NATURAL AND SYNTHETIC COMPOUNDS FOR TREATING CANCER AND OTHER DISEASES

FIELD OF THE INVENTION

This invention provides methods of synthesizing new compounds for pharmaceutical uses.

BACKGROUND OF THE INVENTION

This invention provides methods of synthesizing new compounds for pharmaceutical uses. This invention provides methods, compounds and compositions. This invention provides compounds for up regulating levels of biomarker of Death Receptors including DR1, DR4 and DR5 or alike thereof, for treating cancer, inhibiting cancer invasion, cell invasion, or cancer cell invasion, wherein the cancers comprise breast, leukocytic, liver, ovarian, bladder, prostatic, skin, bone, brain, leukemia, lung, colon, CNS, melanoma, renal, cervical, esophageal, testicular, splenic, kidney, lymphatic, pancreatic, stomach and thyroid cancers.

SUMMARY OF THE INVENTION

This invention provides methods of synthesizing new compounds for pharmaceutical uses. This invention provides compounds, compositions, and methods for treating cancer, inhibiting cancer invasion, cell invasion, macromolecular invasion, cancer cell invasion, and metastasis. This invention provides a use of compounds, compositions, for manufacturing medicament for treating cancer, inhibiting cancer invasion, macromolecular invasion, virus invasion and metastasis. This invention provides compounds for use as mediator or inhibitor of adhesion protein or angiopoietin. This invention provides compounds for use in a method of modulating attachment or adhesion of cells or angiogenesis, by modulating or inhibiting adhesion protein macromolecules, or angiopoietin. The compounds comprise the structures selected from the formulae in the present application, wherein the compounds are synthesized or isolated, wherein the compounds comprise the saponins, triterpenes, pentacyclic triterpenes, and compounds selected from formulae in the present application, wherein the cancers comprise breast, leukocytic, liver, ovarian, bladder, prostatic, skin, bone, brain, leukemia, lung, colon, CNS, melanoma, renal, cervical, esophageal, testicular, splenic, kidney, lymphatic, pancreatic, stomach and thyroid cancers. This invention provides compounds for use as a mediator for cell circulating, cell moving, cell homing and inflammatory diseases. This invention provides compounds for improving blood circulation; soothing stroke; preventing plaque formation; and promoting their dissipation; improving blood viscosity; reducing cardiovascular clotting; reducing cerebrovascular clotting; reducing thrombosis, arteriosclerosis, coronary heart disease, hypertension, diabetes, thrombocytopenia purpura, hemoptysis, and hematemesis; treating blood in the stool, uterine bleeding, traumatic bleeding, abdominal irritation, swelling, fluttering, blood circulation, swelling, pain; treating bronchiectasis, tuberculosis and lung abscess caused by hemoptysis; reducing bleeding, antitussive, expectorant and analgesic effect, and dilating blood vessels; reducing blood pressure and the treatment of cerebral arteriosclerosis; and elevating blood lipids and reducing cholesterol.

This invention provides compounds for up regulating the level of biomarker of Death Receptors including DR1, DR4 and DR5 or alike thereof, for the treatment of cancer, and inhibiting cancer cell growth in a subject. This invention is a method for increasing the agonist activity of anti DR5 antibody. This invention is a method for increasing the level of Death Receptor of cells, up regulating biomarker of Death Receptors including DR1, DR4, DR5 or their alike, in cells, wherein the compounds are selected from this application.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. HPLC profiles of esterification products of E4A with Tigloyl chloride (A) from different times of esterification reaction. Reaction products were obtained from each time of reaction (5 sec, 1 min, 2 min, 5 min, and 10 min) were fractionated by HPLC. The profile is plotted according to HPLC elution time and optical density of fractions. The reaction was performed at room temperature (Top row) and 0° C. (bottom row).

FIG. 2. HPLC profiles of esterification products of E4A with 3,3-dimethylacryloly chloride (B) from different times of esterification reaction. Reaction products were obtained from each time of reaction (5 sec, 1 min, 2 min, 5 min, and 10 min) were fractionated by HPLC. The profile is plotted according to HPLC elution time and optical density of fractions. The reaction was performed at room temperature (Top row) and 0° C. (bottom row).

FIG. 3. MTT cytotoxic activity of times study at room temperature, (A) E4A-Tigloyl(A); (B) E4A- 3,3-dimethylacryloly(B); C: E4A-4-pentenoyl(C).

FIG. 4. MTT cytotoxic activity of times study at 0° C., (A) E4A-Tigloyl(A); (B) E4A-3,3-dimethylacryloly(B); C: E4A-4-pentenoyl(C).

FIG. 5. MTT cytotoxic activity of times study, (A) E4A-cinnamoyl(J); (B) E4A-hexanoyl(D); C: E4A-2-ethylbutyryl(E); and D, controls: Tig control is tigloyl chloride without E4A; AC control is acetyl chloride without E4A; H is acetyl chloride with E4A reaction 1 min.

FIG. 6. MTT cytotoxic activity of times study, (A) E4A-acetyl(H); (B) E4A-crotonoyl(I)

FIG. 7. HPLC profiles of E4A-Tig in 1 min and 2 hours

FIG. 8. MTT cytotoxic activity of times study for E4A-Tig. Results: E4A-Tigs from reaction of 5 sec to 1 min are most active. Activity decreased after 1 min of reaction. Minimal to no activity was obtained at 10 minutes or longer.

FIG. 9. Results of HPLC profiles of E4A-Tigs : E4A, E4A-ASAP (5 sec), E4A-1 min, E4A-2 min, E4A-5 min, E4A-10 min, E4A-30 min.

FIG. 10. Results of Activity order: M, N, O, P, Q, R, S, T, E4A; M = E4A has no activity.

FIG. 11. (A) The IC50 of Tig-S in KB cells is about 4 ug/ml; and the corresponding IC50 in ES2 cells is less than 1 ug/ml; (B) The IC50 of Tig-S in ES2 cells, MTT assay with low doses of Tig-S, the IC50 of Tig-S in ES2 cells is approximately equal to 0.1 ug/ml

FIG. 12. (A) Results: Swiss3T3 cells are mouse normal fibroblast which were used in this experiment to compare with ES2 (human ovarian cancer) in Tig-R cytotoxicity determination. The preliminary results indicate that the IC50 of Tig-R in SW3T3 cells is above 20 ug/ml while the corresponding IC50 in ES2 cells is about 2.8 ug/ml. (B) Effect of Tig-R on Normal human lung fibroblast (WI38). Results: The IC50 of Tig-R in normal human fibroblast cells (WI38) is about 10-15 ug/ml. This IC50 value is 3 times higher than those in ES2 (3 ug/ml).

FIG. 13. (A) Results: Tig-N, -Q, -R, -T -S and -V do not have hemolytic activity up to 20 ug/ml. The graphs results are overlapped at the bottom of the figure. The original compound ES lyse 100% red blood cells (RBC) at 5 ug/ml. (B) Results: compared to Y3, ACH-Y3 is less potent in hemolytic activity. Tig-R has no hemolytic activity.

FIG. 14. (A) Comparison of potency of compound Y in inhibiting growth of ovarian cancer cells. The IC50 for Compound Y is about 1.5 µg/ml. (B) Hemolytic activity of Xanifolia-Y, B-Escin, Xanifolia-X, ACH-Y and AKOH-Y.

FIG. 15. Inhibition of W138 cells with Tig-S (6 days). Results: IC50 = 1.5 ug/ml; At 10 20 ug/ml, about negative 10% cell growth.

FIG. 16. Inhibition of ES2 cells with Tig-S (2 days). Results: IC50 = 0.3 ug/ml; At 5-10 ug/ml, negative 70% cells growth.

FIG. 17. A comparison of non-cancerous WI38 with ES2 cancer cells. MTT OD is proportional to the amount of live cells. Here, the MTT OD in the no drug control represents 100% of cell growth. This study shows that at 10 ug/ml of Tig-S, WI38 cells maintain about 55% of the control cell growth, while ES2 cells have only 10% of the control cell growth.

FIG. 18. A comparison of non-cancerous WI38 with cancer cells. In these results, the MTT OD from cells before and after treated with different concentrations of drug was plotted. The result shows that: For WI38 cells, the IC₁₀₀ value is about 10 ug/ml [IC₁₀₀ is defined as the MTT OD value after the drug-treatment equal to the original OD value before the drug-treatment. At this condition (IC₁₀₀), it indicates there is 100% inhibition of growth, but there is no cell loss. At 20 ug/ml, the OD decrease to about 90% of the original value, indicating there is about 10% cell loss or death. For ES2 cells, the IC₁₀₀ value is about 0.16 -0.3 ug/ml. However, there is a big decrease of OD with higher drug concentrations indicating there is cell death. At 10 ug/ml, the OD is 12% of the original value, indicating over 90% cells cell lost.

FIG. 19. Tig-S induces cell-death by the apoptosis mechanism.

FIGS. 20-21. Leukemia K562 cells were treated with Tig-S for three days. The number inside charts is the Tig-S concentration in ug/ml; The first peak is the intensity of G0/G1 cells. The last peak is the G2/M cells. Between these two peaks represents the intensity of S-phase cells.

FIG. 22. Animal study result shows Group A Mice - Implanted tumor and no drug, died on day 27. Group B Mice - Implanted tumor and with (Tig-S) drug 100 mg/kg, twice daily, 5 days.

FIG. 23. Animal study result shows Group A Mice - Implanted tumor and no drug. Group B Mice - Implanted tumor and with (Tig-R) drug 100 mg/kg, twice daily, 5 days.

FIG. 24. Animal study result shows Group A Mice - Implanted tumor and no drug; Group B Mice - Implanted tumor and with (Tig-V) drug 50 mg/kg, twice daily, 10 days.

FIG. 25. Inhibition of lung H460 cells growth with Tig-S for one day. IC50 = 3.4 ug/ml.

FIG. 26. Inhibition of lung H460 cells growth with Tig-S for 4 days. IC50 = 3 ug/ml.

FIG. 27. Inhibition of Leukemia K562 cells by Tig-S: Tig-S inhibits Leukemia K562 cells growth with IC50 about 0.6 ug/ml. No growth (IC100) was observed beginning on day 2 at 2.5 ug/ml or higher.

FIG. 28. Inhibition of cancer OVCAR3 cells by natural compounds escin, X, Y0, Y1, Y2, Y3, Y7, Y8, Y9, and Y10.

FIG. 29. Comparing the activities of natural compound Y and synthetic compound Tig-S and Tig-R with leukemia cancer. The synthetic compounds increase the potency and decrease the toxicity. (A) Natural compound; (B) synthetic compound Tig-S; C: synthetic compound Tig-R.

FIG. 30. Compare the activities of natural compound Y and synthetic compound Tig-S and Tig-R with prostate cancer. The synthetic compounds increase the potency and decrease the toxicity. (A) Natural compound; (B) synthetic compound Tig-S; C: synthetic compound Tig-R.

. Drug-effects of Tig-R

FIGS. 34-36. Drug-effects of Tig-S

FIG. 37. Inhibition of ovarian cancer (OCAR3) by Tig-s. Tig-S inhibits OCAR3 cells’ growth with an IC₅₀ value of 2.5 ug/ml; and inhibition of pancreas cancer (Capan), Tig-S inhibits Capan cells’ growth with an IC₅₀ value of about 1 ug/ml.

FIG. 38. Inhibition of eye cancer (Y79) Retinoblastoma by Tig-S. The IC50 for Tig-S is0.1 ug/ml.

FIG. 39. Combined effect of anti-DR5 and Tig-R in HepG2 cells.

FIG. 40. Combined inhibition of Jurkat cell by antibody and Tig-R.

FIG. 41. Combined inhibition of Jurkat cell by antibody and Tig-R; Results: Tig-R enhances the anti-DR5 antibody therapy for Leukemia. The combined effect is synergistic.

FIG. 42. Combination effect of Tig-R and anti-DR5 antibody in Capan cells (pancreas cancer). 1, Capan (pancreas cancer) cells were inhibited by Anti-DR5 alone with IC50 about 0.17 ug/ml. 2, With 2.5 ug/ml of Tig-R, the IC50 decreased to about 0.06 ug/ml. With 5 ug/ml of Tig-R, the IC50 decreased to about 0.01 ug/ml (a decrease of 94%). These results indicate that the addition of Tig-R to anti-DR5 antibody in treatment significantly enhances the inhibition effect.

FIG. 43. Drug-R enhances the anti-DR5 antibody therapy for liver cancer HepG2 cells

Results: This is a repeated experiment with 5 and added a higher 7.5 ug/ml concentration of Tig-. The results show that the combined effects of Tig-R and anti-DR5 antibody occurred. The data also suggests a synergistic effect between with these two agents.

DETAILED DESCRIPTION OF THE INVENTION

This invention provides a method of synthesizing new active compounds for pharmaceutical uses. This invention provides an anti-adhesion therapy which uses the compound as a mediator or inhibitor of adhesion proteins and angiopoietins. It inhibits excessive adhesion and inhibits cell viral and macromolecular attachment. It modulates angiogenesis. The compounds are also used as mediators of cell viral and macromolecular adhesion receptor(s).

This invention provides compounds or a composition comprising the compounds provided in the invention for treating cancers; for inhibiting cancer growth; for inhibiting viruses; for preventing cerebral aging; for improving memory; for improving cerebral functions; for curing enuresis, frequent micturition, urinary incontinence; for treating neurodegenerative diseases, dementia, Alzheimer’s disease, autism, brain trauma, Parkinson’s disease or other diseases caused by cerebral dysfunctions; for treating arthritis, rheumatism, poor circulation, arteriosclerosis, Raynaud’s syndrome, angina pectoris, cardiac disorder, coronary heart disease, headache, dizziness, kidney disorder; cerebrovascular disease; for inhibiting NF-kappa B activation; for treating brain edema, severe acute respiratory syndrome, respiratory viral diseases, chronic venous insufficiency, hypertension, chronic venous disease, oedema, inflammation, hemorrhoids, peripheral edema formation, varicose vein disease, flu, post traumatic edema and postoperative swelling; for inhibiting blood clots and ethanol absorption; for lowering blood sugar; for regulating adrenocorticotropin and corticosterone levels. This invention provides a composition for anti-MS, anti-aneurysm, anti-asthmatic, anti-oedematous, anti-inflammatory, anti-bradykinic, anti-capillarihemorrhagic, anti-cephalagic, anti-cervicobrachialgic, anti-eclamptic, anti-edemic, anti-encaphalitic, anti-epiglottitic, anti-exudative, anti-flu, anti-fracture, anti-gingivitic, anti-hematomic, anti-herpetic, anti-histaminic, anti-hydrathritic, anti-meningitic, antioxidant, anti-periodontic, anti-phlebitic, anti-pleuritic, anti-raucedo, anti-rhinitic, anti-tonsilitic, anti-ulcer, anti-varicose, anti-vertiginous, cancerostatic, corticosterogenic, diuretic, fungicide, hemolytic, hyaluronidase inhibitor, lymphagogue, natriuretic, pesticide, pituitary stimulant, thymolytic, vasoprotective, inhibiting leishmaniases, modulating adhesion or angiogenesis of cells, and anti-parasitic; increase the expression of the genes: ANGPT2, DDIT3, LIF and NFKB1Z, and manufacturing an adjuvant composition and venotonic treatment.

This invention provides compounds for up regulating levels of Death Receptors including DR1, DR4 and DR5 or alike thereof, for the treatment of cancer, and inhibiting cancer cells growth in a subject. This invention is a method for up regulating levels of Death Receptors including DR1, DR4, DR5 or their alike in cells for inhibition of cell growth.

This invention is a method for up regulating levels of DR4, DR5, RELL1, RELL2, RELT or TNFR of cells. The present invention provides a DR4, DR5, RELL1, RELL2, RELT or TNFR inducer and/or a DR4, DR5, RELL1, RELL2, RELT or TNFR agonist for use in a method for the treatment of cancer, inhibiting cancer cells growth in a subject, said method comprises administering to a subject an effective amount of the DR4, DR5, RELL1, RELL2, RELT or TNFR inducer and the DR4, DR5, RELL1, RELL2, RELT or TNFR agonist, wherein the DR4, DR5, RELL1, RELL2, RELT or TNFR inducer is a compound selected from this application, wherein the compound includes triterpene derivatives, terpenoid derivatives, and the DR4, DR5, RELL1, RELL2, RELT or TNFR agonist is an antibody or an antigen-binding fragment. In one embodiment, the method further comprises administration of a DR5 agonist and/or a PD-1 antagonist, PD-L1 antagonist. In another embodiment, the cancers comprise breast cancer, leukocytic cancer, liver cancer, ovarian cancer, bladder cancer, prostatic cancer, skin cancer, bone cancer, brain cancer, leukemia cancer, lung cancer, colon cancer, CNS cancer, melanoma cancer, renal cancer, cervical cancer, esophageal cancer, testicular cancer, splenic cancer, kidney cancer, lymphatic cancer, pancreatic cancer, stomach cancer, eye cancer and thyroid cancer. This invention provides a method of treating cancers with compound(s) that mediate death receptors including DR1, DR4, DR5 or their alike.

This invention provides compounds, compositions and methods for treating cancer diseases, inhibiting cancer invasion, inhibiting cancer growth, and inhibiting cancer metastasis, wherein the compounds comprise the structures selected from the formulae of the present application, wherein the compounds can be synthesized or isolated, wherein the compounds comprise the triterpenes, pentacyclic triterpenes, saponins, and compounds selected from formulae in this application, wherein the cancers comprise breast cancer, leukocytic cancer, liver cancer, ovarian cancer, bladder cancer, prostatic cancer, skin cancer, bone cancer, brain cancer, leukemia cancer, lung cancer, colon cancer, CNS cancer, melanoma cancer, renal cancer, cervical cancer, esophageal cancer, testicular cancer, splenic cancer, kidney cancer, lymphatic cancer, pancreatic cancer, stomach cancer, eye cancer and thyroid cancer; wherein the cells comprise breast cell, leukocytic cell, liver cell, ovarian cell, bladder cell, prostatic cell, skin cell, bone cell, brain cell, leukemia cell, lung cell, colon cell, CNS cell, melanoma cell, renal cell, cervical cell, esophageal cell, testicular cell, splenic cell, kidney cell, lymphatic cell, pancreatic cell, stomach cell, eye cell and thyroid cell.

This invention provides compounds for improving blood circulation; soothing stroke; preventing plaque formation and promoting their dissipation; improving blood viscosity; reducing cardiovascular clotting; reducing cerebrovascular clotting; reducing thrombosis, arteriosclerosis, coronary heart disease, hypertension, diabetes, thrombocytopenia purpura, hemoptysis, hematemesis; treating blood in the stool, uterine bleeding, traumatic bleeding, abdominal irritation, swelling, fluttering, blood circulation, swelling, and pain; treating bronchiectasis, tuberculosis and lung abscess caused by too hemoptysis; reducing bleeding, antitussive, expectorant and analgesic effect, and dilated blood vessels; reducing blood pressure and the treatment of cerebral arteriosclerosis; andelevating blood lipids and reducing cholesterol.

This invention shows that the presence of group selected from acetyl, angeloyl, tigloyl, senecioyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, methylbutanoyl, methylpropanoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, alkenylcarbonyl, ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl, C(2-18) Acyl, or sugar moiety substituted with acetyl, angeloyl, tigloyl, senecioyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, methylbutanoyl, methylpropanoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, alkenylcarbonyl, ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl, C(2-18) Acyl, at a pentacyclic triterpene, triterpene, triterpeniod, triterpeniod saponin, terpene, isoprene or compound selected from formulae of the present application, produces inhibition of cancer growth, cancer invasion, cell invasion, cancer cell invasion, cell adhesion, cell circulation or cell attachment, blocking the DNA synthesis of cancer cell, increasing the potency and decreasing the toxicity.

This invention shows that the presence of a group selected from acetyl, angeloyl, tigloyl, senecioyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, methylbutanoyl, methylpropanoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, alkenylcarbonyl, ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl, C(2-18) Acyl at carbon position 21, 22, 24 and/or 28 of a pentacyclic triterpene, triterpene, triterpeniod, triterpeniod saponin or compound selected from formulae of the present application, produces inhibition of cancer growth, cancer invasion, cell invasion, cancer cell invasion or macromolecular cell invasion. In another embodiment, the presence of group(s) selected from acetyl, angeloyl, tigloyl, senecioyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, methylbutanoyl, methylpropanoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, alkenylcarbonyl, ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl, C(2-18) Acyl at carbon position 2, 3, 8, 15, 21, 22, 23, 24 and/or 28 of a pentacyclic triterpene, triterpeniod, triterpeniod saponin or compound selected from formulae of the present application produces activities including inhibition of cancer growth, cancer invasion, cell invasion, cancer cell invasion, cell adhesion, cell attachment or cell circulating wherein the group may attached with an O, S, NH, CH2O to the carbon of triterpene, triterpeniod, triterpeniod saponin or compound selected from formulae of the present application; wherein the group may be selected from group of CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-lsobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl, (CnH2n)O-angeloyl, (CnH2n)O-tigloyl, (CnH2n)O-senecioyl, (CnH2n)O-acetyl, (CnH2n)O-Crotonoyl, (CnH2n)O-3,3-Dimethylacryloyl, (CnH2n)O-Cinnamoyl, (CnH2n)O-Pentenoyl, (CnH2n)O-Hexanoyl, (CnH2n)O-benzoyl, (CnH2n)O-Ethylbutyryl, (CnH2n)O-alkyl, (CnH2n)O-dibenzoyl, (CnH2n)O-benzoyl, (CnH2n)O-alkanoyl, (CnH2n)O-alkenoyl, (CnH2n)O-benzoyl alkyl substituted O-alkanoyl, (CnH2n)O-alkanoyl substituted phenyl, (CnH2n)O-alkenoyl substituted phenyl, (CnH2n)O-aryl, (CnH2n)O-acyl, (CnH2n)O-heterocylic, (CnH2n)O-heteroraryl, (CnH2n)O-alkenylcarbonyl, (CnH2n)O-alkane, (CnH2n)O-alkene and (CnH2n)O-sugar moiety, wherein n is 1 or 2 or 3 or 4 or over 5. In one embodiment, the presence of group at carbon position 24 produces activities. In another embodiment, the presence of group at carbon position 24 and 28 produces activities. In another embodiment, the presence of group at carbon position 24 and 21 produces activities. In another embodiment, the presence of group at carbon position 24, 28 and 21, produces activities. In another embodiment, the presence of group at carbon position 24, 28 and 22 produces activities. In another embodiment, the presence of group at carbon position 24, 28 and 3 produces activities. In another embodiment, the presence of group at carbon position 24, and 3 produces activities. In another embodiment, the presence of group at carbon position 28 and 3 produces activities. In another embodiment, the presence of group at carbon position 3 produces activities. In another embodiment, the presence of group at carbon position 21 and 22 produces activities. In another embodiment, the hemolytic activity of the compound is reduced. In embodiment, the compound is attached a sugar moiety(ies), acid moiety(ies) or alduronic acid. In another embodiment, the presence of group at carbon position 1, produces activities. In another embodiment, the presence of group at carbon position 2, produces activities. In another embodiment, the presence of group at carbon position 3, produces activities. In another embodiment, the presence of group at carbon position 4, produces activities. In another embodiment, the presence of group at carbon position 5, produces activities. In another embodiment, the presence of group at carbon position 6, produces activities. In another embodiment, the presence of group at carbon position 7, produces activities. In another embodiment, the presence of group at carbon position 8, produces activities. In another embodiment, the presence of group at carbon position 9, produces activities. In another embodiment, the presence of group at carbon position 10, produces activities. In another embodiment, the presence of group at carbon position 11, produces activities. In another embodiment, the presence of group at carbon position 12, produces activities. In another embodiment, the presence of group at carbon position 13, produces activities. In another embodiment, the presence of group at carbon position 14, produces activities. In another embodiment, the presence of group at carbon position 15, produces activities. In another embodiment, the presence of group at carbon position 16, produces activities. In another embodiment, the presence of group at carbon position 17, produces activities. In another embodiment, the presence of group at carbon position 18, produces activities. In another embodiment, the presence of group at carbon position 19, produces activities. In another embodiment, the presence of group at carbon position 20, produces activities. In another embodiment, the presence of group at carbon position 21, produces activities. In another embodiment, the presence of group at carbon position 22, produces activities. In another embodiment, the presence of group at carbon position 23, produces activities. In another embodiment, the presence of group at carbon position 24, produces activities. In another embodiment, the presence of group at carbon position 25, produces activities. In another embodiment, the presence of group at carbon position 26, produces activities. In another embodiment, the presence of group at carbon position 27, produces activities. In another embodiment, the presence of group at carbon position 28, produces activities. In another embodiment, the presence of group at carbon position 29, produces activities. In another embodiment, the presence of group at carbon position 30, produces activities. In another embodiment, the compounds are in the form of a powder, crystal, or liquid. In another embodiment, the compounds are in a capsule, or with a pharmaceutically acceptable carrier or diluent. In another embodiment, the activities are for treating cancers; for the inhibition of cancer growth, cancer invasion, cell invasion, cancer cell invasion, cell adhesion, cell attachment, and cell circulating; for treating mad cow disease; for treating prion diseases; for inhibiting viruses; for preventing cerebral aging; for improving memory; for improving cerebral functions; for curing enuresis, for frequent micturition, for urinary incontinence; for dementia, Alzheimer’s disease, autism, brain trauma, Parkinson’s disease or other diseases caused by cerebral dysfunctions or neurodegeneration; for treating arthritis, rheumatism, poor circulation, arteriosclerosis, Raynaud’s syndrome, angina pectoris, cardiac disorder, coronary heart disease, headache, dizziness, and kidney disorder; for cerebrovascular diseases; inhibiting NF-kappa B activation; for treating brain edema, severe acute respiratory syndrome, respiratory viral diseases, chronic venous insufficiency, hypertension, chronic venous disease, oedema, inflammation, hemorrhoids, peripheral edema formation, varicose vein disease, flu, post traumatic edema and postoperative swelling; for inhibiting blood clots, for inhibiting ethanol absorption; for lowering blood sugar; and for regulating adrenocorticotropin and corticosterone levels. This invention provides a composition for anti-MS, anti-aneurysm, anti-asthmatic, anti-oedematous, anti-inflammatory, anti-bradykinic, anti-capillarihemorrhagic, anti-cephalagic, anti-cervicobrachialgic, anti-eclamptic, anti-edemic, anti-encaphalitic, anti-epiglottitic, anti-exudative, anti-flu, anti-fracture, anti-gingivitic, anti-hematomic, anti-herpetic, anti-histaminic, anti-hydrathritic, anti-meningitic, antioxidant, anti-periodontic, anti-phlebitic, anti-pleuritic, anti-raucedo, anti-rhinitic, anti-tonsilitic, anti-ulcer, anti-varicose, anti-vertiginous, cancerostatic, corticosterogenic, diuretic, fungicide, hemolytic, hyaluronidase inhibitor, lymphagogue, natriuretic, pesticide, pituitary stimulant, thymolytic, vasoprotective, inhibiting leishmaniases, modulating adhesion or angiogenesis of cells, and anti-parasitic; increasing the expression of the genes: ANGPT2, DDIT3, LIF and NFKB1Z, and manufacturing an adjuvant composition and venotonic treatment. In another embodiment, the compound is arresting cells in the S-phase and blocking their entering into the G2/M phase of cell cycle. The compound blocks the DNA synthesis of cancer cells.

This invention provides compounds for improving blood circulation; soothing stroke; preventing plaque formation and promoting their dissipation; improving blood viscosity; reducing cardiovascular clotting; reducing cerebrovascular clotting; reducing thrombosis, arteriosclerosis, coronary heart disease, hypertension, diabetes, thrombocytopenia purpura, hemoptysis, and hematemesis; treating blood in the stool, uterine bleeding, traumatic bleeding, abdominal irritation, swelling, fluttering, blood circulation, swelling, and pain; treating bronchiectasis, tuberculosis and lung abscess caused by hemoptysis; reducing bleeding, antitussive, expectorant and analgesic effect, and dilating blood vessels; reducing blood pressure and the treatment of cerebral arteriosclerosis; and elevating blood lipids and reducing cholesterol.

In another embodiment, the compound is arresting cells in the S-phase and blocking their entrance into the G2/M phase of cell cycle. The compound blocks the DNA synthesis of cancer cells. This application produces synthetic compounds, increase,potency and decreasing the toxicity.

In another embodiment, the presence of group at carbon position 1, reduces activities. In another embodiment, the presence of group at carbon position 2, reduces activities. In another embodiment, the presence of group at carbon position 3, reduces activities. In another embodiment, the presence of group at carbon position 4, reduces activities. In another embodiment, the presence of group at carbon position 5, reduces activities. In another embodiment, the presence of group at carbon position 6, reduces activities. In another embodiment, the presence of group at carbon position 7, reduces activities. In another embodiment, the presence of group at carbon position 8, reduces activities. In another embodiment, the presence of group at carbon position 9, reduces activities. In another embodiment, the presence of group at carbon position 10, reduces activities. In another embodiment, the presence of group at carbon position 11, reduces activities. In another embodiment, the presence of group at carbon position 12, reduces activities. In another embodiment, the presence of group at carbon position 13, reduces activities. In another embodiment, the presence of group at carbon position 14, reduces activities. In another embodiment, the presence of group at carbon position 15, reduces activities. In another embodiment, the presence of group at carbon position 16, reduces activities. In another embodiment, the presence of group at carbon position 17, reduces activities. In another embodiment, the presence of group at carbon position 18, reduces activities. In another embodiment, the presence of group at carbon position 19, reduces activities. In another embodiment, the presence of group at carbon position 20, reduces activities. In another embodiment, the presence of group at carbon position 21, reduces activities. In another embodiment, the presence of group at carbon position 22, reduces activities. In another embodiment, the presence of group at carbon position 23, reduces activities. In another embodiment, the presence of group at carbon position 24, reduces activities. In another embodiment, the presence of group at carbon position 25, reduces activities. In another embodiment, the presence of group at carbon position 26, reduces activities. In another embodiment, the presence of group at carbon position 27, reduces activities. In another embodiment, the presence of group at carbon position 28, reduces activities. In another embodiment, the presence of group at carbon position 29, reduces activities. In another embodiment, the presence of group at carbon position 30, reduces activities. In another embodiment the activities include but are not limited to hemolytic activity, cancer activity, arthritis, rheumatism, poor circulation, arteriosclerosis, Raynaud’s syndrome, angina pectoris, cardiac disorder, coronary heart disease, headache, dizziness, kidney disorder; soothing stroke; plaque formation; cardiovascular; cerebrovascular; thrombosis, arteriosclerosis, coronary heart disease, hypertension, diabetes, thrombocytopenia purpura, hemoptysis, hematemesis; blood in the stool, uterine bleeding, traumatic bleeding, abdominal irritation, swelling, fluttering, swelling, pain; bronchiectasis, tuberculosis and lung abscess; bleeding, tussive, expectorant and analgesic effect, blood pressure and cerebral arteriosclerosis; blood lipids and cholesterol.

This invention shows a method of synthesizing active compound by attaching functional group to a core compound, wherein the functional group(s) comprises() a group which is/are selected from ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, pentenoyl, hexanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, crotonoyl, 2-butenoyl, Isobutyryl, methylpropanoyl,2-methylpropanoyl, ethylbutyryl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl, tigloyl, angeloyl, acetyl, crotonoyl, 3,3-Dimethylacryloyl, senecioyl, cinnamoyl, benzoyl, ethylbutyryl, dibenzoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, and heteroraryl, wherein the core compound is a triperpene or 5 ring triterpene. In another embodiment, the core compound is a 4-ring terpene. In another embodiment, the core compound is a 3-ring terpene. In another embodiment, the core compound is a 2-ring terpene. In another embodiment, the core compound is a 1 ring terpene. The compounds provided in the invention are for treating cancers, inhibition of cancer growth, cancer invasion, cell invasion, cancer cell invasion; cell adhesion, cell attachment, and cell circulating; for treating mad cow disease; treating prion diseases; for inhibiting viruses; for preventing cerebral aging; for improving memory; improving cerebral functions; for curing enuresis, frequent micturition, urinary incontinence; dementia, Alzheimer’s disease, autism, brain trauma, Parkinson’s disease or other diseases caused by cerebral dysfunctions or neurodegeneration; for treating arthritis, rheumatism, poor circulation, arteriosclerosis, Raynaud’s syndrome, angina pectoris, cardiac disorder, coronary heart disease, headache, dizziness, kidney disorder; cerebrovascular diseases; inhibiting NF-kappa B activation; for treating brain edema, severe acute respiratory syndrome, respiratory viral diseases, chronic venous insufficiency, hypertension, chronic venous disease, oedema, inflammation, hemorrhoids, peripheral edema formation, varicose vein disease, flu, post traumatic edema and postoperative swelling; for inhibiting blood clotsand ethanol absorption; for lowering blood sugar; and for regulating adrenocorticotropin and corticosterone levels. This invention provides a composition for anti-MS, anti-aneurysm, anti-asthmatic, anti-oedematous, anti-inflammatory, anti-bradykinic, anti-capillarihemorrhagic, anti-cephalagic, anti-cervicobrachialgic, anti-eclamptic, anti-edemic, anti-encaphalitic, anti-epiglottitic, anti-exudative, anti-flu, anti-fracture, anti-gingivitic, anti-hematomic, anti-herpetic, anti-histaminic, anti-hydrathritic, anti-meningitic, antioxidant, anti-periodontic, anti-phlebitic, anti-pleuritic, anti-raucedo, anti-rhinitic, anti-tonsilitic, anti-ulcer, anti-varicose, anti-vertiginous, cancerostatic, corticosterogenic, diuretic, fungicide, hemolytic, hyaluronidase inhibitor, lymphagogue, natriuretic, pesticide, pituitary stimulant, thymolytic, vasoprotective, inhibiting leishmaniases, modulating adhesion or angiogenesis of cells, anti-parasitic; increasing the expression of the genes: ANGPT2, DDIT3, LIF and NFKB1Z, and manufacturing an adjuvant composition and venotonic treatment. In another embodiment, the compound is arresting cells in the S-phase and blocking their entrance into the G2/M phase of cell cycle. The compound blocks the DNA synthesis of cancer cells.

When healthy cells in the eye change — or mutate — and grow too quickly in a disorganized way, they can form a mass of tissue called a tumor. If these problem cells start in the eye, it’s called intraocular cancer, or primary eye cancer. If they spread to eye from another part of body, it’s called secondary eye cancer. Retinoblastoma is the most common type of eye cancer in children. The compounds in this invention inhibit the growth of eye cancers including Retinoblastoma.

Retinoblastoma occurs in approximately 1 in 18,000 live births. Metastases occur most commonly in the CNS, bones, bone marrow and liver. If untreated, the tumors invade locally and then metastasize, causing death within two years. There are three options -surgery, chemotherapy and radiation. Because systemic carboplatin is now commonly used in the treatment of retinoblastoma, concern has been raised about hearing loss related to such therapy.

Experiments presented in this invention showed that the compound AKOH has no effect in inhibiting cancer growth, cancer invasion, cell invasion or cancer cell invasion. AKOH was obtained by removing the angeloyl groups from carbon positions 21 and 22 of the active Xanifolia Y(Y3). This invention shows that the ability for inhibiting cancer invasion, cell invasion or cancer cell invasion of Xanifolia Y(Y3) are lost by removing angeloyl groups from carbon positions 21 and 22.

Experiments presented in this invention showed that the core compound including E4A, E5A, Xanifolia Y-core have no effect in inhibiting cancer growth, cancer invasion, cell invasion or cancer cell invasion. Xanifolia Y-core was obtained by removing the angeloyl groups from carbon positions 21 and 22, and the sugar moieties from carbon 3 of the active Xanifolia Y(Y3). E4A (E IV A) was obtained by removing the groups from carbon positions 3, 21 and 22 of the active Escin. E5A (E V A) was obtained by removing the groups from carbon positions 3, 21 and 22 of the active Escin. This invention showed that the core compound including E4A, E4AY2, E5A, Xanifolia Y-core and AKOH have no hemolytic activity and no anti-cancer activity.

This invention showed that functional groups attached at carbon position 24 of a pentacyclic triterpene did not produce hemolytic activity, which has bio-activities including inhibiting cancer growth, inhibiting cancer invasion, cell invasion or cancer cell invasion. This invention showed that functional groups attached at carbon position 3 of a pentacyclic triterpene did not produce hemolytic activity, which has bio-activities including inhibiting cancer growth, inhibiting cancer invasion, cell invasion or cancer cell invasion. This invention showed that functional group(s) attached at carbon position 3 and 1 or 2 or 3 of carbon position 28, 21, 22, 24 of a pentacyclic triterpene did not produce hemolytic activity, which has bio-activities including inhibiting cancer growth, inhibiting cancer invasion, cell invasion or cancer cell invasion.

This invention showed that functional groups attached at carbon position 2 of a pentacyclic triterpene did not produce hemolytic activity, which has bio-activities including inhibiting cancer growth, inhibiting cancer invasion, cell invasion or cancer cell invasion. This invention showed that function group(s) attached at carbon position 2 and 1 or 2 or 3 of carbon position 28, 21, 22, 24 of a pentacyclic triterpene did not produce hemolytic activity, which has bio-activities including inhibiting cancer growth, inhibiting cancer invasion, cell invasion or cancer cell invasion.

This invention provides a pentacyclic triterpene with reduced hemolytic activity for treating diseases, wherein the triterpene comprising a group(s) attached at its core producing bio-activities. This invention provides a pentacyclic triterpene with reduced hemolytic activity comprising a group(s) attached at carbon position 3, or carbon position 24 and 1 or 2 or 3 of other position(s) of a pentacyclic triterpene, which has bio-activities. This invention provides a triterpene with reduced hemolytic activity comprising a group(s) attached at carbon position 15, 16, 21, 22, 23, 24, 28, 29, 30 and/or 3 of a pentacyclic triterpene, which has bio-activities. This invention provides a composition comprising a triterpene with reduced hemolytic activity comprising a group(s) attached at carbon position 3, or carbon position 24 and 1 or 2 or 3 of other position(s) of a pentacyclic triterpene, which has bio-activities. This invention provides a method for bio-activities treatment including but not limited to treating cancers, comprising administering to said subject an effective amount of compound, wherein the compound is a pentacyclic triterpene with reduced hemolytic activity comprising a group(s) attached at carbon position 3, or carbon position 24 and 1 or 2 or 3 of other position(s) of a pentacyclic triterpene, which has bio-activities.

The compound of the present application can be obtained with the method:

1. Dissolving core compound or triterpenes, hydroxylated triterpenes core in pyridine;
2. Adding acyl chloride;
3. The mixture is stirred for a length of time including 5 sec, 10 sec, 20 sec, 30 sec, 40 sec,1 min, 2 min, 5 min, 10 min, 30 min, 1 hr, 2 hr, 18 hr, 2 days or 3 days at 0° C., 25° C., 50° C. or 75° C.;
4. At the end of reaction, an aqueous solution of acid or base, or water is added to the reaction mixture;
5. The solution is then extracted of ethyl acetate and ethyl acetate is removed by evaporation and lyophilization;
6. Dissolving the reaction product in acetonitrile with Trifluoroacetic acid or DMSO;
7. Testing the reaction product of mixtures and individual fractions with MTT cytotoxic assay;
8. Selecting the HPLC fractions for isolation is according to the cytotoxic activity of the reaction product obtained at a specific reaction time;
9. Purifying the active esterification products with HPLC;
10. Collecting the products; and
11. Testing the products.

The compound of the present application, wherein the core compound is terpene, isoprene, or triterpene core; wherein the core compound is hydroxylated; wherein the core compound was dissolved in pyridine; wherein the acyl chloride including Tigloyl chloride, angeloyl chloride, Acetyl chloride, Crotonoyl chloride, 3,3-Dimethylacryloyl chloride, senecioyl chloride, Cinnamoyl chloride, Pentenoyl chloride, Hexanoyl chloride, benzoyl chloride and Ethylbutyryl chloride; wherein the reaction time for the mixture is stirred for 5 sec, 1 min, 2 min, 5 min, 10 min, 30 min, 1 hr, 2 hr, 18 hr, 2 days or 3 days; wherein the temperature is 0° C., 25° C., 50° C. or 75° C. temperature; wherein the acid including HCl or the base is a weak base including NaHCO3 is added to the reaction mixture; wherein the solution is then extracted 3 times with ethyl acetate and lyophilization; wherein the reaction product is dissolved in 80% acetonitrile - 0.005% Trifluoroacetic acid or DMSO; wherein selecting the HPLC fractions for isolation is according to the cytotoxic activity of the reaction product obtained at a reaction time of 5 sec, 1 min, 2 min, 5 min, 10 min, 30 min, 1 hr, 2 hr, 18 hr, 2 days or 3 days.

This invention showed that functional groups attached at carbon position 23 of a pentacyclic triterpene did not produce hemolytic activity, which has bio-activities including inhibiting cancer growth, inhibiting cancer invasion, cell invasion or cancer cell invasion. This invention showed that function group(s) attached at carbon position 24 and 1 or 2 or 3 of carbon position 28, 21, 22 of a pentacyclic triterpene did not produce hemolytic activity, which has bio-activities including inhibiting cancer growth, inhibiting cancer invasion, cell invasion or cancer cell invasion.

This invention provides a triterpene with reduced hemolytic activity for treating diseases, wherein the triterpene comprising a group(s) attached at its core producing bio-activities. This invention provides a pentacyclic triterpene with reduced hemolytic activity comprising a group(s) attached at carbon position 24, or carbon position 24 and 1 or 2 or 3 of other position(s) of a pentacyclic triterpene, which has bio-activities. This invention provides a composition comprising a pentacyclic triterpene with reduced hemolytic activity comprising a group(s) attached at carbon position 24, or carbon position 24 and 1 or 2 or 3 of other position(s) of a pentacyclic triterpene, which has bio-activities. This invention provides a method for bio-activities treatment including but not limited to treating cancers, comprising administering to said subject an effective amount of compound, wherein the compound is a triterpene with reduced hemolytic activity comprising a group(s) attached at carbon position 24, or carbon position 24 and 1 or 2 or 3 of other position(s) of a pentacyclic triterpene, which has bio-activities.

This invention showed that Tig-N, Tig -Q, Tig -R, Tig-T Tig-S and Tig-V do not have hemolytic activity up to 20 ug/ml. The original compound ES lyse 100% red blood cells (RBC) at 5 ug/ml. Compared to Y3, the ACH-Y3 is less potent in hemolytic activity. Tig-R has no hemolytic activity. This invention showed that Tig-N, Tig -Q, Tig -R, Tig-T Tig-S and Tig-V have anti-cancer activities.

Many saponins and triterpenes have hemolytic characteristic that damage red blood cells. This severe side effect makes people hesitate to use saponins or triterpenes in medicines. This invention produces synthesized saponins and triterpenes with reduced hemolytic characteristic for use as medicament. This invention produces compounds with reduced hemolytic characteristic for use as medicament. The medicament can be used for treating cancer, inhibiting cancer growth, cancer invasion, cell invasion or cancer cell invasion. This application produces synthetic compounds with increased potency and decreased toxicity.

This invention shows that the ability for inhibiting cancer growth, cancer invasion, cell invasion or cancer cell invasion are maintained when the sugar moieties are removed from carbon position 3 of an active compound, triterpene, triterpenoid, or triterpenoid saponin. Experiments presented in this invention showed that the compound ACH-Y3 has the ability to inhibit cancer invasion, cell invasion or cancer cell invasion. The compound ACH-Y3 was obtained by removing the sugar moieties from carbon position 3 of an active Xanifolia Y(Y3). This invention shows that the ability for inhibiting cancer invasion, cell invasion or cancer cell invasion are maintained when the sugar moieties are removed from the carbon position 3 of active Xanifolia Y(Y3).

A compound which has bio-activities including inhibiting cancer growth, inhibiting cancer invasion, cell invasion or cancer cell invasion is called an active compound.

This invention provides a use for compounds, compositions, and methods for manufacturing medicament for treating cancers, inhibition of cancer growth, cancer invasion, cell invasion, cancer cell invasion; cell adhesion, cell attachment, cell circulating, or for inhibiting cancer metastasis, wherein the compounds comprise the structures selected from the formulae of the present application, wherein the compounds can be synthesized or isolated, wherein the compounds comprise the pentacyclic triterpenes, wherein the compounds with reduced hemolytic, wherein the cells comprise cancer cells, wherein the cancers comprise breast cancer, leukocytic cancer, liver cancer, ovarian cancer, bladder cancer, prostatic cancer, skin cancer, bone cancer, brain cancer, leukemia cancer, lung cancer, colon cancer, CNS cancer, melanoma cancer, renal cancer, cervical cancer, esophageal cancer, testicular cancer, splenic cancer, kidney cancer, lymphatic cancer, pancreatic cancer, stomach cancer, eye cancer and thyroid cancer.. The method of inhibiting cancer invasion, cell invasion or cancer cell invasion activities uses non-cytotoxic drug concentrations. The method of inhibiting metastasis uses non-cytotoxic drug concentrations. There is no noticeable change in cell morphology.

This invention provides triterpene(s) with reduced hemolytic activity for treating diseases, wherein the triterpene can be a pentacyclic triterpene comprising a group(s) attached at its core producing bio-activities. This invention provides a pentacyclic triterpene with reduced hemolytic effect, comprising a group(s) attached at carbon position 24, or carbon position 24 and 1 or 2 or 3 of other position(s) of a pentacyclic triterpene, which has bio-activities. This invention provides a composition comprising a triterpene with reduced hemolytic activity comprising a group(s) attached at carbon position 24, or carbon position 24 and 1 or 2 or 3 of other position(s) of a pentacyclic triterpene, which has bio-activities. This invention provides a method for bio-activities treatment including but not limited to treating cancers, comprising administering to said subject an effective amount of compound, wherein the compound is a triterpene with reduced hemolytic activity, comprising a group(s) attached at carbon position 24, or carbon position 24 and 1 or 2 or 3 of other position(s) of a triterpene, which has bio-activities, wherein a compound selected from A1-18, A20-32, B1-18, B20-32, C1-18, C20-32, D1-18, D20-32, D1-18, D20-32, D1-18, D20-32, D1-18, D20-32, D1-18, D20-32, E1-18, E20-32, G1-18, G20-32, H1-18, H20-32, I1-18, I20-32, J1-18, J20-32, K1-18, K20-32, Tig—Sen—n, Tig—Cro—n, Tig—Acy—n, Tig—Pen—n, Tig—Hex—n, Tig—Cin—n, Tig—Ang—n, Tig—Eth—n, Tig—R—Sen—n, Tig—R—Cro—n, Tig—R—Acy—n, Tig—R—Pen—n, Tig—R—Hex—n, Tig—R—Cin—n, Tig—R—Ang—n, Tig—R—Eth—n, wherein n=1 to 6, and a salt, ester, metabolite thereof, and the compounds selected from formulae 2A, and K; wherein the compound is selected from Tig-N, Tig -Q, Tig -R, Tig-T Tig-S and Tig-V. In another embodiment, the compounds are in form of powder, crystal, or liquid. In another embodiment, the compounds are in a capsule, or with pharmaceutically acceptable carrier or diluent.

This invention provides methods for increasing the level of DR5 of cells as a biomarker, treating cancers, inhibition of cancer growth, cancer invasion, cell invasion, cancer cell invasion; cell adhesion, cell attachment, cell circulating, migration, metastasis or growth of cancers, wherein the methods comprise affecting gene expression, wherein the methods comprise stimulating gene expression, or wherein the methods comprise inhibiting the gene expression, or wherein the methods comprise administering to a subject an effective amount of compounds, compositions in this application. In another embodiment, the method comprises contacting said cell with a compound selected from A1-18, A20-32, B1-18, B20-32, C1-18, C20-32, D1-18, D20-32, D1-18, D20-32, D1-18, D20-32, D1-18, D20-32, D1-18, D20-32, E1-18, E20-32, G1-18, G20-32, H1-18, H20-32, I1-18, I20-32, J1-18, J20-32, K1-18, K20-32, Xanifolia Y0, Y1, Y2, Y(Y3), Y5, Y7, Y8, Y9, Y10, Xanifolia (x), M10, Escin(bES), Aescin, ACH-Y(Y3), ACH-Y10, ACH-Y2, ACH-Y8, ACH-Y7, ACH-Y0, ACH-X, ACH-Z4, ACH-Z1, ACH-Escin(bES), ACH-M10, Tig—Sen—n, Tig—Cro—n, Tig—Acy—n, Tig—Pen—n, Tig—Hex—n, Tig—Cin—n, Tig—Ang—n, Tig—Eth—n, Tig—R—Sen—n, Tig—R—Cro—n, Tig—R—Acy—n, Tig—R—Pen—n, Tig—R—Hex—n, Tig—R—Cin—n, Tig—R—Ang—n, Tig—R—Eth—n, wherein n=1 to 6, and a salt, ester, metabolite thereof, and the compounds selected from formulae 2A, and K. In another embodiment, the compounds are in form of powder, crystal, or liquid. In another embodiment, the compounds are in a capsule, or with pharmaceutically acceptable carrier or diluent. In vitro studies show that a compound selected from structure (2A) or (K) inhibits cell adhesion to culture flasks. The compound blocks the function of these adhesive molecules on cells. In another embodiment, the selected compound blocks the function of these adhesive molecules on cells. In another embodiment, the selected compound blocks the function of these adhesive molecules on carcinoma cells. In another embodiment, the selected compound blocks the function of these adhesive molecules on the mesothelial cells. This invention provides an anti-adhesion therapy which uses the compound as a mediator or inhibitor of adhesion proteins and angiopoietins. It inhibits excess adhesion and inhibits cell attachment. This invention provides compounds for use as a mediator for cell circulating, cell moving and inflammatory diseases. In another embodiment, the selected compound binds to the adhesive proteins (by masking) on the membrane and inhibits the interaction of adhesion proteins with their receptors. In another embodiment, the selected compound’s action on the membrane affects adhesion proteins’ function in the membrane. The loss of adhesion activity of cancer cells is result from direct or indirect action of the selected compound on membrane proteins.

This invention provides a use of compounds or methods for inhibiting cancer invasion, cell invasion, cancer cell invasion, macromolecular cell invasion, migration, metastasis or growth of cancers, wherein this invention comprises a process and method for administration of the composition, wherein administration is by intravenous injection, intravenous drip, intraperitoneal injection or oral administration; wherein administration is by intravenous drip: 0.003-0.03 mg/kg body weight of compound dissolved in 250 ml of 10% glucose solution or in 250 ml of 0.9% NaCl solution, or by intravenous injection: 0.003-0.03 mg/kg body weight per day of compound dissolved in 10-20 ml of 10% glucose solution or of 0.9% NaCl solution, or 0.01-0.03 mg/kg body weight of compound dissolved in 250 ml of 10% glucose solution or in 250 ml of 0.9% NaCl solution, or by intravenous injection: 0.01-0.03 mg/kg body weight per day of compound dissolved in 10-20 ml of 10% glucose solution or of 0.9% NaCl solution, or 0.01-0.05 mg/kg body weight of compound dissolved in 250 ml of 10% glucose solution or in 250 ml of 0.9% NaCl solution, or by intravenous injection: 0.01-0.05 mg/kg body weight per day of compound dissolved in 10-20 ml of 10% glucose solution or of 0.9% NaCl solution, or 0.05-0.2 mg/kg body weight of compound dissolved in 250 ml of 10% glucose solution or in 250 ml of 0.9% NaCl solution, or by intravenous injection: 0.05-0.2 mg/kg body weight per day of compound dissolved in 10-20 ml of 10% glucose solution or of 0.9% NaCl solution, or by intravenous drip: 0.1-0.2 mg/kg body weight per day of compound dissolved in 250 ml of 10% glucose solution or in 250 ml of 0.9% NaCl solution, or by intravenous injection: 0.1-0.2 mg/kg body weight per day compound dissolved in 10-20 ml of 10% glucose solution or of 0.9% NaCl solution, or by intraperitoneal injection(I.P.): 2.5 mg/kg body weight per day compound dissolved in 10% glucose solution or of 0.9% NaCl solution, or by oral administration wherein the dosage of mammal is 1-10 mg/kg, 10-30 mg/kg, 30-60 mg/kg, or 60-90 mg/kg body weight of compound, or by intravenous injection or intravenous drip wherein the dosage of mammal is 0.01-0.1 mg/kg body weight, 0.1-0.2 mg/kg, 0.2 - 0.4 mg/kg body weight, or 0.4-0.6 mg/kg body weight of compound, or by intraperitoneal injection (I.P.) wherein the dosage of mammal is 1-3 mg/kg, 3-5 mg/kg, 4-6 mg/kg, or 6-10 mg/kg body weight of compound, or 10-50 mg/kg body weight of compound, or 50-100 mg/kg body weight of compound, or 30-70 mg/kg body weight of compound or 100-150 mg/kg body weight of compound.

This invention provides a use of compounds or methods for treating cancers, inhibition of cancer growth, cancer invasion, cell invasion, cancer cell invasion; macromolecular cell invasion, cell adhesion, cell attachment, cell circulating, migration, metastasis or growth of cancers, infection or re-infection of virus or infectious macromolecules, and cancer cell fusion, wherein the invention comprises a pharmaceutical composition comprising the compound of this invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent, wherein said compound is present in a concentration of 0.01 ug/ml to 65 ug/ml, or wherein said compound is present in a concentration of 0.01 ug/ml to 40 ug/ml, or wherein said compound is present in a concentration of 0.01 ug/ml to 30 ug/ml, or wherein said compound is present in a concentration of 0.01 ug/ml to 10 ug/ml, or wherein said compound is present in a concentration of 0.01 ug/ml to 5 ug/ml, or wherein said compound is present in a concentration of 5 ug/ml to 10 ug/ml, or wherein said compound is present in a concentration of 0.1 ug/ml to 5 ug/ml, or wherein said compound is present in a concentration of 0.1 ug/ml to 7.5 ug/ml, or wherein said compound is present in a concentration of 0.1 ug/ml to 10 ug/ml, or wherein said compound is present in a concentration of 0.1 ug/ml to 15 ug/ml, or wherein said compound is present in a concentration of 0.1 ug/ml to 20 ug/ml, or wherein said compound is present in a concentration of 0.1 ug/ml to 30 ug/ml, or wherein said compound is present in a concentration of 1 ug/ml to 5 ug/ml, or wherein said compound is present in a concentration of 1 ug/ml to 7.5 ug/ml, or wherein said compound is present in a concentration of 1 ug/ml to 10 ug/ml, or wherein said compound is present in a concentration of 1 ug/ml to 15 ug/ml, or wherein said compound is present in a concentration of 1 ug/ml to 20 ug/ml, or wherein said compound is present in a concentration of 1 ug/ml to 30 ug/ml, or wherein said compound is present in a concentration of 3 ug/ml to 5 ug/ml, or wherein said compound is present in a concentration of 3 ug/ml to 7.5 ug/ml, or wherein said compound is present in a concentration of 3 ug/ml to 10 ug/ml, or wherein said compound is present in a concentration of 3 ug/ml to 15 ug/ml, or wherein said compound is present in a concentration of 3 ug/ml to 20 ug/ml, or wherein said compound is present in a concentration of 3 ug/ml to 30 ug/ml, or wherein said compound is present in a concentration of 4 ug/ml to 5 ug/ml, or wherein said compound is present in a concentration of 4 ug/ml to 7.5 ug/ml, or wherein said compound is present in a concentration of 4 ug/ml to 10 ug/ml, or wherein said compound is present in a concentration of 4 ug/ml to 15 ug/ml, or wherein said compound is present in a concentration of 4 ug/ml to 20 ug/ml, or wherein said compound is present in a concentration of 4 ug/ml to 30 ug/ml, or wherein said compound is present in a concentration of 5 ug/ml to 8 ug/ml, or wherein said compound is present in a concentration of 5 ug/ml to 9 ug/ml, or wherein said compound is present in a concentration of 5 ug/ml to 10 ug/ml, or wherein said compound is present in a concentration of 5 ug/ml to 15 ug/ml, or wherein said compound is present in a concentration of 5 ug/ml to 20 ug/ml, or wherein said compound is present in a concentration of 5 ug/ml to 30 ug/ml, or wherein said compound is present in a concentration of 7 ug/ml to 8 ug/ml, or wherein said compound is present in a concentration of 7 ug/ml to 9 ug/ml, or wherein said compound is present in a concentration of 7 ug/ml to 10 ug/ml, or wherein said compound is present in a concentration of 7 ug/ml to 15 ug/ml, or wherein said compound is present in a concentration of 7 ug/ml to 20 ug/ml, or wherein said compound is present in a concentration of 7 ug/ml to 30 ug/ml, or wherein said compound is present in a concentration of 30 ug/ml to 70 ug/ml or wherein said compound is present in a concentration of 70 ug/ml to 100 ug/ml or wherein said compound is present in a concentration of 100 ug/ml to 150 ug/ml.

This invention provides a use of compounds or methods for treating cancers, inhibition of cancer growth, cancer invasion, cell invasion, cancer cell invasion; macromolecular cell invasion, cell adhesion, cell attachment, cell circulating, migration, metastasis or growth of cancers, infection or re-infection of virus or infectious macromolecules, and cancer cell fusion, wherein the invention comprises a pharmaceutical composition comprising the compound of this invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent, wherein said compound is present in a concentration of 0.008 uM to 80 uM, or wherein said compound is present in a concentration of 0.01 uM to 60 uM, or wherein said compound is present in a concentration of 0.01 uM to 50 uM, or wherein said compound is present in a concentration of 0.01 uM to 40 uM, or wherein said compound is present in a concentration of 0.01 uM to 30 uM, or wherein said compound is present in a concentration of 0.01 uM to 20 uM, or wherein said compound is present in a concentration of 0.01 uM to 10 uM, or wherein said compound is present in a concentration of 5 uM to 10 uM, or wherein said compound is present in a concentration of 0.1 uM to 5 uM, or wherein said compound is present in a concentration of 0.1 uM to 7.5 uM, or wherein said compound is present in a concentration of 0.1 uM to 10 uM, or wherein said compound is present in a concentration of 0.1 uM to 15 uM, or wherein said compound is present in a concentration of 0.1 uM to 20 uM, or wherein said compound is present in a concentration of 0.1 uM to 30 uM or wherein said compound is present in a concentration of 0.1 uM to 40 uM, or wherein said compound is present in a concentration of 0.1 uM to 50 uM or wherein said compound is present in a concentration of 0.1 uM to 60 uM, or wherein said compound is present in a concentration of 0.1 uM to 80 uM, or wherein said compound is present in a concentration of 1 uM to 5 uM, or wherein said compound is present in a concentration of 1 uM to 7.5 uM, or wherein said compound is present in a concentration of 1 uM to 10 uM, or wherein said compound is present in a concentration of 1 uM to 15 uM, or wherein said compound is present in a concentration of 1 uM to 20 uM, or wherein said compound is present in a concentration of 1 uM to 30 uM or wherein said compound is present in a concentration of 1 uM to 40 uM, or wherein said compound is present in a concentration of 1 uM to 50 uM or wherein said compound is present in a concentration of 1 uM to 60 uM, or wherein said compound is present in a concentration of 1 uM to 80 uM, or wherein said compound is present in a concentration of 3 uM to 5 uM, or wherein said compound is present in a concentration of 3 uM to 7.5 uM, or wherein said compound is present in a concentration of 3 uM to 10 uM, or wherein said compound is present in a concentration of 3 uM to 15 uM, or wherein said compound is present in a concentration of 3 uM to 20 uM, or wherein said compound is present in a concentration of 3 uM to 30 uM or wherein said compound is present in a concentration of 3 uM to 40 uM, or wherein said compound is present in a concentration of 3 uM to 50 uM or wherein said compound is present in a concentration of 3 uM to 60 uM, or wherein said compound is present in a concentration of 3 uM to 80 uM, or wherein said compound is present in a concentration of 5 uM to 8 uM, or wherein said compound is present in a concentration of 5 uM to 10 uM, or wherein said compound is present in a concentration of 5 uM to 15 uM, or wherein said compound is present in a concentration of 5 uM to 20 uM, or wherein said compound is present in a concentration of 5 uM to 30 uM or wherein said compound is present in a concentration of 5 uM to 40 uM, or wherein said compound is present in a concentration of 5 uM to 50 uM or wherein said compound is present in a concentration of 5 uM to 60 uM, or wherein said compound is present in a concentration of 5 uM to 80 uM. or wherein said compound is present in a concentration of 7 uM to 8 uM, or wherein said compound is present in a concentration of 7 uM to 10 uM, or wherein said compound is present in a concentration of 7 uM to 15 uM, or wherein said compound is present in a concentration of 7 uM to 20 uM, or wherein said compound is present in a concentration of 7 uM to 30 uM or wherein said compound is present in a concentration of 7 uM to 40 uM, or wherein said compound is present in a concentration of 7 uM to 50 uM or wherein said compound is present in a concentration of 7 uM to 60 uM, or wherein said compound is present in a concentration of 7 uM to 80 uM or wherein said compound is present in a concentration of 70 uM to 100 uM, or wherein said compound is present in a concentration of 90 uM to 120 uM.

The invention will be better understood by reference to the Experimental Details which follow, but those skilled in the art will readily appreciate that the specific experiments detailed are only illustrative and are not meant to limit the invention as described herein, which is defined by the claims which follow thereafter.

Throughout this application, various references or publications are cited. Disclosures of these references or publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.

It is to be noted that the transitional term “comprising”, which is synonymous with “including”, “containing” or “characterized by”, is inclusive or open-ended and does not exclude additional, un-recited elements or method steps.

Example 1
Tablet for Dose Containing 10 mg, 20 mg30 mg of Active Compound

Active compound
1 mg
5 mg
10 mg
20 mg
30 mg

Microcrystalline cellulose
20 mg
20 mg
19.75 mg
60 mg
100 mg

Corn starch
29 mg
24.5 mg
19.75 mg
19.25 mg
18.5 mg

Magnesium stearate
0 mg
0.5 mg
0.5 mg
0.75 mg
1.5 mg

The active compound, cellulose, and a portion of the corn starch are mixed and granulated to 10% corn starch paste. The resulting granulation is sieved, dried and blended with the remainder of the corn starch and the magnesium stearate. The resulting granulation is then compressed into tablets containing 1, 5, 10, 20, 30 mg, respectively of active ingredient per tablet.

Example 2
Intravenous Solution Preparation

An intravenous dosage form of the active compound is prepared as follows:

Active compound 1-10 ug
Sodium citrate 5-50 mg
Citric acid 1-15 mg
Sodium chloride 1-8 mg
Water for injection (USP) q.s. to 1 mL

Utilizing the above quantities, the active compound is dissolved at room temperature in a prepared solution of sodium chloride, citric acid, and sodium citrate in water for injection.

Example 3
Intravenous Drip Preparation

0.25-2.5 mg compound dissolved in 250 ml of 10% glucose solution or in 250 ml of 0.9% NaCl solution.

Intravenous drip preparation: 1-2.mg compound dissolved in 250 ml of 10% glucose solution or in 250 ml of 0.9% NaCl solution

Treatment of angelic acid with one of the many standard chlorinating reagents including phosphorus oxychloride, phosphorus trichloride and thionyl chloride produces tigloyl chloride. Oxalyl chloride produces a 2:1 ratio of angeloyl chloride to tigloyl chloride. Treatment of potassium salt in diethyl ether with oxalyl chloride and catalytic DMF for 2 hr at 0° C. produces pure angeloyl chloride.

embedded image

Acid Hydrolysis of the Following Compounds

a) Xanifolia(Y),

or chemical name: 3-O-[β-D-galactopyranosyl (1→2)]-α-L-arabinofuranosy (1→3)-β-D-glucuronopyranosyl-21,22-O-diangeloyl-3β, 15α, 16α, 21β, 22α, 28-hexahydroxyolean-12-ene;

c) Xanifolia (Y2),

or chemical name: 3-O-[β-D-glucopyranosyl-(1→2)]-α-L-arabinofuranosy (1→3)-β-D-glucuronopyranosyl-21,22-O-diangeloyl-3β, 15α, 16α, 21β, 22α, 24β, 28-heptahydroxyolean-12-ene;

d) Xanifolia (Y8),

or chemical name: 3-O-[β-glucopyranosyl (1→2)]-α-arabinofuranosyl (1→3)-β-glucuronopyranosyl-21, 22-O-diangeloyl-3β, 16α, 21β, 22α, 24β, 28-hexahydroxyolean-12-ene;

f) Xanifolia (Y10),

or chemical name: 3-O-[β-galactopyranosyl (1→2)]-α-arabinofuranosyl (1→3)-β-glucuronopyranosyl-21, 22-O-diangeloyl-3β, 16α, 21β, 22α, 28-pentahydroxyolean-12-ene.

j) structure (M10)

m) structure (bES):

After acid hydrolysis of the above, an isolated, purified or synthesized compound is produced having a structure (ACH) selected from following:

embedded image - (ACH-Y)

embedded image - ACH-Y10;

embedded image - ACH-Y2;

embedded image - ACH-bES;

embedded image - ACH-M10;

embedded image - ACH-n2

embedded image - ACH-n3;

embedded image - ACH-n3

The composition comprises bioactive compounds from natural plants or synthesis.

The program is based on our purification methods and biological assays including the MTT assay. See International Application No. PCT/US05/31900, filed Sep. 7, 2005, U.S. Serial No. 11/289,142, filed Nov. 28, 2005, and U.S. Serial No. 11/131551, filed May 17, 2005, and PCT/US2008/002086, 1188-ALA-PCT, filed Feb. 15, 2008, 12/344,682, 1020-B1-US, filed Dec. 29, 2008. The details of Analysis of gene expression of ES2 cells after Y-treatment by Microarray, Data Analysis Methods and Western blot in PCT/US2008/002086, 1188-ALA-PCT, filed Feb. 15, 2008, and the cell invasion experiments methods in International Application PCT/US2010/0042240, filed Jul. 16, 2010.

The Haemolytic Assay

Erythrocytes (RBC) were isolated from human blood (EDTA whole blood, collected randomly). 50 ul of the 10% RBC suspension (in PBS) was added to 2 ml of sample solutions (concentration range from 0.1 ug/ml to 400 ug/ml) in PBS. The mixture was vortexed briefly and sat for 60 min at room temperature. The mixture was spun at 3 K for 10 min and the relative amounts of lysed hemoglobin in the supernatant were measured at 540 nm. The synthetic compounds of present application were tested with this method.

Acid Hydrolysis of Saponin

15 mg Xanifolia-Y was dissolved in 1 ml of methanol. 1 ml of 2N HCl was then added. The mixture was refluxed in 80° C. water bath for 5 hours. The solution was then neutralized by adding 2 ml of 1N NaOH (to final pH 4-6). The aglycone was then extracted with ethylacetate 3 ml × 2. The extracts were collected and pooled. Further isolation of aglycone (ACH-Y) was achieved by HPLC with isocratic elution of 80 -100% acetonitrile. Repeating the experiment with compounds Z4, Y10, Y2, Y8, Y7, Y0, X, M10 and ESCIN (bES) gives the following compounds respectively: ACH-Z4, ACH-Y10, ACH-Y2, ACH-Y8, ACH-Y7, ACH-Y0, ACH-X, ACH-E, ACH-Z5, ACH-M10 and ACH-bES.

Removal of the Acyl Group by Alkaline Hydrolysis

20 mg of Xanifolia-Y was dissolved in 0.5 ml of 1N NaOH. The solution was incubated in 80° C. water bath for 4 hours. It was cooled to room temperature before being neutralized with 0.5 ml 1N HCl (adjust pH to about 3). The mixture was extracted with 2 ml 1-butanol 3 times. The butanol fractions were collected and lyophilized. The hydrolyzed saponin was further purified with HPLC in a C-18 column eluted with 25% acetonitrile.

embedded image - AKOH-Y;

embedded image - AKOH-M10

Compounds AKOH-Y and AKOH-M10 do not show the ability to inhibit cancer growth, cancer invasion, cell invasion or cancer cell invasion.

Core Compound

A core compound or pentacyclic triterpenes, hydroxylated triterpenes is obtained by acid and alkaline hydroysis of saponin from natural sources. A pentacyclic triterpene can also be obtained by synthetic methods. A method for synthesizing the core compound is as follows:

Beta-Escin, compound Y, Y10, Y2, Y8, Y7, Y0, X, or M10 dissolved in 1 M NaOH (20 mg/ml) was incubated at 70° C. for 5 hours. The hydrolyzed solution was neutralized with HCl, and the water was evaporated by lyophilization. The product was dissolved in 50% methanol and 1N HCl. The mixture was incubated at 70° C. for 5 hours. The solution was neutralized with NaOH. The hydrolyzed product was extracted with ethylacetate, which was subsequently removed by evaporation. Further purification of the hydrolyzed product of core compounds including (E4A) were archived with FPLC chromatography in a C18 column equilibrated with 70% acetonitrile/TFA at the flow rate of 1 ml/min. The core compounds are obtained.

The core compounds do not show the ability to inhibit cancer growth, cancer invasion, or cell adhesion. The structures of core compounds:

embedded image - (A)

embedded image - (B)

embedded image - (C)

embedded image - (D1)

embedded image - (D2)

embedded image

also named as bES-core, E IV A, ES4A, E4A or (E);

embedded image - E4AY2;

embedded image - E6A

embedded image

also named as ES V, E5A or (F);

embedded image - (G)

wherein R1, R2, R5, R8 represent OH; R3 represents OH, H or absent; R4, R10 represent CH3 or CH2OH; R9, R11, R12, R13, R14, R15 represent CH3;

embedded image - (H1)

embedded image - (H2)

wherein R1, R2, R5, R8, R17, R18 represent OH; R3 represents OH, H or absent; R9, R11, R12, R13, R14, R15 represent CH3.

embedded image - (J)

A typical numbering 1 to 30 of carbon positions of a pentacyclic triterpene.

embedded image

wherein R1, R2, R5, R8, R17, R18 represent OH; R9, R11, R12, R13, R14, R15 represent CH3, also named E4A or (E).

embedded image - (P1)

embedded image - (P2)

wherein R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14, R15, R16 R17 represent H, OH, CH2OH, COOH, OR CH3, named as (P1), (P2).

This invention provides a method of synthesizing new active compounds. A method of attaching functional groups to the core compounds including but not limited to (A), (B), (C), (D1), (D2), (E), (F), (G), (H1), (H2), (J), E4A, E4A2Y, E6A, (P1), P(2),] involves esterification of core compounds with acyl halide, wherein the halide including chloride, bromide, fluoride and iodide, wherein the acyl halide comprises acyl chloride, wherein acyl chloride including but not limited to Tigloyl chloride, angeloyl chloride, Acetyl chloride, Crotonoyl chloride, 3,3-Dimethylacryloyl chloride, senecioyl chloride, Cinnamoyl chloride, Pentenoyl chloride, Hexanoyl chloride, benzoyl chloride, Ethylbutyryl chloride, Propionyl chloride, 2-Propenoyl chloride, Isobutyryl chloride, Butyryl chloride, (2E)-2-pentenoyl chloride, 4-Pentenoyl chloride, 5-Hexenoyl chloride, Heptanoyl chloride, Octanoyl chloride, Nonanoyl chloride, Decanoyl chloride, Lauroyl chloride, Myristoyl chloride, Oleoyl chloride for 5 sec, 1 min, 2 min, 5 min, 10 min, 30 min, 1 hr, 2 hr, 18 hr, 2 days or 3 days at 0° C., 25° C. or 75° C. temperature. At the end of reaction, 5 ml of 2N HCl or 1 M NaHCO3 is added to the reaction mixture. The solution is then extracted 3 times with 10 ml of ethyl acetate which is then evaporated under vacuum and at 45° C. and lyophilization. The reaction product is dissolved in 80% acetonitrile - 0.005% Trifluoroacetic acid. The active esterification products are purified with HPLC. MTT activity was performed to test the activity of acyl chloride, solution after the reaction, individual fractions, and individual compounds. The core compounds are synthetic, semi synthetic or from natural source. The core compounds are including terpene, isoprene, triterpenes, and hydroxylated triterpenes.

MTT activity of acylation of core compounds in different reaction time period of (ASAP)5 sec, 1 min, 2 min, 5 min, 10 min, 30 min, 1 hr, 2 hr, 18 hr, 2 days or 3 days at 0° C., 25° C. or 75° C. temperature were studied. HPLC profiles of esterification products of core compound E4A with acyl halide, wherein the halide comprise chloride, bromide, fluoride and iodide, wherein the acyl halide comprise acyl chloride, wherein acyl chloride comprise tigloyl chloride, angeloyl chloride, acetyl chloride, crotonoyl chloride, 3,3-Dimethylacryloyl chloride, senecioyl chloride, cinnamoyl chloride, pentenoyl chloride, hexanoyl chloride, benzoyl chloride, ethylbutyryl chloride, propionyl chloride, 2-propenoyl chloride, isobutyryl chloride, butyryl chloride, (2E)-2-pentenoyl chloride, 4-Pentenoyl chloride, 5-hexenoyl chloride, heptanoyl chloride, octanoyl chloride, nonanoyl chloride, decanoyl chloride, Lauroyl chloride, myristoyl chloride, oleoyl chloride show that the compounds vary in composition when the time or temperature of the reaction is changed. See example in FIGS. 1-12 (U.S. Serial No. 14/313080) and Experiments 1-29

The peaks, fractions and compounds are selected according to the activities of times studies and the changes of peaks. Selecting the HPLC fractions for isolation is according to the cytotoxic activity of the reaction product obtained at a specific time. The compounds having strong to weak activities are selected and isolated. Selecting the HPLC fractions for isolation may be according to the cytotoxic activity of times studies and the change of peaks. The anti-cancer activities are the MTT studies of bone (U2OS), lung (H460), bladder (HTB-9), ovary (ES2), colon (HCT116), pancreas (Capan), ovary(OVCAR3), prostate (DU145), skin (SK-Mel-5), mouth (KB), kidney (A498), breast (MCF-7), liver (HepG2), brain (T98G), leukemia (K562), cervix (HeLa).

Esterification of core compound E4A with Tigloyl chloride and isolation of the compounds with HPLC give the following compounds: wherein Tig=Tigloyl

embedded image

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

A1
OH
OH
OH
OH
O—Tig
OH
moderate

A2
OH
OH
OH
OH
OH
O—Tig
moderate

A3
OH
OH
OH
OH
O—Tig
O—Tig
strong

A4
O—Tig
OH
OH
OH
O—Tig
O—Tig
moderate

A5
OH
O—Tig
OH
OH
O—Tig
O—Tig
moderate

A6
OH
OH
O—Tig
OH
O—Tig
O—Tig
moderate

A7
OH
OH
OH
O—Tig
O—Tig
O—Tig
moderate

A8
O—Tig
O—Tig
OH
OH
O—Tig
O—Tig
weak

A9
OH
O—Tig
O—Tig
OH
O—Tig
O—Tig
weak

A10
OH
OH
O—Tig
O—Tig
O—Tig
O—Tig
weak

A11
O—Tig
OH
O—Tig
OH
O—Tig
O—Tig
weak

A12
OH
O—Tig
OH
O—Tig
O—Tig
O—Tig
weak

A13
O—Tig
OH
OH
O—Tig
O—Tig
O—Tig
weak

A14
OH
O—Tig
O—Tig
OH
O—Tig
O—Tig
weak

A15
O—Tig
O—Tig
O—Tig
OH
O—Tig
O—Tig
weak

A16
O—Tig
O—Tig
OH
O—Tig
O—Tig
O—Tig
weak

A17
O—Tig
OH
O—Tig
O—Tig
O—Tig
O—Tig
weak

A18
OH
O—Tig
O—Tig
O—Tig
O—Tig
O—Tig
weak

A19
O—Tig
O—Tig
O—Tig
O—Tig
O—Tig
O—Tig
none

A20
O—Tig
O—Tig
OH
OH
OH
O—Tig
moderate

A21
O—Tig
O—Tig
OH
OH
O—Tig
OH
moderate

A22
O—Tig
O—Tig
OH
O—Tig
OH
OH
moderate

A23
O—Tig
O—Tig
O—Tig
OH
OH
OH
moderate

A24
O—Tig
O—Tig
OH
OH
OH
OH
moderate

A25
O—Tig
OH
OH
OH
OH
O—Tig
moderate

A26
OH
O—Tig
OH
OH
OH
O—Tig
moderate

A27
OH
OH
O—Tig
OH
OH
O—Tig
moderate

A28
OH
OH
OH
O—Tig
OH
O—Tig
moderate

A29
O—Tig
OH
OH
OH
O—Tig
OH
moderate

A30
OH
O—Tig
OH
OH
O—Tig
OH
moderate

A31
OH
OH
O—Tig
OH
O—Tig
OH
moderate

A32
OH
OH
OH
O—Tig
O—Tig
OH
moderate

Esterification of core compound E4A with Angeloyl chloride and isolation of the compounds with HPLC give the following compounds: wherein Ang = Angeloyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

G1
OH
OH
OH
OH
O-Ang
OH
moderate

G2
OH
OH
OH
OH
OH
O-Ang
moderate

G3
OH
OH
OH
OH
O-Ang
O-Ang
strong

G4
O-Ang
OH
OH
OH
O-Ang
O-Ang
moderate

G5
OH
O-Ang
OH
OH
O-Ang
O-Ang
moderate

G6
OH
OH
O-Ang
OH
O-Ang
O-Ang
moderate

G7
OH
OH
OH
O-Ang
O-Ang
O-Ang
moderate

G8
O-Ang
O-Ang
OH
OH
O-Ang
O-Ang
weak

G9
OH
O-Ang
O-Ang
OH
O-Ang
O-Ang
weak

G10
OH
OH
O-Ang
O-Ang
O-Ang
O-Ang
weak

G11
O-Ang
OH
O-Ang
OH
O-Ang
O-Ang
weak

G12
OH
O-Ang
OH
O-Ang
O-Ang
O-Ang
weak

G13
O-Ang
OH
OH
O-Ang
O-Ang
O-Ang
weak

G14
OH
O-Ang
O-Ang
OH
O-Ang
O-Ang
weak

G15
O-Ang
O-Ang
O-Ang
OH
O-Ang
O-Ang
weak

G16
O-Ang
O-Ang
OH
O-Ang
O-Ang
O-Ang
weak

G17
O-Ang
OH
O-Ang
O-Ang
O-Ang
O-Ang
weak

G18
OH
O-Ang
O-Ang
O-Ang
O-Ang
O-Ang
weak

G19
O-Ang
O-Ang
O-Ang
O-Ang
O-Ang
O-Ang
none

G20
O-Ang
O-Ang
OH
OH
OH
O-Ang
moderate

G21
O-Ang
O-Ang
OH
OH
O-Ang
OH
moderate

G22
O-Ang
O-Ang
OH
O-Ang
OH
OH
moderate

G23
O-Ang
O-Ang
O-Ang
OH
OH
OH
moderate

G24
O-Ang
O-Ang
OH
OH
OH
OH
moderate

G25
O-Ang
OH
OH
OH
OH
O-Ang
moderate

G26
OH
O-Ang
OH
OH
OH
O-Ang
moderate

G27
OH
OH
O-Ang
OH
OH
O-Ang
moderate

G28
OH
OH
OH
O-Ang
OH
O-Ang
moderate

G29
O-Ang
OH
OH
OH
O-Ang
OH
moderate

G30
OH
O-Ang
OH
OH
O-Ang
OH
moderate

G31
OH
OH
O-Ang
OH
O-Ang
OH
moderate

G32
OH
OH
OH
O-Ang
O-Ang
OH
moderate

Esterification of core compound E4A with (3,3-Dimethylacryloyl chloride) senecioyl chloride and isolation of the compounds with HPLC give the following compounds: Wherein Sen = senecioyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

B1
OH
OH
OH
OH
-Sen
OH
moderate

B2
OH
OH
OH
OH
OH
O-Sen
moderate

B3
OH
OH
OH
OH
O-Sen
O-Sen
strong

B4
O-Sen
OH
OH
OH
O-Sen
O-Sen
moderate

B5
OH
O-Sen
OH
OH
O-Sen
O-Sen
moderate

B6
OH
OH
O-Sen
OH
O-Sen
O-Sen
moderate

B7
OH
OH
OH
O-Sen
O-Sen
O-Sen
moderate

B8
O-Sen
O-Sen
OH
OH
O-Sen
O-Sen
weak

B9
OH
O-Sen
O-Sen
OH
O-Sen
O-Sen
weak

B10
OH
OH
O-Sen
O-Sen
O-Sen
O-Sen
weak

B11
O-Sen
OH
O-Sen
OH
O-Sen
O-Sen
weak

B12
OH
O-Sen
OH
O-Sen
O-Sen
O-Sen
weak

B13
O-Sen
OH
OH
O-Sen
O-Sen
O-Sen
weak

B14
OH
O-Sen
O-Sen
OH
O-Sen
O-Sen
weak

B15
O-Sen
O-Sen
O-Sen
OH
O-Sen
O-Sen
weak

B16
O-Sen
O-Sen
OH
O-Sen
O-Sen
O-Sen
weak

B17
O-Sen
OH
O-Sen
O-Sen
O-Sen
O-Sen
weak

B18
OH
O-Sen
O-Sen
O-Sen
O-Sen
O-Sen
weak

B19
O-Sen
O-Sen
O-Sen
O-Sen
O-Sen
O-Sen
none

B20
O-Sen
O-Sen
OH
OH
OH
O-Sen
moderate

B21
O-Sen
O-Sen
OH
OH
O-Sen
OH
moderate

B22
O-Sen
O-Sen
OH
O-Sen
OH
OH
moderate

B23
O-Sen
O-Sen
O-Sen
OH
OH
OH
moderate

B24
O-Sen
O-Sen
OH
OH
OH
OH
moderate

B25
O-Sen
OH
OH
OH
OH
O-Sen
moderate

B26
OH
O-Sen
OH
OH
OH
O-Sen
moderate

B27
OH
OH
O-Sen
OH
OH
O-Sen
moderate

B28
OH
OH
OH
O-Sen
OH
O-Sen
moderate

B29
O-Sen
OH
OH
OH
O-Sen
OH
moderate

B30
OH
O-Sen
OH
OH
O-Sen
OH
moderate

B31
OH
OH
O-Sen
OH
O-Sen
OH
moderate

B32
OH
OH
OH
O-Sen
O-Sen
OH
moderate

Esterification of core compound E4A with 4-Pentenoyl chloride and isolation of the compounds with HPLC give the following compounds: wherein Pen = 4-Pentenoyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

C1
OH
OH
OH
OH
O-Pen
OH
moderate

C2
OH
OH
OH
OH
OH
O-Pen
moderate

C3
OH
OH
OH
OH
O-Pen
O-Pen
strong

C4
O-Pen
OH
OH
OH
O-Pen
O-Pen
moderate

C5
OH
O-Pen
OH
OH
O-Pen
O-Pen
moderate

C6
OH
OH
O-Pen
OH
O-Pen
O-Pen
moderate

C7
OH
OH
OH
O-Pen
O-Pen
O-Pen
moderate

C8
O-Pen
O-Pen
OH
OH
O-Pen
O-Pen
weak

C9
OH
O-Pen
O-Pen
OH
O-Pen
O-Pen
weak

C10
OH
OH
O-Pen
O-Pen
O-Pen
O-Pen
weak

C11
O-Pen
OH
O-Pen
OH
O-Pen
O-Pen
weak

C12
OH
O-Pen
OH
O-Pen
O-Pen
O-Pen
weak

C13
O-Pen
OH
OH
O-Pen
O-Pen
O-Pen
weak

C14
OH
O-Pen
O-Pen
OH
O-Pen
O-Pen
weak

C15
O-Pen
O-Pen
O-Pen
OH
O-Pen
O-Pen
weak

C16
O-Pen
O-Pen
OH
O-Pen
O-Pen
O-Pen
weak

C17
O-Pen
OH
O-Pen
O-Pen
O-Pen
O-Pen
weak

C18
OH
O-Pen
O-Pen
O-Pen
O-Pen
O-Pen
weak

C19
O-Pen
O-Pen
O-Pen
O-Pen
O-Pen
O-Pen
none

C20
O-Pen
O-Pen
OH
OH
OH
O-Pen
moderate

C21
O-Pen
O-Pen
OH
OH
O-Pen
OH
moderate

C22
O-Pen
O-Pen
OH
O-Pen
OH
OH
moderate

C23
O-Pen
O-Pen
O-Pen
OH
OH
OH
moderate

C24
O-Pen
O-Pen
OH
OH
OH
OH
moderate

C25
O-Pen
OH
OH
OH
OH
O-Pen
moderate

C26
OH
O-Pen
OH
OH
OH
O-Pen
moderate

C27
OH
OH
O-Pen
OH
OH
O-Pen
moderate

C28
OH
OH
OH
O-Pen
OH
O-Pen
moderate

C29
O-Pen
OH
OH
OH
O-Pen
OH
moderate

C30
OH
O-Pen
OH
OH
O-Pen
OH
moderate

C31
OH
OH
O-Pen
OH
O-Pen
OH
moderate

C32
OH
OH
OH
O-Pen
O-Pen
OH
moderate

Esterification of core compound E4A with Hexanoyl chloride and isolation of the compounds with HPLC give the following compounds: wherein Hex = Hexanoyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

D1
OH
OH
OH
OH
O—Hex
OH
moderate

D2
OH
OH
OH
OH
OH
O—Hex
moderate

D3
OH
OH
OH
OH
O—Hex
O—Hex
strong

D4
O—Hex
OH
OH
OH
O—Hex
O—Hex
moderate

D5
OH
O—Hex
OH
OH
O—Hex
O—Hex
moderate

D6
OH
OH
O—Hex
OH
O—Hex
O—Hex
moderate

D7
OH
OH
OH
O—Hex
O—Hex
O—Hex
moderate

D8
O—Hex
O—Hex
OH
OH
O—Hex
O—Hex
weak

D9
OH
O—Hex
O—Hex
OH
O—Hex
O—Hex
weak

D10
OH
OH
O—Hex
O—Hex
O—Hex
O—Hex
weak

D11
O—Hex
OH
O—Hex
OH
O—Hex
O—Hex
weak

D12
OH
O—Hex
OH
O—Hex
O—Hex
O—Hex
weak

D13
O—Hex
OH
OH
O—Hex
O—Hex
O—Hex
weak

D14
OH
O—Hex
O—Hex
OH
O—Hex
O—Hex
weak

D15
O—Hex
O—Hex
O—Hex
OH
O—Hex
O—Hex
weak

D16
O—Hex
O—Hex
OH
O—Hex
O—Hex
O—Hex
weak

D17
O—Hex
OH
O—Hex
O—Hex
O—Hex
O—Hex
weak

D18
OH
O—Hex
O—Hex
O—Hex
O—Hex
O—Hex
weak

D19
O—Hex
O—Hex
O—Hex
O—Hex
O—Hex
O—Hex
none

D20
O—Hex
O—Hex
OH
OH
OH
O—Hex
moderate

D21
O—Hex
O—Hex
OH
OH
O—Hex
OH
moderate

D22
O—Hex
O—Hex
OH
O—Hex
OH
OH
moderate

D23
O—Hex
O—Hex
O—Hex
OH
OH
OH
moderate

D24
O—Hex
O—Hex
OH
OH
OH
OH
moderate

D25
O—Hex
OH
OH
OH
OH
O—Hex
moderate

D26
OH
O—Hex
OH
OH
OH
O—Hex
moderate

D27
OH
OH
O—Hex
OH
OH
O—Hex
moderate

D28
OH
OH
OH
O—Hex
OH
O—Hex
moderate

D29
O—Hex
OH
OH
OH
O—Hex
OH
moderate

D30
OH
O—Hex
OH
OH
O—Hex
OH
moderate

D31
OH
OH
O—Hex
OH
O—Hex
OH
moderate

D32
OH
OH
OH
O—Hex
O—Hex
OH
moderate

Esterification of core compound E4A with 2-Ethylbutyryl chloride and isolation of the compounds with HPLC give the following compounds: wherein Eth= 2-Ethylbutyryl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

E1
OH
OH
OH
OH
O—Eth
OH
moderate

E2
OH
OH
OH
OH
OH
O—Eth
moderate

E3
OH
OH
OH
OH
O—Eth
O—Eth
strong

E4
O—Eth
OH
OH
OH
O—Eth
O—Eth
moderate

E5
OH
O—Eth
OH
OH
O—Eth
O—Eth
moderate

E6
OH
OH
O—Eth
OH
O—Eth
O—Eth
moderate

E7
OH
OH
OH
O—Eth
O—Eth
O—Eth
moderate

E8
O—Eth
O—Eth
OH
OH
O—Eth
O—Eth
weak

E9
OH
O—Eth
O—Eth
OH
O—Eth
O—Eth
weak

E10
OH
OH
O—Eth
O—Eth
O—Eth
O—Eth
weak

E11
O—Eth
OH
O—Eth
OH
O—Eth
O—Eth
weak

E12
OH
O—Eth
OH
O—Eth
O—Eth
O—Eth
weak

E13
O—Eth
OH
OH
O—Eth
O—Eth
O—Eth
weak

E14
OH
O—Eth
O—Eth
OH
O—Eth
O—Eth
weak

E15
O—Eth
O—Eth
O—Eth
OH
O—Eth
O—Eth
weak

E16
O—Eth
O—Eth
OH
O—Eth
O—Eth
O—Eth
weak

E17
O—Eth
OH
O—Eth
O—Eth
O—Eth
O—Eth
weak

E18
OH
O—Eth
O—Eth
O—Eth
O—Eth
O—Eth
weak

E19
O—Eth
O—Eth
O—Eth
O—Eth
O—Eth
O—Eth
none

E20
O—Eth
O—Eth
OH
OH
OH
O—Eth
moderate

E21
O—Eth
O—Eth
OH
OH
O—Eth
OH
moderate

E22
O—Eth
O—Eth
OH
O—Eth
OH
OH
moderate

E23
O—Eth
O—Eth
O—Eth
OH
OH
OH
moderate

E24
O—Eth
O—Eth
OH
OH
OH
OH
moderate

E25
O—Eth
OH
OH
OH
OH
O—Eth
moderate

E26
OH
O—Eth
OH
OH
OH
O—Eth
moderate

E27
OH
OH
O—Eth
OH
OH
O—Eth
moderate

E28
OH
OH
OH
O—Eth
OH
O—Eth
moderate

E29
O—Eth
OH
OH
OH
O—Eth
OH
moderate

E30
OH
O—Eth
OH
OH
O—Eth
OH
moderate

E31
OH
OH
O—Eth
OH
O—Eth
OH
moderate

E32
OH
OH
OH
O—Eth
O—Eth
OH
moderate

Esterification of core compound E4A with Acetyl chloride (H) and isolation of the compounds with HPLC give the following compounds: wherein Acy = Acetyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

H1
OH
OH
OH
OH
O—Acy
OH
moderate

H2
OH
OH
OH
OH
OH
O—Acy
moderate

H3
OH
OH
OH
OH
O—Acy
O—Acy
strong

H4
O—Acy
OH
OH
OH
O—Acy
O—Acy
moderate

H5
OH
O—Acy
OH
OH
O—Acy
O—Acy
moderate

H6
OH
OH
O—Acy
OH
O—Acy
O—Acy
moderate

H7
OH
OH
OH
O—Acy
O—Acy
O—Acy
moderate

H8
O—Acy
O—Acy
OH
OH
O—Acy
O—Acy
weak

H9
OH
O—Acy
O—Acy
OH
O—Acy
O—Acy
weak

H10
OH
OH
O—Acy
O—Acy
O—Acy
O—Acy
weak

H11
O—Acy
OH
O—Acy
OH
O—Acy
O—Acy
weak

H12
OH
O—Acy
OH
O—Acy
O—Acy
O—Acy
weak

H13
O—Acy
OH
OH
O—Acy
O—Acy
O—Acy
weak

H14
OH
O—Acy
O—Acy
OH
O—Acy
O—Acy
weak

H15
O—Acy
O—Acy
O—Acy
OH
O—Acy
O—Acy
weak

H16
O—Acy
O—Acy
OH
O—Acy
O—Acy
O—Acy
weak

H17
O—Acy
OH
O—Acy
O—Acy
O—Acy
O—Acy
weak

H18
OH
O—Acy
O—Acy
O—Acy
O—Acy
O—Acy
weak

H19
O—Acy
O—Acy
O—Acy
O—Acy
O—Acy
O—Acy
none

H20
O—Acy
O—Acy
OH
OH
OH
O—Acy
moderate

H21
O—Acy
O—Acy
OH
OH
O—Acy
OH
moderate

H22
O—Acy
O—Acy
OH
O—Acy
OH
OH
moderate

H23
O—Acy
O—Acy
O—Acy
OH
OH
OH
moderate

H24
O—Acy
O—Acy
OH
OH
OH
OH
moderate

H25
O—Acy
OH
OH
OH
OH
O—Acy
moderate

H26
OH
O—Acy
OH
OH
OH
O—Acy
moderate

H27
OH
OH
O—Acy
OH
OH
O—Acy
moderate

H28
OH
OH
OH
O—Acy
OH
O—Acy
moderate

H29
O—Acy
OH
OH
OH
O—Acy
OH
moderate

H30
OH
O—Acy
OH
OH
O—Acy
OH
moderate

H31
OH
OH
O—Acy
OH
O—Acy
OH
moderate

H32
OH
OH
OH
O—Acy
O—Acy
OH
moderate

Esterification of core compound E4A with Crotonoyl chloride and isolation of the compounds with HPLC give the following compounds: wherein Cro = Crotonoyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

I1
OH
OH
OH
OH
O—Cro
OH
moderate

I2
OH
OH
OH
OH
OH
O—Cro
moderate

I3
OH
OH
OH
OH
O—Cro
O—Cro
strong

I4
O—Cro
OH
OH
OH
O—Cro
O—Cro
moderate

I5
OH
O—Cro
OH
OH
O—Cro
O—Cro
moderate

I6
OH
OH
O—Cro
OH
O—Cro
O—Cro
moderate

I7
OH
OH
OH
O—Cro
O—Cro
O—Cro
moderate

I8
O—Cro
O—Cro
OH
OH
O—Cro
O—Cro
weak

I9
OH
O—Cro
O—Cro
OH
O—Cro
O—Cro
weak

I10
OH
OH
O—Cro
O—Cro
O—Cro
O—Cro
weak

I11
O—Cro
OH
O—Cro
OH
O—Cro
O—Cro
weak

I12
OH
O—Cro
OH
O—Cro
O—Cro
O—Cro
weak

I13
O—Cro
OH
OH
O—Cro
O—Cro
O—Cro
weak

I14
OH
O—Cro
O—Cro
OH
O—Cro
O—Cro
weak

I15
O—Cro
O—Cro
O—Cro
OH
O—Cro
O—Cro
weak

I16
O—Cro
O—Cro
OH
O—Cro
O—Cro
O—Cro
weak

I17
O—Cro
OH
O—Cro
O—Cro
O—Cro
O—Cro
weak

I18
OH
O—Cro
O—Cro
O—Cro
O—Cro
O—Cro
weak

I19
O—Cro
O—Cro
O—Cro
O—Cro
O—Cro
O—Cro
none

I20
O—Cro
O—Cro
OH
OH
OH
O—Cro
moderate

I21
O—Cro
O—Cro
OH
OH
O—Cro
OH
moderate

I22
O—Cro
O—Cro
OH
O—Cro
OH
OH
moderate

I23
O—Cro
O—Cro
O—Cro
OH
OH
OH
moderate

I24
O—Cro
O—Cro
OH
OH
OH
OH
moderate

I25
O—Cro
OH
OH
OH
OH
O—Cro
moderate

I26
OH
O—Cro
OH
OH
OH
O—Cro
moderate

I27
OH
OH
O—Cro
OH
OH
O—Cro
moderate

I28
OH
OH
OH
O—Cro
OH
O—Cro
moderate

I29
O—Cro
OH
OH
OH
O—Cro
OH
moderate

I30
OH
O—Cro
OH
OH
O—Cro
OH
moderate

I31
OH
OH
O—Cro
OH
O—Cro
OH
moderate

I32
OH
OH
OH
O—Cro
O—Cro
OH
moderate

Esterification of core compound E4A with Cinnamoyl chloride and isolation of the compounds with HPLC give the following compounds: wherein Cin = Cinnamoyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

J1
OH
OH
OH
OH
O—Cin
OH
moderate

J2
OH
OH
OH
OH
OH
O—Cin
moderate

J3
OH
OH
OH
OH
O—Cin
O—Cin
strong

J4
O—Cin
OH
OH
OH
O—Cin
O—Cin
moderate

J5
OH
O—Cin
OH
OH
O—Cin
O—Cin
moderate

J6
OH
OH
O—Cin
OH
O—Cin
O—Cin
moderate

J7
OH
OH
OH
O—Cin
O—Cin
O—Cin
moderate

J8
O—Cin
O—Cin
OH
OH
O—Cin
O—Cin
weak

J9
OH
O—Cin
O—Cin
OH
O—Cin
O—Cin
weak

J10
OH
OH
O—Cin
O—Cin
O—Cin
O—Cin
weak

J11
O—Cin
OH
O—Cin
OH
O—Cin
O—Cin
weak

J12
OH
O—Cin
OH
O—Cin
O—Cin
O—Cin
weak

J13
O—Cin
OH
OH
O—Cin
O—Cin
O—Cin
weak

J14
OH
O—Cin
O—Cin
OH
O—Cin
O—Cin
weak

J15
O—Cin
O—Cin
O—Cin
OH
O—Cin
O—Cin
weak

J16
O—Cin
O—Cin
OH
O—Cin
O—Cin
O—Cin
weak

J17
O—Cin
OH
O—Cin
O—Cin
O—Cin
O—Cin
weak

J18
OH
O—Cin
O—Cin
O—Cin
O—Cin
O—Cin
weak

J19
O—Cin
O—Cin
O—Cin
O—Cin
O—Cin
O—Cin
none

J20
O—Cin
O—Cin
OH
OH
OH
O—Cin
moderate

J21
O—Cin
O—Cin
OH
OH
O—Cin
OH
moderate

J22
O—Cin
O—Cin
OH
O—Cin
OH
OH
moderate

J23
O—Cin
O—Cin
O—Cin
OH
OH
OH
moderate

J24
O—Cin
O—Cin
OH
OH
OH
OH
moderate

J25
O—Cin
OH
OH
OH
OH
O—Cin
moderate

J26
OH
O—Cin
OH
OH
OH
O—Cin
moderate

J27
OH
OH
O—Cin
OH
OH
O—Cin
moderate

J28
OH
OH
OH
O—Cin
OH
O—Cin
moderate

J29
O—Cin
OH
OH
OH
O—Cin
OH
moderate

J30
OH
O—Cin
OH
OH
O—Cin
OH
moderate

J31
OH
OH
O—Cin
OH
O—Cin
OH
moderate

J32
OH
OH
OH
O—Cin
O—Cin
OH
moderate

Esterification of core compound E4A with benzoyl chloride and isolation of the compounds with HPLC give the following compounds: wherein Ben = benzoyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
None

K1
OH
OH
OH
OH
O—Ben
OH
moderate

K2
OH
OH
OH
OH
OH
O—Ben
moderate

K3
OH
OH
OH
OH
O—Ben
O—Ben
strong

K4
O—Ben
OH
OH
OH
O—Ben
O—Ben
moderate

K5
OH
O—Ben
OH
OH
O—Ben
O—Ben
moderate

K6
OH
OH
O—Ben
OH
O—Ben
O—Ben
moderate

K7
OH
OH
OH
O—Ben
O—Ben
O—Ben
moderate

K8
O—Ben
O—Ben
OH
OH
O—Ben
O—Ben
weak

K9
OH
O—Ben
O—Ben
OH
O—Ben
O—Ben
weak

K10
OH
OH
O—Ben
O—Ben
O—Ben
O—Ben
weak

K11
O—Ben
OH
O—Ben
OH
O—Ben
O—Ben
weak

K12
OH
O—Ben
OH
O—Ben
O—Ben
O—Ben
weak

K13
O—Ben
OH
OH
O—Ben
O—Ben
O—Ben
weak

K14
OH
O—Ben
O—Ben
OH
O—Ben
O—Ben
weak

K15
O—Ben
O—Ben
O—Ben
OH
O—Ben
O—Ben
weak

K16
O—Ben
O—Ben
OH
O—Ben
O—Ben
O—Ben
weak

K17
O—Ben
OH
O—Ben
O—Ben
O—Ben
O—Ben
weak

K18
OH
O—Ben
O—Ben
O—Ben
O—Ben
O—Ben
weak

K19
O—Ben
O—Ben
O—Ben
O—Ben
O—Ben
O—Ben
none

K20
O—Ben
O—Ben
OH
OH
OH
O—Ben
moderate

K21
O—Ben
O—Ben
OH
OH
O—Ben
OH
moderate

K22
O—Ben
O—Ben
OH
O—Ben
OH
OH
moderate

K23
O—Ben
O—Ben
O—Ben
OH
OH
OH
moderate

K24
O—Ben
O—Ben
OH
OH
OH
OH
moderate

K25
O—Ben
OH
OH
OH
OH
O—Ben
moderate

K26
OH
O—Ben
OH
OH
OH
O—Ben
moderate

K27
OH
OH
O—Ben
OH
OH
O—Ben
moderate

K28
OH
OH
OH
O—Ben
OH
O—Ben
moderate

K29
O—Ben
OH
OH
OH
O—Ben
OH
moderate

K30
OH
O—Ben
OH
OH
O—Ben
OH
moderate

K31
OH
OH
O—Ben
OH
O—Ben
OH
moderate

K32
OH
OH
OH
O—Ben
O—Ben
OH
moderate

Esterification of core compound E4A with Propionyl chloride and isolation of the compounds with HPLC give the following compounds: wherein Ppi = Propionyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

K1
OH
OH
OH
OH
O—Ppi
OH
moderate

K2
OH
OH
OH
OH
OH
O—Ppi
moderate

K3
OH
OH
OH
OH
O—Ppi
O—Ppi
strong

K4
O—Ppi
OH
OH
OH
O—Ppi
O—Ppi
moderate

K5
OH
O—Ppi
OH
OH
O—Ppi
O—Ppi
moderate

K6
OH
OH
O—Ppi
OH
O—Ppi
O—Ppi
moderate

K7
OH
OH
OH
O—Ppi
O—Ppi
O—Ppi
moderate

K8
O—Ppi
O—Ppi
OH
OH
O—Ppi
O—Ppi
weak

K9
OH
O—Ppi
O—Ppi
OH
O—Ppi
O—Ppi
weak

K10
OH
OH
O—Ppi
O—Ppi
O—Ppi
O—Ppi
weak

K11
O—Ppi
OH
O—Ppi
OH
O—Ppi
O—Ppi
weak

K12
OH
O—Ppi
OH
O—Ppi
O—Ppi
O—Ppi
weak

K13
O—Ppi
OH
OH
O—Ppi
O—Ppi
O—Ppi
weak

K14
OH
O—Ppi
O—Ppi
OH
O—Ppi
O—Ppi
weak

K15
O—Ppi
O—Ppi
O—Ppi
OH
O—Ppi
O—Ppi
weak

K16
O—Ppi
O—Ppi
OH
O—Ppi
O—Ppi
O—Ppi
weak

K17
O—Ppi
OH
O—Ppi
O—Ppi
O—Ppi
O—Ppi
weak

K18
OH
O—Ppi
O—Ppi
O—Ppi
O—Ppi
O—Ppi
weak

K19
O—Ppi
O—Ppi
O—Ppi
O—Ppi
O—Ppi
O—Ppi
none

K20
O—Ppi
O—Ppi
OH
OH
OH
O—Ppi
moderate

K21
O—Ppi
O—Ppi
OH
OH
O—Ppi
OH
moderate

K22
O—Ppi
O—Ppi
OH
O—Ppi
OH
OH
moderate

K23
O—Ppi
O—Ppi
O—Ppi
OH
OH
OH
moderate

K24
O—Ppi
O—Ppi
OH
OH
OH
OH
moderate

K25
O—Ppi
OH
OH
OH
OH
O—Ppi
moderate

K26
OH
O—Ppi
OH
OH
OH
O—Ppi
moderate

K27
OH
OH
O—Ppi
OH
OH
O—Ppi
moderate

K28
OH
OH
OH
O—Ppi
OH
O—Ppi
moderate

K29
O—Ppi
OH
OH
OH
O—Ppi
OH
moderate

K30
OH
O—Ppi
OH
OH
O—Ppi
OH
moderate

K31
OH
OH
O—Ppi
OH
O—Ppi
OH
moderate

K32
OH
OH
OH
O—Ppi
O—Ppi
OH
moderate

Esterification of core compound E4A with 2-propenoyl chloride and isolation of the compounds with HPLC give the following compounds: wherein Ppe = Propenoyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

K1
OH
OH
OH
OH
O-Ppe
OH
moderate

K2
OH
OH
OH
OH
OH
O—Ppe
moderate

K3
OH
OH
OH
OH
O—Ppe
O—Ppe
strong

K4
O—Ppe
OH
OH
OH
O—Ppe
O—Ppe
moderate

K5
OH
O—Ppe
OH
OH
O—Ppe
O—Ppe
moderate

K6
OH
OH
O—Ppe
OH
O—Ppe
O—Ppe
moderate

K7
OH
OH
OH
O—Ppe
O—Ppe
O—Ppe
moderate

K8
O—Ppe
O—Ppe
OH
OH
O—Ppe
O—Ppe
weak

K9
OH
O—Ppe
O—Ppe
OH
—Ppe
O—Ppe
weak

K10
OH
OH
O—Ppe
O—Ppe
O—Ppe
O—Ppe
weak

K11
O—Ppe
OH
O—Ppe
OH
O—Ppe
O—Ppe
weak

K12
OH
O—Ppe
OH
O—Ppe
O—Ppe
O—Ppe
weak

K13
O—Ppe
OH
OH
O—Ppe
O—Ppe
O—Ppe
weak

K14
OH
O—Ppe
O—Ppe
OH
O—Ppe
O—Ppe
weak

K15
O—Ppe
O—Ppe
O—Ppe
OH
O—Ppe
O—Ppe
weak

K16
O—Ppe
O—Ppe
OH
O—Ppe
O—Ppe
O—Ppe
weak

K17
O—Ppe
OH
O—Ppe
O—Ppe
O—Ppe
O—Ppe
weak

K18
OH
O—Ppe
O—Ppe
O—Ppe
O—Ppe
O—Ppe
weak

K19
O—Ppe
O—Ppe
O—Ppe
O—Ppe
O—Ppe
O—Ppe
none

K20
O—Ppe
O—Ppe
OH
OH
OH
O—Ppe
moderate

K21
O—Ppe
O—Ppe
OH
OH
O—Ppe
OH
moderate

K22
O—Ppe
O—Ppe
OH
O—Ppe
OH
OH
moderate

K23
O—Ppe
O—Ppe
O—Ppe
OH
OH
OH
moderate

K24
O—Ppe
O—Ppe
OH
OH
OH
OH
moderate

K25
O—Ppe
OH
OH
OH
OH
O—Ppe
moderate

K26
OH
O—Ppe
OH
OH
OH
O—Ppe
moderate

K27
OH
OH
O—Ppe
OH
OH
O—Ppe
moderate

K28
OH
OH
OH
O—Ppe
OH
O—Ppe
moderate

K29
O—Ppe
OH
OH
OH
O—Ppe
OH
moderate

K30
OH
O—Ppe
OH
OH
O—Ppe
OH
moderate

K31
OH
OH
O—Ppe
OH
O—Ppe
OH
moderate

K32
OH
OH
OH
O—Ppe
O—Ppe
OH
moderate

Esterification of core compound E4A with Isobutyryl chloride and isolation of the compounds with HPLC give the following compounds: wherein Iso= Isobutyryl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

K1
OH
OH
OH
OH
O—Iso
OH
moderate

K2
OH
OH
OH
OH
OH
O—Iso
moderate

K3
OH
OH
OH
OH
O—Iso
O—Iso
strong

K4
O—Iso
OH
OH
OH
O—Iso
O—Iso
moderate

K5
OH
O—Iso
OH
OH
O—Iso
O—Iso
moderate

K6
OH
OH
O—Iso
OH
O—Iso
O—Iso
moderate

K7
OH
OH
OH
O—Iso
O—Iso
O—Iso
moderate

K8
O—Iso
O—Iso
OH
OH
O—Iso
O—Iso
weak

K9
OH
O—Iso
O—Iso
OH
O—Iso
O—Iso
weak

K10
OH
OH
O—Iso
O—Iso
O—Iso
O—Iso
weak

K11
O—Iso
OH
O—Iso
OH
O—Iso
O—Iso
weak

K12
OH
O—Iso
OH
O—Iso
O—Iso
O—Iso
weak

K13
O—Iso
OH
OH
O—Iso
O—Iso
O—Iso
weak

K14
OH
O—Iso
O—Iso
OH
O—Iso
O—Iso
weak

K15
O—Iso
O—Iso
O—Iso
OH
O—Iso
O—Iso
weak

K16
O—Iso
O—Iso
OH
O—Iso
O—Iso
O—Iso
weak

K17
O—Iso
OH
O—Iso
O—Iso
O—Iso
O—Iso
weak

K18
OH
O—Iso
O—Iso
O—Iso
O—Iso
O—Iso
weak

K19
O—Iso
O—Iso
O—Iso
O—Iso
O—Iso
O—Iso
none

K20
O—Iso
O—Iso
OH
OH
OH
O—Iso
moderate

K21
O—Iso
O—Iso
OH
OH
O—Iso
OH
moderate

K22
O—Iso
O—Iso
OH
O—Iso
OH
OH
moderate

K23
O—Iso
O—Iso
O—Iso
OH
OH
OH
moderate

K24
O—Iso
O—Iso
OH
OH
OH
OH
moderate

K25
O—Iso
OH
OH
OH
OH
O—Iso
moderate

K26
OH
O—Iso
OH
OH
OH
O—Iso
moderate

K27
OH
OH
O—Iso
OH
OH
O—Iso
moderate

K28
OH
OH
OH
O—Iso
OH
O—Iso
moderate

K29
O—Iso
OH
OH
OH
O—Iso
OH
moderate

K30
OH
O—Iso
OH
OH
O—Iso
OH
moderate

K31
OH
OH
O—Iso
OH
O—Iso
OH
moderate

K32
OH
OH
OH
O—Iso
O—Iso
OH
moderate

Esterification of core compound E4A with Butyryl chloride and isolation of the compounds with HPLC give the following compounds: wherein But = Butyryl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

K1
OH
OH
OH
OH
O—But
OH
moderate

K2
OH
OH
OH
OH
OH
O—But
moderate

K3
OH
OH
OH
OH
O—But
O—But
strong

K4
O—But
OH
OH
OH
O—But
O—But
moderate

K5
OH
O—But
OH
OH
O—But
O—But
moderate

K6
OH
OH
O—But
OH
O—But
O—But
moderate

K7
OH
OH
OH
O—But
O—But
O—But
moderate

K8
O—But
O—But
OH
OH
O—But
O—But
weak

K9
OH
O—But
O—But
OH
O—But
O—But
weak

K10
OH
OH
O—But
O—But
O—But
O—But
weak

K11
O—But
OH
O—But
OH
O—But
O—But
weak

K12
OH
O—But
OH
O—But
O—But
O—But
weak

K13
O—But
OH
OH
O—But
O—But
O—But
weak

K14
OH
O—But
O—But
OH
O—But
O—But
weak

K15
O—But
O—But
O—But
OH
O—But
O—But
weak

K16
O—But
O—But
OH
O—But
O—But
O—But
weak

K17
O—But
OH
O—But
O—But
O—But
O—But
weak

K18
OH
O—But
O—But
O—But
O—But
O—But
weak

K19
O—But
O—But
O—But
O—But
O—But
O—But
none

K20
O—But
O—But
OH
OH
OH
O—But
moderate

K21
O—But
O—But
OH
OH
O—But
OH
moderate

K22
O—But
O—But
OH
O—But
OH
OH
moderate

K23
O—But
O—But
O—But
OH
OH
OH
moderate

K24
O—But
O—But
OH
OH
OH
OH
moderate

K25
O—But
OH
OH
OH
OH
O—But
moderate

K26
OH
O—But
OH
OH
OH
O—But
moderate

K27
OH
OH
O—But
OH
OH
O—But
moderate

K28
OH
OH
OH
O—But
OH
O—But
moderate

K29
O—But
OH
OH
OH
O—But
OH
moderate

K30
OH
O—But
OH
OH
O—But
OH
moderate

K31
OH
OH
O—But
OH
O—But
OH
moderate

K32
OH
OH
OH
O—But
O—But
OH
moderate

Esterification of core compound E4A with (2E)-2-pentenoyl chloride and isolation of the compounds with HPLC give the following compounds: wherein 2pe = 2-pentenoyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

K1
OH
OH
OH
OH
O—2pe
OH
moderate

K2
OH
OH
OH
OH
OH
O—2pe
moderate

K3
OH
OH
OH
OH
O—2pe
O—2pe
strong

K4
O—2pe
OH
OH
OH
O—2pe
O—2pe
moderate

K5
OH
O—2pe
OH
OH
O—2pe
O—2pe
moderate

K6
OH
OH
O—2pe
OH
O—2pe
O—2pe
moderate

K7
OH
OH
OH
O—2pe
O—2pe
O—2pe
moderate

K8
O—2pe
O—2pe
OH
OH
O—2pe
O—2pe
weak

K9
OH
O—2pe
O—2pe
OH
O—2pe
O—2pe
weak

K10
OH
OH
O—2pe
O—2pe
O—2pe
O—2pe
weak

K11
O—2pe
OH
O—2pe
OH
O—2pe
O—2pe
weak

K12
OH
O—2pe
OH
O—2pe
O—2pe
O—2pe
weak

K13
O—2pe
OH
OH
O—2pe
O—2pe
O—2pe
weak

K14
OH
O—2pe
O—2pe
OH
O—2pe
O—2pe
weak

K15
O—2pe
O—2pe
O—2pe
OH
O—2pe
O—2pe
weak

K16
O—2pe
O—2pe
OH
O—2pe
O—2pe
O—2pe
weak

K17
O—2pe
OH
O—2pe
O—2pe
O—2pe
O—2pe
weak

K18
OH
O—2pe
O—2pe
O—2pe
O—2pe
O—2pe
weak

K19
O—2pe
O—2pe
O—2pe
O—2pe
O—2pe
O—2pe
none

K20
O—2pe
O—2pe
OH
OH
OH
O—2pe
moderate

K21
O—2pe
O—2pe
OH
OH
O—2pe
OH
moderate

K22
O—2pe
O—2pe
OH
O—2pe
OH
OH
moderate

K23
O—2pe
O—2pe
O—2pe
OH
OH
OH
moderate

K24
O—2pe
O—2pe
OH
OH
OH
OH
moderate

K25
O—2pe
OH
OH
OH
OH
O—2pe
moderate

K26
OH
O—2pe
OH
OH
OH
O—2pe
moderate

K27
OH
OH
O—2pe
OH
OH
O—2pe
moderate

K28
OH
OH
OH
O—2pe
OH
O—2pe
moderate

K29
O—2pe
OH
OH
OH
O—2pe
OH
moderate

K30
OH
O—2pe
OH
OH
O—2pe
OH
moderate

K31
OH
OH
O—2pe
OH
O—2pe
OH
moderate

K32
OH
OH
OH
O—2pe
O—2pe
OH
moderate

Esterification of core compound E4A with Octanoyl chloride and isolation of the compounds with HPLC give the following compounds: wherein Oct = Octanoyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

K1
OH
OH
OH
OH
O—Oct
OH
moderate

K2
OH
OH
OH
OH
OH
O—Oct
moderate

K3
OH
OH
OH
OH
O—Oct
O—Oct
strong

K4
O—Oct
OH
OH
OH
O—Oct
O—Oct
moderate

K5
OH
O—Oct
OH
OH
O—Oct
O—Oct
moderate

K6
OH
OH
O—Oct
OH
O—Oct
O—Oct
moderate

K7
OH
OH
OH
O—Oct
O—Oct
O—Oct
moderate

K8
O—Oct
O—Oct
OH
OH
O—Oct
O—Oct
weak

K9
OH
O—Oct
O—Oct
OH
O—Oct
O—Oct
weak

K10
OH
OH
O—Oct
O—Oct
O—Oct
O—Oct
weak

K11
O—Oct
OH
O—Oct
OH
O—Oct
O—Oct
weak

K12
OH
O—Oct
OH
O—Oct
O—Oct
O—Oct
weak

K13
O—Oct
OH
OH
O—Oct
O—Oct
O—Oct
weak

K14
OH
O—Oct
O—Oct
OH
O—Oct
O—Oct
weak

K15
O—Oct
O—Oct
O—Oct
OH
O—Oct
O—Oct
weak

K16
O—Oct
O—Oct
OH
O—Oct
O—Oct
O—Oct
weak

K17
O—Oct
OH
O—Oct
O—Oct
O—Oct
O—Oct
weak

K18
OH
O—Oct
O—Oct
O—Oct
O—Oct
O—Oct
weak

K19
O—Oct
O—Oct
—Oct
—Oct
O—Oct
O—Oct
none

K20
O—Oct
O—Oct
OH
OH
OH
O—Oct
moderate

K21
O—Oct
O—Oct
OH
OH
O—Oct
OH
moderate

K22
O—Oct
O—Oct
OH
O—Oct
OH
OH
moderate

K23
O—Oct
O—Oct
O—Oct
OH
OH
OH
moderate

K24
O—Oct
O—Oct
OH
OH
OH
OH
moderate

K25
O—Oct
OH
OH
OH
OH
O—Oct
moderate

K26
OH
O—Oct
OH
OH
OH
O—Oct
moderate

K27
OH
OH
O—Oct
OH
OH
O—Oct
moderate

K28
OH
OH
OH
O—Oct
OH
O—Oct
moderate

K29
O—Oct
OH
OH
OH
O—Oct
OH
moderate

K30
OH
O—Oct
OH
OH
O—Oct
OH
moderate

K31
OH
OH
O—Oct
OH
O—Oct
OH
moderate

K32
OH
OH
OH
O—Oct
O—Oct
OH
moderate

Esterification of core compound E4A with Decanoyl chloride and isolation of the compounds with HPLC give the following compounds: wherein Dec = Decanoyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

K1
OH
OH
OH
OH
O—Dec
OH
moderate

K2
OH
OH
OH
OH
OH
O—Dec
moderate

K3
OH
OH
OH
OH
O—Dec
O—Dec
strong

K4
O—Dec
OH
OH
OH
O—Dec
O—Dec
moderate

K5
OH
O—Dec
OH
OH
O—Dec
O—Dec
moderate

K6
OH
OH
O—Dec
OH
O—Dec
O—Dec
moderate

K7
OH
OH
OH
O—Dec
O—Dec
O—Dec
moderate

K8
O—Dec
O—Dec
OH
OH
O—Dec
O—Dec
weak

K9
OH
O—Dec
O—Dec
OH
O—Dec
O—Dec
weak

K10
OH
OH
O—Dec
O—Dec
O—Dec
O—Dec
weak

K11
O—Dec
OH
O—Dec
OH
O—Dec
O—Dec
weak

K12
OH
O—Dec
OH
O—Dec
O—Dec
O—Dec
weak

K13
O—Dec
OH
OH
O—Dec
O—Dec
O—Dec
weak

K14
OH
O—Dec
O—Dec
OH
O—Dec
O—Dec
weak

K15
O—Dec
O—Dec
O—Dec
OH
O—Dec
O—Dec
weak

K16
O—Dec
O—Dec
OH
O—Dec
O—Dec
O—Dec
weak

K17
O—Dec
OH
O—Dec
O—Dec
O—Dec
O—Dec
weak

K18
OH
O—Dec
O—Dec
O—Dec
O—Dec
O—Dec
weak

K19
O—Dec
O—Dec
O—Dec
O—Dec
O—Dec
O—Dec
none

K20
O—Dec
O—Dec
OH
OH
OH
O—Dec
moderate

K21
O—Dec
O—Dec
OH
OH
O—Dec
OH
moderate

K22
O—Dec
O—Dec
OH
O—Dec
OH
OH
moderate

K23
O—Dec
O—Dec
O—Dec
OH
OH
OH
moderate

K24
O—Dec
O—Dec
OH
OH
OH
OH
moderate

K25
O—Dec
OH
OH
OH
OH
O—Dec
moderate

K26
OH
O—Dec
OH
OH
OH
O—Dec
moderate

K27
OH
OH
O—Dec
OH
OH
O—Dec
moderate

K28
OH
OH
OH
O—Dec
OH
O—Dec
moderate

K29
O—Dec
OH
OH
OH
O—Dec
OH
moderate

K30
OH
O—Dec
OH
OH
O—Dec
OH
moderate

K31
OH
OH
O—Dec
OH
O—Dec
OH
moderate

K32
OH
OH
OH
O—Dec
O—Dec
OH
moderate

Esterification of core compound E4A with Myristoyl chloride and isolation of the compounds with HPLC give the following compounds: wherein Myr = Myristoyl

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A
OH
OH
OH
OH
OH
OH
none

K1
OH
OH
OH
OH
O—Myr
OH
moderate

K2
OH
OH
OH
OH
OH
O—Myr
moderate

K3
OH
OH
OH
OH
O—Myr
O—Myr
strong

K4
O—Myr
OH
OH
OH
O—Myr
O—Myr
moderate

K5
OH
O—Myr
OH
OH
O—Myr
O—Myr
moderate

K6
OH
OH
O—Myr
OH
O—Myr
O—Myr
moderate

K7
OH
OH
OH
O—Myr
O—Myr
O—Myr
moderate

K8
O—Myr
O—Myr
OH
OH
O—Myr
O—Myr
weak

K9
OH
O—Myr
O—Myr
OH
O—Myr
O—Myr
weak

K10
OH
OH
O—Myr
O—Myr
O—Myr
O—Myr
weak

K11
O—Myr
OH
O—Myr
OH
O—Myr
O—Myr
weak

K12
OH
O—Myr
OH
O—Myr
O—Myr
O—Myr
weak

K13
O—Myr
OH
OH
O—Myr
O—Myr
O—Myr
weak

K14
OH
O—Myr
O—Myr
OH
O—Myr
O—Myr
weak

K15
O—Myr
O—Myr
O—Myr
OH
O—Myr
O—Myr
weak

K16
O—Myr
O—Myr
OH
O—Myr
O—Myr
O—Myr
weak

K17
O—Myr
OH
O—Myr
O—Myr
O—Myr
O—Myr
weak

K18
OH
O—Myr
O—Myr
O—Myr
O—Myr
O—Myr
weak

K19
O—Myr
O—Myr
O—Myr
O—Myr
O—Myr
O—Myr
none

K20
O—Myr
O—Myr
OH
OH
OH
O—Myr
moderate

K21
O—Myr
O—Myr
OH
OH
O—Myr
OH
moderate

K22
O—Myr
O—Myr
OH
O—Myr
OH
OH
moderate

K23
O—Myr
O—Myr
O—Myr
OH
OH
OH
moderate

K24
O—Myr
O—Myr
OH
OH
OH
OH
moderate

K25
O—Myr
OH
OH
OH
OH
O—Myr
moderate

K26
OH
O—Myr
OH
OH
OH
O—Myr
moderate

K27
OH
OH
O—Myr
OH
OH
O—Myr
moderate

K28
OH
OH
OH
O—Myr
OH
O—Myr
moderate

K29
O—Myr
OH
OH
OH
O—Myr
OH
moderate

K30
OH
O—Myr
OH
OH
O—Myr
OH
moderate

K31
OH
OH
O—Myr
OH
O—Myr
OH
moderate

K32
OH
OH
OH
O—Myr
O—Myr
OH
moderate

Esterification of E4A—Tig—N with senecioyl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—N
OH
OH
OH
OH
O-Tig
OH
moderate

Tig—Sen—1
OH
OH
OH
OH
O—Tig
O—Sen
strong

Tig—Sen—2
O—Sen
OH
OH
OH
O—Tig
O—Sen
moderate

Tig—Sen—3
OH
O—Sen
OH
OH
O—Tig
O—Sen
moderate

Tig—Sen—4
OH
OH
O—Sen
OH
O—Tig
O—Sen
moderate

Tig—Sen—5
O—Sen
OH
OH
OH
O—Tig
OH
moderate

Tig—Sen—6
OH
O—Sen
OH
OH
O—Tig
OH
moderate

Esterification of E4A—Tig—N with Crotonoyl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—N
OH
OH
OH
OH
O—Tig
OH
moderate

Tig—Cro—1
OH
OH
OH
OH
O—Tig
O—Cro
strong

Tig—Cro—2
O—Cro
OH
OH
OH
O—Tig
O—Cro
moderate

Tig—Cro—3
OH
O—Cro
OH
OH
O—Tig
O—Cro
moderate

Tig—Cro—4
OH
OH
O—Cro
OH
O—Tig
O—Cro
moderate

Tig—Cro—5
O—Cro
OH
OH
OH
O—Tig
OH
moderate

Tig—Cro—6
OH
O—Cro
OH
OH
O—Tig
OH
moderate

Esterification of E4A—Tig—N with Acetyl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—N
OH
OH
OH
OH
O—Tig
OH
moderate

Tig—Acy—1
OH
OH
OH
OH
O—Tig
O—Acy
strong

Tig—Acy—2
O—Acy
OH
OH
OH
O—Tig
O—Acy
moderate

Tig—Acy—3
OH
O—Acy
OH
OH
O—Tig
O—Acy
moderate

Tig—Acy—4
OH
OH
O—Acy
OH
O—Tig
O—Acy
moderate

Tig—Acy—5
O—Acy
OH
OH
OH
O—Tig
OH
moderate

Tig—Acy—6
OH
O—Acy
OH
OH
O—Tig
OH
moderate

Esterification of E4A—Tig—N with 4-Pentenoyl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—N
OH
OH
OH
OH
O—Tig
OH
moderate

Tig—Pen—1
OH
OH
OH
OH
O—Tig
O—Pen
strong

Tig—Pen—2
O—Pen
OH
OH
OH
O—Tig
O—Pen
moderate

Tig—Pen—3
OH
O—Pen
OH
OH
O—Tig
O—Pen
moderate

Tig—Pen—4
OH
OH
O—Pen
OH
O—Tig
O—Pen
moderate

Tig—Pen—5
O—Pen
OH
OH
OH
O—Tig
OH
moderate

Tig—Pen—6
OH
O—Pen
OH
OH
O—Tig
OH
moderate

Esterification of E4A—Tig—N with Hexanoly chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—N
OH
OH
OH
OH
O—Tig
OH
moderate

Tig—Hex—1
OH
OH
OH
OH
O—Tig
O—Hex
strong

Tig—Hex—2
O—Hex
OH
OH
OH
O—Tig
O—Hex
moderate

Tig—Hex—3
OH
O—Hex
OH
OH
O—Tig
O—Hex
moderate

Tig—Hex—4
OH
OH
O—Hex
OH
O—Tig
O—Hex
moderate

Tig—Hex—5
O—Hex
OH
OH
OH
O—Tig
OH
moderate

Tig—Hex—6
OH
O—Hex
OH
OH
O—Tig
OH
moderate

Esterification of E4A—Tig—N with Cinnamoyl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—N
OH
OH
OH
OH
O—Tig
OH
moderate

Tig—Cin—1
OH
OH
OH
OH
O—Tig
O—Cin
strong

Tig—Cin—2
O—Cin
OH
OH
OH
O—Tig
O—Cin
moderate

Tig—Cin—3
OH
O—Cin
OH
OH
O—Tig
O—Cin
moderate

Tig—Cin—4
OH
OH
O—Cin
OH
O—Tig
O—Cin
moderate

Tig—Cin—5
O—Cin
OH
OH
OH
O—Tig
OH
moderate

Tig—Cin—6
OH
O—Cin
OH
OH
O—Tig
OH
moderate

Esterification of E4A—Tig—N with Angeloyl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—N
OH
OH
OH
OH
O—Tig
OH
moderate

Tig—Ang—1
OH
OH
OH
OH
O—Tig
O—Ang
strong

Tig—Ang—2
O—Ang
OH
OH
OH
O—Tig
O—Ang
moderate

Tig—Ang—3
OH
O—Ang
OH
OH
O—Tig
O—Ang
moderate

Tig—Ang—4
OH
OH
O—Ang
OH
O—Tig
O—Ang
moderate

Tig—Ang—5
O—Ang
OH
OH
OH
O—Tig
OH
moderate

Tig—Ang—6
OH
O—Ang
OH
OH
O—Tig
OH
moderate

Esterification of E4A—Tig—N with 2-Ethylbutyryl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—N
OH
OH
OH
OH
O—Tig
OH
moderate

Tig—Eth—1
OH
OH
OH
OH
O—Tig
O—Eth
strong

Tig—Eth—2
O—Eth
OH
OH
OH
O—Tig
O—Eth
moderate

Tig—Eth—3
OH
O—Eth
OH
OH
O—Tig
O—Eth
moderate

Tig—Eth—4
OH
OH
O—Eth
OH
O—Tig
O—Eth
moderate

Tig—Eth—5
O—Eth
OH
OH
OH
O—Tig
OH
moderate

Tig—Eth—6
OH
O—Eth
OH
OH
O—Tig
OH
moderate

Esterification of E4A—Tig—R with senecioyl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—R
OH
OH
OH
OH
O—Tig
O—Tig
strong

Tig—R—Sen—1
O—Sen
O—Sen
OH
OH
O—Tig
O—Tig
weak

Tig—R—Sen—2
O—Sen
OH
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Sen—3
OH
O—Sen
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Sen—4
OH
OH
O—Sen
OH
O—Tig
O—Tig
moderate

Tig—R—Sen—5
O—Sen
OH
O—Sen
OH
O—Tig
O—Tig
weak

Tig—R—Sen—6
OH
O—Sen
O—Sen
OH
O—Tig
O—Tig
weak

Esterification of E4A—Tig—R with Crotonoyl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—R
OH
OH
OH
OH
O—Tig
O—Tig
strong

Tig—R—Cro—1
O—Cro
O—Cro
OH
OH
O—Tig
O—Tig
weak

Tig—R—Cro—2
O—Cro
OH
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Cro—3
OH
O—Cro
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Cro—4
OH
OH
O—Cro
OH
O—Tig
O—Tig
moderate

Tig—R—Cro—5
O—Cro
OH
O—Cro
OH
O—Tig
O—Tig
weak

Tig—R—Cro—6
OH
O—Cro
O—Cro
OH
O—Tig
O—Tig
weak

Esterification of E4A—Tig—R with Acetyl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—R
OH
OH
OH
OH
O—Tig
O—Tig
strong

Tig—R—Acy—1
O—Acy
O—Acy
OH
OH
O—Tig
O—Tig
weak

Tig—R—Acy—2
O—Acy
OH
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Acy—3
OH
O—Acy
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Acy—4
OH
OH
O—Acy
OH
O—Tig
O—Tig
moderate

Tig—R—Acy—5
O—Acy
OH
O—Acy
OH
O—Tig
O—Tig
weak

Tig—R—Acy—6
OH
O—Acy
O—Acy
OH
O—Tig
O—Tig
weak

Esterification of E4A—Tig—R with 4-Pentenoyl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—R
OH
OH
OH
OH
O—Tig
O—Tig
strong

Tig—R—Pen—1
O—Pen
O—Pen
OH
OH
O—Tig
O—Tig
weak

Tig—R—Pen—2
O—Pen
OH
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Pen—3
OH
O—Pen
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Pen—4
OH
OH
O—Pen
OH
O—Tig
O—Tig
moderate

Tig—R—Pen—5
O—Pen
OH
O—Pen
OH
O—Tig
O—Tig
weak

Tig—R—Pen—6
OH
O—Pen
O—Pen
OH
O—Tig
OH
weak

Esterification of E4A—Tig—R with Hexanoly chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—R
OH
OH
OH
OH
O—Tig
O—Tig
strong

Tig—R—Hex—1
O—Hex
O—Hex
OH
OH
O—Tig
O—Tig
weak

Tig—R—Hex—2
O—Hex
OH
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Hex—3
OH
O—Hex
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Hex—4
OH
OH
O—Hex
OH
O—Tig
O—Tig
moderate

Tig—R—Hex—5
O—Hex
OH
O—Hex
OH
O—Tig
O—Tig
weak

Tig—R—Hex—6
OH
O—Hex
O—Hex
OH
O—Tig
O—Tig
weak

Esterification of E4A—Tig—R with Cinnamoyl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—R
OH
OH
OH
OH
O—Tig
O—Tig
strong

Tig—R—Cin—1
O—Cin
O—Cin
OH
OH
O—Tig
O—Tig
weak

Tig—R—Cin—2
O—Cin
OH
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Cin—3
OH
O—Cin
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Cin—4
OH
OH
O—Cin
OH
O—Tig
O—Tig
moderate

Tig—R—Cin—5
O—Cin
OH
O—Cin
OH
O—Tig
O—Tig
weak

Tig—R—Cin—6
OH
O—Cin
O—Cin
OH
O—Tig
O—Tig
weak

Esterification of E4A—Tig—R with Angeloyl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—R
OH
OH
OH
OH
O—Tig
O—Tig
strong

Tig—R—Ang—1
O—Ang
O—Ang
OH
OH
O—Tig
O—Tig
weak

Tig—R—Ang—2
O—Ang
OH
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Ang—3
OH
O—Ang
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Ang—4
OH
OH
O—Ang
OH
O—Tig
O—Tig
moderate

Tig—R—Ang—5
O—Ang
OH
O—Ang
OH
O—Tig
O—Tig
weak

Tig—R—Ang—6
OH
O—Ang
O—Ang
OH
O—Tig
O—Tig
weak

Esterification of E4A—Tig—R with 2-Ethylbutyryl chloride and isolation of the compounds with HPLC give the following compounds:

R1
R2
R5
R8
R17
R18
Cytotoxicity activity

E4A—Tig—R
OH
OH
OH
OH
O—Tig
O—Tig
strong

Tig—R—Eth—1
O—Eth
O—Eth
OH
OH
O—Tig
O—Tig
weak

Tig—R—Eth—2
O—Eth
OH
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Eth—3
OH
O—Eth
OH
OH
O—Tig
O—Tig
moderate

Tig—R—Eth—4
OH
OH
O—Eth
OH
O—Tig
O—Tig
moderate

Tig—R—Eth—5
O—Eth
OH
O—Eth
OH
O—Tig
O—Tig
weak

Tig—R—Eth—6
OH
O—Eth
O—Eth
OH
O—Tig
O—Tig
weak

Esterification of compound (A), (B), (C), (D1), (D2), (E), (F), (G), (H1), (H2), E4A, E4A2Y, (P1), P(2), terpene, isoprene, triterpenes, hydroxylated triterpenes, with acyl halide, wherein the halide comprise chloride, bromide, fluoride and iodide, wherein the acyl halide comprise acyl chloride, wherein acyl chloride comprise tigloyl chloride, angeloyl chloride, acetyl chloride, crotonoyl chloride, 3,3-Dimethylacryloyl chloride, senecioyl chloride, cinnamoyl chloride, pentenoyl chloride, hexanoyl chloride, benzoyl chloride, ethylbutyryl chloride, propionyl chloride, 2-propenoyl chloride, isobutyryl chloride, butyryl chloride, (2E)-2-pentenoyl chloride, 4-Pentenoyl chloride, 5-hexenoyl chloride, heptanoyl chloride, octanoyl chloride, nonanoyl chloride, decanoyl chloride, Lauroyl chloride, myristoyl chloride, oleoyl chloride. The compounds vary in composition when the time or temperature of the reaction is changed. The peaks, fractions and compounds are selected according to the activities of times studies and the changes of peaks. The compounds having strong to weak activities are selected and isolated. The anti-cancer activities (Cytotoxic Assay) are the MTT studies of bone (U2OS), lung (H460), bladder(HTB-9), ovary (ES2), colon (HCT116), pancreas (Capan), ovary(OVCAR3), prostate (DU145), skin (SK-Mel-5), mouth (KB), kidney (A498), breast (MCF-7), liver (HepG2), brain (T98G), leukemia (K562), cervix (HeLa). The active esterification products are purified with HPLC. The reaction product of mixtures and individual compounds are tested with MTT Cytotoxic Assay. Details of method are in Experiment 3 of the present application. A second esterification of compound can be selected from the above experiment results to produce new active compounds. A partial esterification compound is selected from the above experiments to perform a second or repeated with a third esterification with different acyl chloride in order to produce new active compounds with the experiments in the present application.

A method is 1) Dissolving core compound or triterpenes core, hydroxylated triterpenes core, in pyridine; 2) Adding acyl halide or acyl chloride; 3, The mixture is stirred for length of time including 5 sec, 10 sec, 20 sec, 30 sec, 40 sec, 1 min, 2 min, 5 min, 10 min, 30 min, 1 hr, 2 hr, 18 hr, 2 days or 3 days at different temperature; 4) At the end of reaction, aqueous solution of acid or weak base, or water is added to the reaction mixture; 5) The solution is then extracted of ethyl acetate and ethyl acetate is removed by evaporation and lyophilization; 6) Dissolving the reaction product in acetonitrile with Trifluoroacetic acid or DMSO; 7) Testing the reaction product of mixtures and individual fractions with MTT cytotoxic assay; 8) Selecting the HPLC fractions for isolation is according to the cytotoxic activity of the reaction product obtained at a specific reaction time; 10) Purifying the active esterification products with HPLC; 11) Collecting the products; 12) Testing the products; wherein the core compound is terpene, isoprene, or triterpene core or hydroxylated triterpenes core; wherein the core compound was dissolved in pyridine; wherein the acyl chloride including Tigloyl chloride, angeloyl chloride, Acetyl chloride, Crotonoyl chloride, 3,3-Dimethylacryloyl chloride, senecioyl chloride, Cinnamoyl chloride, Pentenoyl chloride, Hexanoyl chloride, benzoyl chloride, Ethylbutyryl chloride, Propionyl chloride, 2-Propenoyl chloride, Isobutyryl chloride, Butyryl chloride, (2E)-2-pentenoyl chloride, 4-Pentenoyl chloride, 5-Hexenoyl chloride, Heptanoyl chloride, Octanoyl chloride, Nonanoyl chloride, Decanoyl chloride, Lauroyl chloride, Myristoyl chloride, and Oleoyl chloride; wherein the reaction time for the mixture is stirred for 5 sec, 10 sec, 20 sec, 30 sec, 40 sec, 1 min, 2 min, 5 min, 10 min, 30 min, 1 hr, 2 hr, 18 hr, 2 days or 3 days; wherein the temperature is 0C, 25C, 50C or 75C temperature; wherein the acid including HCl or the base including NaHCO3 is added to the reaction mixture; wherein the solution is then extracted 3 times with ethyl acetate and lyophilization; wherein the reaction product is dissolved in 80% acetonitrile - 0.005% Trifluoroacetic acid or DMSO; wherein selecting the HPLC fractions for isolation is according to the cytotoxic activity of the reaction product obtained at a reaction time of 5 sec, 10 sec, 20 sec, 30 sec, 40 sec, 1 min, 2 min, 5 min, 10 min, 30 min, 1 hr, 2 hr, 18 hr, 2 days or 3 days. In another embodiment, the reaction time may be over 3 days. In another embodiment, the experiment may be performed under 0C. In another embodiment, the experiment may be performed over 75C.

embedded image

wherein (R1, R2, or R18) and (R17, R5 or R16) are independently selected from the group of O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, 0-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, O-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl; wherein R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14, R15, R16, R17 are independently selected from the group of H, OH, CH2OH, COOH, OR CH3,

The anti-cancer activities of Tig-R compound: IC50 of bone (U2OS) is 4.5 ug/ml, lung (H460) is 4.8 ug/ml, bladder(HTB-9) is 2.5 ug/ml, ovary (ES2) is 2.8 ug/ml, colon (HCT116) is 5.2 ug/ml, pancreas (Capan) 2.4 ug/ml, ovary(OVCAR3) is 5.8, prostate (DU145) is 3.6 ug/ml, skin (SK-Mel-5) is 5.1 ug/ml, mouth (KB) is 3 ug/ml, kidney (A498) is 3.5 ug/ml, breast (MCF-7) is 4.5 ug/ml, liver (HepG2) is 6 ug/ml, brain (T98G) is 8 ug/ml), leukemia (K562) is 2 ug/ml, cervix (HeLa) is 5 ug/ml.

The anti-cancer activities of Tig-V compound: IC50 of bone (U2OS) is 7 ug/ml, lung (H460) is 6.8 ug/ml, bladder(HTB-9) is 4 ug/ml, ovary (ES2) is 2 ug/ml, colon (HCT116) is 8 ug/ml, pancreas (Capan) 5 ug/ml, ovary(OVCAR3) is 9, prostate (DU145) is 4 ug/ml, skin (SK-Mel-5) is 6 ug/ml, mouth (KB) is 4.5 ug/ml, kidney (A498) is 4.8 ug/ml, breast (MCF-7) is 9 ug/ml, liver (HepG2) is 12 ug/ml, brain (T98G) is 14 ug/ml), leukemia (K562) is 4 ug/ml, cervix (HeLa) is 7 ug/ml.

The anti-cancer activities of Tig-N compound: IC50 of bone (U2OS) is 15 ug/ml, lung (H460) is 13 ug/ml, bladder(HTB-9) is 7.5 ug/ml, ovary (ES2) is 9 ug/ml, colon (HCT116) is 15 ug/ml, pancreas (Capan) 8 ug/ml, ovary(OVCAR3) is 18, prostate (DU145) is 4.8 ug/ml, skin (SK-Mel-5) is 15 ug/ml, mouth (KB) is 9 ug/ml, kidney (A498) is 11 ug/ml, breast (MCF-7) is 13 ug/ml, liver (HepG2) is 18 ug/ml, brain (T98G) is 19 ug/ml), leukemia (K562) is 6 ug/ml, cervix (HeLa) is 15 ug/ml.

The anti-cancer activities of Tig-Q compound: IC50 of bone (U2OS) is 20 ug/ml, lung (H460) is 18 ug/ml, bladder(HTB-9) is 10 ug/ml, ovary (ES2) is 12 ug/ml, colon (HCT116) is 22 ug/ml, pancreas (Capan) 9 ug/ml, ovary(OVCAR3) is 23, prostate (DU145) is 15 ug/ml, skin (SK-Mel-5) is 20 ug/ml, mouth (KB) is 12 ug/ml, kidney (A498) is 13 ug/ml, breast (MCF-7) is 18 ug/ml, liver (HepG2) is 24 ug/ml, brain (T98G) is 29 ug/ml), leukemia (K562) is 6 ug/ml, cervix (HeLa) is 20 ug/ml.

The anti-cancer activities of Tig-T compound: IC50 of bone (U2OS) is 20 ug/ml, lung (H460) is 21 ug/ml, bladder(HTB-9) is 12 ug/ml, ovary (ES2) is 14 ug/ml, colon (HCT116) is 23 ug/ml, pancreas (Capan) 10 ug/ml, ovary(OVCAR3) is 25, prostate (DU145) is 16 ug/ml, skin (SK-Mel-5) is 22 ug/ml, mouth (KB) is 13 ug/ml, kidney (A498) is 15 ug/ml, breast (MCF-7) is 20 ug/ml, liver (HepG2) is 26 ug/ml, brain (T98G) is 26 ug/ml), leukemia (K562) is 9 ug/ml, cervix (HeLa) is 18 ug/ml.

The anti-cancer activities of Tig-S compound: IC50 of bone (U2OS) is 5.2 ug/ml, lung (H460) is 5.6 ug/ml, bladder(HTB-9) is 3.5 ug/ml, ovary (ES2) is 0.1 ug/ml, colon (HCT116) is 6.6 ug/ml, pancreas (Capan) 2.9 ug/ml, ovary(OVCAR3) is 6.5, prostate (DU145) is 4.3 ug/ml, skin (SK-Mel-5) is 5.8 ug/ml, mouth (KB) is 4 ug/ml, kidney (A498) is 4.8 ug/ml, breast (MCF-7) is 6.3 ug/ml, liver (HepG2) is 8.5 ug/ml, brain (T98G) is 9 ug/ml), leukemia (K562) is 4.3 ug/ml, cervix (HeLa) is 7 ug/ml.

The anti-cancer activities of Tig-U compound: IC50 of bone (U2OS) is 23 ug/ml, lung (H460) is 19 ug/ml, bladder(HTB-9) is 15 ug/ml, ovary (ES2) is 17 ug/ml, colon (HCT116) is 26 ug/ml, pancreas (Capan) 9 ug/ml, ovary(OVCAR3) is 27, prostate (DU145) is 15 ug/ml, skin (SK-Mel-5) is 24 ug/ml, mouth (KB) is 16 ug/ml, kidney (A498) is 18 ug/ml, breast (MCF-7) is 25 ug/ml, liver (HepG2) is 23 ug/ml, brain (T98G) is 22 ug/ml), leukemia (K562) is 10 ug/ml, cervix (HeLa) is 17 ug/ml.

The IC50 of Tig-R in normal human fibroblast cells (WI38) is about 10-15 ug/ml. This IC50 value is 3 times higher than those in ovary ES2 (2.8 ug/ml) and lung (H460) is 4.8 ug/ml.

Swiss3T3 cells are mouse normal fibroblast which were used in this experiment to compare with ES2 (human ovarian cancer) in Tig-R cytotoxicity determination. The preliminary results indicate that the IC50 of Tig-R in SW3T3 cells is above 20 ug/ml while the corresponding IC50 in ES2 cells is about 2.8 ug/ml.

This invention provides compounds, methods, or uses of a compound for the manufacture of a medicament, or uses of a compound for medicament selected from formula (2A), increasing the level of DR5 of cells, for treating cancers, inhibition of cancer growth, cancer invasion, cell invasion, cancer cell invasion; cell adhesion, cell attachment, cell circulating; for treating mad cow disease; treating prion diseases; for inhibiting viruses; for preventing cerebral aging; for improving memory; improving cerebral functions; for curing enuresis, frequent micturition, urinary incontinence; dementia, Alzheimer’s disease, autism, brain trauma, Parkinson’s disease or other diseases caused by cerebral dysfunctions or neurodegeneration; for treating retinoblastoma; for treating arthritis, rheumatism, poor blood circulation, arteriosclerosis, Raynaud’s syndrome, angina pectoris, cardiac disorder, coronary heart disease, headache, dizziness, kidney disorder; cerebrovascular diseases; inhibiting NF-kappa B activation; for treating brain edema, severe acute respiratory syndrome, respiratory viral diseases, chronic venous insufficiency, hypertension, chronic venous disease, oedema, inflammation, hemorrhoids, peripheral edema formation, varicose vein disease, flu, post traumatic edema and postoperative swelling; for inhibiting blood clots, for inhibiting ethanol absorption; for lowering blood sugar; for regulating adrenocorticotropin and corticosterone levels; for Anti-MS, anti-aneurysm, anti-asthmatic, anti-oedematous, antiinflammatory, anti-bradykinic, anti-capillarihemorrhagic, anti-cephalagic, anticervicobrachialgic, anti-eclamptic, anti-edemic, anti-encaphalitic, anti-epiglottitic, antiexudative, anti-flu, anti-fracture, anti-gingivitic, anti-hematomic, anti-herpetic, antihistaminic, anti-hydrathritic, anti-meningitic, antioxidant, anti-periodontic, anti-phlebitic, anti-pleuritic, anti-raucedo, anti-rhinitic, anti-tonsilitic, anti-ulcer, anti-varicose, antivertiginous, cancerostatic, corticosterogenic, diuretic, fungicide, hemolytic, hyaluronidase inhibitor, lymphagogue, natriuretic, pesticide, pituitary stimulant, thymolytic, vasoprotective, inhibiting leishmaniases, modulating adhesion or angiogenesis of cells, anti-parasitic; for improving blood circulation; soothing stroke; preventing plaque formation and promote their dissipated; improve blood viscosity; reducing cardiovascular clotting; reducing cerebrovascular clotting; reducing thrombosis, arteriosclerosis, coronary heart disease, hypertension, diabetes, thrombocytopenia purpura, hemoptysis, hematemesis; treating blood in the stool, uterine bleeding, traumatic bleeding, abdominal irritation, swelling, fluttering, blood circulation, swelling, pain; treating bronchiectasis, tuberculosis and lung abscess caused by too hemoptysis; reducing bleeding; antitussive; reducing expectorant; reducing analgesic effect; dilate blood vessels; reducing blood pressure; treatment of cerebral arteriosclerosis; elevating blood lipids; reducing cholesterol; manufacturing an adjuvant composition for treatment, using compounds selected from the following:

embedded image - (2A)

salt thereof,

R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14, R15, R16 are independently selected from the group of O, hydrogen, hydroxyl, methyl, O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, 0-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-Isobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl, alkane, alkene and sugar moiety or derivatives thereof; or wherein the structure (2A) comprises at least 2 groups selected from O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, O-2-butenoyl, O-Isobutyryl, 0-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, 0-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl; or wherein R1 and R2 are selected from O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, 0-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl; or wherein R4 and R10 are selected from CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-Isobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl. In another embodiment, R1 and R2 are attached OH. In another embodiment, R4 and R10 are attached a CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, or CH2O-Ethylbutyryl. In another embodiment, R3 and R8 is hydrogen or hydroxyl. In another embodiment, R9, R11, R12, R13, R14 and R15 are independently attached with a methyl. In another embodiment, R4 represents CH3, CHO, CH₂R6 or COR6, wherein R6 is selected from hydroxyl, O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, O-2-butenoyl, O-Isobutyryl, 0-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl and derivatives thereof. In another embodiment, R3 is H or OH; In another embodiment, wherein R8 is H or OH; In another embodiment, R16 is H, CH3, OH,or R4 and R16 may together form —CH2—X—, CH(OH)—X— or C(═O)—X—, wherein the —X— may be O or NH or S; wherein when the C12-13 of ring 3 of the triterpene has a double bond then R16 is H or absent. In another embodiment, R10 represents CH3, CHO, or CH₂R6, wherein R6 is selected from hydroxyl, O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, 0-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl and derivatives thereof; In another embodiment, R5 is a hydrogen, hydroxyl, heterocyclic or O-sugar moiety(ies), wherein the sugar moiety(ies) is/are selected from a group but not limited of glucose, galactose, rhamnose, arabinose, xylose, fucose, allose, altrose, gulose, idose, lyxose, mannose, psicose, ribose, sorbose, tagatose, talose, fructose, alduronic acid, glucuronic acid, galacturonic acid, and derivatives or combinations thereof; In embodiment, the above structures were attached an O with double bond; wherein R9, R10, R11, R12, R13, R14, R15, R16 are independently attached a group selecting from CH₃, CH₂OH, CHO, COOH, COO-alkyl, COO-aryl, COO-heterocyclic, COO-heteroaryl, CH₂Oaryl, CH₂O- heterocyclic, CH₂O- heteroaryl, alkyls group, hydroxyl, acetyl group; wherein R4 and R16 form a divalent radical of formula CH2O, CH(OR7)O, or COOR7, wherein R7 is hydrogen, alkyl, angeloyl, tigloyl, senecioyl, dibenzoyl, benzoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, aryl, acyl, heterocylic, heteroraryl, and derivatives thereof; wherein at least two of R1, R2 and R6 are attached a group selected from O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, 0-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl,, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, 0-2-propenoyl, O-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, 0-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl and derivatives thereof; or at least one of R1, R2, and R4 is a sugar moiety having at least two groups selected from a group consisting of angeloyl, acetyl, tigloyl, senecioyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, benzoyl, dibenzoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, aryl, acyl, heterocylic, heteroraryl, ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl, O-C(2-18) Acyl and their derivatives thereof; or wherein R4 represents CH₂R6, wherein R6 is selected from hydroxyl, O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl and derivatives thereof; wherein R5 is/are the sugar moiety(ies) selected from the following sugars and alduronic acids: glucose, galactose, rhamnose, arabinose, xylose, fucose, allose, altrose, gulose, idose, lyxose, mannose, psicose, ribose, sorbose, tagatose, talose, fructose, glucuronic acid, galacturonic acid; or their derivatives thereof, In another embodiment, R5 is a hydroxyl, O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl and derivatives thereof. In another embodiment, R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14 or R15 comprise of one or more sugar moieties. In another embodiment, R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14 or R15 comprise of one or more acids. In another embodiment, at least 1, or 2, or 3, or 4 of R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14 and R15 is hydroxyl. In another embodiment, at least 2, or 3, or 4, or 5, or 6, or 7 of R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14 and R15 are independently attached a group selected from the group of O-acetyl, O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, 0-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, alkane, alkene and derivatives thereof, wherein the group is attached to the triterpene directly or by connecting moiety(ies); In another embodiment, at least 1 or 2, or 3, or 4, or 5, or 6, or 7 of R1, R2, R3, R4, R5, R8 and R10 are independently attached a group selected from the group of O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl,O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, 0-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl,CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl,CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl,CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-lsobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl and derivatives thereof, wherein the group is attached to the triterpene directly or by connecting moiety(ies). In embodiment, the compound is attached a sugar moiety(ies), acid moiety(ies) or alduronic acid. In another embodiment, the compounds are in form of powder, crystal, or liquid. In embodiment, the compounds are in capsule, or with pharmaceutically acceptable carrier or diluent.

In one embodiment, the cancers comprise breast cancer, leukocytic cancer, liver cancer, ovarian cancer, bladder cancer, prostatic cancer, skin cancer, bone cancer, brain cancer, leukemia cancer, lung cancer, colon cancer, CNS cancer, melanoma cancer, renal cancer, cervical cancer, esophageal cancer, testicular cancer, splenic cancer, kidney cancer, lymphatic cancer, pancreatic cancer, stomach cancer, eye cancer and thyroid cancer; wherein the cells comprise breast cell, leukocytic cell, liver cell, ovarian cell, bladder cell, prostatic cell, skin cell, bone cell, brain cell, leukemia cell, lung cell, colon cell, CNS cell, melanoma cell, renal cell, cervical cell, esophageal cell, testicular cell, splenic cell, kidney cell, lymphatic cell, pancreatic cell, stomach cell, eye cell and thyroid cell. In another embodiment, the compound is selected from the structure:

embedded image - (K)

salt thereof,

R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14, R15 are independently selected from the group of CH3, CH2OH, COOH, O, hydrogen, hydroxyl, methyl, O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl,O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-alkane, O-alkene, O-sugar moiety, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, O-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O- (E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, 0-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-alkane, CH2O-alkene and CH2O-sugar moiety, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-lsobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl, (CnH2n)O-angeloyl, (CnH2n)O-tigloyl, (CnH2n)O-senecioyl, (CnH2n)O-acetyl, (CnH2n)O-Crotonoyl, (CnH2n)O-3,3-Dimethylacryloyl, (CnH2n)O-Cinnamoyl, (CnH2n)O-Pentenoyl, (CnH2n)O-Hexanoyl, (CnH2n)O-benzoyl, (CnH2n)O-Ethylbutyryl, (CnH2n)O-alkyl, (CnH2n)O-dibenzoyl, (CnH2n)O-benzoyl, (CnH2n)O-alkanoyl, (CnH2n)O-alkenoyl, (CnH2n)O-benzoyl alkyl substituted O-alkanoyl, (CnH2n)O-alkanoyl substituted phenyl, (CnH2n)O-alkenoyl substituted phenyl, (CnH2n)O-aryl, (CnH2n)O-acyl, (CnH2n)O-heterocylic, (CnH2n)O-heteroraryl, (CnH2n)O-alkenylcarbonyl, (CnH2n)O-alkane, (CnH2n)O-alkene and (CnH2n)O-sugar moiety, wherein n is 1 or 2 or 3 or 4 or over 5 or derivatives thereof; or wherein any 1 or 2 or 3 or 4 of R1, R2, R3, R4, R5, R8 and R10 are independently attached an O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl,O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, O-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, 0-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-Isobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl,; or wherein R9, R11, R12, R13, R14, R15 are independently attached a CH3; or wherein R10 is attached an O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl,O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, 0-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O- (E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-lsobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl; or wherein R4 and/or R10 are independently attached an O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, 0-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O- (E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-lsobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl; wherein R3 is OH or H or absent; wherein R1, R2, R3, R5, R8 are OH or H or absent; wherein R9, R11, R12, R13, R14, and R15 are CH3; or wherein R1, R2, R5, R8 represent OH; R3 represents OH, H or absent; or wherein R4, R10 represent CH2Oangeloyl; R9, R11, R12, R13, R14, R15 represent CH3; or wherein R1, R2, R5, R8 represent OH or O-tigloyl; R3 represents OH, H, or absent; or wherein R4, R10 represent CH2O tigloyl; R9, R11, R12, R13, R14, R15 represent CH3; wherein the group attaching to the core compound selected from acetyl, angeloyl, tigloyl, senecioyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, methylbutanoyl, methylpropanoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, alkenylcarbonyl, ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl, C(2-18) Acyl are interchangeable; wherein the attached group can be the same group or in combination thereof; wherein the connecting group between the core compound and attached group may be O, S,S(O), S(O)2, C(O), C(O)O, NH, N-alkyl, CH2 or CH2O. In another embodiment, R4 is attached an O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, O-2-butenoyl, O-Isobutyryl, 0-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-lsobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-c/s-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl. In embodiment, the above structures were attached an O with double bond. In another embodiment, the connecting group between the functional group of angeloyl, tigloyl, senecioyl, acetyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, and alkenylcarbonyl ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl, C(2-18) Acyl can be O, S,S(O), S(O)2, C(O), C(O)O, NH, N-alkyl, CH2 or CH2O. In another embodiment, wherein any 1 or 2 or 3 or 4 or 5 or 6 of R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14, R15 are independently selected from the group of A-B, wherein A can be O, S,S(O), S(O)2, C(O), C(O)O, NH, N-alkyl, CH2 or CH2O; wherein B is selected from the group of acetyl, angeloyl, tigloyl, senecioyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, methylbutanoyl, methylpropanoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, alkenylcarbonyl, ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl and C(2-18) Acyl. In another embodiment, R1 is A-B. In another embodiment, R2 is A-B. In an embodiment, R3 is A-B. In another embodiment, R4 is A-B. In an embodiment, R5 is A-B. In another embodiment, R6 is A-B. In an embodiment, R7 is A-B. In another embodiment, R8 is A-B. In an embodiment, R9 is A-B. In another embodiment, R10 is A-B. In an embodiment, R11 is A-B. In another embodiment, R12 is A-B. In an embodiment, R13 is A-B. In another embodiment, R14 is A-B. In an embodiment, R15 is A-B. In another embodiment, the compound is attached a sugar moiety(ies), acid moiety(ies) or alduronic acid. In another embodiment, the compound is selected from the structure:

embedded image - (3K),

salt thereof,

embedded image - (3K2)

salt thereof, R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14, R15, R16 are independently selected from the group of H, O, OH, CH3, CH2OH, COOH hydrogen, hydroxyl, methyl,O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl,O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-alkane, O-alkene, O-sugar moiety, O-acid moiety, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, O-2-butenoyl, O-Isobutyryl, 0-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, COO-angeloyl, COO-tigloyl, COO-senecioyl, COO-acetyl, COO-Crotonoyl, COO-3,3-Dimethylacryloyl, COO-Cinnamoyl, COO-Pentenoyl, COO-Hexanoyl, COO-benzoyl, COO-Ethylbutyryl, COO-alkyl, COO-dibenzoyl, COO-benzoyl, COO-alkanoyl, CH2O-alkenoyl, COO-benzoyl alkyl substituted O-alkanoyl, COO-alkanoyl substituted phenyl, COO-alkenoyl substituted phenyl, COO-aryl, COO-acyl, COO-heterocylic, COO-heteroraryl, COO-alkenylcarbonyl, COO-alkane, COO-alkene, COO-sugar moiety, COO-acid moiety, COO-ethanoyl, COO-propanoyl, COO-propenoyl, COO-butanoyl, COO-butenoyl, COO-pentanoyl, COO-hexenoyl, COO-heptanoyl, COO-heptenoyl, COO-octanoyl, COO-octenoyl, COO-nonanoyl, COO-nonenoyl, COO-decanoyl, COO-decenoyl, COO-propionyl, COO-2-propenoyl, COO-2-butenoyl, COO-Isobutyryl, COO-2-methylpropanoyl, COO-2-ethylbutyryl, COO-ethylbutanoyl, COO-2-ethylbutanoyl, COO-butyryl, COO- (E)-2,3-Dimethylacryloyl, COO-(E)-2-Methylcrotonoyl, COO-3-cis-Methyl-methacryloyl, COO-3-Methyl-2-butenoyl, COO-3-Methylcrotonoyl, COO-4-Pentenoyl, COO-(2E)-2-pentenoyl, COO-Caproyl, COO-5-Hexenoyl, COO-Capryloyl, COO-Lauroyl, COO-Dodecanoyl, COO-Myristoyl, COO-Tetradecanoyl, COO-Oleoyl, COO-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl,CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-alkane, CH2O-alkene and CH2O-sugar moiety, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl,CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl,CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH20-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl,CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-Isobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl,CH2O-(E)-2-Methylcrotonoyl, CH2O-3-c/s-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl,CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl, (CnH2n)O-angeloyl, (CnH2n)O-tigloyl, (CnH2n)O-senecioyl, (CnH2n)O-acetyl, (CnH2n)O-Crotonoyl, (CnH2n)O-3,3-Dimethylacryloyl, (CnH2n)O-Cinnamoyl, (CnH2n)O-Pentenoyl, (CnH2n)O-Hexanoyl, (CnH2n)O-benzoyl, (CnH2n)O-Ethylbutyryl, (CnH2n)O-alkyl, (CnH2n)O-dibenzoyl, (CnH2n)O-benzoyl, (CnH2n)O-alkanoyl, (CnH2n)O-alkenoyl, (CnH2n)O-benzoyl alkyl substituted O-alkanoyl, (CnH2n)O-alkanoyl substituted phenyl, (CnH2n)O-alkenoyl substituted phenyl, (CnH2n)O-aryl, (CnH2n)O-acyl, (CnH2n)O-heterocylic, (CnH2n)O-heteroraryl, (CnH2n)O-alkenylcarbonyl, (CnH2n)O-alkane, (CnH2n)O-alkene and (CnH2n)O-sugar moiety and (CnH2n)O-acid moiety, wherein n is 1 or 2 or 3 or 4 or over 5 or derivatives thereof; wherein the sugar moiety(ies) is/are selected from a group of glucose, galactose, rhamnose, arabinose, xylose, fucose, allose, altrose, gulose, idose, lyxose, mannose, psicose, ribose, sorbose, tagatose, talose, fructose, alduronic acid, glucuronic acid, galacturonic acid, and derivatives or combinations thereof; or wherein any 1 or 2 or 3 or 4 of R1, R2, R3, R4, R5, R8, R16 and R10 are independently attached an O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl,O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, O-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl,O-(E)-2-Methylcrotonoyl, O-3-c/s-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, 0-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl,CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl,CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-Isobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl,CH2O-(E)-2-Methylcrotonoyl, CH2O-3-c/s-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl,CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl; or wherein R9, R11, R12, R13, R14, R15, are independently attached a CH3; or wherein R10 is attached an O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl,O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, 0-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O- (E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH20-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl,CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-lsobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-c/s-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl; or wherein R4 and/or R10 are independently attached an O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, 0-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O- (E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-lsobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-c/s-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl; wherein R3 is OH or H or absent; wherein R1, R2, R3, R5, R8, R16 are OH or H or absent; or wherein an O were attached to the above structures with double bond; wherein R9, R11, R12, R13, R14, and R15 are CH3; or wherein R1, R2, R5, R8 represent OH; R3 represents OH, H or absent; or wherein R4, R10 represent CH2Oangeloyl; R9, R11, R12, R13, R14, R15 represent CH3; or wherein R1, R2, R5, R8 represent OH or O-tigloyl; R3 represents OH, H, or absent; or wherein R4, R10 represent CH2O tigloyl; R9, R11, R12, R13, R14, R15 represent CH3; wherein the group attaching to the core compound selected from acetyl, angeloyl, tigloyl, senecioyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, methylbutanoyl, methylpropanoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, alkenylcarbonyl, ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl, C(2-18) Acyl are interchangeable; wherein the attached group can be the same group or in combination thereof; wherein the connecting group between the core compound and attached group may be O, S,S(O), S(O)2, C(O), C(O)O, NH, N-alkyl, CH2 or CH2O. In another embodiment, R4 is attached an O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, O-2-butenoyl, O-Isobutyryl, 0-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, 0-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2-Isobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl, In another embodiment, the connecting group between the functional group of angeloyl, tigloyl, senecioyl, acetyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, and alkenylcarbonyl ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl, C(2-18) Acyl can be O, S,S(O), S(O)2, C(O), C(O)O, NH, N-alkyl, CH2 or CH2O. In another embodiment, any 1 or 2 or 3 or 4 or 5 or δ of R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14, R15, R16 are independently selected from the group of A-B, wherein A can be O, S,S(O), S(O)2, C(O), C(O)O, NH, N-alkyl, CH2 or CH2O; wherein B is selected from the group of acetyl, angeloyl, tigloyl, senecioyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, methylbutanoyl, methylpropanoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, alkenylcarbonyl, ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl,hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl and C(2-18) Acyl. In another embodiment, R1 is A-B. In another embodiment, R2 is A-B. In another embodiment, R3 is A-B. In another embodiment, R4 is A-B. In another embodiment, R5 is A-B. In another embodiment, R6 is A-B. In another embodiment, R7 is A-B. In another embodiment, R8 is A-B. In another embodiment, R9 is A-B. In another embodiment, R10 is A-B. In another embodiment, R11 is A-B. In another embodiment, R12 is A-B. In another embodiment, R13 is A-B. In another embodiment, R14 is A-B. In another embodiment, R15 is A-B. In another embodiment, this invention provides compounds of the above to improve blood circulation; soothing stroke; Prevent plaque formation and promote their dissipated; improve blood viscosity; reduce cardiovascular; reduce cerebrovascular; reduce thrombosis, arteriosclerosis, coronary heart disease, hypertension, diabetes, thrombocytopenia purpura, hemoptysis, hematemesis; treating blood in the stool, uterine bleeding, traumatic bleeding, abdominal irritation, swelling, flutter, Blood circulation, swelling, pain; Treating bronchiectasis, tuberculosis and lung abscess caused by too hemoptysis; for treating retinoblastoma, reducing bleeding, antitussive, expectorant and analgesic effect, dilate blood vessels; reducing blood pressure and the treatment of cerebral arteriosclerosis; elevated blood lipids and reduced cholesterol. The R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14, R15, R16 bonds of (3K2) can be alpha or beta. The compounds may be in form of salts. In another embodiment, the compounds are in form of powder, crystal, or liquid. In embodiment, the compounds are in capsule, or with pharmaceutically acceptable carrier or diluent. In another embodiment,R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14, R15, R16 are independently selected from the group of O, hydrogen, hydroxyl, methyl, O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, 0-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O- (E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl; wherein the compounds may be in form of salts.

In embodiment the core having structures:

embedded image - T1 ;

embedded image - T2;

embedded image - T3;

embedded image - T4;

embedded image - T5;

embedded image - T6;

embedded image - T7;

embedded image - T8;

embedded image - T9;

embedded image - T10;

embedded image - T11;

embedded image - T12;

embedded image - T13;

embedded image - T14;

embedded image - T15;

embedded image - T16;

embedded image - T17;

embedded image - T18

embedded image - T19,

embedded image - T20

embedded image - T21;

embedded image - T22;

embedded image - T23;

embedded image - T24;

embedded image - T25;

embedded image - T26;

embedded image - T27

wherein R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14, R15, R16, R17, R18, of T19, T20, T21, T22, T23, T24, T25, T26, T27 represent H, OH, O, CH2OH, COOH or CH3. The bonds can be in forms of alpha or beta or in combinations.

Esterification the above core compound with acyl halide, wherein the halide comprise chloride, bromide, fluoride and iodide, wherein the acyl halide comprise acyl chloride, wherein acyl chloride comprise tigloyl chloride, angeloyl chloride, acetyl chloride, crotonoyl chloride, 3,3-Dimethylacryloyl chloride, senecioyl chloride, cinnamoyl chloride, pentenoyl chloride, hexanoyl chloride, benzoyl chloride, ethylbutyryl chloride, propionyl chloride, 2-propenoyl chloride, isobutyryl chloride, butyryl chloride, (2E)-2-pentenoyl chloride, 4-Pentenoyl chloride, 5-hexenoyl chloride, heptanoyl chloride, octanoyl chloride, nonanoyl chloride, decanoyl chloride, Lauroyl chloride, myristoyl chloride, oleoyl chloride. The compounds vary in composition when the time or temperature of the reaction is changed. The peaks, fractions and compounds are selected according to the activities of times studies and the changes of peaks. The compounds having strong to weak activities are selected and isolated. The anti-cancer activities (Cytotoxic Assay) are the MTT studies of bone (U2OS), lung (H460), bladder(HTB-9), ovary (ES2), colon (HCT116), pancreas (Capan), ovary(OVCAR3), prostate (DU145), skin (SK-Mel-5), mouth (KB), kidney (A498), breast (MCF-7), liver (HepG2), brain (T98G), leukemia (K562), cervix (HeLa). The active esterification products are purified with HPLC. The reaction product of mixtures and individual compounds are tested with MTT Cytotoxic Assay. Details of method are in Experiment 3 of the present application. A second esterification of compound can be selected from the above experiment results to produce new active compounds. A partial esterification compound is selected from the above experiments to perform a second or repeated with a third esterification with different acyl chloride in order to produce new active compounds with the experiments in the present application.

A method is 1) Dissolving core compound or triterpenes core, hydroxylated triterpenes core, in pyridine; 2) Adding acyl halide or acyl chloride; 3, The mixture is stirred for length of time including 5 sec, 10 sec, 20 sec, 30 sec, 40 sec, 1 min, 2 min, 5 min, 10 min, 30 min, 1 hr, 2 hr, 18 hr, 2 days or 3 days at different temperature; 4) At the end of reaction, aqueous solution of acid or weak base, or water is added to the reaction mixture; 5) The solution is then extracted of ethyl acetate and ethyl acetate is removed by evaporation and lyophilization; 6) Dissolving the reaction product in acetonitrile with Trifluoroacetic acid or DMSO; 7) Testing the reaction product of mixtures and individual fractions with MTT cytotoxic assay; 8) Selecting the HPLC fractions for isolation is according to the cytotoxic activity of the reaction product obtained at a specific reaction time; 10) Purifying the active esterification products with HPLC; 11) Collecting the products; 12) Testing the products; wherein the core compound is terpene, isoprene, or triterpene core or hydroxylated triterpenes core; wherein the core compound was dissolved in pyridine; wherein the acyl chloride including Tigloyl chloride, angeloyl chloride, Acetyl chloride, Crotonoyl chloride, 3,3-Dimethylacryloyl chloride, senecioyl chloride, Cinnamoyl chloride, Pentenoyl chloride, Hexanoyl chloride, benzoyl chloride, Ethylbutyryl chloride, Propionyl chloride, 2-Propenoyl chloride, Isobutyryl chloride, Butyryl chloride, (2E)-2-pentenoyl chloride, 4-Pentenoyl chloride, 5-Hexenoyl chloride, Heptanoyl chloride, Octanoyl chloride, Nonanoyl chloride, Decanoyl chloride, Lauroyl chloride, Myristoyl chloride, and Oleoyl chloride; wherein the reaction time for the mixture is stirred for 5 sec, 10 sec, 20 sec, 30 sec, 40 sec, 1 min, 2 min, 5 min, 10 min, 30 min, 1 hr, 2 hr, 18 hr, 2 days or 3 days; wherein the temperature is 0° C., 25° C., 50° C. or 75° C. temperature; wherein the acid including HCl or the base including NaHCO3 is added to the reaction mixture; wherein the solution is then extracted 3 times with ethyl acetate and lyophilization; wherein the reaction product is dissolved in 80% acetonitrile - 0.005% Trifluoroacetic acid or DMSO; wherein selecting the HPLC fractions for isolation is according to the cytotoxic activity of the reaction product obtained at a reaction time of 5 sec, 10 sec, 20 sec, 30 sec, 40 sec, 1 min, 2 min, 5 min, 10 min, 30 min, 1 hr, 2 hr, 18 hr, 2 days or 3 days. In another embodiment, the reaction time may be over 3 days. In another embodiment, the experiment may be performed under 0C. In another embodiment, the experiment may be performed over 75° C. In embodiment, the attachment of sugar moiety(ies) can be biosynthesized.

This invention provide the compound having structures of T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15, T16, T17; T18, T19, T20, T21, T22, T23, T24, T25, T26, T27 wherein the R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14, R15, R16, R17, R18 are independently selected from the group of hydrogen, hydroxyl, methyl, O, O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, 0-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O- (E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl,, O-4-(dimethylamino)-2-methylbut-2-enoyl, O-4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl, O-sugar moiety(ies), O-acid moiety(ies), CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl,CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl,CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl,CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl,CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-Isobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl,CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl,CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl,, CH2O-4-(dimethylamino)-2-methylbut-2-enoyl, CH2O-4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl, alkane, alkene and derivatives thereof. In another embodiment, the compound is attached a sugar moiety(ies), acid moiety(ies) or alduronic acid, wherein the sugar moiety(ies) is/are selected from a group of glucose, galactose, rhamnose, arabinose, xylose, fucose, allose, altrose, gulose, idose, lyxose, mannose, psicose, ribose, sorbose, tagatose, talose, fructose, alduronic acid, glucuronic acid, galacturonic acid, and derivatives or combinations thereof;. In another embodiment, the compound is attached a group selected from (CnH2n)O-angeloyl, (CnH2n)O-tigloyl, (CnH2n)O-senecioyl, (CnH2n)O-acetyl, (CnH2n)O-Crotonoyl, (CnH2n)O-3,3-Dimethylacryloyl, (CnH2n)O-Cinnamoyl, (CnH2n)O-Pentenoyl, (CnH2n)O-Hexanoyl, (CnH2n)O-benzoyl, (CnH2n)O-Ethylbutyryl, (CnH2n)O-alkyl, (CnH2n)O-dibenzoyl, (CnH2n)O-benzoyl, (CnH2n)O-alkanoyl, (CnH2n)O-alkenoyl, (CnH2n)O-benzoyl alkyl substituted O-alkanoyl, (CnH2n)O-alkanoyl substituted phenyl, (CnH2n)O-alkenoyl substituted phenyl, (CnH2n)O-aryl, (CnH2n)O-acyl, (CnH2n)O-heterocylic, (CnH2n)O-heteroraryl, (CnH2n)O-alkenylcarbonyl, (CnH2n)O-alkane, (CnH2n)O-alkene and (CnH2n)O-sugar moiety and (CnH2n)O-acid moiety; wherein the sugar moiety(ies) is/are included but not limited to a group of glucose, galactose, rhamnose, arabinose, xylose, fucose, allose, altrose, gulose, idose, lyxose, mannose, psicose, ribose, sorbose, tagatose, talose, fructose, alduronic acid, glucuronic acid, galacturonic acid, and derivatives or combinations thereof; wherein n is 1 or 2 or 3 or 4 or over 5, wherein the compound may be in form of salts.

This invention provides the compound having structures of T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15, T16, T17; T18, T19, T20, T21, T22, T23, T24, T25. T26 provided for treating cancers, inhibition of cancer growth, cancer invasion, cell invasion, cancer cell invasion; cell adhesion, cell attachment, cell circulating; for treating mad cow disease; treating prion diseases; for treating diabetes; for inhibiting viruses; for preventing cerebral aging; for improving memory; improving cerebral functions; for curing enuresis, frequent micturition, urinary incontinence; dementia, Alzheimer’s disease, autism, brain trauma, Parkinson’s disease or other diseases caused by cerebral dysfunctions or neurodegeneration; for treating arthritis, rheumatism, poor circulation, arteriosclerosis, Raynaud’s syndrome, angina pectoris, cardiac disorder, coronary heart disease, headache, dizziness, kidney disorder; cerebrovascular diseases; inhibiting NF-kappa B activation; for treating brain edema, severe acute respiratory syndrome, respiratory viral diseases, chronic venous insufficiency, hypertension, chronic venous disease, oedema, inflammation, hemorrhoids, peripheral edema formation, varicose vein disease, flu, post traumatic edema and postoperative swelling; for inhibiting blood clots, for inhibiting ethanol absorption; for lowering blood sugar; for regulating adrenocorticotropin and corticosterone levels, for treating retinoblastoma. This invention provides a composition for Anti-MS, anti-aneurysm, antiasthmatic, anti-oedematous, anti-inflammatory, anti-bradykinic, anti-capillarihemorrhagic, anti-cephalagic, anti-cervicobrachialgic, anti-eclamptic, anti-edemic, anti-encaphalitic, anti-epiglottitic, anti-exudative, anti-flu, anti-fracture, anti-gingivitic, anti-hematomic, anti-herpetic, anti-histaminic, anti-hydrathritic, anti-meningitic, antioxidant, anti-periodontic, anti-phlebitic, anti-pleuritic, anti-raucedo, anti-rhinitic, anti-tonsilitic, anti-ulcer, anti-varicose, anti-vertiginous, cancerostatic, corticosterogenic, diuretic, fungicide, hemolytic, hyaluronidase inhibitor, lymphagogue, natriuretic, pesticide, pituitary stimulant, thymolytic, vasoprotective, inhibiting leishmaniases, modulating adhesion or angiogenesis of cells, anti-parasitic; increase the expression of the genes: ANGPT2, DDIT3, LIF and NFKB1Z, and manufacturing an adjuvant composition and venotonic treatment. The compound blocks the DNA synthesis of cancer cell; wherein the cancers comprise breast cancer, leukocytic cancer, liver cancer, ovarian cancer, bladder cancer, prostatic cancer, skin cancer, bone cancer, brain cancer, leukemia cancer, lung cancer, colon cancer, CNS cancer, melanoma cancer, renal cancer, cervical cancer, esophageal cancer, testicular cancer, splenic cancer, kidney cancer, lymphatic cancer, pancreatic cancer, stomach cancer, eye cancer and thyroid cancer. This invention provides compounds to improve blood circulation; soothing stroke; Prevent plaque formation and promote their dissipated; improve blood viscosity; reduce cardiovascular; reduce cerebrovascular; reduce thrombosis, arteriosclerosis, coronary heart disease, hypertension, diabetes, thrombocytopenia purpura, hemoptysis, hematemesis; treating blood in the stool, uterine bleeding, traumatic bleeding, abdominal irritation, swelling, flutter, Blood circulation, swelling, pain; Treating bronchiectasis, tuberculosis and lung abscess caused by too hemoptysis; reducing bleeding, antitussive, expectorant and analgesic effect, dilate blood vessels; reducing blood pressure and the treatment of cerebral arteriosclerosis; elevated blood lipids and reduced cholesterol.

Liposome is artificially prepared vesicles which made up of a lipid bilayer. Certain sizes of liposome can enter tumor sites from blood due to the enhanced permeability and retention effect. While human blood vessels are all surrounded by endothelial cells bound by tight junctions, those tight junctions binding tumor vessels are leakier than those binding other vessels and thus liposomes are able to enter these vessels to enhance the delivery, efficacy, bioavailability and absorption of liposome enclosed drug. This invention provides methods to use liposomes or nanoparticles capsules as a carrier delivering the compound as medicament, wherein the size of liposomes or nanoparticles capsules is less than 200 nm or 100-200 nm or 50-100 nm or 5-50 nm or less than 50 nm, wherein the medicament is included but not limited for treating cancer, inhibiting cancer growth, inhibiting cancer invasion, inhibiting cancer metastasis, modulating cell adhesion, modulating cell attachment, wherein the compound is selected from formula (2A) or formula (K) at the above.

Substitution, deletion and/or addition of any group in the above-described compounds by other group(s) will be apparent to one of ordinary skill in the art based on the teachings of this application. In a further embodiment, the substitution, deletion and/or addition of the group(s) in the compound of the invention does not substantially affect the biological function of the compound is included in the invention.

In an embodiment, the compound is selected from the structures:

embedded image - Compound E4A-Tig-R:

embedded image - Compound E4A-Tig-N:

embedded image - Compound E4A-Tig-Q:

embedded image - Compound E4A-Tig-V:

embedded image - Compound E4A-Tig-T:

embedded image - Compound E4A-Tig-U:

embedded image - Compound E4A-Tig-S:

embedded image - Compound E4A-Ang-R:

embedded image - Compound E4A-Ang-V:

embedded image - Compound E4A-Ang-Q:

embedded image - Compound E4A-Ang-N:

embedded image - Compound E4A-Ang-T:

embedded image - Compound E4A-Ang-U:

embedded image - Compound E4A-Ang-S:

embedded image - Compound E4A-Sen-R:

embedded image - Compound E4A-Sen-V:

embedded image - Compound E4A-Sen-N:

embedded image - Compound E4A-Sen-Q:

embedded image - Compound E4A-Sen-S:

embedded image - Compound E4A-Sen-T:

embedded image - Compound E4A-Sen-U:

embedded image - Compound E4A-Cro-R:

embedded image - Compound E4A-Cro-V:

embedded image - Compound E4A- Cro-N:

embedded image - Compound E4A-Cro-Q:

embedded image - Compound E4A- Cro-S:

embedded image - Compound E4A- Cro-T:

embedded image - Compound E4A-Cro-U:

embedded image - Compound E4A- Acy-R:

embedded image - Compound E4A- Acy-V:

embedded image - Compound E4A- Acy-N:

embedded image - Compound E4A- Acy-Q:

embedded image - Compound E4A- Acy-S:

embedded image - Compound E4A- Acy-T:

embedded image - Compound E4A-Acy-U:

embedded image - Compound E4A-Pen-R:

embedded image - Compound E4A-Pen-V:

embedded image - Compound E4A-Pen-N:

embedded image - Compound E4A-Pen-Q:

embedded image - Compound E4A-Pen-S:

embedded image - Compound E4A-Pen-T:

embedded image - Compound E4A-Pen-U:

embedded image - Compound E4A-Cin-R:

embedded image - Compound E4A-Cin-V:

embedded image - Compound E4A-Cin-N:

embedded image - Compound E4A-Cin-Q:

embedded image - Compound E4A-Cin-S:

embedded image - Compound E4A-Cin-T:

embedded image - Compound E4A-Cin-U:

embedded image - Tig-Sen-1:

embedded image - Tig-Cro-1:

embedded image - Tig-Ang-1:

embedded image - Tig-Pen-1:

embedded image - Tig-Acy-1:

embedded image - Tig-Hex-1:

embedded image - Tig-Eth-1:

In embodiment, sugar moiety(ies) or acid moiety(ies) can be attached to the above compounds. In embodiment, the attachment of sugar moiety(ies) can be biosynthesized; In embodiment, the attachment of acid moiety can be biosynthesized; wherein the sugar moiety(ies) or acid moiety(ies) is/are included but not limited to a group of glucose, galactose, rhamnose, arabinose, xylose, fucose, allose, altrose, gulose, idose, lyxose, mannose, psicose, ribose, sorbose, tagatose, talose, fructose, alduronic acid, glucuronic acid, galacturonic acid, and derivatives or combinations thereof;

This invention provides compounds by esterification of core compound (C) or (D1) with acetyl chloride, angeloyl chloride, tigloyl chloride, senecioyl chloride, Crotonoyl chloride, O-3,3-Dimethylacryloyl chloride, Cinnamoyl chloride, Pentenoyl chloride, Hexanoyl chloride, benzoyl chloride, Ethylbutyryl chloride, and isolation of the compounds with HPLC, for treating cancer, inhibiting cancer growth, inhibiting cancer invasion, inhibiting cancer metastasis, modulating cell adhesion, modulating cell attachment, wherein the core compound selected from the following:

embedded image - (C)

embedded image - (D1)

embedded image - (A)

Esterification of compounds (A), (C), or (D1) with acyl chloride including Tigloyl chloride, angeloyl chloride, Acetyl chloride, Crotonoyl chloride, 3,3-Dimethylacryloyl chloride, senecioyl chloride, Cinnamoyl chloride, Pentenoyl chloride, Hexanoyl chloride, benzoyl chloride, Ethylbutyryl chloride, ethanoyl chloride, propanoyl chloride, propenoyl chloride, butanoyl chloride, butenoyl chloride, pentanoyl chloride, hexenoyl chloride, heptanoyl chloride, heptenoyl chloride, octanoyl chloride, octenoyl chloride, nonanoyl chloride, nonenoyl chloride, decanoyl chloride, decenoyl chloride, propionyl chloride, 2-propenoyl chloride, 2-butenoyl chloride, Isobutyryl chloride, 2-methylpropanoyl chloride, 2-ethylbutyryl chloride, ethylbutanoyl chloride, 2-ethylbutanoyl chloride, butyryl chloride, (E)-2,3-Dimethylacryloyl chloride, (E)-2-Methylcrotonoyl chloride, 3-cis-Methyl-methacryloyl chloride, 3-Methyl-2-butenoyl chloride, 3-Methylcrotonoyl chloride, 4-Pentenoyl chloride, (2E)-2-pentenoyl chloride, Caproyl chloride, 5-Hexenoyl chloride, Capryloyl chloride, Lauroyl chloride, Dodecanoyl chloride, Myristoyl chloride, Tetradecanoyl chloride, Oleoyl chloride, C(2-18) Acyl chloride,

The compounds vary in composition when the time or temperature of the reaction is changed. The peaks, fractions and compounds are selected according to the activities of times studies and the changes of peaks. The compounds having strong to weak activities are selected and isolated. The anti-cancer activities are the MTT studies of bone (U2OS), lung (H460), bladder(HTB-9), ovary (ES2), colon (HCT116), pancreas (Capan), ovary(OVCAR3), prostate (DU145), skin (SK-Mel-5), mouth (KB), kidney (A498), breast (MCF-7), liver (HepG2), brain (T98G), leukemia (K562), cervix (HeLa). The active esterification products are purified with HPLC. The reaction product of mixtures and individual compounds are tested with MTT Cytotoxic Assay. Details of method are in Experiment 3 of the present application. A second esterification of compound can be selected from the above experiment results to produce new active compounds. A partial esterification compound is selected from the above experiments to perform a second or repeated with a third esterification with different acyl chloride in order to produce new active compounds with the experiments in the present application, wherein the compound can be selected from K, (H1) or (H2):

embedded image - (H1)

embedded image - (H2)

embedded image - (K)

R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14, R15, R16, R17, R18 are independently selected from the group of CH3, CH2OH, COOH hydrogen, hydroxyl, methyl, O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, 0-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-alkane, O-alkene, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, O-2-butenoyl, O-Isobutyryl, 0-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl; CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2-Isobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl; CH2O-4-(dimethylamino)-2-methylbut-2-enoyl, CH2O-4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl or wherein any 1 or 2 or 3 or 4 of R1, R2, R3, R4, R5, R8, R10, R16, R17, R18 is/are independently attached an O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl,O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, O-2-butenoyl, O-Isobutyryl, 0-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2-Isobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl, CH2O-4-(dimethylamino)-2-methylbut-2-enoyl, CH2O-4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl; R9, R11, R12, R13, R14, R15 are independently attached a CH3; or wherein R10 is attached an O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl,O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, O-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O- (E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, 0-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl, O-4-(dimethylamino)-2-methylbut-2-enoyl, and O-4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl,CH3, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-Isobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl; CH2O-4-(dimethylamino)-2-methylbut-2-enoyl, CH2O-4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl or wherein R4 and R10 are independently attached an CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH2OH, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl, CH2O-ethanoyl, CH2O-propanoyl, CH2O-propenoyl, CH2O-butanoyl, CH2O-butenoyl, CH2O-pentanoyl, CH2O-hexenoyl, CH2O-heptanoyl, CH2O-heptenoyl, CH2O-octanoyl, CH2O-octenoyl, CH2O-nonanoyl, CH2O-nonenoyl, CH2O-decanoyl, CH2O-decenoyl, CH2O-propionyl, CH2O-2-propenoyl, CH2O-2-butenoyl, CH2O-Isobutyryl, CH2O-2-methylpropanoyl, CH2O-2-ethylbutyryl, CH2O-ethylbutanoyl, CH2O-2-ethylbutanoyl, CH2O-butyryl, CH2O-(E)-2,3-Dimethylacryloyl, CH2O-(E)-2-Methylcrotonoyl, CH2O-3-cis-Methyl-methacryloyl, CH2O-3-Methyl-2-butenoyl, CH2O-3-Methylcrotonoyl, CH2O-4-Pentenoyl, CH2O-(2E)-2-pentenoyl, CH2O-Caproyl, CH2O-5-Hexenoyl, CH2O-Capryloyl, CH2O-Lauroyl, CH2O-Dodecanoyl, CH2O-Myristoyl, CH2O-Tetradecanoyl, CH2O-Oleoyl, CH2O-C(2-18) Acyl; CH2O-4-(dimethylamino)-2-methylbut-2-enoyl, CH2O-4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl or wherein R17 and R18 are independently attached an O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, O-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl,O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl, O-ethanoyl, O-propanoyl, O-propenoyl, O-butanoyl, O-butenoyl, O-pentanoyl, O-hexenoyl, O-heptanoyl, O-heptenoyl, O-octanoyl, O-octenoyl, O-nonanoyl, O-nonenoyl, O-decanoyl, O-decenoyl, O-propionyl, O-2-propenoyl, 0-2-butenoyl, O-Isobutyryl, O-2-methylpropanoyl, O-2-ethylbutyryl, O-ethylbutanoyl, O-2-ethylbutanoyl, O-butyryl, O-(E)-2,3-Dimethylacryloyl, O-(E)-2-Methylcrotonoyl, O-3-cis-Methyl-methacryloyl, O-3-Methyl-2-butenoyl, O-3-Methylcrotonoyl, O-4-Pentenoyl, O-(2E)-2-pentenoyl, O-Caproyl, O-5-Hexenoyl, O-Capryloyl, O-Lauroyl, O-Dodecanoyl, O-Myristoyl, O-Tetradecanoyl, O-Oleoyl, O-C(2-18) Acyl; wherein R3 is OH or H or absent; wherein R1, R2, R3, R5, R8 are OH or H or absent; wherein R9, R11, R12, R13, R14, and R15 are CH3; or wherein R1, R2, R5, R8 represent OH; R3 represents OH, H or absent; R4, R10 represent CH2Oangeloyl; R9, R11, R12, R13, R14, R15 represent CH3; or wherein R1, R2, R5, R8 represent OH or O-tigloyl; R3 represents OH, H, or absent; R4, R10 represent CH2Otigloyl; R9, R11, R12, R13, R14, R15 represent CH3;

wherein the group attaching to the core compound selected from acetyl, angeloyl, tigloyl, senecioyl, Crotonoyl, O-3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, methylbutanoyl, methylpropanoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, alkenylcarbonyl, O-4-(dimethylamino)-2-methylbut-2-enoyl, and O-4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl are interchangeable or replaceable thereof. They can be the same group or in combination thereof. The compounds may be in form of salts. The effect of the compounds of the present application on cells is arresting cells in the S-phase and blocking their entering into the G2/M phase of cell cycle. The compounds block the DNA synthesis of cancer cell. This invention provides compounds and method for improving blood circulation; soothing stroke; Prevent plaque formation and promote their dissipated; improve blood viscosity; reducing cardiovascular; reducing cerebrovascular; reducing thrombosis, arteriosclerosis, coronary heart disease, hypertension, diabetes, thrombocytopenia purpura, hemoptysis, hematemesis; treating blood in the stool, uterine bleeding, traumatic bleeding, abdominal irritation, swelling, flutter, Blood circulation, swelling, pain; Treating bronchiectasis, tuberculosis and lung abscess caused by too hemoptysis; reducing bleeding, antitussive, expectorant and analgesic effect, dilate blood vessels; reducing blood pressure and the treatment of cerebral arteriosclerosis; elevated blood lipids and reduced cholesterol.

A composition comprising an effective amount of compound selected from the above formula or a salt, ester, metabolite or derivative thereof can be used as a medicament for blocking the invasion, migration, metastasis of cancer cells, inhibiting tumor or cancer cell growth and for treating cancer, wherein the cancers comprise breast cancer, leukocytic cancer, liver cancer, ovarian cancer, bladder cancer, prostatic cancer, skin cancer, bone cancer, brain cancer, leukemia cancer, lung cancer, colon cancer, CNS cancer, melanoma cancer, renal cancer, cervical cancer, esophageal cancer, testicular cancer, splenic cancer, kidney cancer, lymphatic cancer, pancreatic cancer, stomach cancer, eye cancer and thyroid cancer.

This invention provides a composition comprising the compounds provided in the invention for treating cancers; for inhibiting viruses; for preventing cerebral aging; for improving memory; improving cerebral functions; for curing enuresis, frequent micturition, urinary incontinence; dementia, Alzheimer’s disease, autism, brain trauma, Parkinson’s disease or other diseases caused by cerebral dysfunctions; for treating arthritis, rheumatism, poor circulation, arteriosclerosis, Raynaud’s syndrome, angina pectoris, cardiac disorder, coronary heart disease, headache, dizziness, kidney disorder; diabetes; cerebrovascular diseases; inhibiting NF-kappa B activation; for treating brain edema, severe acute respiratory syndrome, respiratory viral diseases, chronic venous insufficiency, hypertension, chronic venous disease, oedema, inflammation, hemorrhoids, peripheral edema formation, varicose vein disease, flu, post traumatic edema and postoperative swelling; for inhibiting blood clots, for inhibiting ethanol absorption; for lowering blood sugar; for regulating adrenocorticotropin and corticosterone levels. This invention provides a composition for Anti-MS, anti-aneurysm, anti-asthmatic, anti-oedematous, anti-inflammatory, anti-bradykinic, anti-capillarihemorrhagic, anti-cephalagic, anti-cervicobrachialgic, anti-eclamptic, anti-edemic, anti-encaphalitic, anti-epiglottitic, anti-exudative, anti-flu, anti-fracture, anti-gingivitic, anti-hematomic, anti-herpetic, anti-histaminic, anti-hydrathritic, anti-meningitic, antioxidant, anti-periodontic, anti-phlebitic, anti-pleuritic, anti-raucedo, anti-rhinitic, anti-tonsilitic, anti-ulcer, anti-varicose, anti-vertiginous, cancerostatic, corticosterogenic, diuretic, fungicide, hemolytic, hyaluronidase inhibitor, lymphagogue, natriuretic, pesticide, pituitary stimulant, thymolytic, vasoprotective, inhibiting leishmaniases, modulating adhesion or angiogenesis of cells, anti-parasitic; increase the expression of the genes: ANGPT2, DDIT3, LIF and NFKB1Z, and manufacturing an adjuvant composition and venotonic treatment. The composition blocks the DNA synthesis of cancer cell.

This invention provides a method for increasing the level of DR5 of cells, up regulating the expression of death receptor DR5 as biomarker in cancer cells, comprising contacting said cells with an effective amount of a compound having the structure: (3 K) in this application. This invention provides a method for increasing the agonist activities of anti DR5 antibody, comprising contacting said cells with an effective amount of a compound having the structure selecting from (3 K) in this application.

This invention provides a method of identifying a subject having cancer who is likely to be responsive to a treatment compound for increase the level of DR5, comprising administering the treatment compound to the subject having the cancer; obtaining a sample from the subject; determining the level of a biomarker in the sample from the subject; and diagnosing the subject as being likely to be responsive to the treatment compound if the level of the biomarker DR5 in the sample of the subject increase; wherein the treatment compound is a compound having the structure selecting from (3 K) in this application.

Alkenyl means unsaturated linear or branched structures and combinations thereof, having formula R2C=CR2, one or more double bonds therein. Examples of alkenyl groups include vinyl, propenyl, isopropenyl, butenyl, s- and t-butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, and hexadienyl. An aryl is a functional group of organic molecule derived from an aromatic compound such as benzene, a 6-14 membered carbocyclic aromatic ring system comprising 1-3 benzene rings. If two or more aromatic rings are present, then the rings are fused together, so that adjacent rings share a common bond. Examples include phenyl and naphthyl. The aryl group may be substituted with one or more substitutes independently selected from halogen, alkyl or alkoxy. Acyl is a functional group which can be obtained from an organic acid by the removal of the carboxyl. Acyl groups can be written using the general formula -COR, where there is a double bond between the carbon and oxygen. The names of acyl groups typically end in -yl, such as formyl, acetyl, propionyl, butyryl and benzoyl. Benzoyl is one of the acyls, C₆H₅COR, obtained from benzoic acid by the removal of the carboxyl. A heterocyclic compound is a compound containing a heterocyclic ring which refers to a non-aromatic ring having 1-4 heteroatoms, said ring being isolated or fused to a second ring selected from 3- to 7-membered alicyclic ring containing 0-4 heteroatoms, aryl and heteroaryl, wherein heterocyclic compounds include pyrrolidinyl, pipyrazinyl, morpholinyl, trahydrofuranyl, imidazolinyl, thiomorpholinyl, and the like. Heterocyclyl groups are derived from heteroarenes by removal of a hydrogen atom from any ring atom. Alkanoyl is the general name for an organic functional group RCO-, where R represents hydrogen or an alkyl group. Examples of alkanoyls are acetyl, propionoyl, butyryl, isobutyryl, pentanoyl and hexanoyl. Alkenoyl is an alkenylcarbonyl in which the alkenyl is defined above. Examples are pentenoyl (tigloyl) and pentenoyl (angeloyl). Alkyl is a radical containing only carbon and hydrogen atoms arranged in a chain, branched, cyclic or bicyclic structure or their combinations, having 1-18 carbon atoms. Examples include but are not limited to methyl, ethyl, propyl isopropyl, butyl, s-and t-butyl, pentyl, hexyl, heptyl, octyl, nonyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. Benzoyl alkyl substituted alkanoyl refers to straight or branched alkanoyl substituted with at least one benzoyl and at least one alkyl, wherein the benzoyl is attached to a straight or branched alkyl. An example of a benzoyl alkyl substituted alkanoyl is benzoyl methyl isobutanoyl. A sugar moiety is a segment of molecule comprising one or more sugars or derivatives thereof or alduronic acid thereof.

(Y)Y3, Y and Y3 represent the same compound. YM and (ACH-Y) represent the same compound. Connecting moiety is a substructure or a group of atoms which connect the functional group to a core compound. Example: angeloyl group is connected by a sugar moiety to a triterpene core.

Acetyl = ethanoyl; Propionyl = methylpropanoyl; Crotonoyl = 2-butenoyl; Isobutyryl = 2-methylpropanoyl; 2-Ethylbutyryl =2-Ethylbutanoyl; Butyryl = n-Butyryl = butanoyl = C-4 Acyl; trans-2-Methyl-2-butenoyl = (E)-2,3-Dimethylacryloyl chloride = (E)-2-Methylcrotonoyl = 3-cis-Methyl-methacryloyl =Tigloyl; 3,3-Dimethylacryloyl = 3-Methyl-2-butenoyl = 3-Methylcrotonoyl = Senecioyl; Propionyl chloride = methylpropanoyl ; Hexanoyl = Caproyl; Heptanoyl = Enanthic = Oenanthic; Octanoyl = Capryloyl; Dodecanoyl= Lauroyl; Tetradecanoyl= Myristoyl; C(2-18)Acyl is an acyl group having 2 to 18 carbons.

ethanoyl is a C-2 Acyl, propanoyl is a C-3 Acyl, propenoyl is a C-3 Acyl, propionyl is a C-3 Acyl, butanoyl is a C-4 Acyl, butenoyl is a C-4 Acyl, crotonoyl is a C-4 Acyl, pentanoyl is a C-5 Acyl, pentenoyl is a C-5 Acyl, angeloyl is C-5 Acyl, tigloyl is C-5 Acyl, senecioyl is C-5 Acyl, hexanoyl is a C-6 Acyl, hexenoyl is a C-6 Acyl, heptanoyl is a C-7 Acyl, heptenoyl is a C-7 Acyl, octanoyl is a C-8 Acyl, octenoyl is a C-8 Acyl, nonanoyl is a C-9 Acyl, nonenoyl is a C-9 Acyl, decanoyl is a C-10 Acyl, decenoyl is a C-10 Acyl, lauroyl is a C-12 Acyl, dodecanoyl is a C-12 Acyl, myristoyl is a C-14 Acyl, oleoyl is a C-18 Acyl.

The building blocks used in the invention including triterpenes, hydroxylated triterpenes, acetyl, angeloyl, tigloyl, senecioyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, methylbutanoyl, methylpropanoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, alkenylcarbonyl, ethanoyl, propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl and Oleoyl, 4-(dimethylamino)-2-methylbut-2-enoyl, 4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl or halides thereof, or chloride.thereof.

Acetyl chloride

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angeloyl chloride

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tigloyl chloride

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senecioyl chloride

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Crotonoyl chloride

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0-3,3-Dimethylacryloyl chloride

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Cinnamoyl chloride

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Pentenoyl chloride

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Hexanoyl chloride

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benzoyl chloride

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Ethylbutyryl chloride

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Propionyl chloride

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2-Propenoyl chloride,

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Isobutyryl chloride

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Butyryl chloride

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(2E)-2-pentenoyl chloride

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4-Pentenoyl chloride,

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Hexanoyl chloride

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5-Hexenoyl chloride

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Heptanoyl chloride

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Octanoyl chloride

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Nonanoyl chloride

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Decanol chloride

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Lauroyl chloride

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Myristoyl chloride

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Oleoyl chloride

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; 4-(dimethylamino)-2-methylbut-2-enoyl chloride

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4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl chloride

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Acryloyl chloride [Synonym: 2-propenoly chloride]; Propionyl chloride [Synonym: methylpropanoyl chloride]; Crotonoyl chloride [Synonym: 2-butenoyl chloride]; Isobutyryl chloride [Synonym: 2-methylpropanoyl chloride]; 2-Ethylbutyryl chloride [Synonym: 2-Ethylbutanoyl chloride]; Butyryl chloride (Synonym: n-Butyryl chloride, butanoyl chloride, or C-4 Acyl halide); trans-2-Methyl-2-butenoyl chloride [Synonym: (E)-2,3-Dimethylacryloyl chloride, (E)-2-Methylcrotonoyl chloride, 3-cis-Methyl-methacryloyl chloride, Tigloyl chloride]; 3,3-Dimethylacryloyl chloride [Synonym: 3-Methyl-2-butenoyl chloride, 3-Methylcrotonoyl chloride, Senecioyl chloride]; Hexanoyl chloride [Synonym: Caproyl chloride]; Heptanoyl chloride [Synonym: Enanthic chloride, Oenanthic chloride] Octanoyl chloride [Synonym: Capryloyl chloride].

In the presented experiments, concentrations of drug that inhibit 15% cell-growth or less (i.e., 85% of control or above) as compared to the no-drug control (DMSO) are considered non-cytotoxic concentrations. In another embodiment, the concentrations of drug that inhibit 10% cell-growth or less (i.e., 90% of control or above) as compared to the no-drug control (DMSO) are considered non-cytotoxic concentrations. In another embodiment, the concentrations of drug that inhibit 5% cell-growth or less (i.e., 95% of control or above) as compared to the no-drug control (DMSO) are considered non-cytotoxic concentrations. In another embodiment, the concentrations of drug that inhibit 20% cell-growth or less (i.e., 80% of control or above) as compared to the no-drug control (DMSO) are considered non-cytotoxic concentrations. In another embodiment, the concentrations of drug that inhibit 25% cell-growth or less (i.e., 75% of control or above) as compared to the no-drug control (DMSO) are considered non-cytotoxic concentrations. In another embodiment, the concentrations of drug that inhibit 30% cell-growth or less as compared to the no-drug control (DMSO) are considered non-cytotoxic concentrations. In another embodiment, the concentrations of drug that inhibit 45% cell-growth or less as compared to the no-drug control (DMSO) are considered non-cytotoxic concentrations.

The triterpene compound or compounds selected from this invention can be administered to a subject in need thereof, treating the subject, wherein including preventing cancer, or providing an adjuvant effect to the subject, or inhibiting the initiation or promotion of cancer, or killing the cancer/tumor cells, or inhibiting cancer cell invasion. In another embodiment the compounds inhibit the activation of Nuclear Factor-kB, wherein inhibiting the localization or wherein binding the DNA. In another embodiment the compounds block the DNA synthesis. In another embodiment the compounds induce apoptosis in cancer cells.

Determination of gene expression by Real-time PCR method (Brilliant QPCR, Agilent Technologies): The real-time polymerase chain reactions further confirm the results obtained from microarray analysis. The Real-time PCR results (shown below) confirmed that Compound Y3 and YM increase the expression of the genes: ANGPT2, DDIT3, LIF and NFKB1Z, wherein the results in Table 19-21 disclosed in PCT/US09/34115, filed Feb. 13, 2009.

The saponins are partially hydrolyzed into a mixture of products which can be separated by HPLC. Specific partial hydrolysis of saponins can also be achieved with enzymes. The glycosidases catalyze the hydrolysis of the glycosidic linkage. Galactosidase is an enzyme which catalyzes the hydrolysis of galactosides. Glucosidase is an enzyme which breaks glucose from saponin. Other enzyme examples are xylanases, lactase, amylase, chitinase, sucrase, maltase, and neuraminidase.

The sugar moiety of the triterpenoid saponin (example Xanifolia Y) can be removed by acid hydrolysis. The synthetic compound of ACH-Y is obtained. ACH-Y is a triterpene with acyl groups but no sugar moiety. The acyl group of the saponin (example Xanifolia Y) can be removed by alkaline hydrolysis. The synthetic compound AKOH-Y can be obtained. AKOH-Y is a pentacyclic triterpene with sugar moieties. A pentacyclic triterpene can be obtained by acid and alkaline hydroysis of saponins from natural sources. A pentacyclic triterpene can be obtained by synthetic methods (Reference: Surendra et al., Rapid and Enantioselective Synthetic Approaches to Germanicol and Other Pentacyclic Triterpenes, Journal of the American Chemical Society, 2008, 130(27), 8865-8869). Pentacyclic triterpenes with sugar moieties can also be obtained by synthesis (Reference: Ple et al., Synthesis of L-arabinopyranose containing hederagenin saponins, Tetrahedron 61 (2005) 4347-4362). Acylation is the process of adding an acyl group to a compound. The Friedel-Crafts reaction is an example of this process. An active compound can be obtained by acylating a pentacyclic triterpenes, or hydroxylated triterpenes. In another embodiment, acylating C24, C28, C21 and C22 of a pentacyclic triterpenes, or hydroxylated triterpenes produce compounds for inhibiting cancer growth, cancer invasion, cell invasion, cancer cell invasion, molecular cell invasion, cell attachment adhesion, or cell circulation. In another embodiment, the acyl group(s) may be at C3. In another embodiment, a sugar moiety is at C21, 22, or 28, wherein the sugar moiety is attached with 2 acyl groups. In another embodiment, acylating the compounds of (A), (B), (C), (D1), (D2), (F), (G), (H), produce the compounds for inhibiting cancer invasion, cell invasion or cancer cell invasion; cancer metastasis; or cancer growth. The building blocks in the present application are used to synthesize active saponins. In embodiment, the sugar moiety(ies) is/are included but not limited to a group of glucose, galactose, rhamnose, arabinose, xylose, fucose, allose, altrose, gulose, idose, lyxose, mannose, psicose, ribose, sorbose, tagatose, talose, fructose, alduronic acid, glucuronic acid, galacturonic acid, and derivatives or combinations thereof;

Acylating the compound (G) with angeloyl or tigloyl group gives the following compounds

embedded image - (K)

wherein R1, R2, R5, R8 represent OH or O-angeloyl; R3 represents OH, H or O-angeloyl; R4, R10 represent CH3, CH2OH or CH2Oangeloyl; R3 represents OH, H or O-angeloyl; R9, R11, R12, R13, R14, R15 represent CH3; or wherein R1, R2, R5, R8 represent OH or O-tigloyl; R3 represents OH, H or O- tigloyl; R4, R10 represent CH3, CH2OH or CH2O tigloyl; R9, R11, R12, R13, R14, R15 represent CH3; wherein the compounds inhibit cancer growth, cancer invasion, cell invasion or cancer cell invasion.

Acylating the compound (G) with angeloyl, tigloyl, senecioyl, acetyl, Crotonoyl, 3,3-Dimethylacryloyl, Cinnamoyl, Pentenoyl, Hexanoyl, benzoyl, Ethylbutyryl, alkyl, dibenzoyl, benzoyl, alkanoyl, alkenoyl, benzoyl alkyl substituted O-alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocylic, heteroraryl, CH2O-alkenylcarbonyl, alkane, alkene give the compound (K) wherein R1, R2, R5, R8 represent OH, O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, 0-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl,O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl; R4, R10 represent CH3, CH2OH, CH2O-angeloyl, CH2O-tigloyl, CH2O-senecioyl, CH2O-acetyl, CH2O-Crotonoyl, CH2O-3,3-Dimethylacryloyl, CH2O-Cinnamoyl, CH2O-Pentenoyl, CH2O-Hexanoyl, CH2O-benzoyl, CH2O-Ethylbutyryl, CH2O-alkyl, CH2O-dibenzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoyl alkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocylic, CH2O-heteroraryl, CH2O-alkenylcarbonyl,alkane, alkene; R3 is absent of represents OH, H, O-angeloyl, O-tigloyl, O-senecioyl, O-acetyl, O-Crotonoyl, 0-3,3-Dimethylacryloyl, O-Cinnamoyl, O-Pentenoyl, O-Hexanoyl, O-benzoyl, O-Ethylbutyryl,O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoyl alkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocylic, O-heteroraryl, O-alkenylcarbonyl; wherein R9, R11, R12, R13, R14, R15 represent CH3; wherein the compounds inhibit cancer growth, cancer invasion, cell invasion or cancer cell invasion; wherein the compound for use as mediator or inhibitor of adhesion protein or angiopoietin; wherein the compounds use as mediator modulating the secretion, expression, or synthesis of adhesion protein comprises reducing the fibronectin for inhibiting cell attachment, cell adhesion or cell circulation; wherein the adhesion proteins comprise fibronectin, integrins family, myosin, vitronectin, collagen, laminin, polyglycans, cadherin, heparin, tenascin, CD₅₄, and CAM; the compounds use for anti-adhesion therapy and targeting adhesion molecules for therapy.

Applicant further states that anti-adhesion therapy and targeting adhesion molecules for therapy is a new direction for development of drugs. Some examples of anti-adhesion drugs in clinical trials are Efalizumab, Odulimomab, Alicaforsen, Aselizumab etc., which target varies adhesion proteins. Please see TEXT BOOK, Adhesion Molecules: Function and Inhibition, (Reference 2), edited by Klaus Ley page 289-291, 297.

Adhesion molecules in inflammatory disease, (Reference 4), Abstract, line 7-8 “Blockade of the function of expression of CAM has emerged as a new therapeutic target in inflammatory diseases”. Applicants’ invention is an anti-adhesion therapy which is a new use of the compound as a mediator or inhibitor of adhesion proteins and angiopoietins. It inhibits excess adhesion and inhibits cell attachment.

In the present application, Applicants have used compounds selected from structure (2A) for anti-adhesion therapy, as a mediator or inhibitor of adhesion proteins and angiopoietins, and modulation of the cell attachment, and cell adhesion.

This invention provide a synthetic method to obtain semi-natural compounds by chemically removing functional groups of complex natural products to the basic core structure before de-novo chemically adding on active groups directly or sequentially by reaction with the active group donating chemical under different reaction temperature and time to produce series of different active group modified core structure compounds that can be fractionated and easily structurally determined as well as screening for different bio-active efficacies and toxicities as potential new drug candidates.

The activities of compounds are tested with cancer of leukemia(CCRF-CEM, HL60(TB), K-562, MOLT-4, RPMI8226, SR), lung(A549/ATCC, HOP-62, HOP92, NCI-H226, NCI-H322M, NCI-H460, colon(COLO205, HCC-2998, HCT-116, HCT-15, HT29, KM12, SW-620), CNS(SF-268, SF295, SF539, SNB-19, SNB-75, US51), melanoma(LOX IMVI, MALME-3M, M14, MDA-MB-3M, M14, MDA-MB-435, SK-MEL-2, SK-MEL-28, SK-MEL-5, UACC-257, UACC-62), ovary(ICTOV1, OVCAR-3, OVCAR-4, OVCAR-5, OVCAR-8, NCIADR-RES, SKOV3), renal(786-0, A498, ACHN, CAKI-1, SN12C, TK-10, UO-31), prostate(PC-3, DU-145), breast(MCF7, MDA-MB-231, HS578T, T47D, MDA-MB-468).

The room temperature is 25C in the present application.

An inducer is a molecule or compound that regulates gene expression.

An agonist is a substance which initiates a physiological response when combined with receptor. Example: Antibodies initiate cell death when combined with death receptor.

EXPERIMENTAL DETAILS

Experiment details of herb extraction, analysis of extract components by HPLC, determination of the cell-growth activity effected by Xanifolia Y with cells derived from different human organs using MTT Assay, purification of the bioactive components from plant extract, fractionation of plant extracts with FPLC, isolation of component Ys with preparative HPLC, determination of the chemical structure, cell experiments and animal studying are disclosed in PCT/US05/31900, U.S. Serial No. 11/289142, U.S. Serial 10/906303, U.S. Serial No. 11/131551 and U.S. Serial Nos. 11/683198, filed on Mar. 7, 2007, PCT/US2007/077273, filed Aug. 30, 2007, U.S. Serial No. 60/890380, filed on Feb. 16, 2007, U.S. Nos. 60/947,705, filed on Jul. 3, 2007, PCT/US2008/002086, 1188-ALA-PCT, filed Feb. 15, 2008, App′l No. PCT/US09/34115, filed Feb. 13, 2009, Experiments 1-23 of PCT/US2008/002086, 1188-ALA-PCT, filed Feb. 15, 2008 are incorporated in this application.

Experiment 1: Removal of the Sugar Moiety From Saponin by Acid Hydrolysis

15 mg saponin was dissolved in 1 ml of Methanol. 1 ml of 2N HCl was then added. The mixture was refluxed in 80° C. water bath for 5 hours. The solution was then neutralized by adding 2 ml of 1N NaOH (to final pH 4-6). The aglycone was then extracted with ethylacetate 3 ml × 2. The extracts were collected and pooled. Further isolation of aglycone (sugar-removed saponin) was achieved by HPLC with isocratic elution of 80-100% acetonitrile.

Experiment 2: Removal of the Acyl Group by Alkaline Hydrolysis

Methods: 20 mg of saponin was dissolved in 0.5 ml of 1N NaOH. The solution was incubated in 80° C. water bath for 4 hours. It was cooled to room temperature before neutralized with 0.5ml 1N HCl (adjust pH to about 3). The mixture was extracted with 2 ml 1-butanol 3 times. The butanol fractions were collected and lyophilized. The hydrolyzed saponin with further purified with HPLC in a C-18 column eluted with 25% acetonitrile.

Experiment 3: Adding the Acyl Group to Triterpene by Esterification

Method: 40 mg of triterpene core (fraction IV) was dissolved in 1 ml pyridine in a 50 ml tube. Reaction is started by adding 0.2 ml of acyl chloride (Tigloyl chloride, angeloyl chloride, Acetyl chloride, Crotonoyl chloride, 3,3-Dimethylacryloyl chloride(senecioyl chloride), Cinnamoyl chloride, Pentenoyl chloride, Hexanoyl chloride, benzoyl chloride or Ethylbutyryl chloride). The mixture is stirred for 5 sec, 1 min, 2 min, 5 min, 10 min, 30 min, 1 hr, 2 hr, 18 hr, 2 days or 3 days at 0° C., 25° C. or 75° C. temperature. At the end of reaction, 5 ml of 2N HCl or 1 M NaHCO3 is added to the reaction mixture. The solution is then extracted 3 times with 10 ml of ethyl acetate which is then evaporated under vacuum and at 45C and lyophilization. The reaction product is dissolved in 80% acetonitrile - 0.005% Trifluoroacetic acid or DMSO; and was separated with HPLC. Selecting the HPLC fractions for isolation is according to the cytotoxic activity of the reaction product obtained at a specific reaction time. The active esterification products are purified with HPLC. The reaction product of mixtures and individual compounds are tested with MTT cytotoxic assay. Structures are determined with NMR. See examples FIGS. 1-12(U.S. Serial No. 14/313080)

Experiment 4: Preparation of E4A

1. Beta-Escin dissolved in 1 M NaOH (20 mg/ml) was incubated at 70° C. for 5 hours.

2. The hydrolyzed solution was neutralized with HCl and the water was evaporated by lyophilization.

3. The product was dissolved in 50% methanol and 1N HCl. The mixture was incubated at 70° C. for 5 hours.

4. The solution was neutralized with NaOH.

5. The hydrolyzed product was extracted with ethylacetate, which was subsequently removed by evaporation.

6. Further purification of the hydrolyzed product (E4A) was archived with FPLC chromatography in a C18 column equilibrated with 70% acetonitrile/TFA at the flow rate of 1 ml/min.

Experiment 5: Esterification of E4A With Tigloyl Chloride

1. 50 mg of E4A in 1 ml pyridine, stir gently in a 50 ml tube. Esterification was carried out at 25° C. by adding 200 ul Tigloyl chloride.

2. Stir for 1 minute; then immediately add 5 ml of 2N HCl.

3. Stir for 1 hour and sit at room-Temp overnight.

4. Extract the esterification products with 10 ml ethylacetate.

5. Evaporate the ethylacetate.

6. Dissolve the sample with 1 ml DMSO.

7. Fractionate the reaction products with HPLC.

8. Collect samples.

Experiment 6: Isolation of E4A-Tig Active Compounds With HPLC

1. Column: ZORBAX ODS 9.4x250 mm, 5 um

2. Solvents: A: 45% AN/TFA; B: 100% AN/TFA

3. Chromatography conditions: a) Elution: Solvent A to B in 80 min; then with solvent B for 40 min; b) flow rate: 1 ml/mim. c) Monitor OD: at 207 nm;

Experiment 7: MTT Experiment

Cells. HTB-9 (bladder), HeLa-S3 (cervix), DU145 (prostate), H460 (lung), MCF-7 (breast), K562 (leukemia), HCT116 (colon), HepG2 (liver), U2OS (bone), T98G (brain), SK-MEL-5 (Skin) and OVCAR 3, ES2 (ovary), Pancreas (Capan), Mouth (KB), Kidney(A498).

MTT Assay. The procedure for MTT assay followed the method described by Carmichael et al. (1987) with modifications. The cells were seeded into a 96-well plate at for 24 hours before drug-treatment. The cells were then exposed to the drugs for 48, 72, or 96 hours. After the drug-treatment, MTT (0.5 mg/mL) was added to cultures and incubated for an hour. The formazan (product of the reduction of tetrazolium by viable cells) formed and was dissolved with DMSO and the O.D. at 490 nm, and was measured by an ELISA reader. The MTT level of the cells before drug-treatment was also measured (T0). The % cell-growth (%G) is calculated as: %G = (TD-T0 / TC-T0) x 100(1), where TC or TD represents O.D. readings of control or drug-treated cells. When T0 > TD, then the cytotoxicity (LC) expressed as % of the control is calculated as: %LC = (TD-T0 / T0) × 100(2).

MTT Assay is performed to intermediate and final products from experiments.

Experiment 8: Chemical Synthesis, Isolation and Characterization of E4A-Tig-R Chemical Synthesis of E4A-Tig-R: 1. Preparation of E4A; 2. Esterification of E4A with Tigloyl Chloride; 3. Isolation of E4A-Tig-R with HPLC

Cytotoxic activity determination: 1. MTT assay

Chemical structure determination: 1. NMR analysis; 2. Mass Spectrum analysis Compound E4A-Tig-R: 24,28-O-Tigloyl-3β,16α, 21β, 22a, 24β, 28-hexahydroxyolean-12-ene

Experiment 9: Chemical Synthesis, Isolation and Characterization of E4A-Tig-N Chemical Synthesis of E4A-Tig-N: 1. Preparation of E4A; 2. Esterification of E4A with Tigloyl Chloride; 3. Isolation of E4A-Tig-N with HPLC

Cytotoxic activity determination: 1. MTT assay

Chemical structure determination: 1. NMR analysis; 2. Mass Spectrum analysis

Experiment 10: Chemical Synthesis, Isolation and Characterization of E4A-Tig-Q Chemical Synthesis of E4A-Tig-Q: 1. Preparation of E4A; 2. Esterification of E4A with Tigloyl Chloride; 3. Isolation of E4A-Tig-Q with HPLC

Cytotoxic activity determination: 1. MTT assay

Chemical structure determination: 1. NMR analysis; 2. Mass Spectrum analysis

Experiment 11: Chemical Synthesis, Isolation and Characterization of E4A-Tig-V Chemical Synthesis of E4A-Tig-V: 1. Preparation of E4A; 2. Esterification of E4A with Tigloyl Chloride; 3. Isolation of E4A-Tig-V with HPLC

Cytotoxic activity determination: 1. MTT assay

Chemical structure determination: 1. NMR analysis; 2. Mass Spectrum analysis

Experiment 12: Chemical Synthesis, Isolation and Characterization of E4A-Tig-T Chemical Synthesis of E4A-Tig-T: 1. Preparation of E4A; 2. Esterification of E4A with Tigloyl Chloride; 3. Isolation of E4A-Tig-T with HPLC

Cytotoxic activity determination: 1. MTT assay

Chemical structure determination: 1. NMR analysis; 2. Mass Spectrum analysis

Experiment 13: Chemical Synthesis, Isolation and Characterization of E4A-Tig-U Chemical Synthesis of E4A-Tig-U: 1. Preparation of E4A; 2. Esterification of E4A with Tigloyl Chloride; 3. Isolation of E4A-Tig-U with HPLC

Cytotoxic activity determination: 1. MTT assay

Chemical structure determination: 1. NMR analysis; 2. Mass Spectrum analysis

Experiment 14: Chemical Synthesis, Isolation and Characterization of E4A-Tig-S Chemical Synthesis of E4A-Tig-S: 1. Preparation of E4A; 2. Esterification of E4A with Tigloyl Chloride; 3. Isolation of E4A-Tig-S with HPLC

Cytotoxic activity determination: 1. MTT assay

Chemical structure determination: 1. NMR analysis; 2. Mass Spectrum analysis

Experiment 15: Using Method in Experiment 3, Esterification of E4A with Acetyl, Angeloyl, Tigloyl, Senecioyl, Crotonoyl, Cinnamoyl, Pentenoyl gave the following Compounds

embedded image - Compound E4A-Ang-R:

embedded image - Compound E4A-Ang-V:

embedded image - Compound E4A-Ang-Q:

embedded image - Compound E4A-Ang-N:

embedded image - Compound E4A-Ang-T:

embedded image - Compound E4A-Ang-U:

embedded image - Compound E4A-Ang-S:

embedded image - Compound E4A-Sen-R:

embedded image - Compound E4A-Sen-V:

embedded image - Compound E4A-Sen-N:

embedded image - Compound E4A-Sen-Q:

embedded image - Compound E4A-Sen-S:

embedded image - Compound E4A-Sen-T:

embedded image - Compound E4A-Sen-U:

embedded image - Compound E4A-Cro-R:

embedded image - Compound E4A-Cro-V:

embedded image - Compound E4A- Cro-N:

embedded image - Compound E4A- Cro-Q:

embedded image - Compound E4A- Cro-S:

embedded image - Compound E4A- Cro-T:

embedded image - Compound E4A- Cr:

embedded image - Compound E4A-Acy-R:

embedded image - Compound E4A- Acy-V:

embedded image - Compound E4A- Acy-N:

embedded image - Compound E4A- Acy-Q:

embedded image - Compound E4A- Acy-S:

embedded image - Compound E4A- Acy-T:

embedded image - Compound E4A- Acy-U:

embedded image - Compound E4A-Pen-R:

embedded image - Compound E4A-Pen-V:

embedded image - Compound E4A-Pen-N:

embedded image - Compound E4A-Pen-Q:

embedded image - Compound E4A-Pen-S:

embedded image - Compound E4A-Pen-T:

embedded image - Compound E4A-Pen-U:

embedded image - Compound E4A-Pen-R:

embedded image - Compound E4A-Pen-V:

embedded image - Compound E4A-Cin-N:

embedded image - Compound E4A-Cin-Q:

embedded image - Compound E4A-Cin-S:

embedded image - Compound E4A-Cin-T:

embedded image - Compound E4A-Cin-U:

Experiment 16: Esterification of E4A-Tig-N with Senecioyl chloride Chemical Synthesis of E4A-Tig-Sen-1: 1. Esterification of E4A-Tig-N with Senecioyl Chloride; 3. Isolation of E4A-Tig-Sen-1 with HPLC

Cytotoxic activity determination: 1. MTT assay

Chemical structure determination: 1. NMR analysis; 2. Mass Spectrum analysis

Experiment 17: Esterification of E4A-Tig-N with Angeloyl Chloride, Acetyl chloride, Crotonoyl Chloride, 3,3-Dimethylacryloyl Chloride, Senecioyl Chloride, Cinnamoyl Chloride, Pentenoyl chloride, Hexanoyl Cloride, Benzoyl Chloride or Ethylbutyryl Chloride; Isolation with HPLC; Cytotoxic activity determination; Chemical Structure Determination with the Method of Experiment 8, gave the Following Compounds

embedded image

Experiment 18: Inhibition of Cell Adhesion

Methods and Results. ES2 or Hey8A cells were plated in T25 flasks with medium containing 5 ug/ml of compounds selected from structure (2A) including E4A-Tig-R, E4A-Tig-V, E4A-Tig-S, E4A-Tig-N, E4A-Tig-Q, E4A-Tig-T. Cultures were incubated for 5 hours. Attached cells were removed from flasks by trypsinization and the numbers/amounts were counted. Compared to no drug controls, 80 ± 4% of ES2 cells and 60 ± 4% of Hey8A cells were found attached to flasks under this condition. At 5 ug/ml of above compounds, over 90% of unattached cells are alive as determined by the trypan Blue exclusion assay and by their ability to re-attach to flasks when plating in medium without tested compounds. However, with 10 ug/ml tested compounds, less than 40% of cells attached to flasks and many of them are dead cells. This experiment shows that tested compounds inhibit cells adhesion process.

Experiment 19: Fibronectin Secretion Experiment

Western blot is applied in this invention as a method to detect the specific proteins in treated and untreated cells with compounds in this invention, wherein the cells are bladder, cervix, prostate, lung, breast, leukemia, colon, liver, bone, brain, Skin, ovary, Pancreas(Capan), Mouth(KB), Kidney.

Cells: targeted cells were grown in RPMI 1640 medium. 1.5 million cells were seeded in a T25 flask and grown for 24 hours before drug-treatment.

Drug-treatment: Cells cultures were replaced with fresh RPMI medium containing either 2.5 ul of DMSO (as control) [D]; or 10, 20, 30, 40, 80 ug/ml of tested compounds.

After 24 hours, aliquot of culture medium was taken out for Fibronectin determination (Western blot method).

Cell viability at 24 hours was determined by MTT assay. Cultures were replaced with RPMI medium (5 ml) with MTT and incubated for an hour. The formation of formazan was dissolved in 10 ml of DMSO and OD at 570 nm was measured (MTT units). Western Blot: Spent culture medium was mixed with SDS sample buffer, boiled for 3 minutes before loading to SDS gel. Samples were applied to a 6-10% SDS gel and electrophoresis was conducted with 100 volts for 2 hours. Protein was transferred to a nitrocellulose membrane electrophoretically. The nitrocellulose blot was incubated with the first antibody and second antibody (AP conjugated, Promega S3721). The immuno-bands were developed with BCIP/NBT color development system.

Determination of Western blot band intensity: The band-images of Western blot were captured with a digital camera and the intensity of bands was determined using “Image J” software.

Results show that compounds of E4A-Tig-R, E4A-Tig-V, E4A-Tig-S, E4A-Tig-N, E4A-Tig-Q, E4A-Tig-T inhibit fibronectin secretion from 20-40%.in bladder, cervix, prostate, lung, breast, leukemia, colon, liver, bone, brain, Skin, ovary, Pancreas(Capan), Mouth(KB), Kidney.

Experiment 20: Esterification of E4A With Propionyl Chloride

Methods: 50 mg of E4A in 1 ml pyridine, stir gently in a 50 ml tube. Esterification was carried out at 25C by adding 200 ul Propionyl chloride, and immediately withdrawn 200 ul from the mixture and added to 1 ml of 2N HCl. (ASAP sample). At 1, 2, 5, 10 and 60 minutes afterward; 200 ul of reaction mixture was similarly withdrawn and add to 1 ml of 2N HCl. Mixtures were sit at room-Temp overnight. Extract the esterification products with 2 ml ethylacetate. Evaporate the ethylacetate. Dissolve the sample with DMSO (final concentration of 40 mg/ml). Fractionate the reaction products with HPLC (C18 column, 1 ml/min).

HPLC condition: Column: C18 (9.4 × 250 mm, 5 um); Solvents: 80% Acetonitrile -0.005% TFA; Gradient: isocratic; Flowrate: 1 ml / min; O.D.: 207 nm, AT=1024; Chart speed: 0.1 cm/min; Run time: 120 min; MTT assay (Cytotoxicity determination) condition: Cells: ES2 (ovarian cancer). Cell density: plate1 0 K cells per well over night before addition of drug. Drug incubation time: 2 days.

Experiment 21: Esterification of E4A With Isobutyryl Chloride

Methods: 52 mg of E4A in 1 ml pyridine, stir gently in a 50 ml tube. Esterification was carried out at 25° C. by adding 200 ul of isobutyryl chloride. 2 minute later, 4 ml 2N HCl was added to the reaction mixture. Mixtures were kept at room-Temp overnight. Extract the esterification products with 5 ml ethyl acetate. Evaporate the ethyl acetate. Dissolve the sample with DMSO (final concentration of 40 mg/ml). Fractionate the reaction products with HPLC (C18 column).

MTT assay (Cytotoxicity determination) condition: Cells: ES2 (ovarian cancer); Cell density: plate1 0 K cells per well over night before addition of drug; Drug incubation time: 2 days.

Experiment 22: Esterification of E4A with 3,3-dimethylacryloly chloride from different times of esterification reaction. Reaction products obtained from each time of reaction (5 sec, 1 min, 2 min, 5 min, and 10 min) were fractionated by HPLC. The profile is plotted according to HPLC elution time and optical density of fractions. Reaction was performed at Room temperature and 0° C. Fractionate the reaction products with HPLC (C18 column). Cytotoxic activity is determined with MTT. Chemical structure determined with NMR.

Experiment 23: Esterification of E4A with Pentenoyl chloride-from different times of esterification reaction. Reaction products obtained from each time of reaction (5 sec, 1 min, 2 min, 5 min, and 10 min) were fractionated by HPLC. The profile is plotted according to HPLC elution time and optical density of fractions. Reaction was performed at Room temperature. Fractionate the reaction products with HPLC (C18 column). Cytotoxic activity is determined with MTT. Chemical structure determined with NMR.

Experiment 24: Esterification of E4A with Hexanoly chloride from different times of esterification reaction. Reaction products obtained from each time of reaction (5 sec, 1 min, 2 min, 5 min, and 10 min) were fractionated by HPLC. The profile is plotted according to HPLC elution time and optical density of fractions. Reaction was performed at 0C. Fractionate the reaction products with HPLC (C18 column). Cytotoxic activity is determined with MTT. Chemical structure determined with NMR.

Experiment 25: Esterification of E4A with Acetyl chloride (H) from different times of esterification reaction. Reaction products obtained from each time of reaction (1 min, 2 min, 5 min and 10 min) were fractionated by HPLC. The profile is plotted according to HPLC elution time and optical density of fractions. Reaction was performed at Room temperature. Fractionate the reaction products with HPLC (C18 column). Cytotoxic activity is determined with MTT. Chemical structure determined with NMR.

Experiment 26: Esterification products of E4A with Crotonoyl chloride (l) from different times of esterification reaction. Reaction products obtained from each time of reaction (5 sec, 1 min, 2 min, 5 min and 10 min) were fractionated by HPLC. The profile is plotted according to HPLC elution time and optical density of fractions. Reaction was performed at Room temperature. Fractionate the reaction products with HPLC (C18 column). Cytotoxic activity is determined with MTT. Chemical structure determined with NMR.

Experiment 27: Esterification products of E4A with Cinnamoyl chloride (J) from different times of esterification reaction. Reaction products obtained from each time of reaction (1 min, 1 hour, 2 hours, 18 hours, 18 hours(heat)) were fractionated by HPLC. The profile is plotted according to HPLC elution time and optical density of fractions. Reaction was performed at Room temperature and 75C. Fractionate the reaction products with HPLC (C18 column). Cytotoxic activity is determined with MTT. Chemical structure determined with NMR.

Experiment 28: Esterification products of E4A with pentenoyl, hexanoyl, benzoyl, ethylbutyryl, propionyl, 2-propenoyl, isobutyryl, butyryl, (2E)-2-pentenoyl, 4-Pentenoyl, 5-hexenoyl, heptanoyl, octanoyl, nonanoyl, decanoyl, Lauroyl, myristoyl, from different times of esterification reaction. Reaction products obtained from each time of reaction were fractionated by HPLC. The profile is plotted according to HPLC elution time and optical density of fractions. Fractionate the reaction products with HPLC (C18 column). Cytotoxic activity is determined with MTT. Chemical structure determined with NMR.

Experiment 29: Esterification products of E4A with propanoyl, propenoyl, butanoyl, butenoyl, pentanoyl, hexenoyl, heptanoyl, heptenoyl, octanoyl, octenoyl, nonanoyl, nonenoyl, decanoyl, decenoyl, propionyl, 2-propenoyl, 2-butenoyl, Isobutyryl, 2-methylpropanoyl, 2-ethylbutyryl, ethylbutanoyl, 2-ethylbutanoyl, butyryl, (E)-2,3-Dimethylacryloyl, (E)-2-Methylcrotonoyl, 3-cis-Methyl-methacryloyl, 3-Methyl-2-butenoyl, 3-Methylcrotonoyl, 4-Pentenoyl, (2E)-2-pentenoyl, Caproyl, 5-Hexenoyl, Capryloyl, Lauroyl, Dodecanoyl, Myristoyl, Tetradecanoyl, Oleoyl from different times of esterification reaction. Reaction products obtained from each time of reaction were fractionated by HPLC. The profile is plotted according to HPLC elution time and optical density of fractions. Fractionate the reaction products with HPLC (C18 column). Cytotoxic activity is determined with MTT. Chemical structure determined with NMR.

Experiment 30 A Comparison of Non-cancerous With Cancer Cells

Method: A. Cells

-WI38 is a Normal Lung fibroblast. Cells were grown in MEM medium supplemented with 10% FCS, antibiotics and glutamine. 20 K cells were seeded per well (96-welled plate) for one day before drug-treatment
-ES2 is an Ovary Clear cells carcinoma. Cells were grown in RPMI-1640 medium supplemented with 10% FCS, antibiotics and glutamine. 10 K cells were seeded per well (96-welled plate) for one day before drug-treatment.

B. Drug treatment:

-Drug: Tig-S. stored as 1000x stock solution in DMSO.
-Drug concentration used: from 0.15 - 20 ug/ml.
-Drug-treatment was carried out for 2 days (ES2) or 6 days (WI38). For the 6 days incubation, cells were fed with fresh medium (with drug) on day 3 and 5.

C. At the end of the drug-treatment, cytotoxic test was performed with MTT assay.

Conclusion:

For comparison between cancer cells (ES2) and non-cancerous cells (WI38), it is found that:

1. the IC₅₀ for ES2 cells (0.3 ug/ml) is lower than those of WI38 cells (1.5 ug/ml), and
2. The IC₁₀₀ for ES2 cells (0.15-0.3 ug/ml) is lower than those of WI38 cells (10 ug/ml)
3. With 10 ug/ml of Tig-S, about 90% of ES2 cells died while only 10% of WI38 cells died after the drug-treatment.

Based on these studies, it is concluded that the fast-growing tumor cells (ES2) are more sensitive to Tig-S than the slow growing normal cells (WI38).

Experiment 31: Inhibition of Cell Cycling

1. Method: Cells: Human leukemia cells K562 cultured in RPMI1640 medium.

2. Drug: compound from present application or Tig-S (1000x stock solution in DMSO).

3. Start cells concentration: 500000 /ml.

4. Cells were cultured with drug (0 - 20 ug/ml) for total of three days.

5. Cells were harvested by centrifugation (136xg, 6 minutes); fixed with 70% ethanol and kept in -20° C. before staining.

6. Staining: Fixed cells were stained with Propidium iodide/RNase A/0.1% Triton X-100 in PBS.

7. Flow Cytometry Analysis was performed in Baylor Core Facility with a LSRII instrument.

8. Analysis: Single cell was gated and cell count - FL2-Area histogram were plotted.

9. Cell distribution in different cell-cycle phases (G0/G1, S, G2/M) was analyzed.

Results: Cells with no drug, or with 0.15 ug/ml and 0.3 ug/ml of Tig-S, have a similar (same) cell distribution in the G0/G1, S and G2/M phases of cell- cycle. With higher Tig-S concentrations, starting from 0.6 ug/ml, the cells in G2/M phase decrease. The decrease of G2/M cells correlated with higher drug concentrations (up to 20 ug/ml). These results indicate that drug-treated cells were arrested in the S-phase and unable to enter into the G2/M phase of the cell cycle.

Conclusion: The drug-effect of Tig-S on human leukemia K562 cells is arresting cells in the S-phase and blocking their entering into the G2/M phase of cell cycle. The Compound Tig-S block the DNA synthesis of cancer cell.

Experiment 32: Inhibition of H460 Cells Growth With Tig-S

Methods:

A. Cells
- -- H460 cells are derived from a Human Lung large cell carcinoma. Cells were grown in RPMI 1640 medium supplemented with 10% FCS, antibiotics and glutamine. 5000 cells were seeded in a well (96-welled plate) for one day before drug-treatment
B. Drug treatment:
- -- Drug: Tig-S (stored as a 1000x stock solution in DMSO) was used.
- -- Drug concentration used: from 0.15 - 20 ug/ml.
- -- Drug-treatment was carried out for 1, 2 and 4 days. For the 4 days incubation, cells were fed with fresh medium (with drug) on day 2.
C. At the end of the drug-treatment, cytotoxic test was performed with MTT assay. Results:
- 1. Tig-S inhibits H460 cells’ growth with the IC50 of drug 3 ug/ml.
- 2. Minimum cells growth inhibition was observed beyond drug 5 ug/ml.
- 3. No dead cell was found at drug concentration in 20 ug/ml.

The results indicate that Tig-S inhibits the H460 cell’s growth, but is not killing cells at high concentrations. Therefore, Tig-S is an effective drug for inhibition of cancer growth but has low toxicity.

Experiment 33: Inhibition of Human Leukemia K562 Cells by Tig-S

Method:

1. Cells: Human leukemia cells K562 in RPMI1640 medium.
2. Drug: Tig-S (1000x stock solution in DMSO).
3. Start cells concentration: 50×10 K per ml (500000/ml).
4. Cells were cultured with or without drug for total of four days.
5. Cell number is doubled after 2 days of incubation. Fresh medium (equal volume, with or without drug) was then added to culture.
6. Cells were counted every day.

Conclusion:

Tig-S inhibits Leukemia K562 cells growth with IC50 about 0.6 ug/ml.

No grow (IC100) was observed beginning on day 2 at 2.5 ug/ml or higher.

Experiment 34: Apoptosis in K562 Cells Induced by Tig-S

1. Cells: Human leukemia cells K562 cultured in RPMI1640 medium.

2. Drug: Tig-S (1000x stock solution in DMSO).

3. Start cells concentration: 500000/ml.

4. Cells were cultured with drug (0 - 20 ug/ml) for Two days.

5. Collect cells from culture (1 - 4 million) by centrifugation (136xg for 5 min. remove supernatant).

6. Wash cells with 1 ml of cold PBS, collect cells by centrifugation (136xg, 5 min, remove supernatant).

7. Re-suspend cells in 200 ul Binding buffer (10 mM HEPES, 140 mM NaCl, 2.5 mM CaCl2, pH 7.4).

8. Stained cells by adding 10 ul Annexin V (Alexa Fluor 488, Invitrogen cat# A13201), and 2 ul Propidium iodide (1 mg/ml in water) to cell suspension. Mix.

9. Sit at Room temperature for 15 min.

10. Wash cells with 1 ml binding buffer. Collect cells by centrifugation.

11. Re-suspend cells with in 1 ml of binding buffer for analysis by flow cytometry.

12. Flow Cytometry analysis was performed in Baylor Core Facility with a LSRII instrument.

13. Analysis: Control cells stained with PI or Annexin V-488 (Annexin V-FITC) alone were served as references. About 50000 cells were analyzed for each sample.

14.Cell distribution in following groups: live, early apoptosis, late-apoptosis, total apoptosis and dead cells were determined.

15. The percentage of cells in these groups is presented in the FIG. 19.

Results:

The background apoptosis level of K562 cells (no-drug control) is about 10-15%. After drug-treatment with Tig-S, the apoptotic cells population increased (from 15% to 27%) with increased drug concentration (from 2.5 ug/ml to 20 ug/ml).

Similarly, the dead cell population was also increase with the drug concentration.

Conclusion:

Tig-S induces cell-death by the apoptosis mechanism (not necrosis mechanism).

Experiment 35: The Haemolytic Assay

Experiment 36: Animals Experiments
Methods

Athymic Nu/Nu mice are divided into two groups (A, and B).

On day 0, mice of group A and B were transplanted intra-peritoneally (i.p.) with ES2 (human ovarian cancer) cells.

-On day 1 to 5, mice from A group received daily administration of 127 solvent by i.p. route.

On days 1 to 5, mice from B group received daily drug administration (tested drug Tig-S in 127 solvent) by i.p. route at dose of 100 mg/kg, twice daily.

Result in FIG. 22
Experiment 37: Animals Experiments
Methods

Athymic Nu/Nu mice are divided into two groups (A, and B).

On day 0, mice of group A and B were transplanted intra-peritoneally (i.p.) with ES2 (human ovarian cancer) cells.

On day 1 to 5, mice from A group received daily administration of 127 solvent by i.p. route.

On days 1 to 5, mice from B group received daily drug administration (tested drug Tig-R in 127 solvent) by i.p. route at dose of 100 mg/kg, twice daily.

-Result in FIG. 23

Experiment 38: Animals Experiments
Methods

Athymic Nu/Nu mice are divided into two groups (A, and B).

On day 0, mice of group A and B were transplanted intra-peritoneally (i.p.) with ES2 (human ovarian cancer) cells.

On day 1 to 5, mice from A group received daily administration of 127 solvent by i.p. route.

On days 1 to 10, mice from B group received daily drug administration (tested drug Tig-V in 127 solvent) by i.p. route at dose of 50 mg/kg, twice daily.

Result in FIG. 24(U.S. Serial No. 14/313080)

Experiment 39: Inhibition of Capan Cells Growth by Tig-S

A. Cells: Capan cells are derived from Human pancreas carcinoma (pancreas). Cells were grown in RPMI 1640 medium supplemented with 10% FCS, antibiotics and glutamine. 10000 cells were seeded in a well (96-welled plate) for one day before drug-treatment

B. Drug treatment:
- -- Drug: Tig-S (stored as a 1000x stock solution in DMSO) was used.
- -- Drug concentration used: from 0.15 - 20 ug/ml.
- -- Drug-treatment was carried out for 3 days.

C. At the end of the drug-treatment, cytotoxic test was performed with MTT assay. The percentage of drug-treated cells’ growth as compared to those of the no drug control is determined. See FIG. 37.

Experiment 40: Using method in Experiment 3, esterification of E4A2Y with acetyl, angeloyl, tigloyl, senecioyl, Crotonoyl, Cinnamoyl, Pentenoyl, 4-(dimethylamino)-2-methylbut-2-enoyl, and 4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl gave the following compounds:

embedded image

wherein:

1) R1, R2, R3, R5, R8 are OH; R17, R18 are O-acetyl;
2) R1, R2, R3, R5, R8 are OH; R17, R18 are O-angeloyl
3) R1, R2, R3, R5, R8 are OH; R17, R18 are O-tigloyl
4) R1, R2, R3, R5, R8 are OH; R17, R18 are O-senecioyl
5) R1, R2, R3, R5, R8 are OH; R17, R18 are O-Crotonoyl
6) R18, R2, R3, R5, R8 are OH; R17, R18 are O-Cinnamoyl
7) R18, R2, R3, R5, R8 are OH; R17, R18 are O-Pentenoyl
8) R18, R2, R3, R5, R8 are OH; R17, R18 are O-4-(dimethylamino)-2-methylbut-2-enoyl
9) R18, R2, R3, R5, R8 are OH; R17, R18 are O-4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl
10) R18, R2, R3, R5, R8 are OH; R17, R1 are O-acetyl;
11) R18, R2, R3, R5, R8 are OH; R17, R1 are O-angeloyl
12) R18, R2, R3, R5, R8 are OH; R17, R1 are O-tigloyl
13) R18, R2, R3, R5, R8 are OH; R17, R1 are O-senecioyl
14) R18, R2, R3, R5, R8 are OH; R17, R1 are O-Crotonoyl
15) R18, R2, R3, R5, R8 are OH; R17, R1 are O-Cinnamoyl
16) R18, R2, R3, R5, R8 are OH; R17, R1 are O-Pentenoyl
17) R18, R2, R3, R5, R8 are OH; R17, R1 are O-4-(dimethylamino)-2-methylbut-2-enoyl
18) R18, R2, R3, R5, R8 are OH; R17, R1 are O-4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl
19) R2, R3, R5, R8 are OH; R1, R17, R18 are O-acetyl;
20) R2, R3, R5, R8 are OH; R1, R17, R18 are O-angeloyl
21) R2, R3, R5, R8 are OH; R1, R17, R18 are O-tigloyl
22) R2, R3, R5, R8 are OH; R1, R17, R18 are O-senecioyl
23) R2, R3, R5, R8 are OH; R1, R17, R18 are O-Crotonoyl
24) R2, R3, R5, R8 are OH; R1, R17, R18 are O-Cinnamoyl
25) R2, R3, R5, R8 are OH; R1, R17, R18 are O-Pentenoyl
26) R2, R3, R5, R8 are OH; R1, R17, R18 are O-4-(dimethylamino)-2-methylbut-2-enoyl
27) R2, R3, R5, R8 are OH; R1, R17, R18 are O-4-[(2-methoxyethyl)amino]-2-methyl-4oxobut-2-enoyl

Experiment 41: Inhibition of Eye Cancer Cells (retinoblastoma)

Methods

Cells: Human Eye cancer cells Y79 (retinoblastoma) are cultured in RPMI1640 medium supplemented with 20% FCS, glutamine and antibiotics.

Starting cells concentration: 400 K per ml in a 24-well plate.

Drug: Tig-S and Tig-R (100× stock solution in 10%DMSO-90% RPMI-1640).

Drug-treatment: Cells were cultured with or without drug (DMSO control) for 2 days. After Drug-treatment, the quantity of live cells is determined with MTT assay*. With this method, the percentage of cell growth with drugs was determined by comparing to those of no-drug (DMSO) control.

MTT assay. To measure the amount of live cells, MTT (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) (0.5 mg/ml) was added to cultures and incubated for an hour. Cells were then collected by centrifugation. The formation of formazan (proportion to live cells activity) in the cells was dissolved with DMSO and the O.D. at 490 nm was measured by an ELISA reader. The MTT level of cells before drug-treatment (T0) and after the drug-treatment (TD) was measured. The % cell-growth (%G) is calculated as: %G = (TD-T0) / (TC-T0) × 100 where TC or TD represent O.D. readings of control or drug-treated cells.

When T0 > TD, then the negative growth (cytotoxicity) is expressed as - % of the control and is calculated as: - %G = (TD-T0) / T0) × 100.

Results: Both Tig-S and Tig-R inhibit the Y79 cell’s growth.

The IC50 for Tig-S is 0.1 ug/ml; the IC50 for Tig-R is 7.5 ug/ml.

Experiment 42: Induction of DR5 in ES2 Cells by Tig-R Treatment

1). Cells and drug-treatment: ES2 cells (Ovarian cancer cells) were cultured in RPMI1640 medium with 10% FBS, antibiotics (Pen Strep) and L-Glutamine in a 37° C., 5% CO2 incubator. Cells in log-growth phase were treated with Tig-R (10 ug/ml), or DMSO (as control) for 2 days.

2). Total cell protein extraction: Drug-treated cells were collected, pelleted with centrifugation. The cell pellet was extracted with SDS-Sample buffer (containing PMSF and protease inhibitors cocktail). About 0.1 gram cell pellet was extracted in 1 ml of extraction buffer. The cell extract was then incubated in boiling water bath for 6 minutes with occasionally vortexing. The protein extract was immediately frozen in -80° C. before used.

3). SDS gel electrophoresis: 10-20 ug of protein sample was loaded into each lane of a 10% SDS gel. Electrophoresis was conducted with 100 volt constant for 2 hours.

4). Western Blot: Separated proteins from SDS gel were transferred to a nitrocellulose membrane, which was blocked with 5% non-fat dry milk. The membrane was then exposed to 1st antibody (anti-TNFRSF10B) over night, then washed. The membrane was then incubated in 2nd antibody (anti-mouse-AP conjugated) for an hour, then washed. The immuno-bands were developed with BCIP/NBT substrate.

5. Another SDS gel with same protein loading was stained with Coomassie Brilliant Blue (CBB) for protein staining.

Results:

Treatment of ES2 cells (ovarian cancer) with Tig-R increase DR5 at least 2 folds. Results indicate Tig-R induce DR5 in ES2 cells and cause apoptosis.

Experiment 43: Combined Effect of anti-DR5 and Tig-R in HepG2 Cells (human Liver carcinoma)

Cells Culture: HepG2 cells (liver cancer cells) were cultured in DMEM medium supplemented with 10% FBS, antibiotics (Pen Strep) and L-Glutamine. Cells were incubated in a 37° C., 5% CO2 incubator.

Antibody: Antibody against Human TRAIL R2/TNFRSF 10B, (from R&D Systems cat# MAB631-100).

Drug: Tig-R (5 and 7.5 ug/ml in culture medium).

Treatment. Cells in log-growth phase were used. Cells were plated in 96-welled plate with 10 K/well over night before treatment.

Antibodies are serially diluted with medium (started from 1 ug/ml) before adding to cell culture and incubated for 2 days.

For combination studies, Tig-R (5 ug/ml or 7.5 ug/ml) were added together with antibody. After a treatment for 2 days, the quantity of live cells is determined with MTT assay*. The percentage of cell growth with drugs was determined by comparing to those of no-drug (DMSO) control.

MTT assay. Cell growth activity was determined by MTT assay (Carmichael et al.,). Cells were seeded in a 96-wells plate and were exposed to drug for 2 or 3 days. After the drug-treatment, MTT (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) (0.5 mg/ml) was added to cultures for an hour. The formation of formazan was dissolved with DMSO and the O.D. at 490 nm was measured by an ELISA reader. The MTT level of cells before drug-treatment was also measured (T0). The % cell-growth (%G) is calculated as: %G = (TD-T0 / TC-T0) × 100 (1) where TC or TD represent O.D. readings of control or drug-treated cells.

When T0 > TD, then the negative growth (cytotoxicity) is expressed as - % of the control and is calculated as: - %G = (TD-T0 / T0) × 100.

Results: RD anti-DR5 antibody inhibits HepG2 cell’s growth in a dose dependent manner started from 0.1 ug/ml (blue curve). Tig-R alone (5 ug/ml) inhibits about 15-20% HepG2 cells’ growth (at 0 ug/ml antibody concentration). When combining with a fixed amount of Tig-R (5 ug/ml) to various amount of antibody in cultures, we found an increase the inhibition of cell’s growth was observed. The degree of inhibition is synergistic.

Experiment 44: Agonist activity of BD anti-DR5 antibody with Tig-R in Jurkat cells Jurkat cells (Human Acute T-cell leukemia) were cultured in RPMI1640 medium with 10% FBS, antibiotics (Pen Strep) and L-Glutamine in a 37C, 5% CO2 incubator. Cells in log-growth phase (about 0.5 million cells per ml) were treated with antibody (AB) or/and Tig-R. The antibody is Human TRAIL R2/TNFRSF 10B Antibody from R&D (MAB631-100). Treatment time is 2 days. Inhibition of cell growth after treatment was determined by MTT assay.

Conditions for combination studies: Tig-R (2.5 ug/ml) was added together with AB (2-8 ng/ml). Cells

Results: Antibody alone with concentrations of 2, 4 or 8 ng/ml, inhibit 38, 54 or 87%, respectively, of cell growth. Tig-R (2.5 ug/ml) inhibits 8% cell growth. Adding Tig-R (2.5 ug/ml) to 2, 4 or 8 ng/ml antibody, increase its inhibition to 60, 79 or 115%, respectively. With Tig-R, about 20% increase of inhibition was observed. The results indicate that there is a combined synergistic inhibition effect to Jurkat cells by BD Antibody and Tig-R.

Experiment 45: Agonist Activity of BD anti-DR5 Antibody with Tig-R in Capan Cells (Pancreas Carcinoma)
Cells Culture:

Capan cells (pancreas carcinoma) were cultured in RPMI1640 medium supplemented with 10% FBS, antibiotics (Pen Strep) and L-Glutamine. Cells were incubated in a 37° C., 5% CO2 incubator.

Antibody: Antibody against Human TRAIL R2/TNFRSF 10B, (from R&D Systems cat# MAB631-100).

Drug: Tig-R

Treatment. Cells in log-growth phase were used. Cells were plated in 96-welled plate with 10 K/well 1-2 days before treatment.

Antibody are serially diluted with medium before adding to culture and incubated for 2 days.

After a treatment for 2 days, the quantity of live cells is then determined with MTT assay*. The percentage of cell growth with drugs was determined by comparing to the no-drug (DMSO) control.

Results:

1. Capan (pancreas cancer) cells were inhibited by Anti-DR5 alone with the IC50 about 0.17 ug/ml.
2. With 2.5 ug/ml of Tig-R, the IC50 of combined effect decrease to about 0.06 ug/ml (a 65% decrease). With 5 ug/ml of Tig-R, the IC50 of combined effect decrease to about 0.01 ug/ml (a decrease of 94%).
3. These results indicate that addition of Tig-R to anti-DR5 antibody significantly increase the antibody inhibition effect.

The following tables summarized the analysis of synergistic effect of the combined agents. The tables list the inhibition effect of antibody (Ab), Tig-R (R) individually and their combination. As shown in the table (from two experiments), the combined inhibition effect from is higher than the theoretical combined effects of each agent (Ab or R) alone, these results indicate that the combined effect is synergistic.

Treatment with
% Inhibition*
Inhibition from Experimental Compare to Theoretical

Experimental
Theoretical

R
Ab (0.12 ug/ml)
40

2.5 ug/ml
R (2.5 ug/ml)
20

Ab + R
78
60
Increase

Ab (0.25 ug/ml)
66

R (2.5 ug/ml)
20

Ab + R
94
86
Increase

R
Ab (0.062 ug/ml)
33

5 ug/ml
R (5 ug/ml)
29

Ab + R
90
62
Increase

Ab (0.12 ug/ml)
40

R (5 ug/ml)
29

Ab + R
125
69
Increase

Ab (0.25 ug/ml)
66

R (5 ug/ml)
29

Ab + R
167
95
Increase

Experiment 46: Western Blot Method

A. Cells and drug-treatment: HepG2 cells (liver cancer cells), CAPAN cells, ES2 cells (Ovary cell) were cultured in DMEM medium with 10% FBS, antibiotics (Pen Strep) and L-Glutamine in a 37° C., 5% CO2 incubator. Cells in log-growth phase were treated with Tig-R (10 ug/ml), Tig-S (4 ug/ml) or DMSO (as control) for 2 days.

B. Total cell protein extraction: Drug-treated cells were collected, pelleted with centrifugation. The cell pellet was extracted with SDS-Sample buffer (containing PMSF and protease inhibitors cocktail). About 0.1 gram cell pellet was extracted in 1 ml of extraction buffer. The cell extract was then incubated in boiling water bath for 6 minutes with occasionally vortexing. The protein extract was immediately frozen in -80° C. before used.

C. SDS gel electrophoresis: 10-20 ug of protein sample was loaded into each lane of a 10% SDS gel. Electrophoresis was conducted with 100 volt constant for 2 hours.

D. Western Blot: Separated proteins from SDS gel were transferred to a nitrocellulose membrane, which was blocked with 5% non-fat dry milk. The membrane was then exposed to 1st antibody (anti-TNFRSF10B) over night, then washed. The membrane was then incubated in 2nd antibody (anti-mouse-AP conjugated) for an hour, then washed. The immuno-bands were developed with BCIP/NBT substrate.

E. Another SDS gel with same sample loading was stained with Coomassie Brilliant Blue (CBB) for protein staining.

F. Determination of Western blot band intensity: The band-images of Western blot were captured with a digital camera and the intensity of bands was determined using “Image J” software.

HepG2

DMSO
R

1
% of DMSO
15804
19291

100
122

2
% of DMSO
DMSO
R

11093
22435

100
202

3
% of DMSO
DMSO
R

21555
33645

100
156

4
% of DMSO
DMSO
R

16387
31752

100
194

Average (%)
169

SD
37

Results: In liver cancer HepG2 cells, Tig-R -treatment increase 69% of DR5 over DMSO control cells.

CAPAN

DMSO
R

1
% of DMSO
19264
29128

100
151

Results: In pancreatic cancer CAPAN cells, Tig-R -treatment increase 51% of DR5 over DMSO control cells.

ES2

DMSO
R 7.5

1
% of DMSO
19033
29902

100
157

2

DMSO
R

% of DMSO
11608
17133

100
148

3
% of DMSO
DMSO
R

16961
19157

100
113

4

DMSO
R

% of DMSO
13578
25103

100
185

5
% of DMSO
DMSO
R

8988
17941

100
200

Average

(%)
161

Results: In ovarian cancer ES2 cells, Tig-R -treatment increase 61% of DR5 over DMSO control cells.

U2OS

DMSO
R

1
8744
18269

% of DMSO
100
209

2
8348
16969

% of DMSO
100
203

Average (%)
206

Results: In bone cancer U2OS cells, Tig-R -treatment increase 106% of DR5 over DMSO control cells.

	Number	Date	Country
	62345743	Jun 2016	US
	61226043	Jul 2009	US

	Number	Date	Country
Parent	14313080	Jun 2014	US
Child	15287411		US
Parent	13259480	Sep 2011	US
Child	14020099		US

	Number	Date	Country
Parent	16792429	Feb 2020	US
Child	17975790		US
Parent	15612152	Jun 2017	US
Child	16792429		US
Parent	15287411	Oct 2016	US
Child	15612152		US
Parent	14233031	Feb 2014	US
Child	14313080		US
Parent	14020099	Sep 2013	US
Child	14313080		US
Parent	PCT/US2011/044233	Jul 2011	WO
Child	PCT/US2012/046716		WO
Parent	12856322	Aug 2010	US
Child	PCT/US2011/044233		WO
Parent	PCT/US2010/042240	Jul 2010	WO
Child	PCT/US2011/044233		WO
Parent	12541713	Aug 2009	US
Child	PCT/US2010/042240		WO

NATURAL AND SYNTHETIC COMPOUNDS FOR TREATING CANCER AND OTHER DISEASES

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