Table 2. Fucosterol and known inhibitors binding energies and key interacting amino acid residues with selected pro-inflammatory proteins

Inflammatory proteins/PDB ID Ligands Binding Energy (kcal/mol) Key interacting amino acid residues
IL-1β (8C3U) Test compound: fucosterol –7.4 VAL3, PHE46, VASL47, LYS55 PRO57, VAL58, ALA59, MET95, LYS92, LYS93, LYS94, LYS97, VAL100, ASN102, ALA115, PRO118
Known inhibitor: anakinra –6.2 VAL3, ARG4, SER5, LEU6, ASN7, VAL41, PHE42, SER43, SER45, LEU62, LYS63, GLU64, LYS65, PRO91, SER152
Known inhibitor: ustekinumab –6.7 ALA1, VAL3, SER45, PHE46, VAL47, GLU50, PRO57, VAL58, ALA59, PRO91, LYS92, LYS93, LYS94, MET95, VAL100
Co-crystallized ligand: T9C –10.0 VAL3, SER45, PHE46, VAL47, GLU50, LYS55, PRO57, VAL58, ALA59, PRO91, LYS92, LYS93, LYS94, MET95, LYS97, VAL100, ASN102, ALA115
TXA2 (6IIV) Test compound: fucosterol –6.7 PHE30, ALA31, PHE34, CYS35, GLY77, LEU78, THR81, GLY82, VAL85, VAL86, HIS89, MET112, PHE115, GLY116, TRP258, LEU294, ARG295, THR298, GLN301
Known inhibitor: BM-573 –8.8 PHE30, PHE34, CYS35, GLY77, LEU78, THR81, GLY82, VAL85, MET108, MET112, PHE115, SER181, TRP182, LEU291, LEU294, ARG295, THR298, TRP299, GLN301,
TXA2 (6IIV) Known inhibitor: picotamide –9.7 PHE30, ALA31, PHE34, LEU78, THR81, VAL85, MET108, MET112, PHE115, GLY116, SER181, TRP182, TRP258, LEU291, LEU294, ARG295, ALA297, THR298, ALA299, GLN301
Known inhibitor: terutroban –11.2 PHE30, ALA31, PHE34, CYS35, GLY77, LEU78, THR81, GLY82, VAL85, MET112, PHE115, GLY116, PRO179, SER181, TRP182, PHE184, PHE200, TRP258, LEU291, LEU294, ARG295, ALA297, THR298
Co-crystallized ligand: daltroban –9.2 ALA31, GLY77, LEU78, THR81, VAL85, HIS89, MET112, PHE115, GLY116, PRO179, SER181, PHE184, PHE200, TRP258, LEU291, LEU294, ARG295, ALA297, THR298, GLN301
IL-6 (1ALU) Test compound: fucosterol –6.3 LYS66, MET67, PHE74, GLN75, SER76, GLU172, SER176, ARG179
Known inhibitor: methotrexate –4.0 ASP26, ARG30, LEU33, GLN175, SER176, LEU178, ARG179, ARG182
Known inhibitor: oxaprozin –5.5 ARG30, LEU33, GLN175, SER176, LEU178, ARG179, ARG182
Co-crystallized ligand: tartaric acid –3.8 GLN175, SER176, LEU178, ARG179, ARG182
COX-2 (5KIR) Test compound: fucosterol –4.6 VAL89, LEU93, VAL116, ARG120, VAL349, LEU352, SER353, TYR355, PHE357, LEU359, PHE381, LEU384, TYR385, TRP387, PHE518, MET522, VAL523, GLY526, ALA527, SER530, LEU531
Known inhibitor: celecoxib –11.5 HIS90, THR94, VAL116, ARG120, GLN192, VAL349, LEU352, SER353, GLY354, TYR355, LEU359, LEU384, TRP387, ARG513, ALA516, ILE517, PHE518, MET522, VAL523, GLY526, ALA527, SER530, LEU531,
Known inhibitor: aspirin –6.5 TYR348, VAL349, LEU352, SER353, PHE381, LEU384, TYR385, TRP387, PHE528, MET522, VAL523, GLY526, ALA527, SER530, LEU531
Known inhibitor: isoxicam –7.2 GLN192, GLU346, ASP347, ASN350, HIS351, GLY354, SER579, PHE580, SER581,
Known inhibitor: ketorolac –8.8 VAL116, ARG120, VAL349, LEU352, SER353, TYR355, LEU359, PHE381, TRP387, PHE518, MET522, VAL523, GLY526, ALA527, SER530, LEU531
Co-crystallized ligand: rofecoxib –10.1 HIS90, GLN192, LEU351, SER353, VAL349, LEU352, TYR355, TRP387, ARG513, ALA516, ILE517, PHE518, MET522, VAL523, GLY526, ALA527, SER530
TGF-β (1PY5) Test compound: fucosterol –3.4 ILE211, GLY212, VAL219, ALA230, VAL231, LYS232, PHE234, GLU245, LEU260, LEU278, SER280, ASP281, TYR282, HIS283, GLU284, GLY286, SER287, LYS337, LEU340
Known inhibitor: tranilast –7.1 ILE211, GLY212, LYS213, GLY214, VAL219, ALA230, LYS232, LEU260, SER280, ASP281, TYR282, HIS283, GLY286, SER287, LYS337, LEU340, ALA350, ASP351
Known inhibitor: imatinib mesylate –3.1 VAL219, LYS232, GLU245, LEU260, LEU340, ALA350, ASP351, LEU352
Known inhibitor: MG-132 0.9 ILE211, GLY212, LYS213, GLY214, VAL219, ALA230, VAL231, LYS232, TYR249, SER280, ASP281, TYR282, GLY286, ALA350, LEU260, HIS283, SER287, LYS337, LEU340, ASP351
Co-crystallized ligand: PY1 –7.2 ILE211, VAL219, ALA230, VAL231, LYS232, LEU260, LEU278, SER280, ASP281, TYR282, HIS283, GLY286, LEU340, ASP351
LOX-3 (1NO3) Test compound: fucosterol –3.1 THR274, LEU277, GLN514, HIS518, TRP519, HIS523, ILE557, LEU560, ALA561, LEU565, VAL571, ILE572, TYR575, PHE576, GLN716, ILE765, ASP766, VAL769, ILE770, LEU773, ILE857,
Known inhibitor: zileuton –2.6 GLN514, HIS518, TRP519, HIS523, ILE557, ALA561, LEU565, VAL571, ILE572, THR575, PHE576, GLN716, ASP766, VAL769, ILE770, LEU773, ILE857
LOX-3 (1NO3) Known inhibitor: REV-5901 0.1 LEU277, GLN514, LEU515, HIS518, TRP519, HIS523, ILE557, ALA561, LEU565, VAL566, VAL571, ILE572, THR575, PHE576, ILE765, ASP766, VAL769, ILE770, LEU773, ILE857,
Co-crystallized ligand: 4-Nitrocatechol –4.9 LEU277, GLN514, HIS518, TRP519, HIS523, ILE557, ASN558, ALA561, LEU565, VAL566, ILE572, LEU773, ILE857
COX, cyclooxygenase; IL, interleukin; LOX, lipoxygenase; PDB, protein data bank; TGF, transforming growth factor; TXA, thromboxane.