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(12) Patent Application Publication (10) Pub. No.: US 2015/0010506 A1 Jansen Et Al

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(12) Patent Application Publication (10) Pub. No.: US 2015/0010506 A1 Jansen Et Al US 2015 0010506A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0010506 A1 Jansen et al. (43) Pub. Date: Jan. 8, 2015 (54) THERAPEUTIC PLACENTAL Publication Classification COMPOSITIONS, METHODS OF MAKING AND METHODS OF USE (51) Int. Cl. A 6LX 35/50 (2006.01) (71) Applicant: OSIRIS THERAPEUTICS, INC., A638/2I (2006.01) Columbia, MD (US) A638/17 (2006.01) A638/9 (2006.01) (72) Inventors: Timothy Jansen, Baltimore, MD (US); A638/8 (2006.01) Samson Tom, Basking Ridge, NJ (US); A638/22 (2006.01) Alla Danilkovitch, Columbia, MD (US); (52) U.S. Cl. Dana Yoo, Falls Church, MD (US); CPC ............. A61K 35/50 (2013.01); A61 K38/1825 Jaime Zerhusen, Columbia, MD (US); (2013.01); A61 K38/1808 (2013.01); A61 K Gabriele Putz. Todd, Elkridge, MD 38/1866 (2013.01); A61 K38/1841 (2013.01); A61K 38/1858 (2013.01); A61 K38/1833 (US); Amy Elizabeth Johnson, (2013.01); A61 K38/22 (2013.01); A61 K Baltimore, MD (US) 38/1754 (2013.01); A61K 38/193 (2013.01): A6 IK38/212 (2013.01) (73) Assignee: OSIRIS THERAPEUTICS, INC., USPC ......... 424/85.7; 424/93.7; 424/85.1435/374 Columbia, MD (US) (57) ABSTRACT This invention provides a therapeutic placental composition (21) Appl. No.: 14/272,339 comprising placental cells and other placental components derived from placental tissue. A cryopreserved placental composition is also provided. The placental compositions can (22) Filed: May 7, 2014 be used to stimulate and promote angiogenesis, reduce inflammation, and to reduce scar formation, among others. Related U.S. Application Data The placental tissue can optionally be an amnion, chorion, a trophoblast-depleted chorion, umbilical cord, Wharton's (63) Continuation-in-part of application No. 14/056,101, jelly, placental cotyledon, and/or maternal decidua. The pla filed on Oct. 17, 2013, which is a continuation of cental composition of the present invention is useful in treat application No. 13/030,580, filed on Feb. 18, 2011, ing a patient with a tissue injury (e.g., wound or burn) by now abandoned. applying the placental composition to the injury or in close (60) Provisional application No. 61/338,464, filed on Feb. proximity. Placental compositions may also be used to pro 18, 2010, provisional application No. 61/338,489, mote or increase regeneration of tissue. Similar application is filed on Feb. 18, 2010, provisional application No. useful with ligament and tendon repair and for engraftment 61/369,562, filed on Jul 30, 2010. procedures such as bone engraftment. Patent Application Publication Jan. 8, 2015 Sheet 1 of 28 US 201S/0010506 A1 Figure 1 Analysis of Live Cells/Gram of Chorion Patent Application Publication Jan. 8, 2015 Sheet 2 of 28 US 201S/0010506 A1 Analysis of Cells Recovered foram of Tissue Digested from a Pool of Donors .888 . 38.33 33-33 :::::::::8 3.33:38 3.388-338 Patent Application Publication Jan. 8, 2015 Sheet 3 of 28 US 201S/0010506 A1 Figure 3 Patent Application Publication Jan. 8, 2015 Sheet 4 of 28 US 201S/0010506 A1 Figure 4 Analysis of Digestion with Collagenase II 1.6OE-05 1.40E--05 120E--05 1.00E--05 8,00E--04 6.00E--04 4.00E--04 2.00E--04 0.00E--00 Digested Undigested Patent Application Publication Jan. 8, 2015 Sheet 5 of 28 US 201S/0010506 A1 H-OSS H-OSS 99/00 G?un61– |0|01 H-OSS H-OSS Patent Application Publication Jan. 8, 2015 Sheet 6 of 28 US 201S/0010506 A1 Figure 6 Analysis of Cryoprotectants D1385%DMSOinSa?ine Plasma-LyteA D13810%DMSO,5%HSAin Plasma-LyteA D13910%DMSO,5%HSAin Plasma-LyteA D1395%DMSO,HSAin Plasma-LyteA D14010%DMSO,5%HSAin D14010%G?yerco?inSabine Patent Application Publication Jan. 8, 2015 Sheet 7 of 28 US 201S/0010506 A1 S. Control eggage x5% DMSO *****.*.*.*.*.*. 5% DMSO 1%HSA o-10% DMSO 5%HSA D 3 20% ic O% O% ------------------------------------------------------------------------------------------- Baseline 1 Month 2 Months 3 Months Patent Application Publication Jan. 8, 2015 Sheet 8 of 28 US 201S/0010506 A1 Figure 8 3.33:838. 838 38.33 X. &3:38 Patent Application Publication Jan. 8, 2015 Sheet 9 of 28 US 201S/0010506 A1 Figure 9 is as fire of sixties 3.88 3.88:33 8:::8. 3. &. 8.8:48:S 3..................., Patent Application Publication Jan. 8, 2015 Sheet 10 of 28 US 201S/0010506 A1 Figure 10 A Expression of bFGF in Placental Product Derived from Chorion 3500 3OOO 25OO - 2OOO 500 OOO 14 days in culture $88: 83.8:38: Patent Application Publication Jan. 8, 2015 Sheet 11 of 28 US 201S/0010506 A1 Figure 11 Response to Hypoxia 250 :- 8 Normoxia : hypoxia Placental Product Patent Application Publication Jan. 8, 2015 Sheet 12 of 28 US 201S/0010506 A1 Figure 12 VEGF Content of Fresh v. Cryopreserved Placental Composition 500 :- & Fresh 8 Cryopreserved AO s 3OO : S. is 200 irrx:rx:r OO O ------------~-------------------~--------- D144. D45 D46 B bFGF Content of Fresh v. Cryopreserved Placental Composition 7500 ------------------------------------------------------------------------------------------------------------------------------------------ & Fresh 8: Cryopreserved 6OOO ........................................................&................ : 5 4500 i.ek...r. 3. 3OOO 1500 :- Patent Application Publication Jan. 8, 2015 Sheet 13 of 28 US 201S/0010506 A1 Figure 13 Expression of IFN-2a in placental composition derived from chorion 15 O 5 FN-2a (ne2) Expression of TGF-33 in placental compsition derived from Chorionic membrane 2 O 4.68OOOOO 2 O O TGF-33 (n=4) Patent Application Publication Jan. 8, 2015 Sheet 14 of 28 US 201S/0010506 A1 Figure 14 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - TGF-33 Content of Placental Compositions CM53 CM154 CM55 CM56 Patent Application Publication Jan. 8, 2015 Sheet 15 of 28 US 201S/0010506 A1 Figure 15 A Expression of bone reparative proteias in piaceata product derived from chorion 8:::: Patent Application Publication Jan. 8, 2015 Sheet 16 of 28 US 201S/0010506 A1 Figure 16 Patent Application Publication Jan. 8, 2015 Sheet 17 of 28 US 201S/0010506 A1 Figure 17 A Quantity of Viable Cells in 29 lots of Placental Composition 400000 --------------------------------------------------------------------------------------------------------------------------------------------------------------------- 35OOOO ~ 300000 - 250000 - - - - 200000 i . 50000 a mm a a a a a a a a a m OOOOO i . Ill. 2 3 4 5 6 8 9 O 2 3 4 5 6.7 1892O 21 22 23 24 25 26 27 28 29 Lot Nurnber % Viability of 29 lots of Placental Composition OO 90 80 70 60 50 40 30 20 O 2 3 4 5 6 7 8 9 O 1 2 3 4 5 16 7 1819 20 21, 22 23 24 25 26 27 28 29 lot Nurnber Patent Application Publication Jan. 8, 2015 Sheet 18 of 28 US 201S/0010506 A1 Figure 18 -etw. Digested Piacenta Product Patent Application Publication Jan. 8, 2015 Sheet 19 of 28 US 201S/0010506 A1 Figure 19 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - Elastase inhibition by Minced Placental Composition | Patent Application Publication Jan. 8, 2015 Sheet 20 of 28 US 201S/0010506 A1 Figure 20 VEGF Content in Minced & Digested 300 Placental Composition 2SO 200 150 OO - 50 --- Patent Application Publication Jan. 8, 2015 Sheet 21 of 28 US 201S/0010506 A1 Figure 21 VEGF Content in Minced & Digested Placental 2800 Composition after Guanidine HCl lysis Vinced lot Digested Lot 1. Vinced Lot2 Digested Lot 2 Patent Application Publication Jan. 8, 2015 Sheet 22 of 28 US 201S/0010506 A1 Figure 22 bFGF Content of Minced Placental Composition Minced lot Minced lot 2 Patent Application Publication Jan. 8, 2015 Sheet 23 of 28 US 201S/0010506 A1 Figure 23 Sustained Growth Factor Release in Minced & Digested Placental Product 2500 -O-Minced -HDigested 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Days in Culture Patent Application Publication Jan. 8, 2015 Sheet 24 of 28 US 201S/0010506 A1 Figure 24 Placental Composition after 14 Days in Culture 20min Serva Collagenase Digest 20min Worthington Collagenase Digest YY & Patent Application Publication Jan. 8, 2015 Sheet 25 of 28 US 201S/0010506 A1 Figure 25 Patent Application Publication Jan. 8, 2015 Sheet 26 of 28 US 201S/0010506 A1 Figure 26 issue Piece Sizes of Chorion Minced with a Mezzaluna Patent Application Publication Jan. 8, 2015 Sheet 27 of 28 US 201S/0010506 A1 FIGURE 27A LPS-induced TNF-o, secretion by Placental Tissues 600 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 400 -- - - us - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - g 1200 ... S. 1000 - - - - - - - - - - - - - - - - - - - - - - - - - - - - Medium St 800 ... LPS s 600 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -- 400 + - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 2OO -- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - O o . m . ...., ...a -- AM Placental issues Positive Control FIGURE 27B LPS-induced TNF-a Secretion by Placental Tissues 600 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 400 - - - - - - - - - - - - - - 1200 --------------- 5. 000 - - - - - - - - - - - - - - Medium St 800 . +LPS i 600
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