Ficus extracts having angiogenesis inhibiting activity and methods of isolating and using the same

A process for obtaining a Ficus extract having angiogenesis inhibiting activity in a human or animal, the process comprising:

• a) preparing an aqueous extract from a latex-containing portion of a Ficus variant;
• fractionating the aqueous extract;
• and
• b) selecting as the Ficus extract having angiogenesis inhibiting activity in a human or animal, a substantially protein-free fraction containing one or more compounds having a molecular weight of about 200 Da to about 2,000 Da, wherein the Ficus extract exhibits angiogenesis inhibiting activity.

A method of screening a Ficus extract having angiogenesis inhibiting activity, the method comprising:

• a) isolating a Ficus extract having angiogenesis inhibiting activity by a process comprising:
• mixing a latex-containing portion of a Ficus variant with a polar solvent to provide a liquid extract;
• b) washing the liquid extract with an organic solvent to provide an aqueous extract;
• and fractionating the aqueous extract to provide an extract having angiogenesis inhibiting activity;
• c) applying the Ficus extract having angiogenesis inhibiting activity to a tumor;
• and measuring neovessel growth from the tumor.

A Ficus extract having angiogenesis inhibiting activity and having a chromatograph having peaks at about 4.9 minutes, about 6.0 minutes, about 21.3 minutes, about 22.5 minutes, about 35.0 minutes, and about 49.7 minutes, when subjected to HPLC using a C18 reverse phase column having an internal diameter of about 4.6 mm and a length of about 250 mm;

• a) wherein a first eluent is acetonitrile and a second eluent is water containing about 0.3% phosphoric acid and about 2.5% acetonitrile, the concentration of the first eluent and second eluent is 100% by volume, and from 0 minutes to about 20 minutes the first eluent increases linearly from 0% to about 10% by volume, from about 20 minutes to about 50 minutes the first eluent increases linearly from about 10% to about 20% by volume, from about 50 minutes to about 65 minutes the first eluent increases linearly from about 20% to about 40% by volume, and from about 65 minutes to about 80 minutes the first eluent increases linearly from about 40% to about 60% by volume;
• and
• b) wherein the column temperature is about 25° C.;
• the injection volume is about 10 μL;
• the flow rate is about 1 mL/minute;
• from 0 minutes to about 80 minutes the pressure increases linearly from about 1,000 psi to about 3,000 psi;
• and detection is at about 254 nm.

A Ficus extract having angiogenesis inhibiting activity and having a chromatograph substantially in accordance with FIG. 16 when subjected to HPLC using a C18 reverse phase column having an internal diameter of about 4.6 mm and a length of about 250 mm;

• a) wherein a first eluent is acetonitrile and a second eluent is water containing about 0.3% phosphoric acid and about 2.5% acetonitrile, the concentration of the first eluent and second eluent is 100% by volume, and from 0 minutes to about 20 minutes the first eluent increases linearly from 0% to about 10% by volume, from about 20 minutes to about 50 minutes the first eluent increases linearly from about 10% to about 20% by volume, from about 50 minutes to about 65 minutes the first eluent increases linearly from about 20% to about 40% by volume, and from about 65 minutes to about 80 minutes the first eluent increases linearly from about 40% to about 60% by volume;
• and
• b) wherein the column temperature is about 25° C.;
• the injection volume is about 10 μL;
• the flow rate is about 1 mL/minute;
• from 0 minutes to about 80 minutes the pressure increases linearly from about 1,000 psi to about 3,000 psi;
• and detection is at about 254 nm.