# US 2017/0010255 A1 · Methods for identifying modulators of membrane potentials in bipolar disorder and attention deficit hyperactivity disorder > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2017/0010255 A1 - Document ID: US20170010255A1 - Status: Published application - Kind: A1 - Application number: 15/276,088 - Filed: 2016-09-26 - Published: 2017-01-12 - Earliest claimed: 2014-04-29 - Assignees: PsychNostics, LLC - Inventors: Alagu P. THIRUVENGADAM - Web page: https://patentgrep.com/patent/US20170010255A1 ## CPC - G01N 33/5041: {involving analysis of members of signalling pathways} - G01N 33/5008: {for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics} - G01N 33/502: {for testing non-proliferative effects} - G01N 33/6872: {Intracellular protein regulatory factors and their receptors, e.g. including ion channels} - G01N 2333/912: transferring phosphorus containing groups, e.g. kinases (2.7) - G01N 2333/91215: with a definite EC number (2.7.1.-) - G01N 2800/304: Mood disorders, e.g. bipolar, depression - G01N 2800/305: Attention deficit disorder; Hyperactivity - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract The present invention provides methods to modulate key elements along the DAG signaling pathway as well as a diagnostic assay, device and methods of using the same to diagnose bipolar disorder (BD) and attention deficit hyperactivity disorder (ADHD). Methods to identify diagnostic markers and drug targets for BD and ADHD. Methods of identifying effective compounds responsible for membrane potentials and excitabilities influencing bipolar disorder (BD) and attention deficit hyperactivity disorder (ADHD). Methods of identifying an effective compound that modulates the activity of Ca2+/CaM enzyme and compounds involved in changing the K+ gradient across the plasma membrane thereby increasing or decreasing the membrane potential ratio (MPR™) values. The invention provides methods of identifying a compound that modulates the activity of PKC which is an important protein of the DAG signaling pathway. Methods of identifying a compound that modulates DAG and its related enzymes along the DAG signaling pathway are provided. These compounds decrease or increase the membrane potential ratio (MPR™) in BD and ADHD patients. ## Claims ### Claim at position 1 1-12. (canceled) ### Claim 13 A method of identifying an agent for treatment of attention deficit hyperactivity disorder (ADHD) comprising: combining a first population of human ADHD patient cells comprising cells that express human calcium-activated potassium-channels hSK4, and with a test agent suspected of altering human calcium-activated potassium channels hSK4 activity; obtaining a test ratio of a mean membrane potential of the first population of human ADHD patient cells incubated in vitro in the presence of the test agent and in the absence of K+, to a mean membrane potential of a second population of the human patient cells incubated in vitro in the absence of the test agent and presence of K+; comparing the test ratio to (a) and/or (b): (a) a control ratio of a mean membrane potential from control human cells known to not have ADHD comprising cells that express human calcium-activated potassium-channels hSK4, incubated in vitro in the presence of an agent that alters human calcium-activated potassium channels hSK4 activity and in the absence of K+, to a mean membrane potential of the control human cells incubated in vitro in the absence of the agent that alters human calcium-activated potassium channels hSK4 activity and in the presence of K+, (b) an ADHD control ratio of a mean membrane potential from ADHD control human cells known to have ADHD comprising cells that express human calcium-activated potassium-channels hSK4, incubated in vitro in the presence of the agent that alters human calcium-activated potassium channels hSK4 activity and in the absence of K+, to a mean membrane potential of the ADHD control human cells incubated in vitro in the absence of the agent that alters human calcium-activated potassium channels hSK4 activity and in the presence of K+; wherein when: (1) the test ratio is not significantly different from the control ratio of (a), (2) the test ratio is decreased towards the control ratio of (a) in comparison to (b), or (3) the test ratio is decreased in comparison to the ADHD control ratio of (b), the test agent modulates the mean membrane potential in ADHD patients. ### Claim 14 (depends on 13) The method of claim 13, wherein the agent is selected from the group consisting of a calcium-calmodulin (Ca2+/CaM) kinase inhibitor, a diacylglycerol kinase inhibitor, and a protein kinase C inhibitor. ### Claim 15 (depends on 14) The method of claim 14, wherein the agent is a PKC inhibitor selected from the group consisting of phorbol 12-myristate 13-acetate (PMA), 3-(1H-indol-3-yl)-4-[2-(4-methylpiperazin-1-yl)quinazolin-4-yl]pyrrole-2,5-dione (Sotrastaurin or AEB07), 13-hydroxyoctadecadienoic acid (13-HODE), aprinocarsen, bisindolylmaleimide, bryostatin-1, butein, calphostin C, 7,8-dihydroxycoumarin, 4′-demethylamino-4′-hydroxystaurosporine, rottlerin, ruboxistaurin, staurosporine, and verbascoside. ### Claim 16 (depends on 14) The method of claim 14, wherein the agent is a calcium-calmodulin (Ca2+/CaM) kinase inhibitor. ### Claim 17 (depends on 16) The method of claim 16, wherein the calcium-calmodulin (Ca2+/CaM) kinase inhibitor is autocamtide-2-related inhibitory peptide (AIP). ### Claim 18 (depends on 14) The method of claim 14, wherein the agent is a diacylglycerol kinase inhibitor. ### Claim 19 (depends on 18) The method of claim 18, wherein the diacylglycerol kinase inhibitor is 6-[2-[4-[(4-fluorophenyl)phenylmethylenel-1-piperidinyl]ethyl]-7-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one (ALX). ### Claim at position 9 20-26. (canceled) ### Claim 27 A method of identifying an agent that modulates diacylglycerol signaling for the treatment of attention deficit hyperactivity disorder (ADHD) comprising: combining a first population of human ADHD patient cells and a test agent suspected of altering diacylglycerol signaling; obtaining a test ratio of a mean membrane potential of the first population of human ADHD patient cells incubated in vitro in the presence of the test agent and in the absence of K+, to a mean membrane potential of a second population of the human ADHD patient cells incubated in vitro in the absence of the test agent and the presence of K+ or absence of K+; comparing the test ratio to (a) and/or (b): (a) is a control ratio of a mean membrane potential from control human cells known to not have ADHD incubated in vitro in the presence of an agent that alters diacylglycerol signaling and in the absence of K+, to a mean membrane potential of the control human cells known incubated in vitro in the absence of the agent that alters diacylglycerol signaling and in the presence of K+ or absence of K+, (b) is an ADHD control ratio of a mean membrane potential from ADHD control human cells known to have said ADHD incubated in vitro in the presence of the agent that alters diacylglycerol signaling and in the absence of K+, to a mean membrane potential of the ADHD control human cells incubated in vitro in the absence of the agent that alters diacylglycerol signaling and in the presence of K+ or absence of K+; wherein when: (1) the test ratio is not significantly different from the control ratio of (a), (2) the test ratio is decreased towards the control ratio of (a) in comparison to (b), or (3) the test ratio is decreased in comparison to the ADHD control ratio of (b), the test agent modulates the mean membrane potential in ADHD patients. ### Claim 28 (depends on 27) The method of claim 27, wherein the human cells are selected from the group consisting of red blood cells and lymphoblasts. ### Claim 29 (depends on 27) The method of claim 27, wherein the agent is selected from the group consisting of a calcium-calmodulin (Ca2+/CaM) kinase inhibitor, a diacylglycerol kinase inhibitor, and a protein kinase C inhibitor. ### Claim 30 (depends on 29) The method of claim 29, wherein the agent is a PKC inhibitor selected from the group consisting of phorbol 12-myristate 13-acetate (PMA), 3-(1H-indol-3-yl)-4-[2-(4-methylpiperazin-1-yl)quinazolin-4-yl]pyrrole-2,5 dione (Sotrastaurin or AEB07), 13-hydroxyoctadecadienoic acid (13-HODE), aprinocarsen, bisindolylmaleimide, bryostatin-1, butein, calphostin C, 7,8-dihydroxycoumarin, 4′-demethylamino-4′-hydroxystaurosporine, rottlerin, ruboxistaurin, staurosporine, and verbascoside. ### Claim 31 (depends on 29) The method of claim 29, wherein the agent is a calcium-calmodulin (Ca2+/CaM) kinase inhibitor. ### Claim 32 (depends on 31) The method of claim 31, wherein the calcium-calmodulin (Ca2+/CaM) kinase inhibitor is autocamtide-2-related inhibitory peptide (AIP). ### Claim 33 (depends on 29) The method of claim 29, wherein the agent is a diacylglycerol kinase inhibitor. ### Claim 34 (depends on 33) The method of claim 33, wherein the diacylglycerol kinase inhibitor is 6-[2-[4-[(4-fluorophenyl)phenylmethylenel-1-piperidinyl]ethyl]-7-methyl-5H-thiazolo[3,2-a]pyrimidin-5-one (ALX). ### Claim at position 18 35-36. (canceled) ### Claim 37 (depends on 27) A method of identifying an agent that modulates diacylglycerol signaling for the treatment of attention deficit hyperactivity disorder (ADHD) of claim 27, wherein: the first population of human patient cells is combined with a test agent suspected of altering PKC activity, the control human cells and the bipolar control human cells are incubated in vitro in the presence of phorbol 12-myristate 13-acetate (PMA), and the human cells are selected from the group consisting of red blood cells and lymphoblasts. ### Claim at position 20 38-43. (canceled)