US009855225B2 (12 ) United States Patent ( 10 ) Patent No. : US 9 ,855 ,225 B2 Eisenstein et al. ( 45) Date of Patent : Jan . 2 , 2018 I ( 54 ) CANNABINOID RECEPTOR TREATMENTS A61K 31/ 404 (2006 .01 ) A61K 31 /415 (2006 .01 ) ( 75 ) Inventors : Toby K . Eisenstein , Wyndmoor, PA A61K 31/ 454 ( 2006 . 01 ) (US ) ; Rebecca R . Hartzell , Indian A61K 39 / 395 ( 2006 .01 ) Rocks Beach , FL (US ) ; Martin W . A61K 45 /06 ( 2006 .01 ) Alder , Warrington , PA (US ) ; Joseph J . COZK 16 / 24 ( 2006 .01 ) Meissier, Ardmore , PA (US ) (52 ) U . S . Cl. ( 73 ) Assignee : TEMPLE UNIVERSITY OFFICE OF CPC . .. A61K 31 /09 (2013 . 01 ) ; A61K 31/ 095 TECHNOLOGY TRANSFER , ( 2013 .01 ) ; A61K 31 /35 (2013 . 01 ) ; A61K Philadelphia , PA (US ) 31 /352 ( 2013 .01 ) ; A61K 31 /404 ( 2013 .01 ) ; A61K 31/ 415 ( 2013 . 01 ) ; A61K 31/ 454 ( * ) Notice : Subject to any disclaimer, the term of this ( 2013 . 01 ) ; A61K 39 /3955 ( 2013 .01 ) ; A61K patent is extended or adjusted under 35 45 /06 ( 2013. 01 ) ; CO7K 16 /244 ( 2013 .01 ) U . S . C . 154 ( b ) by 101 days . (58 ) Field of Classification Search CPC .. .. .. .. .. A61K 31 /09 ; A61K 31/ 095 (21 ) Appl. No. : 14 /239 , 413 See application file for complete search history. (22 ) PCT Filed : Aug. 17 , 2012 (56 ) References Cited ( 86 ) PCT No. : PCT/ US2012 / 051330 U . S . PATENT DOCUMENTS $ 371 (c ) ( 1 ) , 2005/ 0159449 Al * 7 / 2005 Martin .. .. CO7D 303 / 10 ( 2 ) , ( 4 ) Date : Feb . 26 , 2016 514 /317 2008/ 0139635 A1* 6 / 2008 Martin .. .. A61K 31 /045 (87 ) PCT Pub. No. : W02013 /025984 514 / 397 PCT Pub . Date : Feb . 21, 2013 * cited by examiner (65 ) Prior Publication Data Primary Examiner — Yong Chong US 2017 /0014355 A1 Jan . 19 , 2017 ( 74) Attorney , Agent, or Firm — Duane Morris LLP Related U . S . Application Data (57 ) ABSTRACT (60 ) Provisional application No . 61/ 524 , 883, filed on Aug. The present invention includes methods and compositions 18 , 2011 . for treating a transplant recipient by administration of a CB2 receptor agonist either alone or in combination with one or (51 ) Int . Ci. more active pharmaceutical ingredients to block the rejec A61K 31 / 09 ( 2006 . 01 ) tion of foreign tissue and prolong grafted organs, tissues and A61K 31/ 095 ( 2006 .01 ) cells . A61K 31/ 35 ( 2006 .01 ) A61K 31 / 352 ( 2006 .01 ) 6 Claims, 16 Drawing Sheets U . S . Patent Jan . 2 , 2018 Sheet 1 of 16 US 9 ,855 , 225 B2 FIGURE 1A FIGURE 1B 14W Work Ooooo ! USU * * *HANS . SYNET W* with SuppressionIndex#SD ** - Artikel in SuppressionIndexSD Unawe engooszt nem tuumarket Wwwwwwwwwwwwwww ... - 4 .tami** reiman w i sh wwwy Concentration 0 - 1966 (UM ) Concentrati JWH - C . mytom JWH -015 1444 0 . 1966 (CB , antagonist) (C8 , antagonist) Ethanol Vehicle { 0 . 5 % 3 .2 x 10 + M m . 3 . 2 * 10 - $ _ M & Eth??? ??ce in 1 .6 x 10 - 1 , 6 x 10M 6 . 4 x 10 M 6 , 4 x 10 -5M NNmet n 32 X 10 .5M . .. 3 .2x10 -SM U . S . Patent Jan . 2 , 2018. Sheet. 2 of 16 . US 9 ,855 , 225 B2 No treatment ????? 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WORYna wa n aoty XYMAXNo treatment 00 Sexo opaguodoogooooooo w Ethanol Vehicle IL-2(pg/mL) * w JWH -015 w tooteggere 2 4 16 32 Concentration JWH -015 ( M ) 6007PODCOCOCO ogP C * * o hraneifrom ti 400 ) INanomat WT: DMSO Vehicle IL-2(pg/mL) KO : 0 - 1966 * atau * en on KO : DMSO Vehicle * * * * montering WT: 0 - 1966 cococacoooooooooooooooooooooooooooooduonossono be 1 2 4 8 16 32 ConcenConcentration 0 - 1966 ( UM ) FIGURE 4B U . S . 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Patent Jan . 2 , 2018 Sheet 7 of 16 US 9 ,855 , 225 B2 0 - 1966 * IL-10(pg/ml) * o n . to v . wwwa Wa vo memea with.www wat wes * apiyeA * p < p apun * ** p < 00 . 0101 2 4 8 16 32 Concentration 0 - 1966 (UM ) Figure 7 U . S . Patent Jan . 2 , 2018 Sheet 8 of 16 US 9 ,855 , 225 B2 PercentTregs oba * p < 0 .01 0 24 48 II Time (hours ) No TreatmentTreatme go Vehicle 0 -1966 ($ 32 PM ) Figure 8 U . S . Patent Jan . 2 , 2018 Sheet 9 of 16 US 9 ,855 , 225 B2 ooo0000000000000000 poooooooooooooooooooooooooooooooooooo o oooo o o va Vehicles Exer * * 2 . Y a p , R299 * * * * * * * No Treatment P ASSALAMU WANN A LA ' up ve asXapusuoisselddns * ANN * withNNNNNNN ? XXXX * * A WWW on and . pomarawwwwwwwwwwwwwtocoLALO vetene anti L 10 ( 1 uglmL ) * No antibody O. BA 32 Concentration 0 - 1966 (UM ) Figure 9 U . S . Patent Jan . 2 , 2018 Sheet 10 of 16 US 9 ,855 ,225 B2 PercentTregs WWW Wooowoooo No Vehicle 0 . 1966 Q . 1966 O 1966 Treatment 32 um 16uM 8AM No Antibody 1 . 0 ug mL/ 1 . 6 ugml/ Figure 10 enU . S . Patentmoet wowJan . 2 , 2018 Sheet. 11. of 16 . US 9 ,855 ,225 B2 XX X * * * 4 ???????????10 12 14 Day (Post operative ) gent 0 - 1966 Vehicle Figure 11 0 atent Jan . 2 , 2018 Sheet 12 of 16 US 9 , 855 ,225 B2 000000AAAAAAAA A INCA 0000000000000000A ? MeancPMx103SD woooowwook Sinatia. ? * * 0 . 00 Vehicle 1966 Figure 12 . U . S . Patent Jan . 2 , 2018 Sheet 13 of 16 US 9 ,855 ,225 B2 ooooooo XXXXXXXX000000odds Foxp3*cells(%ofCD4+Cells) oooooooooooooooooooooo0000000oooooooooooooooo SAXXXXXXX qöboooooooooooooo * p < 0 .01 Vehicle 0 . 1966 Figure 13 atent Jan . 2 , 2018 Sheet 14 of 16 US 9 ,855 ,225 B2 1494D DDDDD ws %Tregs 4 WWW 0 . 01 Vehicle 0 - 1966 Figure 14 atent Jan . 2 , 2018 Sheet 15 of 16 US 9 , 855 ,225 B2 8 mmmmmm |_.sowwuxiaomusion Assong * : ?? :??? ?? Seedassales : ???? 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For under 35 U . S . C . $ 371 of International Application No .: example , one of the drawbacks of using marijuana , a CB , PCT/ US2012 /051330 , which was filed Aug . 17 , 2012 , and CB2 receptor agonist, is the resulting psychotropic effects which claimsthe benefit of the filing date of U . S . Provisional due to CB2 receptor activation . Cannabinoids that selec Application No . 61 / 524 ,883 , which was filed on Aug . 18 , " tively target CB , receptors , and thus , the immune system , 2011. For the purpose ofany U . S . application that may claim have great clinical potential as therapeutic agents in the the benefit of U . S . Provisional Application No . 61/ 524 , 883 , treatment of graft rejection since they are expected to have the contents of that earlier filed application are hereby little or no CB , receptor activity ( i. e . psychotropic effects ) . incorporated by reference in their entirety . Current immunosuppressant therapies used to prevent or 15 block tissue rejection in organ transplantation are associated U . S . GOVERNMENT RIGHTS with significant untoward effects . For example, toxicity with chronic tacrolimus use is associated with post - transplanta This invention was supported in part by United States tion diabetes mellitus (PTDM ) due to the death of pancreatic Government Grant No . DA13429 , DA06650 and T32 - 20 islet cells , and also with kidney damage . Tacrolimus and DA07237 awarded by National Institutes of Drug Abuse .