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(12) Patent Application Publication (10) Pub. No.: US 2012/0083445 A1 Tseng Et Al

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(12) Patent Application Publication (10) Pub. No.: US 2012/0083445 A1 Tseng Et Al US 20120083445A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2012/0083445 A1 Tseng et al. (43) Pub. Date: Apr. 5, 2012 (54) COMPOSITIONS CONTAINING HC-HA Publication Classification COMPLEX AND METHODS OF USE THEREOF (51) Int. Cl. A 6LX 38/57 (2006.01) (75) Inventors: Scheffer Tseng, Pinecrest, FL (US); A6IPCI2P 2L/027/02 30:2006.O1 8: Hua He, Miami, FL (US) A6IP 9/00 (2006.01) A6IP37/06 2006.O1 (73) Assignee: TissueTech, Inc., Miami, FL (US) C07K I4/8 308: A6IP 29/00 (2006.01) (21) Appl. No.: 13/262,725 (52) U.S. Cl. ... 514/13.3: 530/380: 514/20.3: 435/69.2: 514/18.6 (22) PCT Filed: Apr. 26, 2010 (57) ABSTRACT (86). PCT No.: PCT/US 10/32452 Disclosed herein, in certain embodiments, is an HCHA com S371 (c)(1) plex comprising hyaluronan and a heavy chain of ICI. (2), (4) Date: Dec. 16, 2011 wherein the transfer of the heavy chain of ICI is catalyzed by s 9 TSG-6. Further disclosed herein, in certain embodiments, is O O an HCHA complex comprising hyaluronan and a heavy Related U.S. Application Data chain of ICI, wherein the transfer of the heavy chain of ICI is (60) Provisional application No. 61/267,776, filed on Dec. catalyzed by the TSG-6 like protein. Additionally, disclosed 8, 2009, provisional application No. 61/172,621, filed herein are methods of manufacturing said complex and meth on Apr. 24, 2009. ods of use thereof Patent Application Publication Apr. 5, 2012 Sheet 1 of 14 US 2012/0083445 A1 A A: Ex3 . O O.O2 OS C, OH 8 9 10 1 12 :::::::::::::::::::::: :--Rigi k3 a 53- re 88 8-all- st F-S i i E 48 S : , . FIGURE 1 Patent Application Publication Apr. 5, 2012 Sheet 2 of 14 US 2012/0083445 A1 FIGURE 1 D Patent Application Publication Apr. 5, 2012 Sheet 3 of 14 US 2012/0083445 A1 A. B 3. s: 2. A. s SS- S-S SS S- 85-8 . a. s S. & . S.Š...S. : 33. -------- -S-8 :S-3s: S& ssssss R. SS Y-3's - ::::::::::::: FIGURE 2 Patent Application Publication Apr. 5, 2012 Sheet 4 of 14 US 2012/0083445 A1 ?:y:fr->--~~~~~~~~~~~~~~~~) ·[Ti a si: E ?;----L? FIGURE 3 Patent Application Publication Apr. 5, 2012 Sheet 5 of 14 US 2012/0083445 A1 j, FIGURE 4 Patent Application Publication Apr. 5, 2012 Sheet 6 of 14 US 2012/0083445 A1 A -- HA--TSG-3 o E e 15 3. 8 .5 25, 5th 2.) HA Concentration giml) B reconstitution of HC-HA (rchCHA) in vitro M Oil None HAase gCH 1 2 3 4 5 6 7 8 9 O 11 HCeHA O. HC bikin HCs FIGURE 5 Patent Application Publication Apr. 5, 2012 Sheet 7 of 14 US 2012/0083445 A1 A TGF-31 Promoter Activity SCO S. t 20000. : S log Ctrl HA HA HATSG-6 HCHA HCHA B MTT Assay O.8 k : E O.S c g 0.4 .2 S. 0 - K Ctrl HA Afd HATSG-E HCHA HCHA FIGURE 6 Patent Application Publication Apr. 5, 2012 Sheet 8 of 14 US 2012/0083445 A1 S. S. s . i. s FIGURE 7 Patent Application Publication Apr. 5, 2012 Sheet 9 of 14 US 2012/0083445 A1 FIG. 8 Patent Application Publication Apr. 5, 2012 Sheet 10 of 14 US 2012/0083445 A1 & S S & S.X S Ayyyy S S S Š Š Š Š S Š S Š S S Š S. S. FIG. 9 Patent Application Publication Apr. 5, 2012 Sheet 11 of 14 US 2012/0083445 A1 r S 3. L. Fraction No. 8.8rn: frarrior Fraction: No. 3.8m ffrartsot: Protein • Iril Q Donor 1 - 3 E. & E is 8 ' E L E 3 4 5. 0.3milraction Figure 10 Patent Application Publication Apr. 5, 2012 Sheet 12 of 14 US 2012/0083445 A1 '''''''' assassassissassissasssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssss. FIGURE 11 Patent Application Publication Apr. 5, 2012 Sheet 13 of 14 US 2012/0083445 A1 s 4. Y - Y - Y Y. YY Y- C-33A 3. Y Y Y V V V v N. Y. V. iCEE f : th........... -0.4 is is - - - - Sr. FIGURE 12 Patent Application Publication Apr. 5, 2012 Sheet 14 of 14 US 2012/0083445 A1 i,4 Š & \ w & -C-A, Š Y, AFAE 8 : \ ^ f \ y Š CA to *, N C}} E. 0.2 i. - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -0.20.01 . S -0.4 log HCHA concentration (agfin FIGURE 13 US 2012/0083445 A1 Apr. 5, 2012 COMPOSITIONS CONTAINING HC-HA inflammatory agent. In some embodiments, the method fur COMPLEX AND METHODS OF USE ther comprises administering an additional antibiotic agent. THEREOF 0007 Disclosed herein, in certain embodiments, is a method of reducing or preventing angiogenesis comprising CROSS-REFERENCE administering an HCHA complex of any of claims 1-6 to an individual in need thereof. In some embodiments, the HCHA 0001. This application claims the benefit of U.S. Provi complex is produced by contacting (a) hyaluronan, (b) a sional Application No. 61/172,621, filed 24 Apr. 2009, and heavy chain of ICI, and (c) TSG-6. In some embodiments, the U.S. Provisional Application No. 61/267,776, filed 8 Dec. HCHA complex is produced by contacting (a) hyaluronan, 2009 both of which are incorporated herein by reference. (b) HC1 and HC2 of ICI, and (c) TSG-6. In some embodi ments, the HCHA complex is produced by contacting (a) BACKGROUND OF THE INVENTION hyaluronan, (b) a heavy chain of ICI, and (c) the TSG-6 like protein. In some embodiments, the HCHA complex is pro 0002 The amniotic membrane (AM) is the innermost duced by contacting (a) hyaluronan, (b) HC1 and HC2 of ICI, membrane enwrapping the fetus in the amniotic cavity. The and (c) the TSG-6 like protein. In some embodiments, the AM consists of a simple epithelium, a thick basement mem method further comprises co-administering an additional brane, and an avascular stroma. chemotherapeutic agent. 0008 Disclosed herein, in certain embodiments, is a SUMMARY OF THE INVENTION method of preventing transplant rejection comprising con tacting a tissue or a plurality of cells with an HCHA complex 0003 Disclosed herein, in certain embodiments, is an of any of claims 1-6. In some embodiments, the method HCHA complex comprising hyaluronan and a heavy chain further comprises contacting the tissue or plurality of cells of ICI, wherein the transfer of the heavy chain of ICI is with reperfusion solution. In some embodiments, the HCHA catalyzed, at least in part, by TSG-6, recombinant TSG-6, complex is produced by contacting (a) hyaluronan, (b) a TSG-6 like protein, recombinant TSG-6 like protein, or a heavy chain of ICI, and (c) TSG-6. In some embodiments, the combination thereof. In some embodiments, the HCHA HCHA complex is produced by contacting (a) hyaluronan, complex comprises HC1 and HC2 of ICI. In some embodi (b) HC1 and HC2 of ICI, and (c) TSG-6. In some embodi ments, the HCHA complex has a purity of at least 75%. ments, the HCHA complex is produced by contacting (a) 0004 Disclosed herein, in certain embodiments, is an hyaluronan, (b) a heavy chain of ICI, and (c) the TSG-6 like HCHA complex comprising hyaluronan and a heavy chain protein. In some embodiments, the HCHA complex is pro of ICI, wherein the transfer of the heavy chain of ICI is duced by contacting (a) hyaluronan, (b) HC1 and HC2 of ICI, catalyzed by the TSG-6 like protein and/or a recombinant and (c) the TSG-6 like protein. In some embodiments, the TSG-6 like protein. In some embodiments, the HCHA com method further comprises co-administering an additional plex comprises HC1 and HC2 of ICI. In some embodiments, immuno-Suppressive agent. the HCHA complex has a purity of at least 75%. 0009 Disclosed herein, in certain embodiments, is a 0005 Disclosed herein, in certain embodiments, is a method of manufacturing an HCHA complex comprising, method of reducing or preventing inflammation, comprising contacting (a) HA; (b) HC1 and HC2 of ICI, wherein at least administering an HCHA disclosed herein to an individual in one of HC1 and HC2 is optionally recombinant; and (c) need thereof. In some embodiments, the HCHA complex is TSG-6 or TSG-6 like protein, wherein the TSG-6 or TSG-6 produced by contacting (a) hyaluronan, (b) a heavy chain of like protein is optionally recombinant. In some embodiments, ICI, and (c) TSG-6. In some embodiments, the HCHA com the method further comprises a bioreactor. In some embodi plex is produced by contacting (a) hyaluronan, (b) HC1 and ments, the method further comprises a plurality of cells HC2 of ICI, and (c) TSG-6. In some embodiments, the wherein the cells are engineered to constitutively express HCHA complex is produced by contacting (a) hyaluronan, TSG-6 or TSG-6 like protein. In some embodiments, the (b) a heavy chain of ICI, and (c) the TSG-6 like protein. In method further comprises a plurality of cells wherein the cells some embodiments, the HCHA complex is produced by are engineered to constitutively express HC1, HC2, or both. contacting (a) hyaluronan, (b) HC1 and HC2 of ICI, and (c) 0010 Disclosed herein, in certain embodiments, is a the TSG-6 like protein. In some embodiments, the method method of isolating HCHA from amniotic material compris further comprises administering an additional anti-inflamma ing: (a) processing the amniotic material Such that it is Suit tory agent. In some embodiments, the method further com able for extraction of an HCHA complex; and (b) extracting prises administering an additional antibiotic agent.
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