BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 depicts the chemical structure of TEMPOL and the seven hemigramicidin derivative compounds.
FIG. 1A shows TEMPOL.
FIG. 1B shows a dipeptidic TEMPO analog—XJB-5-208.
FIG. 1C shows a hemigramicidin-TEMPO conjugate—XJB-5-125.
FIG. 1D shows a hemigramicidin compound that does not have the TEMPO moiety—XJB-5-127.
FIG. 1E shows a hemigramicidin-TEMPO conjugate—XJB-5-131.
FIG. 1F shows a hemigramicidin compound that does not have the TEMPO moiety—XJB-5-133.
FIG. 1G shows a hemigramicidin-TEMPO conjugate—XJB-5-197.
FIG. 1H shows a hemigramicidin compound that does not have the TEMPO moiety—XJB-5-194.
FIG. 2 depicts an example of a synthetic pathway for the TEMPO-hemigramicidin conjugates.
FIG. 3 shows an EPR-based analysis of integration and reduction of nitroxide Gramicidin S peptidyl-TEMPO conjugates in MECs.
FIG. 4 shows an flourescein isothiocyanate-dextran (FD4) read-out which reflects the effect of Gramicidin-S TEMPO conjugates on rat ileal mucosal permeability following profound hemorrhagic shock. Data are expressed as a percentage of the change permeability relative to that observed in simultaneously assayed control segments loaded during shock with normal saline solution.
FIG. 4A shows an FD4 read-out of TEMPOL which is used as a “positive control” for the gut mucosal protection assay.
FIG. 4B shows an FD4 read-out of TEMPO conjugate XJB-5-208 reflecting gut mucosal protection.
FIG. 4C shows an FD4 read-out of XJB-5-125 which has the TEMPO payload, but fails to provide protection against gut barrier dysfunction induced by hemorrhage.
FIG. 4D shows an FD4 read-out of XJB-5-127 which lacks the TEMPO payload and fails to provide protection against gut barrier dysfunction induced by hemorrhage.
FIG. 4E shows an FD4 read-out of TEMPO conjugate XJB-5-131 reflecting gut mucosal protection.
FIG. 4F shows an FD4 read-out of XJB-5-133 which lacks the TEMPO payload even though it possesses the same hemigramicidin mitochondria targeting moiety as the most active compound, XJB-5-131.
FIG. 4G shows an FD4 read-out of XJB-5-197 which has the TEMPO payload, but fails to provide protection against gut barrier dysfunction induced by hemorrhage.
FIG. 4H shows an FD4 read-out of XJB-5-194 which lacks the TEMPO payload and fails to provide protection against gut barrier dysfunction induced by hemorrhage.
FIG. 5 shows graphical representations of the effect of nitroxide conjugates on ActD-induced apoptosis.
FIG. 5A is a graphical representation of superoxide production based upon mean fluorescence intensity from 10,000 ileal cells.
FIG. 5B is a graphical representation of phosphatidylserine (PS) externalization as indicated by the percentage of annexin V-positive cells.
FIG. 5C is a graphical representation of caspase-3 activity as indicated by amount of its specific substrate present, Z-DVED-AMC, in nmol/mg protein.
FIG. 5D is a graphical representation of DNA fragmentation as indicated by propidium iodide fluorescence.
FIG. 5E is a graphical representation of PS externalization at different concentrations of the compound 5a.
FIG. 5F is a graphical representation of adenosine triphosphate (ATP) levels in mitochondria in the presence or absence of 5a or 2-deoxyglucose.
FIG. 6 illustrates the effects of intraluminal XJB-5-131 on hemorrhage-induced peroxidation of phospholipids in intestinal mucosa.
FIG. 6A is a graphical representation of the peroxidation of phosphatidylcholine (PC).
FIG. 6B is a graphical representation of peroxidation activity with respect to phosphatidylethanolamine (PE).
FIG. 6C is a graphical representation of peroxidation activity with respect to phosphatidylserine (PS).
FIG. 6D is a graphical representation of peroxidation activity with respect to cardiolipin (CL).
FIG. 7 is a graphical representation of caspase 3 and 7 activity that illustrates the effects of intraluminal XJB-5-131.
FIG. 8 is a graphical representation of permeability of XJB-5-131 with respect to Caco-2 BBe human enterocyte-like monolayers subjected to oxidative stress. The permeability of the monolayers is expressed as a clearance (pL·h−1·cm−2).
FIG. 9A is a graphical representation of the effects of intravenous treatment with XJB-5-131 on MAP (mean arterial pressure, mm Hg) of rates subjected to volume controlled hemorrhagic shock.
FIG. 9B is a graphical representation of the effects of intravenous treatment with XJB-5-131 on survival probability of rates subjected to volume controlled hemorrhagic shock.
Patent Application
20070161573
