Frank Greenway, M.D.Pennington Center

March 3, 2009

Angiogenesis

• Angiogenesis, growing new blood vessels, is a basic biological process

• Angiogenesis in adults is used for menses, fetal development and healing wounds.

• Inhibition of angiogenesis in adults should have minimal side effects or toxicity.

• Cancer cannot grow past 2mm without angiogenesis and fat needs angiogenesis too.

Human Angiogenesis Assay

• Discs are cut from the veins of placentas (after births). These placentas are left over after the delivery of a baby and would be discarded. The discs are put in a clot-like gel.

• The discs sprout new blood vessels (initiation) and the new vessels continue to gro over a 2-week period (angiogenic index from 1-16)

Human Tissue Predicts Activity

Cmpd Human Animal Clinical Use

Steroid Yes Yes Yes Psoriasis

Heparin Yes Yes Yes Tumors

Noni Yes Yes Herb Tumors

Taxol Yes Yes Yes Tumors

Endost. No 10-5m Yes Yes Tumors

Angiost. No 10-4m Yes Yes Tumors

Jung SP et al. Angiogenesis. 4(3):175-86, 2001.

Hornick CA et al. Angiogenesis. 6(2):143-9, 2003.

Sweet Leaf Tea

• This is a different tea than tea from the plant Camilla, the tea consumed in Britain, the US (black) or in Chinese restaurants (green)

• Sweet leaf tea is sweet due to a chemical made by the plant called Ruboside that is a non-caloric sweetener.

• Sweet leaf tea is consumed as part of the diet in an area in China and is related to blackberry, a plant called Rubus.

Black Raspberry

Inhibition of Angiogenesis by Sweet Leaf Tea in placental vein assay

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Fat is an Angiogenic Tissue

• Fat grows and recedes throughout life and is composed basically of fat and vessels.

• TNP-470 is a fungus-derived angiogenic inhibitor that makes people very sick.

• TNP-470 and other angiogenic inhibitors have been shown to reverse obesity in rodent models of obesity and it does not make the rodents sick.

Intermittent Treatment with TNP-470, 10 mg/kg/d

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Human Angiogenesis Assay in Fat Tissue

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Sweet Leaf Tea Inhibits Angiogenesis in Human Fat Tissue

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Figure 6. Anti-angiogenic property of the crude and purified sweet leaf tea extracts assayed in a human adipose tissue angiogenesis model. Crude extract showed a partial inhibitory activity against overall angiogenic response (left) whereas the purified extract at the same concentration of 0.5 mg/ml completely inhibited angiogenic responses. The purified extract also totally prevented the initiation of new angiogennicblood vessels (right) compared to partial activity of the crude extract.

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Figure 6. Anti-angiogenic property of the crude and purified sweet leaf tea extracts assayed in a human adipose tissue angiogenesis model. Crude extract showed a partial inhibitory activity against overall angiogenic response (left) whereas the purified extract at the same concentration of 0.5 mg/ml completely inhibited angiogenic responses. The purified extract also totally prevented the initiation of new angiogennicblood vessels (right) compared to partial activity of the crude extract.

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Partially Purified Extract with 13 Chemicals

Purification and Standardization

• Purification gets rid of the plant chemicals and other plant components that do not inhibit angiogenesis. Purification resulted in 5 chemicals important for effectiveness

• Standardization is the identification of the important chemicals and being sure that every batch of the purified plant has the full amount of the effective chemicals. Standardization is necessary for a consistent product.