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C12) United States Patent (IO) Patent No.: US 9,534,212 B2 Turner (45) Date of Patent: Jan.3,2017

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C12) United States Patent (IO) Patent No.: US 9,534,212 B2 Turner (45) Date of Patent: Jan.3,2017 IIIIII IIIIIIII Ill lllll lllll lllll lllll lllll lllll lllll lllll 111111111111111111 US009534212B2 c12) United States Patent (IO) Patent No.: US 9,534,212 B2 Turner (45) Date of Patent: Jan.3,2017 (54) HEAT-INACTIVATED COMPLEMENT FOREIGN PATENT DOCUMENTS FACTOR B COMPOSITIONS AND METHODS WO W02004060817 * 7/2004 (71) Applicant: WILLIAM MARSH RICE WO W02011/058286 * 5/2011 UNIVERSITY, Houston, TX (US) OTHER PUBLICATIONS (72) Inventor: Nancy A. Turner, Houston, TX (US) WHO Technical Report, Guidelines on viral inactivation and (73) Assignee: WILLIAM MARSH RICE removal procedures intended to assure the viral safety of human UNIVERSITY, Houston, TX (US) blood plasma products, Series No. 924, 2004.* Zheng et al., Effect of plasma exchange on plasma ADAMTS13 metalloprotease activity, inhibitor level, and clinical outcome in ( *) Notice: Subject to any disclaimer, the term ofthis patients with idiopathic and nonidiopathic thrombotic patent is extended or adjusted under 35 thrombocytopenic purpura, Blood, Jun. 1, 2004 vol. 103, No. 11.* U.S.C. 154(b) by O days. Soltis, R. et al. "The Effect of Heat Inactivation of Serum on Agregation of Immunoglobulins." 1979. Immunology, vol. 36, pp. (21) Appl. No.: 14/631,263 37-45; p. 37, Right Colunm, Second Paragraph; p. 38, Left Colunm, Third Paragraph. Belogrudov, G. et al. "Factor B and Mitochondrial ATP Synthase (22) Filed: Feb. 25, 2015 Complex." Dec. 14, 2001. Journal of Biological Chemistry; vol. 277, pp. 6097-6103; p. 6098, Left Colunm, Fourth and Sixth (65) Prior Publication Data Paragraphs. DOI: 10.1074.BC.Mlll256200. US 2015/0175994 Al Jun. 25, 2015 Le, G. et al. "Profiling the Enzymatic Properties and Inhibition of Human Complement Factor B." Oct. 5, 2007. Journal of Biological Chemistry. vol. 282, pp. 34809-34816. Abstract. DOI: 10.1074/JBC. Related U.S. Application Data M705646200. Chapin J, Weksler B, Magro C, Laurence J. Eculizurnab in the (63) Continuation of application No. Treatment of Refractory Idiopathic Thrombotic Thrombocytopenic PCT/US2013/056609, filed on Aug. 26, 2013. Purpura. [Letter]. BR J Haernatol. 2012;157 (6):772-774. Waters AM, Licht C. Ahus Caused by Complement Dysregulation: (60) Provisional application No. 61/693,633, filed on Aug. New Therapies on the Horizon. Pediatric Nephrology. 2011;26 27, 2012. (1):41-57. Hellstern P, Solheim BG. The Use of Solvent/Detergent Treatment in Pathogen Reduction of Plasma. Transfus Med Hemother. 2011 ;38 (51) Int. Cl. (1):65-70. C12N 9/50 (2006.01) A61K 38/00 (2006.01) * cited by examiner C12N 9/64 (2006.01) A61K 38/48 (2006.01) Primary Examiner - Suzanne M Noakes (52) U.S. Cl. Assistant Examiner - Stephen Chong CPC ................. C12N 9/50 (2013.01); A61K 38/00 (74) Attorney, Agent, or Firm - Robert R. Riddle; Reed (2013.01); A61K 38/482 (2013.01); C12N Smith LLP 9/6424 (2013.01); Cl2Y 304/21047 (2013.01) ( 58) Field of Classification Search (57) ABSTRACT None The present disclosure is directed to compositions compris­ See application file for complete search history. ing heat-inactivated complement factor B and methods of using the same to treat thrombotic or complement-mediated (56) References Cited inflammatory disorders. U.S. PATENT DOCUMENTS 6 Claims, 36 Drawing Sheets 2010/0120665 Al* 5/2010 Kaleko et al. .................... 514/8 (26 of 36 Drawing Sheet(s) Filed in Color) U.S. Patent Jan.3,2017 Sheet 1 of 36 US 9,534,212 B2 FIGURE IA 10 CF! (A C2 0,001 CFD C1QB OJlOOI U.S. Patent Jan.3,2017 Sheet 2 of 36 US 9,534,212 B2 FIGURE 1B Hi {a) ... .7.hrs ......24 hrs .. 14 {) C!QB C4 C2 0 CFB CFO C5 CFH CFI CFP VWF AD U.S. Patent Jan.3,2017 Sheet 3 of 36 US 9,534,212 B2 FIGURE lC - l eh }') -l i :1 :: 1 L :~_cj_LI_ 30 min I hr 3 hrs 5 hrs 7 hrs 24 hrs 48 hrs 72 hrs 96 hrs U.S. Patent Jan.3,2017 Sheet 4 of 36 US 9,534,212 B2 FIGURE2A (:?. 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U.S. Patent Jan.3,2017 Sheet 12 of 36 US 9,534,212 B2 FIGURES A H.lJVEC-re1ea8e<l Complement Components ':i ( .•.J FB FD C5 Fl FH FP J-i C!q C4 }--1 0 l5 20 25 JO 35 4(l 50 'h, of the Lrni:~ih ofULVWF String ,viih Ea('.b (\,mp!emeni C:,;nponem Atrnched U.S. Patent Jan.3,2017 Sheet 13 of 36 US 9,534,212 B2 FIGURE 5 (cont) B HUVEC-released Complement Components plus Heated Serum FB .,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,:,, FD :=:Jh F! .:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:i:id fH ::::::::::::::::::::::::::::::::::::::::::::::::::::::::::·}-------------- : fl' '1mmm11,,.....-- C!q / .......................................................... l·····························································' C4 r, n 50 U.S. Patent Jan.3,2017 Sheet 14 of 36 US 9,534,212 B2 F1GURE6 B 1¥) 1Stl JJO 2$0 JJO J~O :tK~ tftH;i:r:{~;, i:.:t{i:s U.S. Patent Jan.3,2017 Sheet 15 of 36 US 9,534,212 B2 FIGURE 6 (cont) I) U.S. Patent Jan.3,2017 Sheet 16 of 36 US 9,534,212 B2 FIGURE7 U.S. Patent Jan.3,2017 Sheet 17 of 36 US 9,534,212 B2 FIGURE 7 (cont) U.S. Patent Jan.3,2017 Sheet 18 of 36 US 9,534,212 B2 FIGURE SA U.S. Patent Jan.3,2017 Sheet 19 of 36 US 9,534,212 B2 :FIGURE SA (cont) U.S. Patent Jan.3,2017 Sheet 20 of 36 US 9,534,212 B2 FIGURE 8B U.S. Patent Jan.3,2017 Sheet 21 of 36 US 9,534,212 B2 FIGURE 8B (cont) .,_,,'»~ 3-5-0:.) -~ 3.0(X) ~ :fil 2~--c~.;: ~ J.O{XJ t 1S-OO >.-. g 1000 M. S-00· {.) {l 4 U.S. Patent Jan.3,2017 Sheet 22 of 36 US 9,534,212 B2 FIGURE8C U.S. Patent Jan.3,2017 Sheet 23 of 36 US 9,534,212 B2 FIGURE SC (cont) I ;~ C l(i 10 JC 4 U.S. Patent Jan.3,2017 Sheet 24 of 36 US 9,534,212 B2 FIGURE 9 A FH >· 3::itY.) _..~ 1c-0n }~:=~~ ::: ;,:, .r 2::;00 ~ ?.f~K> "» ~ 1$()!-.~ ]" JO(;(} u.. ~:lK> 0 lc'~Jl~i :s 4 U.S. Patent Jan.3,2017 Sheet 25 of 36 US 9,534,212 B2 FIGURE 9 (cont) B Fl U.S. Patent Jan.3,2017 Sheet 26 of 36 US 9,534,212 B2 FIGURE 9 (cont) U.S. Patent Jan.3,2017 Sheet 27 of 36 US 9,534,212 B2 FIGURE 10 A C3 U.S. Patent Jan.3,2017 Sheet 28 of 36 US 9,534,212 B2 FIGURE 10 (cont) ................·lJlVW!' 500 () 5 40 4 U.S. Patent Jan.3,2017 Sheet 29 of 36 US 9,534,212 B2 FIGURE 10 (cont) B cs "':350{> M t. 3()00 .fil"' 250(> C 50 4 U.S. Patent Jan.3,2017 Sheet 30 of 36 US 9,534,212 B2 FIGURE 10 (cont) U.S. Patent Jan.3,2017 Sheet 31 of 36 US 9,534,212 B2 FIGURE 11 U.S. Patent Jan.3,2017 Sheet 32 of 36 US 9,534,212 B2 FIGURE 11 (cont) E 12 3 4 5 6 7 ,\{!(;(!+ + + + + + + 0 w 40 U.S. Patent Jan.3,2017 Sheet 33 of 36 US 9,534,212 B2 FIGURE 12 U.S. Patent Jan.3,2017 Sheet 34 of 36 US 9,534,212 B2 FIGURE 12 (cont) U.S. Patent Jan.3,2017 Sheet 35 of 36 US 9,534,212 B2 FIGURE 13 7000 Complement component intensity C: per micron of ULVWF string -~2 6000 · E.... (D 5000 0. >- -·w 4000 · C: (l) C: -...., 3000 C: tD 0 r{)_ 2000 ,_tD 0 ::::, 1000 .. w... 0 C3 FB FD C5 FH Fl FP C4 U.S. Patent Jan.3,2017 Sheet 36 of 36 US 9,534,212 B2 FIGURE 14 7000 ··•·•• HUVEC-re!eased C: E 6000 fl HUVEC ·t NS 0 .E •y· 1'.. 5000 ~ Q. ~ ·; 4000 e: ,S I -...t: 3000 C: I;)> (.; e{JJ 2000 .a0 u.
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