CHANNEL MODULATORS
The inventions relate to compositions and articles of manufacture comprising connexin modulators, pannexin modulators, gap junction modulators, hemichannel modulators, and pannexin channel modulators and their use, alone or in combination, in treating ocular and other disorders.
1 - 73 . (canceled)
74 . A method for estimating the dose of a benzoylamino benzopyran test compound for use in modulating connexin hemichannel opening, the method comprising: a) providing one or more benzoylamino benzopyran test compounds and a connexin 43 hemichannel opening inhibition test system; b) selecting a positive test compound based on its inhibition of connexin hemichannel opening in the test system, at one or more tested amounts; and c) selecting one or more doses of said positive test compound for modulating connexin 43 hemichannel opening based on the one or more tested amounts.
75 . The method of claim 74 , wherein the benzoylamino benzopyran test compound is a compound of Formula I.
76 . The method of claim 74 , wherein the benzoylamino benzopyran test compound is a compound of Formula II.
77 . The method of claim 74 , wherein the method is used to identify a range of dosages of a positive test compound for modulation of connexin 43 hemichannel opening.
78 . The method of claim 74 , further comprising the step of determining an IC50 for said test compound.
79 . The method of claim 74 , wherein the method is carried out in vitro.
80 . The method of claim 74 , wherein the method is carried out in vivo.
81 . The method of claim 79 , wherein the connexin 43 hemichannel opening inhibition test system comprises a scrape load assay and the in vitro method quantifies scrape-loaded dye spread.
82 . The method of claim 79 , wherein the in vitro method is a whole-cell voltage clamp assay.
83 . The method of claim 79 , wherein the connexin 43 hemichannel opening inhibition test system comprises an in vitro ischemia injury-reperfusion ATP release assay and the in vitro method quantifies total extracellular ATP release.
84 . The method of claim 83 , wherein the in vitro method comprises human cerebral microvascular endothelial (hCMVEC) cells.
85 . The method of claim 83 , wherein the connexin 43 hemichannel opening inhibition test system comprises a retinal ischemia-reperfusion model
86 . The method of claim 80 , wherein the connexin 43 hemichannel opening inhibition test system comprises an in vivo rodent animal model.
87 . The method of claim 74 , wherein the connexin hemichannel is selected from the group consisting of connexin 30, connexin 31.1, connexin 36, connexin 37, connexin 40, connexin 43, connexin 45, connexin 50, and connexin 57 hemichannels.
88 . The method of claim 74 , wherein the connexin hemichannel is a connexin 43 hemichannel.
89 . The method of claim 74 , wherein the connexin hemichannel is a connexin 45 hemichannel.
90 . The method of claim 74 , wherein the connexin hemichannel is a connexin 40 hemichannel.
91 . The method of claim 74 , wherein the connexin hemichannel is a connexin 37 hemichannel.
92 . The method of claim 74 , further comprising the step of testing said positive benzoylamino benzopyran test compound dose for inhibition of NLRP3 inflammasome activity.
93 . A method according to either of claim 74 or 101 , wherein the dose is estimated for use in humans.
94 . A method for evaluating the therapeutic efficacy of a compound of Formula II for inhibition of connexin hemichannel opening, the method comprising: a) providing one or more test compounds of Formula II and a connexin hemichannel opening inhibition test system; and b) selecting positive test compounds based on their ability to inhibit connexin hemichannel opening in the test system, thereby identifying the therapeutic efficacy of a test compound for the ability to inhibit connexin hemichannel opening.
95 . The method of claim 94 , wherein the connexin hemichannel is a connexin 43 hemichannel.
96 . The method of claim 95 , wherein the connexin hemichannel is a connexin 43 hemichannel.
97 . The method of claim 95 , wherein the connexin hemichannel opening inhibition test system is an in vitro test system.
98 . The method of claim 97 , wherein a positive test compound is selected based on the ability to inhibit ATP release from connexin hemichannels.
99 . The method of claim 97 , wherein a positive test compound is selected based on the ability to inhibit inflammasome activation.
100 . The method of claim 97 , wherein NLRP3 inflammasome activation is inhibited.
101 . A method for estimating the dose of a benzoylamino benzopyran test compound for use in preventing, inhibiting, and/or reducing the function or activity of a connexin 43 hemichannel, the method comprising: a) providing one or more benzoylamino benzopyran test compounds and a connexin 43 hemichannel inhibition test system; b) selecting a positive test compound based on its ability to prevent, inhibit, and/or reducing the function or activity of a connexin 43 hemichannel in the test system; and, (c) identifying a dose of said positive test compound for its ability to prevent, inhibit, and/or reducing the function or activity of a connexin 43 hemichannel.
102 . A method according to claim 101 , wherein the dose for preventing, inhibiting, and/or reducing the function or activity of a connexin 43 hemichannel is estimated for use in humans.
103 . The method of claim 101 , further comprising the step of testing said positive benzoylamino benzopyran test compound dose for inhibition of NLRP3 inflammasome activity.
104 . A method for estimating the dose of a benzoylamino benzopyran test compound for use in preventing, inhibiting, and/or reducing the function or activity of an inflammasome, the method comprising: a) providing one or more benzoylamino benzopyran test compounds and an inflammasome inhibition test system; b) selecting a positive test compound based on its ability to prevent, inhibit, and/or reducing the function or activity of an inflammasome in the test system; and, (c) identifying a dose of said positive test compound for its ability to prevent, inhibit, and/or reducing the function or activity of an inflammasome.
105 . The method of claim 104 , wherein the inflammasome is an NLRP3 inflammasome.
106 . A method for estimating the dose of a connexin 43 hemichannel modulation compound for use in preventing, inhibiting, and/or reducing the function or activity of an inflammasome, the method comprising: a) providing one or more connexin 43 hemichannel modulation test compounds and an inflammasome inhibition test system; b) selecting a positive test compound based on its ability to prevent, inhibit, and/or reducing the function or activity of an inflammasome in the test system; and, (c) identifying a dose of said positive test compound for its ability to prevent, inhibit, and/or reducing the function or activity of an inflammasome.
107 . The method of claim 106 , wherein the connexin 43 hemichannel modulation compound is a benzoylamino benzopyran.
108 . The method of claim 107 , wherein the benzoylamino benzopyran is a compound according to Formula I.
109 . The method of claim 106 , wherein the inflammasome is an NLRP3 inflammasome.