sixty-fourth embodiments.

[00201] In a sixty-sixth embodiment, is hydrogen, methyl, or propyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the sixty-fifth embodiments.

[00202] In a sixty-seventh embodiment, R^b js hydrogen or methyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the sixty-sixth embodiments.

[00203] In a sixty-eighth embodiment, js selected from indolyl, pyrrolo[2,3-£,]pyridmyi, quinolinyl, mdazolyl, and naphthyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the sixty-seventh embodiments.

[00204] In a sixty-nineth embodiment, R^c js selected from indolyl, and pyrrolo[2,3-b]pyridmyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the sixty-eighth embodiments.

[00205] In a seventieth embodiment, each Rl-^ independently is selected from fluoro, chloro, bromo, iodo, methyl, ethyl, propyl, isopropyl, carboxy, carboxymethyl, methoxy, difluoromethoxy, trifluoromethoxy, and ethoxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the sixty-nineth embodiments.

[00206] In a seventy-first embodiment, each R^^ independently is selected from methyl, chloro, and methoxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the seventieth embodiments.

[00207] In a seventy-second embodiment, each R14a independently is selected from amino, hydroxy, A-methylamino, AtyV-dimethylamino, A’-ethylamino. methoxy, and ethoxy, and the other groups are as provided in the general Formula (I) above, or as in the first through the seven ty -fi rst embodi ments.

[00208] In a seventy-third embodiment, each R^a independently is amino, and the other groups are as provided in the general Formula (1) above, or as in the first through the seventy-second embodiments.

[00209] In a seventy-fourth embodiment, each independently is selected from hydrogen, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl. isobutyl, w-pentyl, isopentyl, neopentyl, trifluoromethyl, trifluoroethyl, phenyl, benzyl, chlorophenvl, dichlorophenyl, fluorophenyl, difluorophenyl, bromophenyl, iodophenyl, chlorobenzyl, dichlorobenzyl, fluorobenzyl, difluorobenzyl, bromobenzyl, iodobenzyl, naphthyl, naphthylmethyl, pyrazolyl, pyrazolylmethyl, indolyl. indolylmethyl. imidazolyl, imidazolylmethyl, pyridyl, and pyridylmethyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the seventy-third embodiments.

[00210] In a seventy-fifth embodiment, each R^b independently is selected from hydrogen, methyl, ethyl, and phenyl, and the other groups are as provided in the general Formula (I) above, or as in the first through the seventy-fourth embodiments. ]00211 ]   In certain embodiments of the methods provided herein, the compound is a compound of Formula (II) or a pharmaceutically acceptable salt thereof: (II) wherein: R1' is hydrogen, Ci-to alkyl, monocycloalkyl, monoheterocycloalkyl, monocyclic aryl, or monocyclic heteroaryk R2' is cycloalkyl, heterocycloalkyl, aryl, or heteroaiyl; R’'1 is hydrogen or Ct-3 alkyl; R3d is hydroxy(Ci-6 alkyl); R3a and RJ'b together with the carbon and nitrogen atom to which they are attached form a 5- or 6-niembered heterocycloalkyl; R3c is hydrogen or Ci-3 alkyl; R4' is hydrogen or Ci-6 alkyl; R’’a is hydrogen or C1-6 alkyl; R5'&is hydrogen or carboxy(Ci-6 alkyl); or R5'a and R5'b together with the carbon and nitrogen atom to which they are attached form a 5- or 6-membered heterocycloalkyl; Rz is monocyclic aryl or monocyclic heteroaryl; Rs' is bicyclic aryl or bicyclic heteroaryl; R9' is hydrogen or Ci-4 alkyl, R10 is hydrogen. Ci-e alkyl, -OH. or-COOH; Rtr is monocyclic aryl or monocyclic heteroaryl; RL-i'a is hydrogen or Ci-6 alkyl; R,?,b is hydrogen or Ci~4 alkyl; R'3’c is hydrogen. Ci-s alkyl, Ci-6 alkoxy; and wherein the asterisk (*) represents attachment to the compound at the carbonyl carbon bound to the linker, wherein the w avy line (wow*) represents attachment to the nitrogen bound to the linker; and wherein: m is selected from 1 to 4; n is selected from 1 to 4; wherein each of the C1-3 alkyl, Ci-4 alkyl, Cue alkyl, C1-6 alkoxy, cycloalky I, heterocycloalkyl, aryl, or heteroaiyl are optionally substituted with 0.1, or 2 halo, hydroxyl, carboxy (-COOH). or aminocarbonyl (-CONH2) groups.

[00212] In Formula (II), R1 may be, in any embodiment of any method disclosed herein, hydrogen, C1-10 alkyl, C3-6 monocycloalkyl, C3-5 monoheterocycloalkyl, C5-6 monocyclic aryl, or C5-6 monocyclic heteroaiyl. In certain embodiments, R1 is Ci-6 alkyd or C5-6 monocyclic aryl. In certain embodiments, R1 is C2-4 alkyl or optionally substituted phenyl. In certain embodiments, Rr is propyl, butyl, or phenyl.

[00213] In some embodiments of the method disclosed herein, the linker of the compound of X O / * Formula (II) is: O              . In some embodiments of the methods disclosed herein, the 0 rVs! n h2 O Ht< linker is               *     . In some embodiments of the method disclosed herein, the linker

[00214] In certain embodiments of the methods disclosed herein, the compound that binds to TNFR1 is a compound of Formula (II), wherein R2‘ is substituted or unsubstituted pyridmyl or phenyl. In certain embodiments, R2‘ is substituted or unsubstituted 4-pyridinyl or phenyl. In some embodiments, R2' is 4-pyridinyl. In certain embodiments, R2 is hydroxy-substituted phenyl. In certain embodiments, R2' is 4-hydroxy-phenyl.

[00215] In certain embodiments of the methods disclosed herein, the compound that binds to TNFR1 is a compound of Formula (II), wherein R2% is hydroxy(Ci-6 alkyl). In certain embodiments, R?" is hydroxymethyl.

[00216] In certain embodiments of the methods disclosed herein, the compound that binds to TNFR1 is a compound of Formula (II), wherein R3‘a and Rjb together with the carbon and nitrogen atom to which they are attached form an optionally substituted 5- or 6-membered heterocycloalkyl having 0 or I additional heteroatoms selected from the group consisting of N, O, and S. In certain embodiments, R3a and R3'b together with the carbon and nitrogen atom to which they are attached form an optionally substituted 5- or 6-membered helerocycloalkyl, x kX wherein the an optionally substituted 5- or 6-membered heterocycloalkyl is         ' or ' , wherein the wavy line (■✓wv') represents attachment to adjacent atoms in the compound.

[00217] In certain embodiments of the methods disclosed herein, the compound that binds to TNFRi is a compound of Formula (II), wherein R4' is hydrogen or Ci-6 alkyl. In certain Attorney Ref. No. 14463-061 -228 embodiments, R4' is hydrogen or C1-3 alkyl. In certain embodiments, R4’ is hydrogen or methyl. In certain embodiments, R4' is hydrogen. In certain embodiments, R4' is methyl.

[00218] In certain embodiments of the methods disclosed herein, the compound that binds io TNF.R1 is a compound of Formula (II), wherein R3 a is hydrogen or Ci-s alkyl. In certain embodiments, R3'a is hydrogen or methyl. In certain embodiments, R3a is methyl.

[80219] In certain embodiments of the methods disclosed herein, the compound that binds to TNFR1 is a compound of Formula (IT), wherein R5b is hydrogen or carboxy(C 1-6 alkyl). In certain embodiments, Ri bis carboxytCm alkyd). In certain embodiments, R5bis hydrogen or-(CH2)2COOH. In certain embodiments, R3b is hydrogen. In certain embodiments. R5b is -(CH2)2COOH.

[00220] In certain embodiments of the methods disclosed herein, the compound that binds to TNFR1 is a compound of Formula (II), wherein R5’a and R5 b together with the carbon and nitrogen atom to which they are attached form an optionally substituted 5- or 6-membered heterocycloalkyl having 1 or 2 heteroatoms selected from the group consisting of N, O, and S. In certain embodiments, Ria and R3b together with the carbon and nitrogen atom to which they are attached form an optionally substituted 5-membered heterocycloalkyl. In certain embodiments, R,a and R5b together with atoms to which they are attached form a 5- or 6-membered vW X \ heterocycloalkyl which is          wherein the wavy line (owv') represents attachment to the adjacent atoms of the compound.

[00221] In certain embodiments of the methods disclosed herein, the compound that binds to TNFRI is a compound of Formula (II), wherein R7' is substituted or unsubstituted pyridinyl or phenyl. In certain embodiments. R7’ is substituted or unsubstituted 4-pyridinyl or phenyl. In some embodiments, R4 is 4-pyridinyl. In certain embodiments, R7 is hydroxy-substituted phenyl. In certain embodiments, R7' is 4-hydroxy-phenyl. In certain embodiments, R7 is carboxy-substituted phenyl. In certain embodiments, R7' is 4-carboxy-phenyl.

[00222] In Formula (II). R8' may be. in any embodiment of any method disclosed herein, substituted, or unsubstituted Cs-io bicyclic aryl or 8-, 9-, or 10-membered bicyclic heteroaryl. In some embodiments of the methods disclosed herein. R8’ is substituted or unsubstituted naphthyl. In certain embodiments. R8' is substituted or unsubstituted indolyl. In certain embodiments, R8 is Ml J J "'N R8a or H . wherein Rsa is hydrogen, C1-3 alkyl, or halo. In certain RSa is chloro. In certain embodiments, RSa is hydrogen.

[00223] In certain embodiments of the methods disclosed herein, the compound that binds to TNFRI is a compound of Formula (II), wherein R9' is hydrogen or Cur alkyl. In certain embodiments, R9' is hydrogen. In certain embodiments, R9' is C1-4 alkyl. In certain embodiments, R9 is methyl.

[00224] In certain embodiments of the methods disclosed herein, the compound that binds to TNFRI is a compound of Formula (II), wherein R10' is hydrogen, C1-6 alkyl, -OH, or - COOH. In certain embodiments, R10' is -OH or-COOH. In certain embodiments, R10' is -OH. In certain embodiments, R10' is -COOH.

[00225] In certain embodiments of the methods disclosed herein, the compound that binds to TNFR1 is a compound of Formula (II), R11 is monocy clic aryl or heteroary l. In certain embodiments, Rir is substituted monocyclic aryl. In certain embodiments, R11' is unsubslituted monocyclic aryl. In certain embodiments, R'r is substituted monocyclic heteroaryl. In certain embodiments, Rn' is unsubstituted monocyclic heteroaryl. In certain embodiments, R11’ is substituted or unsubstituted imidazolyl or phenyl. In certain embodiments, R“ is substituted with hydroxyl and -CONH2. In certain embodiments, R19 is

[00226] In certain embodiments of the methods disclosed herein, the compound that binds to TNFRI is a compound of Formula (II), wherein R13'3 is hydrogen or C1-3 alkyl. In certain embodiments, Rld'a is hydrogen or methyl. In certain embodiments, R!3a is hydrogen. In certain embodiments, R139’ is methyl. [G0227] In certain embodiments of the methods disclosed herein, the compound that binds to TNFRI is a compound of Formula (II), wherein R9?’' is hydrogen or C1-4 alkyl. In certain embodiments, R13h is hydrogen or Ci-4 alkyl. In certain embodiments. R13b is C1-4 alkyl. In certain embodiments, R!, b is methy l. In certain embodiments, Rb’b is hydrogen.

[00228] In certain embodiments of the methods disclosed herein, the compound that binds to TNFRI is a compound of Formula (II), wherein R1S’° is hydrogen or C1-6 alkoxy. In certain embodiments, R1j'' is hydrogen or methoxy. In certain embodiments, R)jt is hydrogen. In certain embodiments, Rlj c is methoxy .

[00229] In certain embodiments of the methods disclosed herein, the compound that binds to TNFRI is a compound of Formula (II): R9 is propyl, butyl, or phenyl; R3a is hydrogen; R3'1’ is hydroxymethyl; or RJ'a and R3'b together with the carbon and nitrogen atom to which they are attached form a 6-membered heterocycloalkyl; R8' is substituted or unsubstituted naphthyl or indolyl; R9 is H or methyl; and R;o is -OH or -COOH,

[00230] In certain embodiments of the methods disclosed herein, the compound that binds to TNFR1 is a compound of Formula (II), wherein each of the alkyd, alkoxy, cycloalkyd, heterocycloalkyl, aryl, or heteroand may be optionally and independently substituted with one or more of halo, hydroxyl, carboxy (-COOH). or aminocarbonyl (-CONH2) groups. [0023.1] In certain embodiments of the methods disclosed herein, the compound that binds to TNFR1 is Compound .4: Compound A, or a pharmaceutically acceptable salt thereof As will be recognized by one of skill in the art. Compound A may also be described by SEQ ID NO:6.

[00232] In certain embodiments of the methods disclosed herein, the compound that binds to TNFR1 is Compound B: Compound B, or a pharmaceutically acceptable salt thereof. As will be recognized by one of skill in the art, Compound B may also be described by SEQ ID N0:7.

[00233] In certain embodiments of the methods disclosed herein, the compound that binds to or a pharmaceutically acceptable salt thereof. As will be recogni zed by one of skill in the art. Compound C may also be described by SEQ ID N0:8.

[00234] In certain embodiments of the methods disclosed herein, the compound that binds io TNFR1 is Compound D: Compound D, or a pharmaceutically acceptable salt thereof. As will be recognized by one of skill in the art, Compound D may also be described by SEQ ID N0:9.

[00235] The mechanism of action (MOA) of anti-TNFa biologies such as infliximab, adalimumab, golimumab and certolizumab, involves binding to the cytokine TNFa and thus, inhibition of engagement with both TNFR1 and tumor-necrosis factor receptor 2 (TNFR2). While not being bound by any specific theory, the compounds that bind to the TNFR1 binding pocket as provided herein selectively inhibit TNFR1 to specifically attenuate the proinflammatory activities of TNFa-mediated TNFR1 signaling, and to spare / passively enable TNFa-TNFR2 pro-homeostatic signaling, which may confer better therapeutic efficacy than the standard of care anti-TNFa biologies. Accordingly, provided herein, in certain aspects, are compounds and compositions comprising the same that bind to the TNFR1 binding pocket provided herein. Also provided are methods of modulating TNFR1 -mediated biological activity in subject, such as inflammation, apoptosis, necroptosis, cell differentiation, cell survival, and cell proliferation. 8. EXAMPLES 8.1. EXAMPLE I: METHODS OF SYNTHESIS 8.1.1. General Procedures to Access Building Blocks and Monomers

[00236] The compounds of the present invention can be prepared according to the procedures of the following schemes and specific examples, or modifications thereof, using readily available starting materials, appropriate materials and reagents and conventional synthetic procedures and are further exemplified by the following specific examples. In these reactions, it is also possible to make use of variants which are themselves known to those of ordinary skill in this art but are not mentioned in greater detail. The general procedures for making the compounds claimed in this invention can be readily understood and appreciated by one skilled in the art from viewing the following schemes. The examples also include methods for testing such compounds in biophysical, biochemical, and cellular assays. The compounds illustrated in the examples are not, however, to be construed as forming the only genus that is considered as the disclosure.

[00237] Unless otherwise specifically indicated, all reagents are commercially available, known in the literature, or readily synthesized by one skilled in the art. lire general route applied to the synthesis of compounds of Formula (II) (e.g, Compounds A, B, C, D) is described in the Schemes that follow. In some instances, the order of carrying out the reaction steps in the schemes may be varied to facilitate the reaction or to avoid unwanted reaction products. Additionally, various protecting group strategies familiar to one skilled in the art of organic synthesis and solid phase peptide synthesis may be employed to facilitate the reaction, to improve yield and purity, or to avoid unwanted reaction products.

[00238] All reagents and solvents were purchased from commercial sources and used without further purification unless otherwise noted. All temperatures are in degrees Celsius (°C), and ambient temperature or room temperature (RT) is 20 °C. Most compounds were purified by reversed-phase preparative high-performance liquid chromatography (HPLC) or mediumpressure liquid chromatography (MPLC) on silica gel. The course of the reactions was followed by liquid chromatography / mass spectrometry (LC-MS) or Ultra performance liquid chroma.iogra.phy / mass spectrometry (UPLC-MS); electrospray ionization (ESI); UV detection at 254 nm). Proton, fluorine, and carbon magnetic resonance (’H, 19F and BC NMR) spectra were recorded on a 300, 400, 500, or 600 MHz Varian or Bruker spectrometer, and chemical shifts are reported in parts per million (ppm) relative to tetramethylsilane and referenced to residual solvent. Coupling constants are reported in hertz. 5H NMR data are reported as given here: chemical shift (multiplicity [singlet (s), doublet (d), triplet (0. quartet (q), doublet of doublets (dd), doublet of triplets (dt), triplet of doublets (td), triplet of triplets (tt), doublet of doublet of doublets (ddd), multiplet (m), and broad singlet (br. s)], coupling constant [Hz] and integration). Reactions sensitive to moisture or air were performed under nitrogen or argon using anhydrous solvents and reagents. The progress of reactions was determined by either analytical thin layer chromatography (TLC) usually performed with pre-coated TLC plates (E. Merck, Darmstadt, Germany), silica gel 60F-254, layer thickness 0.2.5 mm or liquid chromatography-mass spectrometry (LC-MS).

[00239] Unless otherwise indicated, when ratios of compounds (such as for examples solvents) are given, the ratio is on a volume-to-volume basis. For example, solvent gradient ranging from 100% hexanes to 50% EtOAc / hexanes means a gradient starting from a mixture of 100 parts by volume of hexanes varying to mixture of 50 parts by volume ethyl acetate to 50 parts by volume of hexanes.

[00240] The term "‘w / w' means weight of compound to total weight. For example, NaH 60% w / w means 60 parts by weight NaH to 100 parts total weight. [00241 ] The following examples are provided so that the invention might be more fully understood. These examples are illustrative only and should not be construed as limiting the invention in any way. Wherein a racemic mixture is produced, the enantiomers may be separated using SFC reverse or normal phase chiral resolution conditions either after isolation of the final product or at a suitable Intermediate, followed by processing of the single isomers individually. It is understood that alternative methodologies may also be employed in the synthesis of these key intermediates and examples. Asymmetric methodologies (e.g., chiral catalysis, auxiliaries, biocatalytic process) may be used where possible and appropriate. The exact choice of reagents, solvents, temperatures, and other reaction conditions depends upon the nature of the intended product.

[00242] The amino acids used to synthesize the Final Compounds as described herein may be prepared by the following methods and are also commercially available. 8.L2. Intermediates and Monomers Syntheses Synthetic Scheme 1 Step 1 PfTrpB-7E6, PLP phosphate buffer (pH - 8.0) DMSO Step 2 Na2CO3, FmocOSu, THF Fmoc-SbMeW-OH Method of Making (25’3<S)-2-((((9 / f-fiuoren-9-yl)inethoxy)carbonyI)ainino)-3-(lff-mdol-3-yi)butanoic acid (Fmoc-SbMeW-OH):

[00243] Step 1: Into a 500 mL 4-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen, was placed DMSO (20 mL, 4 V), indole (5,00 g, 42 mmol), (2S',3 / / )-2-aniino-3-hydroxybutanoic acid (10.17 g, 85 mmol, 2 equiv), 0.2 M potassium phosphate buffer (230 mL, 46 V, pH =: 8.0)- The resulting solution was stirred at 65 °C. PLP (46 mg, 0.18 mmol, 0.0041 equiv) in H2O (2 mL) and PfTrpB 7E6 (LOO g, 20 wt. %) in 0.2 M potassium phosphate buffer (2 mL, pH = 8) was added into the solution. The resulting solution was stirred for 18 h at 65 °C. The reaction progress was monitored by LC-MS. Upon completion, the resulting solution was used directly in next step.

[00244] Step 2: Into the solution was added NaaCOs (9.05 g, 85 3 mmol, 2.0 equiv), FmocOSu (21.58 g, 64.0 mmol, 1.5 equiv) and THF (100 mL) at 0 °C. The resulting solution was stirred for 3 h at RT. The solution was adjusted to pH = 3 with aq. 6 N HC1. The mother liquor was collected by filtration and extracted with EtOAc (3 x 200 mL). The combined organic layers were washed with DI water (2 * 400 mL) and brine (400 mL). dried over NasSCX filtered, and concentrated under vacuum. The crude product was triturated with (DCM / w-heptane " L I; v / v), and the solids were collected by filtration to afford (2S,3S)-2-((((9#-fluoren-9-y])methoxy)carbonyI)amino)~3~(lH-indol-3-yr)butanoic acid. MS ESI calculated for C27H24N2O4 [M + H]+ 441, found 441. JH NMR (400 MHz. DMSO-oU 6 10.88 (s. 1H), 7.87 (d. . / 7.5 Hz, 2H), 7.69 7 56 (m, 3H), 7.39 11., / 7.4 Hz, 2H), 7.38-7.21 (m, 4H), 7.18 (d.. / 2.1 Hz. 1H), 7.05 (ddd, J - 8.1, 6.9, 1.2 Hz, 1H), 6.96 (td, J- 7.4, 7.0, 1.1 Hz, 1H), 4.43-4.32 (m, IH), 4.21 (d, 7= 10.0 Hz, IH), 4.16 (t, 7= 5.3 Hz, 2H), 3.50 (q. 7= 7.1 Hz, IH), 1.33 (d.7=7.1 Hz. 3H). Synthetic Scheme 2 Step 1 PHrpB-7E6, PLP phosphate buffer (pH = 8.0) DMSO Step 2 Na2CO3, FmocOSu, THF MeO' NHFmoc Fmoc-SbMeW7OMe-OH Method of Making (2S^S)-2-((((9H41uoren-9-yl)methoxy)carbonyl)aniiiio)-3-(7-methoxy-lfMndoK5-yl)butanoic add (Finoc-SbMeW7OMe-OH):

[00245] Step I: Into a 5 L 4-necked round-bottom flask, purged and maintained with an inert atmosphere of nitrogen, was placed DMSO (640 mL, 4 V), 7-methoxy-l / / -indole (50 g, 0.34 mol, 1 equiv), (2$37?)-2-amino-3-hydroxybutanoic acid (81 g, 0.68 mol, 2 equiv) and 0.2 M potassium phosphate buffer (2300 mL, pH = 8.0, 46 V). The resulting solution was heated to 65 °C. PLP (5 g, 0.02 mol. 0.056 equiv) and PflrpB 7E6 (50 g, 100 wt. %) were added into the solution. The reaction mixture was stirred overnight at 65 °C. Upon completion, the reaction mixture was used directly in the next step.

[00246] Step 2: Into the reaction mixture were added NazCOs (72 g, 0.68 mol, 2.0 equiv), Fmoc-OSu (230 g, 0.68 mol. 2 equiv), and THF (500 mL) at 0 °C. The reaction was stirred for 3 h at RT. The solution was then adjusted to pH = 5-6 with an aq. 6 N HC1 solution. The mixture was then filtered, and the filtrate was extracted with EtOAc (3 x 1 L). The combined organic layers were washed with DI water (2 x 1 L) and brine (1 L). dried over NaaSOr, filtered, and concentrated under vacuum. The residue was purified by trituration with DCM / heptane (5 V:5 V) for 1 h to yield (2S.35)-2-((((97 / -fluoren-9-yl)methoxy)carbonyl)amino)-3-(7-methoxy-l^ indoL3-yl)butanoic acid. MS ESI calculated for C28H26.N2O5 [M + H]~ 471. found 471. ’H NMR (400 MHz, DMSO-Ts): S 12.63 (s, 1H), 10.95 (d, J = 2.6 Hz, IH). 7.87 (dt. 7= 7.6. 1.0 Hz. 2H), 7.68--7.61 (m, 2H), 7.40 (tt, 7 = 7.5, 1.1 Hz, 2H), 7.28 (tdd, 7= 7.5, 3.4, 1.1 Hz, 3H), 7.20 (d, 7 = 8.0 Hz, IH), 7.10 (d, J- 2.5 Hz, IH), 6.89 (1,7= 7.8 Hz, IH), 6.66-6.59 (m, IH), 4.36 (dd. 7 8.5, 7.2 Hz, IH). 4.27-4.07 (m, 3H). 3.89 (s, 3H). 3.44 (q, 7= 7.1 Hz, IH). 1.33 (d, 7= 7.1 Hz, 3H). 8.13. Preparation of Final Compounds A. Generalized Procedure for Synthesizing Linear Peptide Precursors

[00247] Peptides in Tables 1 and 2 were synthesized using standard solid-phase synthesis using Fmoc / rBu chemistry as exemplified in Chan, W.C.: White. P.D. “Fmoc Solid-Phase Synthesis: a Practical Approach”, Oxford University7 Press, Oxford. 2000; Steward. J.; Young, J. “Solid Phase Peptide Synthesis”, Pierce Chemical Company, Rockford, 1984.; Benoiton. N.L. “Chemistry of Peptide Synthesis”, CRC Press, New York, 2006; and Lloyd-Williams, P.; Albericio, F.; Giralt. E. “Chemical Approaches to the Synthesis of Peptides and Proteins”. CRC Press. New York, 1997.

[00248] During peptide chain elongation, the a-amino group of each amino acid was protected with a 9#-fluoren-9-ylmethoxycarbony1 group (Fmoc). To avoid any side reactions during the chain elongation steps, any reactive amino acid side chains also carry' acid-labile protecting groups, effectively masking the reactive groups until removal upon treatment with strong acid. After completion of each coupling step, the Fmoc group of the TV-terminal amino acid was removed with piperidine or 4-methylpiperidine and the resin was thoroughly washed to prepare for the coupling of the subsequent Fmoc-protected ammo acid derivative. |00249] The side chain protecting groups used were: 1. terf-butyl (?Bu) for a-Me-t.-Ser (aMeS), L-Ser (S), L-Tyr, (Y) PhedCOOH, A-Me-t-Glu (NMeE); 2. trityl (Til) for L-Asn (N), L-Cys (C); 3. rerLbutoxy-carbonyl (Boc) forL-Trp (W). a-Me-L-Trp (aMeW), L-His (H). Pip2c.

[00250] Fmoc-protected amino acids were typically obtained from vendors such as Sigma-Aldrich® (Millipore Sigma, St. Louis, MO), Novabiochem® (Millipore Sigma, St. Louis, MO), Chem-lmpex (Wood Dale, IL), Combi-Blocks (San Diego, CA), Ambeed (Arlington Hts, IL), AstaTech Inc. (Bristol. PA), Iris Biotech (Marktredwitz, Germany), Acrotein BioChem Inc (Hoover. AL), Amatek (Berwyn. PA), ChemScene LLC (Monmouth Junction. NJ), BLD Pharmatech Co., Limited (Cincinnati, OH), AchemBlock (Hayward, CA), AA Blocks LLC (San Diego, CA), abcr GmbH (Karlsruhe, Germany), Enamine Ltd. (Kyiv, Ukraine), Chern Shuttle (Burlingame, CA), or PharmaBlock (USA), Inc. (Hatfield, PA). B. Synthetic Procedures used to Prepare Thioether Cyclic Peptides Synthetic Scheme 3 MeO Release from solid support and side chain deprotection TFA / TIS / H2O / DTT, 38 °C 1. Procedure A a) Solid Phase Synthesis of Peptides (SPPS) |00251] Peptides were synthesized on a Liberty Blue™ synthesizer from CEM Corporation (Matthews, NC), using standard solid-phase synthesis using Fmoc / rBu chemistry as summarized above in Synthetic Scheme 3.

[00252] Reactions were topically performed at a 50 pmol scale using Rink Amide MBHA LL resin (100-200 mesh, 0.4 mmol / g loading, 1% DVB cross-linked polystyrene, Novabiochem*). All the amino acids were dissolved at a 0.2 M concentration in anh. DMF orNMP. The amino acids were activated with equimolar amounts of Oxyma Pure solution (0.5 M in anh, DMF), and a 2-fold molar excess of DIC solution (0.5 M in anh. DMF). |00253] Every synthesis cycle included: (1) Fmoc amino acid deprotection by 20% v / v piperidine or V-methylmorpholine (NMM) in anh. DMF (90 °C microwave assisted healing, 1 min); (2) Coupling (potentially repeated twice for difficult couplings) with Fmoc-protected amino acid / DIC / Oxyma (5, 5, and 10 equiv, respectively, 90 °C microwave assisted heating. 2 or 4 ram). Cycles of Fmoc deprotection and Fmoc-protected amino acid coupling were repeated with the desired monomers until the full linear peptide was formed: (3) ’The peptide .V-terminus was eventually capped with the required 2-chloroacetyl acetol moiety by coupling with chloroacetic acid and DIC (5 and 10 equiv, respectively, 90 °C microwave assisted heating, 2 min, repeated twice). b) Cleavage and Deprotection

[00254] After completion of the synthesis, the linear resin-bound peptide was transferred into a fritted plastic column. The resin was washed with DCM then, dried under negative pressure on a Razor* peptide cleavage system from CEM Corporation (Matthews. NC). The peptide was deprotected and cleaved from the solid support by treatment with TFA / HzO / TlS / DODT (92.5:2.5:2.5:2.5, v / v / v / v; 10 mL) at 38 °C for 30 min. The solution was collected into a 50 mL centrifuge tube. The resin washed with TFA (2 x 2 mL). The cleavage solution was partially concentrated to a volume of ~5 mL under vacuum. The crude linear peptide was precipitated from the TFA cleavage solution using chilled tert-butyl methyl ether (MTBE; 40 mL) and collected by centrifugation (4000 rpm). The supernatant was removed. Additional cold MTBE (30 mL) was added to the peptide pellet and clarified by centrifugation. The resulting crude peptide pellet was blown dried by a stream of nitrogen gas. c) Peptide Cyclization

[00255] The crude solids were dissolved in 1:1 (v / v) mixture of DI water / MeCN (15 mL). The pH was adjusted to pH 8 by addition of an aq. 0.2 M ammonium bicarbonate solution (3 mL.). The solution was shaken at RI for approximately 2 h and monitored by UPLC-MS. After the reaction was complete, the reaction solution was quenched by addition of TFA (200 uL), frozen and lyophilized. d) HPLC Purification

[00256] The crude residue was then dissolved in DMSO and purified by preparative reversed-phase high performance liquid chromatography on a Waters™ SunFire Prep Cl 8 OBD column (IOOA, 5 um, column size 19 * 150 mm, Milford. MA) using an Agilent MS-Directed Preparative HPLC-MS system. Mobile phase: (A) 0.1% TFAm HPLC-grade water and (B) 0.1% TFA in HPLC-grade acetonitrile; flow rate: 35 mL / min; UV wavelength L - 215 nm; gradient: gradient: 0.5%B / min for 40 min starting at 5%B less than the calculated %B required for compound elution. In some instances, better separation was obtained by 0.1 % NH4OH as modifier.

[00257] UV absorbing fractions containing the target m / z ions were collected and the fractions containing the desired product were combined, concentrated in vacuo, and freeze-dried to afford the cyclized peptide as a solid.

[00258] Confirmation of identity and purity assessment of final compounds w ere performed by UPLC-MS, which was measured by a reverse phase Waters™ ACQUTTY UPLC-MS system. Column: Waters™ Cortecs C18+ Column (90 A, 1.6 pm, column size 2.1 x 100 mm). Mobile phase: (A) 0.1% TFA in HPLC-grade water and (B) 0.1% TFA in HPLC-grade acetonitrile; injection volume: 0.5 pL; flow rate: 0.7 mL / min; Column Temperature: 60 °C; UV wavelength X = 2.15 nm; gradient: 2-35% in 9.1 min. Synthetic Scheme 4 Solid-phase peptide synthesis 1) NMP (20% in DMF) 2) HATU / DIPEA, 50 or 75 °C OMe Cyclization DI PEA, DMSO, RT Release from solid support and side chain deprotection TFA / TIS'HjO / DTT, RT 2. Procedure B a) Solid Phase Synthesis of Peptides (SPPS)

[00259] Peptides were synthesized on a Biotage® Syro II peptide synthesizer (Biotage Corporation, Boston, MA) using standard solid-phase synthesis using Fmoc / tBu chemistry as summarized above in Synthetic Scheme 6.

[00260] Reactions were typically performed at the 10 pmol or 25 nmol scale using Rink Amide MBHA LL resin (100--200 mesh. 0.4 mmol / g loading, Novabiochem'*). Solutions of the reagents and amino acid monomers were prepared as follows: Fmoc-protected amino acids (0.2 M or 0.4 M in anh. DMF, NMP, or DMSO). HATU (0.5 M in anh. DMF), DIPEA (2.0 M in anh. NMP), (CLAchO (0.4 M in anh. DMF). 4-methylpiperidine (20% v / v in anh. DMF). |00261] Every synthesis cycle included: (I) Single or double coupling with Fmoc-protected amino acid / HATU / DIPEA (4, 4 and 8 equiv, respectively: at 50 °C or 75°C; 15 min for single coupling or 15 min + 30 min for double coupling). After the coupling reaction was complete the mixture was filtered, and the peptidyl resin was washed with DMF; (2) Fmoc deprotection (repeated three times) using 20% (v / v) 4-methylpiperidine in DMF (RT; 3 min). The mixture was filtered, and the peptidyl resin was washed with DMF. Cycles of Fmoc deprotection and Fmoc-protected amino acid coupling were repeated with the desired monomers until the full linear peptide was formed, (3) The peptide A-termmus was capped by the addition of (ClAcjzO and DIPEA (10 and 20 equiv, respectively; repeated twice) in anh. DMF. then the peptidyl resin was washed with DCM. b) Cleavage and Deprotection

[00262] After completion of the synthesis, the linear resin-bound peptide was washed with DCM then dried under positive nitrogen pressure on a peptide cleavage system from Biotage Corporation, Boston, MA

[00263] The peptide was deprotected and cleaved from the solid support by treatment with TFA / HzO / TIS / DODT (92.5:2.5:2.5:2.5, v / v / v / v; 2 mL) at RT for 1 h. The solution was collected into a 50 mL centrifuge tube. The resin washed with TFA (1 mL). The crude linear peptide was precipitated from the TFA cleavage solution using chilled EtzO (30 mL) and collected by centrifugation (4000 rpm). The supernatant was removed. Additional cold Et?O (30 mL) was added to the peptide pellet and clarified by centrifugation. The resulting crude peptide pellet was blown dried by a stream of nitrogen gas. c) Peptide Cyclization.

[00264] The crude solids were re-dissolved in DMSO (1.5 mL). The pH was adjusted by addition of 2.0 M DIPEA inNMP solution (0.5 mL). The solution was shaken at RT for approximately 12 h and monitored by UPLC-MS. After the reaction was complete, the cyclization mixture was acidified by the addition of 40 pL glacial acetic acid and the resulting solutions were filtered and submitted for RP-purification. d) HPLC Purification

[00265] Purification was performed by preparative reversed-phase high performance liquid chromatography (RP-HPLC) on Waters™ X-Bndge Prep Cl8 OBD Prep column (130 A, 5 pm, column size 19 x 100 mm) using a Waters™ MS-Directed AutoPurification HPLC-MS system. Mobile phase: (A) 0.16% TFA in HPLC-grade water and (B) 0.16% TFA in HPLC-grade acetonitrile; flow rate: 25 mL / min; UV wavelength a = 215 nm; gradient: 25-50% B over 5 min. Alternatively purification was performed on Waters™ CSH-C18 Column (19 x 250 mm, 5 pm) using an Agilent, with 1290 infinity II preparative LC system and LC-MSD XT mass spectrometer. Mobile phase: (A) 0.1% formic acid in HPLC-grade water and (B) 0.1% formic acid in HPLC-grade acetonitrile; flow’ rate: 25 mL / min; UV wavelength X = 215 nm; gradient: 20% B over 2.5 min, then increasing to 55% B over the next 17.5 min. UV absorbing fractions containing the target m / z ions were collected and the fractions containing product were confirmed by LC-MS.

[00266] Confirmation of identity and purity assessment of final compounds w ere performed by UPLC-MS, which was measured by a reverse phase Waters™ ACQUTTY UPLC-MS system. Column: Waters™ XSelect CSH C18 Column (130 A, 2.5 um, column size 2.1 x 50 mm). Mobile phase: (A) 0.05% TFA in HPLC-grade water and (B) 0.05% TFA in HPLC-grade acetonitrile; injection volume: 1 gL; flow rate: 1 mL / min; UV wavelength X = 215 nm: gradient: 5 --100% B in 5 min. Lyophilization of combined fractions containing pure peptide resulted in the final cyclized product as a powder. C. Synthetic Procedures used to Prepare Macrolactam Cyclic Peptides Synthetic Scheme 5 Solid-phase peptide synthesis 1) Piperidine (20% in DMF) 2) DIC / Oxyma. 90 °C Macrolacramization and side chain deprotection 1) HATU, HOAt. DiPEA, THr. RT 2) TFATn$ / H2O (+ pheno: or DTI), RT Release of protected peptide from solid support 30% HF!P in DOW. RT 3. Procedure C a) Solid Phase Synthesis of Peptides (SPPS)

[00267] Peptides were synthesized on a Liberty Blue™ synthesizer from CEM Corporation, using standard solid-phase synthesis using Fmoc / fBu chemistry' as summarized above in Synthetic Scheme 9.

[00268] Reactions were typically performed at a 100 pmol scale using 2-chlorotrityl chloride resin (100-200 mesh, 1% DVB, 1.71 mmol / g loading, Novabiochem®) or 2-chlorotrityl chloride resin preloaded with the starting ammo acid (0.66 mmol / g loading, Novabiochem®), based on the synthetic strategy.

[00269] The loading of the initial C-terminal amino acid to the 2CTC resin was performed in a fritted plastic syringe by treating with 1.0 millimolar excess of Fmoc-protected amino acid solution (0.2 M in a 4; 1 (v / v) DCM / NMP mixture) and 5-fold molar excess of DIPEA at RT for 45 min. MeOH (0.4 mL) was added and the slurry was shaken for 15 min at RT. The resin was drained and washed with DMF. The Fmoc-protecting group was removed by treating with piperidine (20% v / v in DMF) at RT for 10 min. This process was repeated once more (twice total). The resin was washed with DMF, MeOH and DCM raid dried under vacuum. [00270J Evety synthesis cycle included: (1) Fmoc amino acid deprotection by 20% v / v piperidine in anh. DMF (50 °C microwave assisted heating, 10 min; repeated twice); (2) Coupling (potentially repeated twice tor difficult couplings) with Fmoc-protected amino acid / HATU / DIPEA (5, 5, and 10 equiv. respectively; 50 °C microwave assisted heating, 10 min). Cycles of Fmoc deprotection and Fmoc-protected amino acid coupling were repeated with the desired monomers until the full linear protected peptide was formed; (3) The peptide A-terminus was deprotected using 20% v / v piperidine in DMF (50 °C microwave assisted heating, 10 ram; repeated twice). b) Cleavage of Protected Linear Peptide

[00271] After completion of the synthesis, the linear resin-bound peptide was transferred into a fritted plastic column. The resin was washed w ith DCM then the linear peptide w as cleaved off the solid support by treatment with HFIP / DCM (3:7, v / v; 5 mL) at RT for 5 min. The solution was collected into a round bottom flask. This process was repeated twice more (three times total), and the cleavage solution was combined. The solvents were removed under reduced pressure. c) Solution Phase Macroiactamization

[00272] The crude protected linear peptide was resuspended in anh. THF (1.5 mg / mL). Anhydrous DMF (500 pL) was added. DIPEA (5 equiv), HOAt (0.8 equiv) and HATU (1.2 equiv) were added to the peptide solution and the solution stirred at RT. The reaction wus monitored by UPLC-MS and stopped after complete conversion of the starting material. The reaction solution was then concentrated under reduced pressure to dryness. d) Deprotection of the Protected Cyclic Peptide

[00273] The cyclic peptide was deprotected by treatment with TFAThO / TIS / Phenol (90:2:4:4, v / v / v / w; 15 mL) at RT for 1 h. The crude cyclic peptide was precipitated from the TFA cleavage solution using chilled MTBE (50 mL) and collected by centrifugation. The supernatant was removed. Additional cold MTBE (40 ml.) was added to the peptide pellet and clarified by centrifugation. This process was repeated once more (twice total). The resulting crude peptide pellet was dissolved in 1:1 (v / v) mixture of DI water / MeCN (10 mL) and 5% (v / v) of TFA was added to the solution to remove the tiyptophan adduct. The solution was frozen and lyophilized. e) IIPLC Purification

[00274] The crude residue was then dissolved in DMSO and purified by preparative reversed-phase high performance liquid chromatography on a Waters™ Xbridge Protein BEH C4 OBD prep column (300A, 5 pm. column size 30 x 250 mm) using a Waters™ 2545 HPLC system equipped with Waters™ 2489 UV / Visible detector. Mobile phase: (A) 0,1% TFA in HPLC-grade water and (B) 0.1% TFA in HPLC-grade acetonitrile; flow rate: 50 mL / min; UV wavelength a = 214 nm; gradient: 0.5%B / min starting at 5% B less than the calculated %B required for compound elution over 20 min. UV absorbing fractions containing the target were collected and the fractions containing the desired product were combined, concentrated in vacuo and freeze-dried to afford the cyclized peptide as a solid.

[00275] Confirmation of identity and purity assessment of final compounds were performed by UPLC-MS, which was measured by a reverse phase Waters™ ACQUITY UPLC-MS system. Column: Waters™ ACQUITY UPLC Protein BEH C4 (300 A, 1.7 pm. column size 2..1 x 100 mm, Milford, MA). Mobile phase: (A) 0.1% TFA in HPLC-grade water and (B) 0.1% TFA in HPLC-grade acetonitrile; injection volume: 0.5 pL, flow rate: 0.4 mL / min, Column 'Temperature: 45 °C; UV wavelength X = 214 nm; gradient: 20-20% B in 1 min and 20-40% in 3 inin. Synthetic Scheme 6 Maprolactamizauon and side chain deprotection 1) HATU, HOAt DIPEA, THE RT 2) TFA / TIS / H2O phenol or DTT), RT Reiesse of protected peptide from solid support 30% HFiP in DCM, RT D. Synthesis of Specific Compounds SEQ ID NO:4 [00276J The TR-FRET probe (ClAc*-F-Y-S-W-Sar-N-Y-W-S-Y-Y-Sar-W-C*-G-Aea- K(bsotin)-NH2; SEQ ID NO:4) was prepared using Synthetic Procedure A starting with Fmoc-Lys(biotin)-OH (Novabiochem®’. cat. # 852097) and adding a Gly-Aea spacer (SEQ ID NO:5) attached to the carboxy moiety of the C-termmal cysteine. MS ESI calculated for C121H145C1N25O28S2[M + 2H]24 1231.51, found 1231.5. 8.2. EXAMPLE 2: BIOLOGICAL ASSAYS 8,2.1. Procedure For Human STNFRl-Peptide Displacement Assay [00277| The affinity was determined using LanthaScreen Eu-based time-resolved fluorescence resonance energy transfer (TR-FRET)-based binding assay.

[00278] A TR-FRET assay measuring the displacement of biotinylated peptide SEQ ID NO:4 from human sTNFRl in a pre-formed complex by a macrocyclic peptide yields the affinity CKf'pp) of the cyclic peptide for sTNFRl. The reagents for the assay include: the C -tenmnally 6His-iagged sTNFRl (residues 1-211) protein purified from a S / 21 baculovirus expression system, the biotin-tagged B0372 peptide described in section '‘Preparation of Final Compounds’’, part E, the LanthaScreen™ anti-His Tag Europium donor (Life Technologies Corporation (San Diego, CA), cat. ft PV5597), and a streptavidin-conjugate Alexa Fluor™ 647 acceptor (Invitrogen (Waltham, MA), cat. # S32357). Time-resolved TR-FRET is achieved when sTNFRl is bound to biotinylated peptide SEQ ID NO:4 through coupling of the donor and acceptor FRET pair.

[00279] 100 nM biotinylated peptide SEQ ID NO:4 and 0.25 nM sTNFRl -6His are added to a binding buffer (lx DPBS, Gibed™, cat. # 20012-027 with 0.1% BSA and 0.05% Tween® 20) to form the protein-peptide complex. 100 nM biotinylated peptide SEQ ID NO:4 was chosen as it reflects the ~Kd for this complex. After 30 min at ambient temperature, each test peptide (2 mM stock in DMSO) is titrated in a 20 point, ~3-foId discrete dose-response using an acoustic-dispense liquid handler. Specifically, 80 nt of peptide / DMSO is transferred into a ProxiPlate-384 Plus 384-udi assay plate (PerkinElmer, cat. # 6008289). To this plate, 8 uL of the previously formed sTNFRl - biotiny lated peptide SEQ ID NO:4 complex is added and allowed to reach binding equilibrium with the test peptide at ambient temperature for 90 min. For TR-FRET detection. 8 uL of 0.25 nM of LanthaScreen™ anti-His Tag Europium and 25 nM of streptavidin-conjugate Alexa Fluor™ 647 in the same binding buffer are added to the assay plate and read on an EnVision® plate reader (PerkinElmer (Boston, MA)) after a 90 min final incubation.

[00280] The TR-FRET signal is measured with the following EnVision'® settings: excitation laser at 337 nm: emission! = 615 nm, emission2 = 665 nm, LANCE / DELFIA dichroic mirror, delay time = 100 ms. The signal of each well is determined as the ratio of the emission at 665 nm to that at 615 nm. Percent displacement is determined by normalization to control wells containing DMSO (0%) or a saturating concentration of a displaceable peptide (100%). The %-effect as a function of peptide concentration is fit to a five-parameter logistic fit based on the Morrison equation to account for tight binding kinetics. The inflection point of the 5-parameter fit is the measured Adapp reported in Table 3. It should be noted that peptides with weaker binding affinity where tight binding was not observed under these assay conditions were titrated with 10 pts titration range and dose-response curves were analyzed using a 4-parameter logistic model to calculate TCso values (Table 3). sTNFRl sequence (SEQ ID NO:15): LVPHLGDREKRDSVCPQGKYIHPQNNSICCTKCHKGTYLYNDCPGPGQDTDCRECESGSFTASENHL RHCLSCSKCRKEMGQVEISSCTVDRDTVCGCRKNQYRHYWSENLFQCFNCSLCLNGTVHLSCQEKQN TVCTCHAGFFLRENECVSCSNCKKSLECTKLCLPQIENVKGTEDSGTTGGGLNDIFEAQKIEWHERE VLFQGPHHHHHHGSWSHPQFEK 8.2.2. Procedure for Binding Affinity' and Selectivity Measured by SPR |00281] Affinity and selectivity for hTNFRl and TNFR2 was determined using SPR. Human TNFR1 or TNFR2 genes (extracellular domain only) were synthesized with a C-terminal Avi- tag, HRV 3C protease cleavage site followed by a 6His-tag for affinity purification. The final gene product was cloned into pBAC™-l vector (Millipore Sigma) for expression testing in insect cells. Constructs were transfected into Sf21 cells for baculovirus generation followed by protein expression analysis in Sf21 and Tni cell lines Large scale expression in Sf21 cells was performed using baculovirus infected insect cells (BIICs) infected at a MOI = 2.0 grown for 72 h. Tarset proteins secreted into the media were harvested via centrifugation and concentrated using di afi I trail on. TNFR was purified by immobilized metal ion affinity chromatography (IMAC) using a Ni-Sepharose HisTrap™ column (Cytiva™). Isolated TNFR had His tag removed using HRV 3C protease followed by additional purification using Ni-NTA chromatography. Biotinylation was performed on Ni-NTA flow-through fractions concentrated to 45 p.M and treated with BirA ligase (1:100, pg / pg ratio) in 50 mM Tris pH 7.5, 150 mM NaCl, 50 pM biotin. 10 mM magnesium acetate, 10 mM ATP at 4 °C for 20 h. Final purification of biotin-TNFR and buffer exchange into 20 mM HEPES pH 7.5, 150 mM NaCl was performed using aHiLoad® Superdex® 200 26 / 60 column (Cytiva™). Highly purified protein was aliquoted and stored at -80 °C. Human TNFR1 construct sequence (SEQ ID NO:16): MGLSTVPDLLLPLVLLELLVGIYPSGVIGLVPHLGDREKRDSVCPQGKYIHPQNNSICCTKCHKGTY LYNDCPGPGQDTDCRECESGSFTASENHLRHCLSCSKCRKEMGQVEISSCTVDRDTVCGCRKNQYRH YWSENLFQCFNCSLCLNGTVHLSCQEKQNTVCTCHAGFFLRENECVSCSNCKKSLECTKLCLPQIEN VKGTEDSGTTGGGLNDIFEAQKIEWHELEVLFQGPHHHHHHGSWSHPQFEK Human TNFR2 construct sequence (SEQ ID NO:17): MAPVAVWAALAVGLELWAAAHALPAQVAFTPYAPEPGSTCRLREYYDQTAQMCCSKCSPGQHAKVFC TKTSDTVCDSCEDSTYTQLWWVPECLSCGSRCSSDQVETQACTREQNRICTCRPGWYCALSKQEGC RLCAPLRKCRPGFGVARPGTETSDWCKPCAPGTFSNTTSSTDICRPHQICNWAIPGNASMDAVCT STSPTRSMAPGAVHLPQPVSTRSQHTQPTPEPSTAPSTSFLLPMGPSPPAEGSTGDGGGLNDIFEAQ KIEWHELEVLFQGPHHHHHHGSWSHPQFEK

[00282] The SPR assay uses a Cytiva™ 8K or 8K+ biosensor instrument to measure the kinetics of peptide analytes binding to captured human TNFR1 extracellular domain (ECD) protein; peptide analytes are also evaluated for any off-target binding to human TNFR2 ECD protein in parallel. Recombinant human TNFR1 ECD and human TNFR2 ECD proteins, biotinylated via engineered C-termmal avHags, are immobilized to a regenerable streptavidin biosensor chip surface (Cytiva™ Biotin CAPture kit, Series S; cat. # 28920234). Peptide analytes binding to immobilized human TNFR1 ECD protein and their subsequent dissociation are conducted in a buffer of 10 mM HEPES pH 7.4. 150 mM NaCl. 3 mM EDTA, 0.05% P20. 2% DMSO under laminar flow conditions and measured in response units (RU), which are directly proportional to the accumulated mass on the surface of the biosensor chip; the biosensor chip surface is regenerated via Cytiva™ proprietary methods between the evaluation of each peptide analyte. The binding and dissociation of peptide analytes are displayed as sensorgrams in real time and recorded for subsequent determination of binding kinetics The SPR assay determines the association and dissociation rate constants, ko-. and kotr, for each peptide analyte and, from these values, calculates the binding affinity, Ka. reported in Table 3. 8.2.3. Procedure for THP-1 Lucia NF-kB cells

[00283] The inhibition of TNFa-induced NF-kB signal transduction pathways was evaluated in THP-1 cells. THP-1 cells harboring NF-icB-inducible Lucia™ reporter gene (InvivoGen, cat. # thpl-nfkb) designed for monitoring NF-kB signal transduction pathway was employed to assess the inhibitory effect macrocyclic peptides on TNFR1 signaling. The cells were cultured in media containing RPMI1640, 2 mM L-glutamine. 25 mM HEPES, 10% heat-inactivated fetal bovine serum with appropriate selection antibiotics. Assays were performed in media that was devoid of selection antibiotics. For profiling, 125 nL of peptides in DMSO (10 mM) were dispensed into Falcon® 384-well Optilux white / clear flat bottom TC-treated microtest microplate (Coming, cat. # 353963) using Echo® 555. The peptides were tested in a dose response to determine EC50’s [10 or 1 pM top concentration and 1:3 dilutions). A suspension of 1 million cells / mL was dispensed al 20 pL / well in a 384-well plate to a density of 20,000 cells / we1l and pre-incubated with peptides at 37 °C for 1 h. Subsequently. 5 pl., of TN.F containing media (recombinant human TNFa; R&D Systems USA, cat. # 210-TA-005 / CF) or TNF-free media (for controls) was added to the wells. The final concentration of TNFa in the assay in 10 ng / mL. After overnight incubation at 37 °C (16-18 h), 5 pL of supernatant was aspirated and transferred to a 384-welI microplate (PerkinElmer, cat. # 6007680) for measurement of secreted luciferase activity with 25 pL of QUANTI-Luc™ luminescence assay reagent (InvivoGen, cat. # rep-qlcl) in each well. The luminescence was measured in EnVision® Multimode Plate Reader (PerkinElmer). Dose response curves were analyzed using a 4-parameter logistic model to calculate ICso values, reported in Table 3, using TIBCO® Spotfire® software employing appropriate Minimum and Maximum controls.

[00284] The amino acid sequences, synthetic procedure used to synthesize and purify the macrocyclic peptides, calculated monoisotopic masses, molecular formulas, calculated molecular weights and mass spectral data for Compounds A-D (SEQ ID NOS: 6. 7, 8, and 9) are provided below in Table 2. Table 2: Compounds A-D (SEQ ID NOs: 6-9) Compound SEQ ID NO. Sequence Procedure Mol. Weight (g / mol) A 6 c\ do(C!Ac-Ahp-4Pal-S-SbMeW-dP-N-Phe4COOH-2Nal-aMeS-Phe4COOH-H-Sar-aMeW-(bNib B 2023.22 B 7 cyclo(bhcLeu-Nle-4Pal-Pip2c~ W-NMeE-N-Phe4COOH-Trp7Cl-aMeS-Phe4COOH-Y-Sar-SbMeW7OMe) C 2096.68 C 8 cyclo(bhcLeu-Nle-4Pal-Pip-W-NMeE-N-4Pd-Trp7Cl-aMeS-Phe4COOH-Phe3CONH2-Sar-SbMeW7OMe) c 2079.70 D 9 cydo(ClAc-F-Y-S-W-Sar-N- Y-W-S-Y-Y-Sar-W-C)-G-E- E-NH2 A 2264.38 Molecular Formula Exact mass (Da) Purity (%) Measured Mass (Da) Observed Ion C102HH9N21O22S 2021.86 91.0 1012.0 [M+2H]2+ C106H123C1N20O24 2094.87 100.0 2097.0 pH Hi- C106H124C1N21O22 2077.89 97.7 1040.4 iM-2Hi2 C111H126N22O29S 2262.88 95.0 1132.9 [NR2H]2+ WO 2025 / 166198                                   PCT / US2025 / 014081

[00285] The biological activities (binding ICso and Xda!’p in the displacement assay, affinity Kd, and cell activity in THP-1 cells) of Compounds A-D (SEQ ID NOS: 6, 7, 8, and 9 as disclosed herein) are provided below in Table 3. Table 3: Biological Activities of Compounds Nos. A-D (SEQ ID NOs: 6-9) Compound SEQ ID NO. TR-FRET, 1C5S (nM) TR- FRET, KdaPP (nM) SPR, hTNFRl Kd (nM) SPR, hTNFRl Kd (nM) THP-1, IC5« (nM) A 6 13.0 20.8 B 7 0.20 0.11 0.10 >1000 4.2 C 8 0.31 0.14 >1000 1.0 D 9 9.87 7.43 >1000 36.4 8.3. EXAMPLE 3: CHARACTERIZATION OF THE COMPOUND / TNFA / STNFR1 COMPLEX 8.3.1. Expression And Purification of TNFR1 and TNT Alpha

[00286] Various constructs of the extracellular domain of TNFR1 or complexed TNFa / extracellular domain of TNFRI were recombinantly expressed and purified by chromatography. TNFRI_V7 was expressed using Baculovirus cells and purified from the growth medium by nickel-affmity chromatography followed by size exclusion chromatography. Pure protein was concentrated to 6 mg / mL. TNFR1_V 14, VI and V10 were expressed using E. coll cells, the protein was solubilized and isolated from the inclusion bodies, refolded and purified by Ion-exchange and size exclusion chromatography. Pure protein was concentrated to 18 mg / mL. Human TNFa was expressed and purified from E. coll cells with an N-terminal HIS-tag. The protein was purified by affinity and size exclusion chromatography steps including remssi'sboMie HIS-tag. 8.3.2. Crystallography Protocol

[00287] Co-crystals were grown by the hanging drop method of vapor diffusion in 96-welI format. Crystallization trials were set up for each of the peptides at a protein: peptide molar ratio 1:1.5 at both 4 and 18°C using the respective constructs. Broad screening of crystallization conditions was performed using commercially available screens. Screening set-up was performed using a Mosquito (TTP Labtech) dispensing 200 nL of sample solution in two separate sub-wells, to which 100-200 nL of precipitant solution was added. The mixture was incubated at 18 °C m a Rockimager (Formulatrix). Crystals appeared between a period of 2 to 45 days. For flashfreezing and data collection, a cryoprotectant solution at a pH and a precipitant concentration matching the crystallization conditions and augmented with 20% v / v glycerol was added to the crystallization drop. The crystals were then harvested with a LithoLoop (Molecular Dimensions) and flash-frozen m liquid nitrogen.

[00288] The X-ray data sets were collected at a wavelength of 1 A either at the Industrial Macromolecular Consortium Association (IMCA) beamline at the Advanced Photon Source (APS), or at the macromolecular crystallography beam line at the Canadian Light Source (CLS). Data, sets were indexed and scaled using the autoPROC (Global Phasing) software with calls to XDS, POINTLESS (CCP4) and STARANISO (Global Phasing) for integration, space group determination and scaling, respectively. The structure was solved by molecular replacement using the MOLREP program (CCP4). Structures were refined using COOT (CCP4) and autoBUSTER (Global Phasing). The constructs that were co-crystallized with each compound is shown below in Table 4. Table 4: TNFR1 Constructs for Crystallization Studies Compound Binding Compound TNFR1 Construct Amino Acid Sequence None (unbound) TNFRi v!4 (SEQ ID NO: Hi MECESGSFTA SENHLRHCLS CSKCRKEMGQ VEISSCTVDR DTVCGCRKNQ YRHYWSENLF QCFNCSLCLN GTVHLSCQEK QNTVCTCHAG FFLRENECVS C A (SEQ ID NO: 6) TNFRvlO (SEQ ID NO: 10) MECESGSFTA SENHLRHCLS CSKCRKEMGQ VEISSCTVDR DTVCGCRKNQ YRHYWSENLF QCFNCSLCLN GTVHLSCQEK QNTVCTCHA B (SEQ ID NO: 7) TNFRv 14 (SEQIDNO:!!) MECESGSFTA SENHLRHCLS CSKCRKEMGQ VEISSCTVDR DTVCGCRKNQ YRHYWSENLF Q C FNCSLCLN GTVHLSCQE K QNTVCTCHAG FFLRENECVS C C (SEQ ID NO: 8) TNFalpha TNFRlv7 (SEQ ID NO :12) MVRSSSRTPS DKPVAHWAN PQAEGQLQWL NRRANALLAN GVELRDNQLV VPSEGLYLIY SQVLFKGQGC PSTHVLLTHT ISRIAVSYQT KVNLLSAIKS PCQRETPEGA EAKPWYEPIY LGGVFQLEKG DRLSAEINRP DYLDFAESGQ VYFGIIALLV PHLGDREKRD SVCPQGKYIH PQNNSICCTK CHKGTYLYND CPGPGQDTDC RECESGSFTA SENHLRHCLS CSKCRKEMGQ VEISSCTVDR DTVCGCRKNQ YRHYWSENLF QCFNCSLCLN GTVHLSCQEK QNTVCTCHAG FFLRENECVS CSNCKKSLEC TKLCLPQIEN VKGTEDSGTT D (SEQ ID NO: 9) TNFalpha TNFRI vl (SEQ ID NO: 13) MVRSSSRTPS DKPVAHWAN PQAEGQLQWL NRRANALLAN GVELRDNQLV VPSEGLYLIY SQVLFKGQGC PSTHVLLTHT ISRIAVSYQT KVNLLSAIKS PCQRETPEGA EAKPWYEPIY LGGVFQLEKG DRLSAEINRP DYLDFAESGQ VYFGIIALMS VCPQGKYIHP QNNSICCTKC HKGTYLYNDC PGPGQDTDCR ECESGSFTAS ENHLRHCLSC SKCRKEMGQV EISSCTVDRD TVCGCRKNQY RHYWSENLFQ CFNCSLCLNG TVHLSCQEKQ NTVCTCHAGF FLRENECVSC Compound TNFR I Construct Amino Add Sequence D (SEQ ID NO: 9) TNFR1v1 (SEQ ID NO: 14) MDSVCPQGKY IHPQNNSICC TKCHKGTYLY NDCPGPGQDT DCRECESGSF TASENHLRHC LSCSKCRKEM GQVEISSCTV DRDTVCGCRK NQYRHYWSEN LFQCFNCSLC LNGTVHLSCQ EKQNTVCTCH AGFFLRENEC VSC

[00289] After 1 h incubation, the sample was divided into 2 aliquots, and a solution of 125 mM zinc chloride solution was added to one of the aliquots to a final concentration of 1 mM. Broad screening of crystallization conditions was performed using commercially available screens at 18 °C. Screening set-up was performed using a Mosquito (TTP Labtech) dispensing 200 nL of sample solution with or without exogenous zinc in two separate sub-wells, to which 100-200 nL of precipitant solution was added. The mixture was incubated at 18 °C in a Rockimager (Formulatrix). Crystals appeared between a time period of 2 to 45 days. For flashfreezing and data collection, a cryoprotectant solution at a pH and a precipitant concentration matching the crystallization conditions and augmented with 20% v / v glycerol was added to the crystallization drop. The crystals were then harvested with a LithoLoop (Molecular Dimensions) and flash-frozen in liquid nitrogen. [0029()] Data collection was performed either at the Industrial Macromolecular Consortium Association (IMCA) beamline at the Advanced Photon Source (APS), or at the macromolecular crystallography beam line at the Canadian Light Source (CLS). Data were processed using the autoPROC (Global Phasing) software with calls to XDS, POINTLESS (CCP4) and STARANISO (Global Phasing) for integration, space group determination and scaling, respectively. The structure wras solved by molecular replacement using the MOLREP program (CCP4). Structures were refined using COOT (CCP4) and autoBUSTER (Global Phasing). 8.3.3. Cryo-EM Grid Preparation and Data Collection

[00291] TNFa, TNFR V7 construct, and Compound B were mixed in the final buffer composition w?as 20 mM Tris, pH 8.0, and 100 mM NaCI at a molar ratio of 1:3:6 and incubated for an hour in ice. Aliquots (3 pL) of purified complex were applied to glow-discharged (20 s on carbon side) C-flat 20 nm thickness holey carbon-on-gold grids (300 mesh, RI .2 / 1.3). The grids were blotted for 3 s at 95% humidity and plunge-frozen into liquid ethane using a Vitrobol Mark IV (Thermo Fisher Scientific). Grids were imaged on a 300 keV Titan Krios cryo-electron microscope (Thermo Fisher Scientific) equipped with an energy filter (Galan GIF BioQuantum) and a post-GIF Gatan K3 Summit direct electron detector. Images were taken on the K3 camera in dose-fractionation mode at a calibrated magnification of 105000. corresponding to 0.84 A per physical pixel (0.42 A per super-resolution pixel). The dose fractionation on the specimen was set to be 1.0625 electrons per A2 per frame and the total number of frames was 40, resulting in a total dose of 42.5 electrons per A2. An energy slit with a width of 20 eV was used during data collection. Fully automated data collection was carried out using Latitude in Gatan Imaging Suite (Gatan Inc.) with a nominal defocus range set from -0.6 to -2.0 pm. Image Shift was used with nine exposure groups per stage shift to improve the throughput of data collection. A total of 12000 movies were collected for the sample. 8.3.4. Cryo-EM Data Processing

[00292] The cryoSPARC Live application of cryoSPARC v2 (Structure Biotechnology') was used to streamline the movie processing, CTF estimation, particle picking, and 2D classification. Preprocessing involved anisotropic motion correction and local CTF estimation. The data were curated by keeping only data with better than 6 A determined by CTF fit resolution. Particle picking started with blob picking —150 A in diameter. Once a small set of particles was extracted and 2D class averages were obtained, the good 2D classes were used as templates for picking on the entire dataset. Then ab initio 3D reconstruction was carried out in cryoSPARC v2. Homogenous refinement coupled with nonuniform and global CTF refinements gave the final 3D reconstructions at 3.2 A. The structure was built starting from atomic models of close homologs. Manual adjustments to the models were performed using Coot and automated refinement was performed using Phenix. Coordinates and restraints for peptides were generated using Grade (Global Phasing Ltd.) and was manually fitted into the density' using real-space refinement in Coot and further refined using Phenix. Model quality was assessed using Molprobity as implemented in Phenix.

[00293] FIGs. 3A-3B depict two-dimensional representations of a fragment of Compound B bound in the binding pocket of the TNFR1 showing interactions between the compound and the pocket. FIGs. 4A-4B depict (A) a three-dimensional top view and (B) a three-dimensional bottom view of the TNFR1 binding pocket when bound to Compound A. Compound B, Compound C, and Compound D of the disclosure, as determined by X-ray crystallography and presented as an overlay. A three-dimensional cartoon representation of Compound B bound in the binding pocket, of the TNFR1 is shown in FIGs. 5A (binding pocket not shown) and SB (binding pocket shown). FIG. 6 is a superimposed imaged depicting the displacement of the CDR4 of the TNFR1 upon binding of Compound B. The green portion of FIG. 6 represent the unbound TNFR1 and red represents TNFR1 bound to Compound B. This displacement is seen with each of Compounds A, B, C, and D as disclosed herein.

Claims

What is claimed is;1. A method of inhibiting a tumor necrosis factor receptor 1 (TNFRl)-mediated activity comprising contacting a TNFR1 with a compound that binds to TNFR1 at a binding pocket between CRD3 and CRD4 of TNFR1.

2. The method of claim 1 or 2, wherein the TNFR1 is a membrane-bound TNFR1,3.     The method of any one of claims 1-3, wherein the TNFR1 comprises an amino acidsequence according to SEQ ID NO: 1.

4. The method of any one of claims 1-3, wherein the binding pocket of the TNFR1 comprises ammo acid residues Asnl34, Thrl35, Cys 137, and Asnl48 of SEQ ID NO: 1.

5. The method of any one of claims 1-4, wherein the binding pocket of the TNFR1 comprises or further comprises one or more of ammo acid residues Seri 18, Lent 19, Cysl20, Leul21, and Asnl22 of SEQ ID NO:1.

6. The method of any one of claims 1-5, wherein the binding pocket of the TNFR1 comprises or further comprises one or more of amino acid residues Thr 138, Cys 139, Leu 145, Asnl48, Glut 49, and Cysl 50 of SEQ ID NO: 1.

7. The method of any one of claims 1-6, wherein the binding pocket of the TNFR1 is defined by amino acid residues Seri 18, Leu 1'19, Cysl20, Leu 121, Asnl22, Cysl 37, Thrl38. Cysl39, Leul45, Asnl48. Glul49, and Cysl50 of SEQ ID NO: 1.8 The method of am one of claims 1-7, wherein the compound binds to the TNFR1 at one or more of residues Asnl 34, Thrl35, Cysl37, and Asnl 48 of SEQ ID NO:1.

9. The method of any one of claims 1-8, wherein the compound compri ses at l east one moiety selected from:a) an H-bond interaction moiety capable of donating an H-bond to residue Asnl 48 of theTNFRl;b) an H-bond interaction moiety' capable of accepting an H-bond from residueCysl 37 of the TNFR1; andc) an H-bond interaction moiety capable of donating an H-bond to residue Cyst 37 of theTNFRl.

10. The method of claim 9. wherein the compound further comprises at least one moiety selected from:a) an H-bond interaction moiety capable of donating an H-bond to residue Thrl35 of the TNFR1: andb) an H-bond interaction moiety capable of accepting an H-bond from residue Asnl34 of the TNFR1.

11. The method of any one of claims 1-10. wherein the compound comprises a peptide.

12. The method of claim 11, wherein the compound is a cyclic peptide.

13. The method of claim 12, wherein the compound comprises 12-18 amino acid residues.

14. The method of any one of claims 1-13, wherein the compound comprises an ammo acid sequence, and wherein the compound binds to the binding pocket of TNFR1 via an aliphatic side chain on the first ammo acid in the sequence.

15. The method of any one of claims 1-14, wherein the compound comprises an amino acid sequence, and wherein the compound binds or further binds to the binding pocket of TNFR1 via a bicyclic aryl or heteroary l moiety7 on the eighth ammo acid in the sequence.

16. The method of claim 15, wherein the bicyclic aryl or heteroaryl moiety is naphthyl or indolyl.

17. The method of any one of claims 1-16, wherein the compound comprises an amino acid sequence, and wherein the compound binds or further binds to the binding pocket of TNFR1 via a monocyclic aryl moiety' on the tenth ammo acid of the sequence.

18. The method of claim 17, wherein the monocyclic aryl moiety7 is phenyl.

19. The method of claims 1-18. wherein the compound comprises an amino acid sequence, and wherein the compound binds or further binds to the binding pocket of TNFR1 via a hydroxyl moiety or an amine moiety on the third amino acid in the sequence.

20. The method of claim 19. wherein the hydroxyl moiety' or an amine moiety' is comprised within an hydroxymethyl moiety, a piperidinyl moiety, or a piperazinyl moiety.

21. The method of claims 1-20, wherein the compound comprises an amino acid sequence, and wherein the compound binds or further binds to the binding pocket of TNFR1 via a hydroxyl moiety' on the ninth amino acid in the sequence.

22. The method of claim 21, wherein the hydroxyl moiety is comprised within an hydroxymethyl moiety.

23. The method of any one of claims 14-21, wherein the eighth amino acid binds to CysI37 of the TNFR1 of SEQ ID NO: 1.

24. The method of any one of claims 14-23, wherei n the tenth amino acid binds to Asn 134 of the TNFRI of SEQ ID NO: 1.

25. The method of any one of claims 14-24. wherein the third amino acid binds to Asnl48 of the TNFRI of SEQ ID NO: 1.

26. The method of any one of claims 14-24, wherein the ninth amino acid binds to Thrl35 of the TNFRI of SEQ ID NO: 1.

27. The method of any one of claims 14-26, wherein the first amino acid is (S)-2-aminoheptanoic acid (Ahp), L-norleucine (Nie), or L-phenylalanine (F).

28. The method of any one of claims 14-27, wherein the third amino acid is (5)-piperazine-2-carboxydic acid (Pip2c), (S)-piperidine-2-carboxylic acid (Pip), or L-serine (S).

29. The method of any one of claims 14-28, wherein the eighth amino acid is 3-(2-naphthyl)~ (.-alanine (2Nal), (5>2-amino-3-(7-chloro-l / 7-indo1-3-yl)propanoic acid (Trp7Cl), orL-try ptophan (W).

30. The method of any one of claims 14-29. wherein the ninth amino acid is a-methyl-L-serine (aMeS) or L-Serine (S).

31. The method of any one of claims 14-30, wherein the tenth amino acid is L-tyrosine (Y) or L-phenvlalanine-4-carboxyIic acid (Phe4COOH).

32. The method of any one of claims 1-31, wherein the compound has a molecular weight offrom about 1200 Dato about 3000 Da. from about 1500 Dato 2500 Da. or from about 1750 Dato about 2250 Da.

33. A method of inhibiting a TNFR1-mediated activity, comprising contacting the TNFR1 with a first compound that competes with a second compound for binding to TNFR1 al a binding pocket between CRD3 and CRD4 of TNFR1, wherein the second compound comprises or consists of a sequence selected from:a)    Cl Ac*-Ahp-4Pal-S-SbMeW -dP-N-Ph e4COOH-2Nal-aMeS-Phe4 COOH-H-S ar-aMeW-C AH- (SEQ ID NO:6);b)    Nle*-4Pal-Pip2c-W-NMeE-N-Phe4COOH-Trp7Cl-aMeS-Phe4COOH-Y-Sar-SbMeW7OMe-bhcLeu* (SEQ ID NO:7);c)    Nle*-4Pal-Pip-W-NMeE-N-4Pal-Trp7Cl-aMeS-Phe4COOH-Phe3CONH2-Sar-SbMeW7OMe-bhcLeu* (SEQ ID NO:8); ord)    ClAc*-F-Y-S-W-Sar-N-¥-W-S-Y-Y-Sar-W-C*-G-E-E-NH2 (SEQ ID NO:9);or a pharmaceutically acceptable salt thereof, wherein * indicates attachment to form a cyclic peptide.

34. A method of inhibiting a TNFRl-mediated acii wn. comprising contacting the TNFR1 with a first compound that competes with a second compound for binding to TNFR1 at a binding pocket between CRD3 and CRD4 of TNFR1, wherein the second compound is Compound A, B, C, or D:Compound ACompound C‘OHCompound D or a pharmaceutically acceptable salt thereof.

35. The method of any one of claims 1-34, wherein the TNFR1 -mediated activity is selected from the group consisting of inflammation, cytokine signaling, apoptosis, necroptosis, cell proliferation, cell survival, cell differentiation, MAPK signaling, NF-kB signaling, and anv combination thereof.

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