US 2011 01 65096A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0165096 A1 Liu et al. (43) Pub. Date: Jul. 7, 2011 (54) FICUS EXTRACTS HAVING ANGIOGENESIS Publication Classification HIBITING ACTIVITY AND METHODS OF (51) Int. Cl. SOLATING AND USING THE SAME A6IR 9/12 (2006.01) A636/00 (2006.01) CI2O 1/02 (2006.01) (75) Inventors: Zhijun Liu, Baton Rouge, LA A6IP35/00 (2006.01) (US); Eugene A. Woltering, A6IP3L/00 (2006.01) Kenner, LA (US) A6IP3L/04 (2006.01) A6IP37/00 (2006.01) A6IP27/02 (2006.01) (73) Assignee: ULLMAN MEDICAL, INC., A6IP 9/02 (2006.01) Silver Spring, MD (US) A6IP3/10 (2006.01) A6IP 7/02 (2006.01) A6IP 7/06 (2006.01) A6IP3/04 (2006.01) (21) Appl. No.: 13/058,957 A6IP4/00 (2006.01) A6IP 9/12 (2006.01) A6IP 9/00 (2006.01) (22) PCT Filed: Aug. 14, 2009 A6IPI/00 (2006.01) (52) U.S. Cl. .............................. 424/45; 424/725; 435/29 (86). PCT No.: PCT/USO9/53860 (57) ABSTRACT The present invention is directed to methods for extracting S371 (c)(1), and isolating extracts having angiogenesis inhibiting activity (2), (4) Date: Mar. 17, 2011 from a latex-containing portion of a Ficus variant, pharma ceutical and nutraceutical compositions comprising the extracts, methods of administering the extracts to treat angio Related U.S. Application Data genesis-dependent diseases and to reduce or inhibit neovessel growth in a Subject in need thereof, and the use of the extracts (60) Provisional application No. 61/088.924, filed on Aug. in the manufacture of a composition for reducing or inhibiting 14, 2008. neovessel growth. Patent Application Publication Jul. 7, 2011 Sheet 1 of 23 US 2011/O165096 A1 100 FIG. 1 Patent Application Publication Jul. 7, 2011 Sheet 2 of 23 US 2011/O165096 A1 Discard Solids S/S 22 225 M liquid Extract 24 are - P -- a Organic Fraction (F) Y242 Aqueous Extract (F) \ 43 Diluted Aqueous Extract Column Fractionation Eluted with DE HO (Fo) 27 Eluted with EtOH:HO (Fass) \)72 FIG. 2 Patent Application Publication Jul. 7, 2011 Sheet 3 of 23 US 2011/O165096 A1 300 312 31 FIG. 3A FIG. 3B Patent Application Publication Jul. 7, 2011 Sheet 4 of 23 US 2011/O165096 A1 4OO Mean Angiogenic Index = 1+1+1+1 = 4 FIG. 4A Mean Angiogenic Index = 4+4+4+4 = 16 FIG. 4B Patent Application Publication Jul. 7, 2011 Sheet 5 of 23 US 2011/O165096 A1 Mean Angiogenic Index (Ficus carica L., variety kadota, day 14) (series 1A) DExcluding Zeros DIncluding Zeros d s 3 3 38 g 3 E 3 E st S S SS2 S. S. C. S. C. S. w CN o S CN o S (NE (NE da o da C) 8 C N). C C N C C N1 N1 a E a E as CD go 8 cc E S C8 d). g 3- it c. it c. d d Treatment FIG. 5A Angiogenesis Initiation (%) 80 (Ficus carica L., variety kadota, day 14) (series 1A) 70 – 60 - 9. 50 – S. 30 — is 20 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 10 – O 9 £ 35, 35 S.F S.F D - - s s - N N CN E CN E cN SC) cN EC. CC N4 W- N1 cy a N1 Na N1 O CD o o O) Co Oc C. v . C. c. 3-es ifC S ifC S H- O H- C C O Treatment FIG. 5B Patent Application Publication Jul. 7, 2011 Sheet 6 of 23 US 2011/O165096 A1 Mean Angiogenic Index 11 (Ficus carica L., variety kadota, day 14) (series 1B) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - DIncluding(IDExcluding Zeros Zeros g 3 - S - 3 3 S 3 s 58 C w CN o S CN o S C E N CN c es CD go 8 cc E o O3 d g 3. in c. in c. O O Treatment F.G. 6A Angiogenesis initiation (%) 70 ----------------------------- (Ficus carica L., variety kadota, day 14) (series 1B). 60 - Initiati to 50 40 30 - S. 20 - 10 O – s- St3 E- S3 2 - S.3 SS. SR3 5 SESS SS38 8 CC N2 R S S2 C. S & N1 N2 E S2 es CD go 8 cc odd O3 g . o st- it c ic O C Treatment F.G. 6B Patent Application Publication Jul. 7, 2011 Sheet 7 of 23 US 2011/O165096 A1 Mean Angiogenic Index (Ficus carica L., variety kadota, day 14) (series 1C) DExcluding Zeros DIncluding Zeros 5 C CC CC cd CO - CO - 9 "as 9 - 9 - - & E SE O E O E C c 8 CN go S CN go S S S cN 9) cN 9) 3 N. C. S & C S N) Y S. E S. E as go E go E is .95 g St. CD in c. - CS C C Treatments Angiogenesis Initiation (%) (Ficus carica L., variety kadota, day 14) (series 1C) Initiation 9 C5 SCO SCO SE SE go(O -8 goCO -8 E < e SS E S 3 S 2 S 2 SS, SS 3 S. S N. C. raS. Na Q S.9 N)S. Y.SS N2 E & E St.ea CD go g g os 23cs g g g Treatment FIG. 7B Patent Application Publication Jul. 7, 2011 Sheet 8 of 23 US 2011/O165096 A1 Mean Angiogenic Index (Ficus carica L., variety kadota, day 14) (series 1D) 3 |Excluding Zeros DIncluding Zeros 2 1 O d s InS S SE L.SE SE SE t E 9 W- Scy O Gs O Gs O S. O S. w- & S & SS Sg Sg N. O. N to 8 a. a S. g. a g a N2 E Na E as (5 E () E (p & D8 (p (p 2 3t - C - O C C Treatments FIG. 8A Angiogenesis initiation (%) 30 (Ficus carica L., variety kadota, day 14) (series 1D) 25 - Initiation 20- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 15 - d 10 al O s g g E gE SE SE SE S SS, SS SS SS SS SS O - cN S CN S E SE Na E & E C k N. W. M cy) - cy) - cy 1. CD isOS isC 9. it9 cC. isop C it 2 itS2' Ye' C P Treatments FI G Patent Application Publication Jul. 7, 2011 Sheet 9 of 23 US 2011/O165096 A1 Mean Angiogenic Index (Ficus carica L., variety kadota, day 14) (series 1E) T Excluding Zeros DIncluding Zeros 234.5671 O C C CC CC C - CC - 9 C C as E as 8 S E S E w-C - &92 - Sc. &92 38cyd SgC is SgC is CNo Sto CNo Sto 8--a CDCC YgDo goa 2 gE gasa opY 8 S^p. S N.p S2 3t - - C C C C Treatments FIG. 9A Angiogenesis initiation (%) 7 O (Ficus carica L., variety kadota, day 14) (series 1E) 1234.56 O O s SSto E SScd E cdSS cdSS S.CO -S S.CO -S OOOOOO K- C cod C cod S. S. S as o St. o St. 3 < a - Yoo N1 Sa E Sa E --- CD CDS (DS 9 Sv 9 S 9 9 ar' - CS- - CS Co - C Treatments FIG. 9B Patent Application Publication Jul. 7, 2011 Sheet 10 of 23 US 2011/O165096 A1 Mean Angiogenic Index (Ficus carica L., variety gold, day 14) (series 2A) (IDExcluding Zeros O Including Zeros C CC CO O CO CO - (O - 9E "as &3 - S- &3 38- SSN 9 SSN 2 &SS ds &SS S. OO E 8 CD r 2 CD 2 s CD S CD CD E CD E as is E E SC 92 3. CD c. c. C C Treatments FIG. 10A Angiogenesis Initiation (%) 70 (Ficus carica L., variety gold, day 14) (series 2A) 60 - 50 - 9. S 40 E 30 9 20 10 - O o g8 g8 SS SE SE SE w K- c) O O) O C S.N s. OnS as cco asto CNC - OS E2 D E NDg E o E S 8E Y . C2E 1. s pa pa ics in c. C ics Ya-Y CD - c. - c. Treatments FIG. 10B Patent Application Publication Jul. 7, 2011 Sheet 11 of 23 US 2011/O165096 A1 Mean Angiogenic Index Patent Application Publication Jul. 7, 2011 Sheet 12 of 23 US 2011/O165096 A1 Mean Angiogenic Index 7 (Ficus carica L., variety gold, day 14) (series 2C) |Excluding Zeros DIncluding Zeros c s 8 8 - - - sts v- CDcD 3S S c 3C S dCD3E S, 93ECD S, OSSS OSS) O CDk S2us c 8s is2 c. 89 LS2 SO 89 LS 8 asN E on5 E E S S CN CN Treatments FIG. 12A Angiogenesis Initiation (%) 40 - (Ficus carica L., variety gold, day 14) (Series 2C) o 3 SE SE g E SE SE SE E - S2 S 2 se S 5.5, 55. 3 is CD S. O O CD2 c. CD(2 v . CD2. c. 1. 2 C. 9 c. CD - C - O Y- - c. - c.5 O L. C. - - Treatments FIG. 12B Patent Application Publication Jul. 7, 2011 Sheet 13 of 23 US 2011/O165096 A1 Mean Angiogenic Index 10 -------------------------(Ficus carica L., variety magnolia, day 14) (series 3A) |Excluding Zeros DIncluding Zeros cE s > 3d > 38 E SEe- as8- > S SE > 3 SE o C (DS, CDS, CD se S CD so 9s 92 9. CD S S. L. S. 8 LS 8 S. S. -- C C CN CN Ye- CN CN Treatments FIG. 13A Angiogenesis initiation (%) 70 - (Ficus carica L., variety magnolia, day 14) (Series 3A) 60 50 9. 40 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - D - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 30 - 9.is 20 - 10 – O 9 5d Scd S Scd S SCO -S SCO S S(O -E SE(O - 8E aCw &> 2- & co2 &> 2 2& 2 &> 82 SB> 8 3.es CD CD 3s (D3C.