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(12) Patent Application Publication (10) Pub. No.: US 2007/0078113 A1 Roth Et Al

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(12) Patent Application Publication (10) Pub. No.: US 2007/0078113 A1 Roth Et Al US 20070078113A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2007/0078113 A1 Roth et al. (43) Pub. Date: Apr. 5, 2007 (54) METHODS, COMPOSITIONS AND Publication Classification ARTICLES OF MANUFACTURE FOR ENHANCING SURVIVABILITY OF CELLS, (51) Int. Cl. TISSUES, ORGANS, AND ORGANISMS A6II 3/66 (2006.01) A 6LX 3/555 (2006.01) (76) Inventors: Mark B. Roth, Seattle, WA (US); Mike A6II 3/85 (2006.01) Morrison, Seattle, WA (US); Eric A6II 3 L/65 (2006.01) Blackstone, Seattle, WA (US); Dana A 6LX 3L/275 (2006.01) Miller, Seattle, WA (US) A6II 3L/26 (2006.01) A 6LX 3L/095 (2006.01) Correspondence Address: A6II 3L/21 (2006.01) FULBRIGHT & UAWORSK LLP. (52) U.S. Cl. .......................... 514/114: 514/184: 514/706; 6OO CONGRESS AVE. 514/126; 514/553: 514/514; SUTE 24 OO 514/526; 514/621 AUSTIN, TX 78701 (US) (57) ABSTRACT (21) Appl. No.: 11/408,734 The present invention concerns the use of oxygen antago (22) Filed: Apr. 20, 2006 nists and other active compounds for inducing stasis or pre-stasis in cells, tissues, and/or organs in vivo or in an organism overall, in addition to enhancing their Survivabil Related U.S. Application Data ity. It includes compositions, methods, articles of manufac ture and apparatuses for enhancing Survivability and for (60) Provisional application No. 60/673,037, filed on Apr. achieving stasis or pre-stasis in any of these biological 20, 2005. Provisional application No. 60/673,295, materials, so as to preserve and/or protect them. In specific filed on Apr. 20, 2005. Provisional application No. embodiments, there are also therapeutic methods and appa 60/713,073, filed on Aug. 31, 2005. Provisional appli ratuses for organ transplantation, hyperthermia, wound heal cation No. 60/731,549, filed on Oct. 28, 2005. Pro ing, hemorrhagic shock, cardioplegia for bypass Surgery, visional application No. 60/762.462, filed on Jan. 26, neurodegeneration, hypothermia, and cancer using the 2006. active compounds described. US 2007/0078113 A1 “?IJII eq9A3 ua) ed Patent Application Publication Apr. 5, 2007 Sheet 2 of 40 US 2007/0078113 A1 00 BO 70, 60 SO, 40. 3O, 20, O o, N2 0.0 kPa 02/2 o.OSkip of N2 O. kP O2/N2. OS kPa O2/N2 FG, 2. Patent Application Publication Apr. 5, 2007 Sheet 3 of 40 US 2007/0078113 A1 70's 60% 0. 30 20 10% D CO OD5 kP can2 0.05 kPa orco FIG. 3 Patent Application Publication Apr. 5, 2007 Sheet 4 of 40 US 2007/0078113 A1 Patent Application Publication Apr. 5, 2007 Sheet 5 of 40 US 2007/0078113 A1 –64/i>SZH ZZ/S––SZH 94/9•SZH 84/z-----SZH 64/º----ZN ?–ZZ/SZN 94/9----ZN –84/ZZN 00:0T00:II 00:6 00:9 00:4 00:9 00:S 00:5 00:2 00:[ Patent Application Publication Apr. 5, 2007 Sheet 6 of 40 US 2007/0078113 A1 eSeOnpu?SºHeq?eXII-uomeuele?s (obec) eneiadual Patent Application Publication Apr. 5, 2007 Sheet 7 of 40 US 2007/0078113 A1 ?ep?nsuê6oupÁH]u?douGºin?eleduuelquepueded ºsnowegoºjn?eleduje1.ÁpogeJoo 00:200:1,00:000:900:3900:$00:9 (unou)eu,1 09 92 02 G?. (O fed) enqeuedue Patent Application Publication Apr. 5, 2007 Sheet 8 of 40 US 2007/0078113 A1 ºIn?eledujeLÁpogaloojsewo (O 6ed) ejneedu.a. Patent Application Publication Apr. 5, 2007 Sheet 9 of 40 US 2007/0078113 A1 elmeleduuelquêpuedaqdoud Od “?IH8-V8 uleod 6nfeoud Patent Application Publication Apr. 5, 2007 Sheet 10 of 40 US 2007/0078113 A1 ??. Patent Application Publication Apr. 5, 2007 Sheet 11 of 40 |-ULI||| 90 Patent Application Publication Apr. 5, 2007 Sheet 12 of 40 US 2007/0078113 A1 ‘5)IJIII 90%.|||||90 GS)#2), Patent Application Publication Apr. 5, 2007 Sheet 13 of 40 US 2007/0078113 A1 “?IHZI Patent Application Publication Apr. 5, 2007 Sheet 14 of 40 US 2007/0078113 A1 Provide mixture of One of - 202 more breathable gas SOUCeS Add a predetermined 204 amount of active agent to the gas mixture Administer gas mixture to subject 2O6 Ventilate exhaled gases 208 FIG, 13 Patent Application Publication Apr. 5, 2007 Sheet 15 of 40 US 2007/0078113 A1 Position chamber over 402 treatment area Reduce pressure in 4O4 Chamber Apply predetermined amount of active agent to 406 chamber FIG. 14 Patent Application Publication Apr. 5, 2007 Sheet 17 of 40 US 2007/0078113 A1 y 2 D () C 9 0.5m 5m 15m 30 m CO, N, only only FIG. 16 OCO, pretreatment no pretreatment 13.5 h 15h 18 h 18h (trial 2) length of anoxic exposure FIG, 17 Patent Application Publication Apr. 5, 2007 Sheet 18 of 40 US 2007/0078113 A1 O. 7 no pretreatmerit DHSICO, pretreatment O. 2 O. length of anoxic exposure . FIG. 18 Patent Application Publication Apr. 5, 2007 Sheet 19 of 40 US 2007/0078113 A1 FIG. 19 Patent Application Publication Apr. 5, 2007 Sheet 20 of 40 US 2007/0078113 A1 Body core temperature of rat in 300ppm H2S - incubator - rat 150.00 350.00 550.00 minutes from H2S exposure (H2S on at 0, off at 360 minutes) FIG. 20A Body core temperature of mouse in 15% CO2 - incubator OUSe ' ' r - we e-n 180.00 380.00 580.00 780.00 minutes from beginnig of CO2 exposure (CO2 on at 0 off at 360 minutes) FIG. 20B Patent Application Publication Apr. 5, 2007 Sheet 21 of 40 US 2007/0078113 A1 mouse H2S80ppm NOUSeHe -e- TOUSe H2S-CO2+He O2 rath2S--CO2 dog CO2 race N / \ / rE - ratheSCO2. He C2 dogF2S CO2 3.He -- dog H2S-CO2+He; C2 FIG 21 Patent Application Publication Apr. 5, 2007 Sheet 22 of 40 US 2007/0078113 A1 500. 54 522 508 520 s 518 504 2 512 510 FIG. 22A Patent Application Publication Apr. 5, 2007 Sheet 23 of 40 US 2007/0078113 A1 00. v-amalam 514 502 524 508 516 518 506 N 504 - T 512 510 FG, 22B Patent Application Publication Apr. 5, 2007 Sheet 24 of 40 US 2007/0078113 A1 Survival in 5% O2 Pretreatments 30 min (no HS) vs. 30 min (last 20 min whS) 100% 80% 60% 40% 20% O% O 10 20 30 40 50. 60 Time (minutes) FIG, 23 Patent Application Publication Apr. 5, 2007 Sheet 25 of 40 US 2007/0078113 A1 Patent Application Publication Apr. 5, 2007 Sheet 26 of 40 US 2007/0078113 A1 Metabolic Rate of a Mouse in 5% O. 3000 2500 2000 1500 OOO 500 O:00 1:OO 2:00 3:00 4:OO 5:00 8:OO 7:OO 8:00 Tirne (hrs) FIG. 25 Patent Application Publication Apr. 5, 2007 Sheet 27 of 40 US 2007/0078113 A1 i I CD V) N al a. g O O y Patent Application Publication Apr. 5, 2007 Sheet 28 of 40 US 2007/0078113 A1 g Patent Application Publication Apr. 5, 2007 Sheet 30 of 40 US 2007/0078113 A1 %09 %09 %08 uolonpold OO u effueuo Patent Application Publication Apr. 5, 2007 Sheet 31 of 40 US 2007/0078113 A1 S. B () X cy d E. o res C cy 2 d d s 0. C O gd Na FIG. 30 Patent Application Publication Apr. 5, 2007 Sheet 32 of 40 US 2007/0078113 A1 resistance to heat Stress 10 0.8 e wn house air -o- HS S 0.6 9 Ol S 5 0.4 CO 0.2 0.0 O 1O 20 30 40 50 60 7.O Time at 35 C (h) FIG. 31 Patent Application Publication Apr. 5, 2007 Sheet 33 of 40 US 2007/0078113 A1 lifespan 10 0.8 ammo House Air a-O- H2S 0.6 0.4 O2 O.O O 5 10 15 20 Time (days after L4) FIG. 32 Patent Application Publication Apr. 5, 2007 Sheet 34 of 40 US 2007/0078113 A1 40 35 30 CO2/hypoxia/Helium - - - - - - H2S 25 20 time (hours) FIG. 33 Mouse Core Body Temperature During 2 Hour H2Se Treatment 45 40 35 30 25 2O 15 10 O 1 2 3 4 5 Time (Hours) FIG. 34 Patent Application Publication Apr. 5, 2007 Sheet 35 of 40 US 2007/0078113 A1 40 35 30 25 Rat CBT 20 - - - - -/- Ambient Temp. 15 10 5 - -- -- T -T- - 6/23/05 6/2305 6/23/05 6/23/05 6/23/05 6/23/05 6/23/05 6/23/05 6:00 7:12 8:24 9:36 10:48 12:00 13:12 14:24 Time FIG. 35 40 39 38 37 - 36 35 34 6/29/05 6/29/05 6/29/05 6/29/05 6/29/05 6/29/05 6/29/05 8:24 9:36 10:48 12:OO 13:12 14:24 15:36 Time FIG. 36 Patent Application Publication Apr. 5, 2007 Sheet 36 of 40 US 2007/0078113 A1 15 II-ii 385 12.3 385 12.4 385 12.5 385 12.6 385 12.7 385 12.8 38.512.9 38.513 Time FIG. 37 5 O 6/16/05 6/16/05 6/16/05 6/16/05 6/16/05 7:12 9:36 12:OO 14:24 16:48 Time F.G. 38 Patent Application Publication Apr. 5, 2007 Sheet 37 of 40 US 2007/0078113 A1 core temp of dog in 9% CO2, 71%. He, 20% O2 room air off, CO2, He off, CO2, He on room air on 2OOO 40.00 6OOO 80.00 time (min) FIG.
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