US 2006O1783 07A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2006/0178307 A1 Bartlett et al. (43) Pub. Date: Aug. 10, 2006 (54) MODULATION OF NMDA RECEPTOR Publication Classification CURRENTS VIA OREXN RECEPTOR AND/OR CRF RECEPTOR (51) Int. Cl. A6II 38/22 (2006.01) (75) Inventors: Selena Bartlett, Berkeley, CA (US); A6II 3L/495 (2006.01) Antonello Bonci, San Francisco, CA A61K 31/4745 (2006.01) (US); Stephanie Borgland, San A61K 31/4706 (2006.01) Francisco, CA (US); Howard Fields, A6II 3/17 (2006.01) Berkeley, CA (US); Sharif Taha, (52) U.S. Cl. ...................... 514/12: 514/255.01: 514/300; Berkeley, CA (US) 514/313; 514/585; 514/595 (57) ABSTRACT Correspondence Address: QUINE INTELLECTUAL PROPERTY LAW This invention pertains to the discoveries that orexin and/or GROUP, PC. CRF increase NMDAR (N-methyl-D-aspartate receptor)- PO BOX 458 mediated currents at excitatory synapses onto a Subset of dopamine cells in the ventral tegmental area (VTA) in the ALAMEDA, CA 94501 (US) mammalian brain. The orexin effect can be blocked by an (73) Assignee: The Regents of the University of Cali orexin receptor type 1 (OXR1). The CRF effect can be blocked by a CRF receptor 2 (CRF-R2) antagonist or by an fornia inhibitor of the CRF-binding protein (CRF-BP). Methods (21) Appl. No.: 11/343,259 are provided that exploit these discoveries to modulate NMDAR-mediated currents in vivo and in vitro and to (22) Filed: Jan. 25, 2006 screen for modulators (upregulators or downregulators) of NMDA-mediated currents. In vivo methods include the use Related U.S. Application Data of modulators of the orexin and CRF pathways to of mitigate a symptom of Substance abuse. The invention also provides (60) Provisional application No. 60/647,748, filed on Jan. methods and compositions for co-administration of modu 26, 2005. lators that act via the orexin and CRF pathways. Patent Application Publication Aug. 10, 2006 Sheet 1 of 14 US 2006/0178307 A1 4. 200 150 00 SO O O 10 20 30 40 50 60 10 100 Time (nin) Orexin A nM 00 75 100 nM orexin A F 1 M SB 334867 50 25 O 10 20 30 4. 50 SO O 10 20 30 40 60 Time (min) Tine (nir) F Biocytin-TR H-FITC Merge 200 75 150 125 100 7 5 100 nM orexin A 0 10 20 30 40 50 60 70 80 Time (min) Fig. 1 Patent Application Publication Aug. 10, 2006 Sheet 2 of 14 US 2006/0178307 A1 A B C g M chellerythrin 1M U73122 ia. ia. S. so 100 sissiliig, g a. 50 0 exi A : g e n 2 in s 2 3 4 so SO 70 O 1. 3. O S. Time (min) Time (min) Tirna (min) D F 100 Mrp-cAMPS 20 all PKI 3. s sis g Oncrek A Y 5 a. e a 2 to so 3 S 6 Tirne trir) line (min) Tim a min) Fig. 2 Patent Application Publication Aug. 10, 2006 Sheet 3 of 14 US 2006/0178307 A1 A B 0.4 MNVP-AAMO77 20 15 o 100 nM orexin a Ho 0.4 MNVP-AAM077 O O 20 30 40 50 60 O 10 20 30 40 50 60 Ting (nir Tine (rnin} C. D o 3 Mifeprodil 150 Eo 125 Zn M 25 100 AM. 300 nM 100 """"" SS 0.O a 75 7s 50 E 10 nM orexin A 00 nMorexin A e c a s 0 10 20 30 4 SO SO 70 80 a Time (min) Time (min) E as 0.4 MNVP-AAMO77 28 0.4 MNVF. AAM077 E 1 25 s 3 Mifenprodil d 3 Miferprodil e R d 100 100 1 MPPDA s s a 75 a 75 100 nM orexin A a. 50 So 25 25 a S O O 10 20 30 40 50 50 2s 0 0 20 30 40 50 60 70 80 Time (min) Time (min) Fig. 3 Patent Application Publication Aug. 10, 2006 Sheet 4 of 14 US 2006/0178307 A1 A - - - - - - - - B no statio C on no stimulation 10 Mt. K-801 O mR as 10 am MK-8O1 S. 20 P g r St s 15 3. Es 1. e E e so e s 2 a 1 20 so a so is 70 Time (min) Time (min) C D hostinator an PO nostitulation 2 S 20 it MMK-8 + 500 MNLA SO o P g 10 MMK-801 + 500 M MMDA 1 wwat M s g 100 E s 50 2 5 e 100 nM orexin A. s 2 0 20 30 O 20 30 40 50 so 70 Time (min) Time (min) Fig. 4 Patent Application Publication Aug. 10, 2006 Sheet 5 of 14 US 2006/0178307 A1 A COCaine COCaine + SB 334867 1.O. S. O.8 O O.6 s2 O. 5. 0.2 g O. saline cocaine saline cocaine SB 334867 B Control 15 min 3-4 hours 1,O O. 2 O, O control 5 in 3-4 hours Fig. 5 Patent Application Publication Aug. 10, 2006 Sheet 6 of 14 US 2006/0178307 A1 Control 15 min 3-4 hours control 15 min 3-4 hours APW control 15 mr 3-4 hours APW 1.0 1. 0. 2. t r al a coffo al m C. to s 0.5 15 min S. 0.5 -w 15 mi s wim 3-4 hours -3-4 hours m 3-4 hours + APW 2 -a 3-hours 4 APW U , 0. 0 10 20 so 40 50 60 O 2 3 4. S 6 nEFSC Amplitude (pA) inter-evont linterval (s) H Time (min) 2.5 5.0 75 100 e (min) 0- it S-100 -200 c .300 Patent Application Publication Aug. 10, 2006 Sheet 7 of 14 US 2006/0178307 A1 Control 15 in 3-4 hours D E g m St 3 g es s . e le; : 3 3 l E E e O contro 1S in 3-4 hours contro Smin 3-4 ours F G 1, 1. cer e . o d - control 0.5 it. g 0.5 or 15 min reme as ours s -o-, 3-4 hours are sE w . 0. 1. 2. O 40 1 2 s 4. H mEPSC Amplitudo pa) inter-event intervals 400 300 k 2 20 100 e s. s Time (min) Time (min) Patent Application Publication Aug. 10, 2006 Sheet 8 of 14 US 2006/0178307 A1 -- cocaine + wahicle -- cocaine + SB 334867 soooo-o-Safine * Wehicle 40000 -e-saline + S8 334867 9. 5 3000 g s Cs i 20000 8 is 100 t O 0 1 2 3 4 s 6 7 8 Days C D 70000 ama 6000 5 5000 40000 au h 3000 2 20000 s amb 0-F 10000 0. Oay 1 Day 7 day 1 day 7 tra-WA Wuicle intra-WTASB 334867 Patent Application Publication Aug. 10, 2006 Sheet 9 of 14 US 2006/0178307 A1 0.75 0.50 0.25 Fig. 9 Patent Application Publication Aug. 10, 2006 Sheet 10 of 14 US 2006/0178307 A1 Patent Application Publication Aug. 10, 2006 Sheet 11 of 14 US 2006/0178307 A1 300 Naive Vehicle Vehicle S334867 Naive Wehicle Weice SB 33436 25 20 1o s O Naive Weice Wehicle SB3348S Naive Wehicle Weice S333,867 Fig. 11 Patent Application Publication Aug. 10, 2006 Sheet 12 of 14 US 2006/0178307 A1 A B 7 6 on 5 8 3 s - 2 O Vehicle Vehicle SB334867 Naive Wehicle Vehicle SB334867 200 10 O Vehicle Vehicle S8334367 Naive Vehicle Vehicle S3334867 Fig. 12 Patent Application Publication Aug. 10, 2006 Sheet 13 of 14 US 2006/0178307 A1 0.01 0 1 1.0 CRF concentration (M) Fig. 13 Patent Application Publication Aug. 10, 2006 Sheet 14 of 14 US 2006/0178307 A1 A B as S 50 5 40 3. S. St 3 30 ()2 20 C. 2 C 10 Ze 0 1 10 100 Orexin A nM Time (min) C D 225 g 150 125 s 175176 3 is 100 125- Yua d 75 1 nM orexin A is 75 al 50 10 nM CRF 1 nM orexin A 25 1OM CRF a 25 O o 20 30 40 50 SO O 10 20 30 40 50 SO Time (min) Time (min) Fig. 14 US 2006/0178307 A1 Aug. 10, 2006 MODULATION OF NMDA RECEPTOR CURRENTS abuse and plays a key role in stress-induced relapse to drug VIA OREXIN RECEPTOR AND/OR CRF taking (Sarnyai et al. (2001) Pharmacol. Rev. 53: 209-243). RECEPTOR The cellular effects of CRF are mediated via two receptors (CRF-R1 and CRF-R2) (Dautzenberg and Hauger (2002) CROSS-REFERENCE TO RELATED Trends Pharmacol. Sci. 23, 71-77); CRF also binds to a APPLICATIONS binding protein (CRF-BP), which is thought to inactivate 0001) This application claims the benefit of U.S. Provi free CRF (Kemp et al. (1998) Peptides 19: 1119-1128). It sional Application No. 60/647,748, filed Jan. 26, 2005, has been suggested that CRF-BP inhibitors, which elevate which is hereby incorporated by reference in its entirety. free CRF levels, may provide potential treatments for disorders where CRF levels are depressed, such as Alzhe STATEMENT AS TO RIGHTS TO INVENTIONS imer's disease and Parkinson's disease (Behan et al. (1995) MADE UNDER FEDERALLY SPONSORED Nature 378: 284-287). It is notable that many of these RESEARCH AND DEVELOPMENT disorders involving elevated CRF levels are also thought to involve elevated dopamine levels and that CRF increases 0002 This invention was made with government support dopamine release in both limbic and cortical projection areas under grant no.