US 201302241 10A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2013/0224110 A1 Bynoe (43) Pub. Date: Aug. 29, 2013 (54) USE OF ADENOSINE RECEPTOR Publication Classification SIGNALING TO MODULATE PERMEABILITY OF BLOOD-BRAIN (51) Int. Cl. BARRIER A 6LX3 L/7076 (2006.01) A 6LX3/59 (2006.01) A6II 45/06 (2006.01) (75) Inventor: Margaret S. Bynoe, Ithaca, NY (US) A613 L/437 (2006.01) (52) U.S. Cl. CPC ........... A6 IK3I/7076 (2013.01); A61 K3I/437 (73) Assignee: CORNELL UNIVERSITY, Ithaca, NY (2013.01); A61 K3I/519 (2013.01); A61 K (US) 45/06 (2013.01) USPC ............ 424/1.49; 514/303: 514/45: 514/267; (21) Appl. No.: 13/823,266 424/649; 424/94.5; 424/133.1: 514/17.7: 514/8.6; 424/85.4; 424/143.1; 424/141.1; (22) PCT Fled: Sep. 16, 2011 424/147.1:435/375 (86) PCT NO.: PCT/US 11/51935 (57) ABSTRACT The present invention relates to a method of increasing blood S371 (c)(1), brain barrier (“BBB) permeability in a subject. This method (2), (4) Date: May 14, 2013 involves administering to the Subject an agentoragents which activate both of the A1 and A2A adenosine receptors. Also disclosed is a method to decrease BBB permeability in a Related U.S. Application Data Subject. This method includes administering to the Subject an agent which inhibits or blocks the A2A adenosine receptor (60) Provisional application No. 61/383,628, filed on Sep. signaling. Compositions relating to the same are also dis 16, 2010. closed. Patent Application Publication Aug. 29, 2013 Sheet 1 of 27 US 2013/02241.1.0 A1 xWWWWWWWWWW sw. sy s s: FIG. I. 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S. 2: tire after wehicle: NECA injection h; -oirs after vehicle/NECA injection FIG. 20A t s igtss afterweisei-ECA injection FIG. 20B Patent Application Publication Aug. 29, 2013 Sheet 19 of 27 US 2013/02241.1.0 A1 3. s 3. 2 - - - - - - C3 FIG. 2 IA Brain AR Expression on Brain AR Expression of Endothetial Celis (IF) Erdothetia Ceis (FISH) 3. FIG 21B FIG. 21C Primary Brain Endotheia Ces AAR AAR FIG. 2 ID Patent Application Publication Aug. 29, 2013 Sheet 20 of 27 US 2013/02241.1.0 A1 - G.25 a 9.35 s ON E S S. G.3 Yr 2 - . S S 8.25 C S - C - s w se - .35 . a - rw F - 5 - t { 0.8 - k- - .' -- , As FIG 21E FIG. 2 IF e 3.25 . 8wehicle 5, 0.20 NECA 3.5 5 g O. - 0.05 : goo x WT A' AA- A s SC-5828: FIG 21G Patent Application Publication Aug. 29, 2013 Sheet 21 of 27 US 2013/02241.1.0 A1 0.4 S s is .3 * i. 3 S 5 (.2 c. s --- Q.: - a 0 out s s: - 3x &, a 8x <is fei:cis 3.45 : 54S. S. 4, 4 S 45 EA S.S. c. : GS 2. 3.8:3 CGS 21880 mg/kg FIG. 2 IIH 3. 34 - ..3. 8 E33 . 3. 8.2 : 3. 3. eigs 3: 38, 8.33 3.8 33 Ea &FA isg ( : FIG. 2 II s . S. 2. 3. 5 s S. siege E.A. &S Ra, 8 sis RS W S - C3 & & CCSScerA + FIG. 21. Patent Application Publication Aug. 29, 2013 Sheet 22 of 27 US 2013/02241.1.0 A1 r — 2.0 - E.8 s: a. s s E & 5. 3. 4. is a 1. s - 8 : . os r -s .5 S. C. s “ 0.0 & S 3 as3.2 S v SSS SSS S$ 3 coor Wasgia . S.C. 3x x exiscar (mgikg) FIG. 22A c 25 - 2S 20 s g 20 s s -- has s a s a g Q V G2. G & S. x - C, s S y ickirgki) is--- less s&Ss C. : S 5. 1. :ECA:gkg exsica, gkg ingikg; FIG. 22B Patent Application Publication Aug. 29, 2013 Sheet 23 of 27 US 2013/02241.1.0 A1 a s E g s 3. g 8 as a. s vehicle 1.org Lexiscani,w, FIG. 22C injection 5 min 33 s s 5 s: ).2 frt E 0.4 t e is 0.3 s C. is *- 0.1 i 0.0 O SO 100 w 150 2}} SC) O Si 23 Time after injection min} Time after injectioi (min) . 8.05 - 0.04 3 - 0.02 , - . encies;Se: S: 82 FIG. 22F Patent Application Publication Aug. 29, 2013 Sheet 24 of 27 US 2013/02241.1.0 A1 APP Motse Brains NECA Vehicle Anti-f-amyloid antibody (given i.v. Post-porten anti-3-amyloid staining L. Fros. 234-23 s i s s FIG. 23E Patent Application Publication Aug. 29, 2013 Sheet 25 of 27 US 2013/02241.1.0 A1 CD31 and B-amyloid Stained Art Mouse Brains FIGS. 23F-23G & - S. 3 is S. 20 is sk. t 5. NECA vehicle FIG. 23H Patent Application Publication Aug. 29, 2013 Sheet 26 of 27 US 2013/0224110 A1 A 8enc, 3 Ce:s 3CO S. 3200a & 2 GOs a 8 sasaas rassaaaaaaaaaa-assassarasaasaasaaraasa. i 38 SE 38 3 8 - As www.www.A A. As ClF3 isc, post-treatienti. - x - c.min} - - Eer,3 Cei. Atti. Uptake Šedia Ya3la NECA textsca: e3 Weis & E.8 sys&38. F-acti Staining 3 it 3 nir 8&ia -: stics C88sis EA Cicci ièxiscat FIGS 24A-24Y Patent Application Publication Aug. 29, 2013 Sheet 27 of 27 US 2013/02241.1.0 A1 i A or A2A i. Basal Conditions agonism Š. S S & S. S S Š ar. CNS S & S S Tight BBB Increased BBB permeability iii. A and A2A agon Sm S S. ces S oieces &S Adhesion diecies 7.9 FOd in Creased BBB permeability v. Aga antagonism S $. S FIG. 25 Š recreased BBB permeability US 2013/02241 10 A1 Aug. 29, 2013 USE OF ADENOSINE RECEPTOR cells are also different in that they have few pinocytic vesicles SIGNALING TO MODULATE which in other tissues allow somewhat unselective transport PERMEABILITY OF BLOOD-BRAIN across the capillary wall. Also lacking are continuous gaps or BARRIER channels running between the cells which would allow unre 0001. This application claims the benefit of U.S. Provi stricted passage. sional Patent Application Ser. No. 61/383,628, filed Sep. 16, 0006. The blood-brain barrier functions to ensure that the 2010, which is hereby incorporated by reference in its environment of the brain is constantly controlled.