KR101873607B1 - New compounds for treating cancer and other diseases - Google Patents

Abstract

The present invention provides a method of synthesizing a novel active compound for pharmaceutical use comprising treatment of cancer wherein the cancer is selected from the group consisting of breast, leukocyte, liver, ovary, bladder, prostate, skin, bone, brain, leukemia, lung, Colon, CNS, melanoma, kidney, cervix, esophagus, testis, spleen, kidney, lymphatic, pancreas, stomach and thyroid cancers. The present invention is an anti-adherent therapy using the compound as an agent or an inhibitor of adhesion proteins and angiopoietin. This inhibits excessive attachment and inhibits cell adhesion. This regulates angiogenesis. The present compounds are also used as mediators of cell adhesion receptors, cell cycle, cell migration and inflammatory diseases.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to compounds for treating cancer and other diseases,

This application claims priority from International Patent Application No. PCT / US2010 / 0042240, filed on July 16, 2010, and US Serial No. 12 / 856,322, filed on August 13, 2010. This application also claims priority to U.S. Serial No. 12 / 541,713, filed August 14, 2009, and U.S. Serial No. 61 / 226,043, filed on July 16, 2009, as priority. This application claims priority from International Patent Application No. PCT / US09 / 34115, filed on February 13, 2009. This application claims the benefit of US Serial No. 61 / 038,277, filed March 20, 2008, US Serial No. 61 / 054,308, filed May 19, 2008, and International Patent Application No. PCT / US2008 / 002086 and International Patent Application No. PCT / US2007 / 077273 filed on August 30, 2007, US Serial No. 60 / 890,380, filed February 16, 2007, 2007 U.S. Serial No. 60 / 947,705, filed on July 3, and U.S. Serial No. 11 / 683,198, filed on Mar. 7, 2007, Serial No. 60 / 795,417, Serial No. 60 / 841,727, filed September 1, 2006, Serial No. 60 / 890,380, filed February 16, 2007, and Application filed on April 27, 2006 , Which claims priority to: (1) U.S. Serial No. 11/289142, filed on November 28, 2005, and U.S. Patent Application Serial No. 11 / U.S. Serial No. 11 / 267,523, filed November 4, 2005; (2) International Application No. PCT / US05 / 31900, filed September 7, 2005, which is a continuation-in-part of U.S. Serial No. 60 / 617,379 filed October 8, 2004, filed September 27, 2004 U.S. Serial No. 60 / 613,811, and U.S. Serial No. 60 / 607,858, filed September 7, 2004); (3) U.S. Serial No. 11 / 131,551, filed May 17, 2005; And (4) U.S. Serial No. 11 / 117,760, filed April 27, 2005. This application is also related to U.S. Serial No. 11 / 412,659, filed April 27, 2006, U.S. Serial No. 10 / 906,303, filed February 14, 2005, and U.S. Serial No. 10 / U.S. Serial No. 12 / 344,682 as priority. The contents of these applications are incorporated herein by reference in their entirety.

Field of invention

The present invention provides compounds, compositions, extracts, and methods for inhibiting cancer invasion, cell invasion, or cancer cell invasion.

The present invention provides a method for synthesizing a novel compound for pharmaceutical use. The present invention provides methods, compounds and compositions for treating cancer by inhibiting cancer invasion, cell invasion or cancer cell invasion wherein the cancer is selected from the group consisting of breast, white blood cells, liver, ovary, bladder, prostate, , Brain, leukemia, lung, colon, CNS, melanoma, kidney, cervix, esophagus, testes, spleen, kidney, lymphatic, pancreas, stomach and thyroid.

The present invention provides a method for synthesizing a novel compound for pharmaceutical use. The present methods provide compounds, compositions and methods for treating cancer by inhibiting cancer invasion, cell invasion, cancer cell invasion and metastasis. The present methods provide the use of compounds, compositions for the manufacture of medicaments for the treatment of cancer by inhibiting cancer invasion and metastasis. The present invention provides compounds for use as mediators or inhibitors of adhesion proteins or angiopoietins. The present invention provides a compound for use in a method for controlling adhesion or adhesion or angiogenesis of a cell by regulating or suppressing adherent protein or angiopoietin. The present compounds comprise a structure selected from the formulas of the present application wherein the present compounds are synthesized or separated and wherein the compound comprises a compound selected from the group consisting of saponins, triterpenes, pentacyclic triterpenes, The present invention relates to a method for treating a cancer of the breast, a white blood cell, a liver, an ovary, a bladder, a prostate, skin, bone, brain, leukemia, lung, colon, CNS, melanoma, kidney, cervix, esophagus, Cancer. The present invention provides compounds for use as a mediator for cell circulation, cell migration and inflammatory diseases.

Figure 1 is the HPLC profile of the esterification product of E4A and tigloyl chloride (A) from different esterification reaction times. The reaction products obtained from each reaction time (5 seconds, 1 minute, 2 minutes, 5 minutes, and 10 minutes) were fractionated by HPLC. The profile is plotted according to the HPLC elution time and the optical density of the fraction. The reaction was carried out at room temperature (upper column) and 0 ° C (lower column).
Figure 2 is an HPLC profile of the esterification product of E4A and 3,3-dimethyl acryloyl chloride (B) from different esterification reaction times. The reaction products obtained from each reaction time (5 seconds, 1 minute, 2 minutes, 5 minutes, and 10 minutes) were fractionated by HPLC. The profile is plotted according to the HPLC elution time and the optical density of the fraction. The reaction was carried out at room temperature (upper column) and 0 ° C (lower column).
Figure 3 is an HPLC profile of the esterification product of E4A and 4-pentenoyl chloride (C) from different esterification reaction times. The reaction products obtained from each reaction time (5 seconds, 1 minute, 2 minutes, 5 minutes, and 10 minutes) were fractionated by HPLC. The profile is plotted according to the HPLC elution time and the optical density of the fraction. The reaction was carried out at room temperature (upper column) and 0 ° C (lower column).
Figure 4 is the HPLC profile of the esterification product of E4A and hexanoyl chloride (D) from different esterification reaction times. The reaction products obtained from each reaction time (5 seconds, 1 minute, 2 minutes, 5 minutes, and 10 minutes) were fractionated by HPLC. The profile is plotted according to the HPLC elution time and the optical density of the fraction. The reaction was carried out at room temperature (upper column) and 0 ° C (lower column). It is also the HPLC profile of the esterification product of E4A and 2-ethylbutyryl chloride (E) from different esterification reaction times. The reaction products obtained from each reaction time (5 seconds, 1 minute, 2 minutes, 5 minutes, and 10 minutes) were fractionated by HPLC. The profile is plotted according to the HPLC elution time and the optical density of the fraction. The reaction was carried out at room temperature (upper column) and 0 ° C (lower column).
Figure 5 is the HPLC profile of the esterification product of E4A and acetyl chloride (H) from different esterification reaction times. The reaction products obtained from each reaction time (5 seconds, 1 minute, 2 minutes, 5 minutes, and 10 minutes) were fractionated by HPLC. The profile is plotted according to the HPLC elution time and the optical density of the fraction. The reaction was carried out at room temperature.
Figure 6 is the HPLC profile of the esterification product of E4A and crotonoyl chloride (I) from different esterification reaction times. The reaction products obtained from each reaction time (5 seconds, 1 minute, 2 minutes, 5 minutes, and 10 minutes) were fractionated by HPLC. The profile is plotted according to the HPLC elution time and the optical density of the fraction. The reaction was carried out at room temperature.
Figure 7 is the HPLC profile of the esterification product of E4A and cinnamoyl chloride (J) from different esterification reaction times. The reaction products obtained from each reaction time (5 seconds, 1 minute, 2 minutes, 5 minutes, and 10 minutes) were fractionated by HPLC. The profile is plotted according to the HPLC elution time and the optical density of the fraction. The reaction was carried out at 75 < 0 > C.
Figure 8 is the HPLC profile of the esterification product of E4A and benzoyl chloride (K) from different esterification reaction times. The reaction products obtained from each reaction time (5 seconds, 1 minute, 2 minutes, 5 minutes, and 10 minutes) were fractionated by HPLC. The profile is plotted according to the HPLC elution time and the optical density of the fraction. The reaction was carried out at 0 < 0 > C.
9 shows A: E4A-Tigloyl at room temperature; B: E4A-3,3-dimethyl acryloyl; C: Time study of MTT cytotoxic activity against E4A-4-pentenoyl.
10 shows A: EA: E4A-Tigloyl at 0 [deg.] C; B: E4A-3,3-dimethyl acryloyl; C: Time study of MTT cytotoxic activity against E4A-4-pentenoyl.
FIG. 11 is a graph showing the spectra of J: E4A-cinnamoyl; D: E4A-hexanoyl; E: E4A-2-ethylbutyryl; And a control (the Tig control is tigloyl chloride without E4A; the AC control is acetyl chloride without E4A); H: Time study of MTT cytotoxic activity on acetyl chloride with 1 minute E4A response.
Figure 12 shows H: E4A-acetyl; I: Time study of MTT cytotoxic activity against E4A-crotonoyl.
Figure 13 is a time study of MTT cytotoxic activity against E4A-Tig at 1, 15, 30, 1 and 2 hours.
Figure 14 is the HPLC profile of E4A-Tig at 1 and 2 hours.
Figure 15 is a time study of MTT cytotoxic activity against E4A-Tig. Results: E4A-Tig from 5 seconds to 1 minute of reaction was most active. Activity decreases after 1 minute of reaction. Minimal activity or no activity at 10 minutes or longer.
Figure 16 shows the results of HPLC profiles of E4A-Tig: E4A, E4A-ASAP (5 sec), E4A-1min, E4A-2min, E4A-5min, E4A-10min and E4A-30min.
Figure 17 shows the result of the sequence of activities: M, N, O, P, Q, R, S, T, E4A; M = E4A has no activity.
Figure 18 is the result of MTT cytotoxic activity of E4A-Tig-R in cancer cells of different organs: A, bone (U20S) IC50 = 4.5 [mu] g / ml; B, bladder (TB9): IC50 = 2.5 [mu] g / ml; C, lung (H460): IC50 = 4.8 [mu] g / ml; D, ovary (ES2): IC50 = 2.8 占 퐂 / ml
Figure 19 shows the results of MTT cytotoxic activity of E4A-Tig-R in cancer cells of different organs: E, colon (HCT116) IC50 = 5.2 [mu] g / ml; F, pancreas (Capan) IC50 = 2.4 [mu] g / ml; G, ovary (OVCAR3) IC50 = 5.8 [mu] g / ml; H, breast (MCF-7) IC50 = 4.5 [mu] g / ml
Figure 20 shows the results of MTT cytotoxic activity of E4A-Tig-R in cancer cells of different organs: I, prostate (DU145) IC50 = 3.6 占 퐂 / 5.1 [mu] g / ml; K, mouth (KB) IC 50 = 3 쨉 g / ml; L, kidney (A498) IC 50 = 3.5 쨉 g / ml
Figure 21 is a result of MTT cytotoxic activity of E4A-Tig-R in cancer cells of different organs: M, liver (HepG2) IC50 = 6 [mu] g / ml; N, brain (T98G) IC50 = 8 [mu] g / ml; P, Leukemia (K562) IC 50 = 2 [mu] g / ml; Q, cervical (HeLa) IC 50 = 5 / / ml
22 (a): Tig-N, -Q, -R, -T -S, and -V do not have hemolytic activity up to 20 μg / ml. Essentially compound ES dissolves 100% red blood cells (RBC) at 5 μg / ml. (b) Results: Compared with Y3, ACH-Y3 is less potent in hemolytic activity. Tig-R does not exhibit hemolytic activity.
Figure 23 (a) Results of HPLC profiles of reaction products. Several fractions were obtained. Individual fractions were combined for further study. (b) Results of purification of E4A-Tig-R.
24 shows the results of MTT assay of E4A-Tig-R on bone U20S cells.
25 shows the HNMR results of E4A-Tig-R.
Figure 26 shows the CNMR results of E4A-Tig-R.
27 shows the HMQC result of E4A-Tig-R.
Figure 28 shows the HMBC results of E4A-Tig-R.
29. The mass spectrum of Tig-R (M + H) is 671.4509. The mass is the same as the proposed structure.
FIG. 30 shows the chemical structure of E4A-Tig-R, 24,28-0-Tigloyl-3 ?, 16 ?, 21 ?, 22 ?, 24 ?, 28-hexahydroxy olean- 670.91548
31 (a). Results of HPLC profiles of reaction products. Several fractions were obtained. Individual fractions were combined for further study. (b) Results of purification of E4A-Tig-N. (c) Results of purification of E4A-Tig-S. (d) Results of purification of E4A-Tig-T.
Figure 32 (a) MTT assay of E4A-Tig-N on bone U20S cells; (b) Results of MTT assay of E4A-Tig-S on bone U20S cells.
Figure 33 (a) MTT assay of E4A-Tig-T on bone U20S cells; (b) shows fmf as a result of MTT assay of E4A-Tig-V on ovarian ES2 cells. IC50 = 2 [mu] g / ml; (c) shows the result of purification of E4A-Tig-Vdml.
34 shows the HNMR results of E4A-Tig-V.
35 shows the HMQC result of E4A-Tig-V.
36 shows the HMBC result of E4A-Tig-V.
37 shows the mass spectrum results of E4A-Tig-V. Tig-R (M + H) mass is 753.4924, which is consistent with the proposed formula (C45H6809).

The present invention provides a method for synthesizing novel active compounds for pharmaceutical use. The present invention provides anti-adhesion therapy using compounds as an agent or inhibitor of adhesion proteins and angiopoietin. This inhibits excessive adhesion and inhibits cell adhesion. This regulates angiogenesis. The compounds are also used as mediators of cell adhesion receptors.

The present invention provides a method for treating cancer; For inhibiting cancer growth, for inhibiting virus; For preventing cerebral aging; To improve memory; For improving cerebral function; Nocturnal enuresis, frequent micturition, urinary incontinence; Dementia, Alzheimer's disease, autism, brain trauma, Parkinson's disease, or other diseases caused by cerebral dysfunction; Arthritis, rheumatism, poor circulation, arteriosclerosis, Raynaud's syndrome, angina pectoris, heart disease, coronary heart disease, headache, dizziness, kidney disease; For treating cerebrovascular disease; For inhibiting NF-kappa B activation; Severe acute respiratory syndrome, viral respiratory disease, chronic venous insufficiency, hypertension, chronic venous disease, edema, inflammation, hemorrhoids, peripheral edema, varicose veins, , Flu, post-traumatic edema and post-operative bookkeeping; For inhibiting thrombosis, for inhibiting ethanol absorption; To lower blood sugar; The compounds provided herein and compositions comprising such compounds for modulating adrenocorticotropin and corticosterone levels are provided. The present invention relates to the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment or prophylaxis of an anti-MS, antianeurysm, antiepileptic, antiepileptic, antiexemptive, antifluic, antifracture, antigingivitic, antihematomic, antiepileptic, antiepileptic, antiepileptic, antiepileptic, antiepileptic, antiepileptic, antihypertensive, antiherpetic, antihistatic, antihydrathritic, antimeningitic, antioxidant, antiperiodic, antiphlebitic, antipleuritic, antirauced, anti-salivary, antirheumatic, antitonsilitic, antiulcer, antivaricose, antivertiginous, cancerostatic, corticosterogenic, diuretic, fungicide, hemolytic, ), Hyaluronidase inhibition, the present invention relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a disease selected from the group consisting of lymphagogue, sodium natriuretic, insecticide, pituitary stimulant, thymolytic, vasoprotective, inhibition of leishmaniases, Antiparasitic; An increase in expression of the genes ANGPT2, DDIT3, LIF and NFKB1Z, and compositions for the preparation of adjuvant compositions and for venotonic treatment.

The present invention provides compounds, compositions and methods for treating cancer diseases, inhibiting cancer infiltration, inhibiting cancer growth or inhibiting cancer metastasis, said compounds being selected from the group consisting of the structures selected from the formulas of the present invention Wherein the compound is synthesized or isolated and the compound comprises a compound selected from the group consisting of triterpenes, pentacyclic triterpenes, saponins, and the formula of the invention, wherein the cancer is selected from the group consisting of breast cancer, , Cancer of the ovary, bladder cancer, prostate cancer, skin cancer, bone cancer, brain cancer, leukemia cancer, lung cancer, colon cancer, CNS cancer, melanoma cancer, kidney cancer, cervical cancer, Wherein said cells are selected from the group consisting of breast cells, leukocyte cells, liver cells, ovarian cells, bladder cells, prostate cells, skin cells, bone cells, brain cells, pancreatic cancer, And a blood clot cell, lung cell, colon cell, CNS cells, melanoma cells, kidney cells, cervical cells, esophageal cells, testicular cells, spleen cells, kidney cells, lymphatic cells, spleen cells, stomach cells, and thyroid cells.

The present invention also relates to the use of a compound selected from the group consisting of pentaacyl triterpenes, triterpenes, triterpenoids, triterpenoid saponins, or compounds of the present invention in combination with tigloyl, angeloyl, acetyl, crotonoyl, Alkanoyl, alkanoyl, alkanoyl, substituted alkanoyl, alkanoyl, substituted alkanoyl, alkenoyl substituted alkanoyl, alkanoyl, substituted alkanoyl, The presence of a sugar moiety substituted with a phenyl group, aryl, acyl, heterocyclic, heteroaryl, sugar moiety, or dianiline group is useful for cancer growth, cancer invasion, cell invasion, cancer cell invasion, cell adhesion, Lt; / RTI > Dimethylacryloyl is a synonym for senesia oil.

The present invention relates to a process for the preparation of a compound selected from the group consisting of pentaacyl triterpenes, triterpenes, triterpenoids, triterpenoid saponins or compounds selected from the formula of the present invention at carbon positions 21, 22, 24 and / Substituted or unsubstituted alkyl, substituted or unsubstituted alkyl, cycloalkyl, cycloalkyl, cycloalkyl, cycloalkyl, cycloalkyl, heterocyclyl, The presence of sugar moieties substituted with alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocyclic, heteroaryl, sugar moieties, or dianergoyl groups has been associated with cancer growth, cancer invasion, Or to inhibit cancer cell infiltration. In one embodiment, at the carbon position 3, 8, 15, 21, 22, 24 and / or 28 of the compound selected from triterpenes, triterpenoids, triterpenoid saponins or the formula of the present invention, Alkanoyl, alkenoyl, benzoylalkyl, substituted benzoyl, acetoacetyl, crotonoyl, 3,3-dimethyl acryloyl, semenoxy, cinnamoyl, pentenoyl, hexanoyl, benzoyl, ethylbutyryl, dibenzoyl, The presence of the group (s) selected from the group consisting of alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocyclic, heteroaryl, and sugar moieties can be used for cancer growth, cancer invasion, Infiltration, cell adhesion, cell adhesion or inhibition of cell cycle. In an embodiment, the presence of a group at carbon position 24 forms activity. In embodiments, the presence of a group at carbon positions 24 and 28 forms activity. In embodiments, the presence of a group at carbon positions 24 and 21 forms activity. In embodiments, the presence of a group at carbon positions 24, 28, and 21 forms activity. In embodiments, the presence of a group at carbon positions 24, 28, and 22 forms activity. In embodiments, the presence of a group at carbon positions 24, 28, and 3 forms activity. In embodiments, the presence of a group at carbon positions 24 and 3 forms activity. In embodiments, the presence of a group at carbon positions 28 and 3 forms activity. In embodiments, the presence of a group at carbon position 3 forms activity. In an embodiment, the presence of a group at carbon positions 21 and 22 forms activity.

The present invention represents a method of synthesizing an active compound by conjugating a functional group to a core compound wherein the functional group (s) is selected from the group consisting of tigloyl, angeloyl, acetyl, crotonoyl, 3,3-dimethyl acryloyl, Alkanoyl, alkanoyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, alkanoyl, alkanoyl, alkanoyl, alkanoyl, alkanoyl, Heterocyclic, and heteroaryl, and the core compound is 5-ring triterpenes. In an embodiment, the core compound is 4-ring terpene. In an embodiment, the core compound is a 3-ring terpene. In an embodiment, the core compound is a bicyclic terpene. In embodiments, the core compound is a single ring terpene. The compounds provided herein can be used to treat cancer, to treat cancer growth, cancer invasion, cell invasion, cancer cell invasion; Inhibits cell adhesion, cell adhesion, cell cycle, inhibits virus, prevents brain aging, improves memory, improves brain function, enuresis, frequent urination, urinary incontinence; Dementia, Alzheimer's disease, autism, brain trauma, Parkinson's disease or other dementia caused by brain dysfunction and is useful for the treatment and prevention of arthritis, rheumatism, poor blood circulation, arteriosclerosis, Raynaud's syndrome, angina, heart disease, coronary heart disease, Headache, dizziness, kidney disease; Treating cerebrovascular disease; Inhibits NF-kappaB activation and is useful for the treatment of brain edema, severe acute respiratory syndrome, viral respiratory disease, chronic venous insufficiency, hypertension, chronic venous disease, edema, inflammation, hemorrhoids, peripheral edema, varicose veins, Treat edema and post-operative swelling; Inhibit blood clotting, inhibit ethanol absorption, lower blood glucose levels, and regulate adrenocorticotropin and corticosterone levels. The present invention relates to a pharmaceutical composition for preventing or treating an anti-MS, anti-aneurysm, anti-asthma, anti-edema, inflammation, anti-Bradykin, hematocrit, anti headache, Anti-inflammatory agents, anti-inflammatory agents, anti-inflammatory agents, anti-inflammatory agents, anti-inflammatory agents, anti-inflammatory agents, antiinflammatory agents, anti-inflammatory agents, antiinflammatory agents, antiinflammatory agents, antihypertensive agents, antihistamines, antihypertratics, Anti-varicose, anti-dizziness, cancer suppression, corticosteroid source, diuretic, fungicide, hemolyzing agent, hyaluronidase inhibition, lymphatic material, sodium diuretic, insecticide, pituitary stimulant, thymus tissue dissolution, vascular protection, , Cell adhesion or angiogenesis regulation, antiparasitics; A composition for suppressing the expression of genes, ANGPT2, DDIT3, LIF and NFKB1Z, and a fishbone composition and a vein incision.

In the experiments described in the present invention, it has been shown 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 alkoyl group from the carbon positions 21 and 22 of the active Xanifolia Y (Y3). The present invention shows that the ability to inhibit cancer infiltration, cell infiltration, or cancer cell infiltration of Zonifolia Y (Y3) is lost by eliminating the angelogroup from carbon positions 21 and 22.

The experiments described in the present invention showed that the core compound comprising E4A, E5A, and Zonifolia Y-core had no effect in inhibiting cancer growth, cancer invasion, cell invasion or cancer cell invasion. The Zonifolia Y-core is obtained by removing the anuroyl group from the carbon positions 21 and 22 of the active Znophiol Y (Y3) and removing the sugar moiety from the carbon position 3. E4A (E IV A) is obtained by removing the group from carbon positions 3, 21 and 22 of the active Escin. E5A (E V A) is obtained by removing the group from carbon positions 3, 21 and 22 of the active Escin.

The present invention shows that core compounds comprising E4A, E5A, Zonifolia Y-core and AKOH do not have hemolytic activity and anticancer activity.

The present invention shows that Tig-N, Tig-Q, Tig-R, Tig-T Tig-S and Tig-V do not have hemolytic activity up to 20 占 퐂 / ml. Essentially, compound ES dissolves 100% red blood cells (RBC) at 5 μg / ml. Compared with Y3, ACH-Y3 is less potent in hemolytic activity. Tig-R does not have hemolytic activity. The present invention shows that Tig-N, Tig-Q, Tig-R, Tig-T, Tig-S and Tig-V have anticancer activity.

The present invention relates to a method for inhibiting cancer growth, cancer invasion, cell invasion or cancer cell infiltration when the sugar moiety is removed from the carbon position 3 of the active compound, triterpene, triterpenoid, or triterpenoid saponin . In the experiments described in the present invention, the compound ACH-Y3 has the ability to inhibit cancer invasion, cell invasion or cancer cell invasion. The compound ACH-Y3 is obtained by removing the sugar moiety from the carbon position 3 of the active Znophiol Y (Y3). The present invention demonstrates that the ability to inhibit cancer invasion, cell invasion or cancer cell invasion is maintained when the sugar moiety is removed from the carbon position 3 of the active Znophylia Y (Y3).

A bioactive compound, including inhibiting cancer growth and inhibiting cancer invasion, cell invasion or cancer cell invasion, is referred to as an active compound.

The present invention relates to a method of treating cancer or treating cancer, cancer growth, cancer infiltration, cell infiltration, cancer cell infiltration, There is provided a use of compounds, compositions and methods for the manufacture of a medicament for inhibiting cell adhesion, cell attachment, cell cycle, or inhibiting cancer metastasis, wherein said compound comprises a structure selected from the formulas of the present invention Said compound being capable of being synthesized or isolated, said compound comprising pentacyclic triterpene, said cell comprising cancer cells, said cancer being selected from the group consisting of breast cancer, leukocyte cancer, liver cancer, ovarian cancer, bladder cancer , Cancer of the prostate, skin cancer, bone cancer, brain cancer, leukemia cancer, lung cancer, colon cancer, CNS cancer, melanoma cancer, kidney cancer, cervical cancer, esophageal cancer, testicular cancer, , Pancreatic cancer, gastric cancer, and thyroid cancer. Methods for inhibiting cancer invasion, cell invasion or cancer cell invasion activity use non-cytotoxic drug concentrations. Methods for inhibiting metastasis use non-cytotoxic drug concentrations. Changes in cell morphology are not clear.

The present invention provides a method of treating cancer, treating cancer, cancer growth, cancer invasion, cancer cell invasion, A method of inhibiting cell adhesion, cell adhesion, cell cycle, or inhibiting the movement, metastasis or growth of a cancer, said method comprising affecting gene expression, or said method comprising stimulating gene expression, Wherein the method comprises inhibiting gene expression, or the method comprising administering to the subject an effective amount of a compound of the invention, or composition. In one embodiment, the method comprises contacting the cell with a compound selected from the group consisting of 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, DM8, D20-32, E1-18, E20-32, G1-18, G20-32, H1-18, (X), Y1, Y2, Y2, Y3, Y5, Y7, Y8, Y9, Y10, ACH-Y1, ACH-Y2, ACH-Y8, ACH-Y7, ACH-Y0, ACH-X, ACH-Z4, ACH-Z1, ACH- -Escin (bES), ACH-M10 and salts, esters, metabolites thereof, and compounds from Formulas 2A and K.

In vitro studies indicate that compounds selected from structure (2A) or (K) inhibit cell adhesion to culture flasks. These compounds block the function of these adhesion molecules on the cells. In one embodiment, the selected compound blocks the function of this adhesion molecule on the cell. In one embodiment, the selected compound blocks the function of such attachment molecules on carcinoma cells. In one embodiment, the selected compound blocks the function of such an adhesion molecule on mesothelial cells. The present invention provides an anti-adherent therapy using a compound as a mediator or inhibitor of adhesion proteins and angiopoietin. This inhibits excessive adhesion and inhibits cell adhesion. The present invention provides compounds for use as mediators for cell cycle, cell migration and inflammatory diseases. In one embodiment, the selected compounds bind (via masking) an attachment protein on the membrane and inhibit the interaction of the attachment protein with their receptor. In one embodiment, the action of the selected compound on the membrane affects the function of the adherent protein in the membrane. Loss of adhesion activity of cancer cells results in the direct or indirect action of selected compounds on the membrane protein.

(The purification method and biological assay of the present invention are described in International Application No. PCT / US05 / 31900, filed September 7, 2005, US Serial No. 11/289142, filed November 28, 2005, US Serial No. 11/131551, filed on March 17, and MTT test on PCT / US2008 / 002086 and 1188-ALA-PCT filed on February 15, 2008, International Application No. PCT / US2010 / 0042240, filed July 16, 2006, the contents of which are incorporated herein by reference).

The present invention provides the use of a compound or method for inhibiting cancer invasion, cell invasion, cancer cell invasion, cancer migration, metastasis or growth, wherein the invention comprises a process and a method of administering a composition, Administration of a compound of 0.003-0.03 mg / kg body weight dissolved in 250 ml of 10% glucose solution or 250 ml of 0.9% NaCl solution by intravenous infusion, intravenous infusion, intraperitoneal injection or oral administration , A compound of 0.003-0.03 mg / kg body weight dissolved in 10-20 ml of 10% glucose solution or 0.9% NaCl solution per day by intravenous injection or 250 ml of 10% glucose solution or 250 ml of 0.9% NaCl solution 0.0 > mg / kg < / RTI > body weight, or by intravenous injection, 10-20 ml of 10% glucose solution or 0.01-0.03 mg / kg body weight dissolved in 0.9% NaCl solution, or 50 ml of 10% glucose Solution or 250 ml 0.0 > mg / kg < / RTI > body weight dissolved in a 0.9% NaCl solution of 0.9% NaCl solution, or 10-20 ml of 10% glucose solution or 0.9% A compound of body weight or a compound of 0.05-0.2 mg / kg body weight dissolved in 50 ml of 10% glucose solution or 250 ml of 0.9% NaCl solution, or 10-20 ml of a 10% glucose solution per day by intravenous injection or 0.05-0.2 mg / kg body weight dissolved in 0.9% NaCl solution, or 0.1-0.2 mg / kg body weight dissolved in 250 ml of 10% glucose solution or 250 ml of 0.9% NaCl solution per day by intravenous drip A compound of body weight may be administered or a compound of 0.1-0.2 mg / kg body weight dissolved in 10-20 ml of 10% glucose solution or 0.9% NaCl solution per day by intravenous injection may be administered or intraperitoneally injected (IP) may be administered Dissolved in 10% glucose solution or 0.9% NaCl solution per day A compound of 2.5 mg / kg body weight, or a dosage of mammal by oral administration is 1-10 mg / kg, 10-30 mg / kg, 30-60 mg / kg, or 60-90 mg / kg body weight, The dosage of the mammal by intravenous injection or intravenous drip is a compound of 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, Intraperitoneal injection (I. P.) is a compound of 1-3 mg / kg, 3-5 mg / kg, 4-6 mg / kg, or 6-1 Omg / kg body weight.

The present invention relates to a method of treating cancer or treating cancer, cancer growth, cancer infiltration, cell infiltration, cancer cell infiltration, The invention provides the use of a compound or method for inhibiting cell adhesion, cell adhesion, cell cycle, cancer migration, metastasis or growth, wherein the invention provides a compound of the invention, or a pharmaceutically acceptable salt thereof, Wherein the compound is present in a concentration from 0.01 μg / ml to 65 μg / ml, or the compound is present in a concentration from 0.01 μg / ml to 40 μg / ml, The compound is present at a concentration of 0.01 to 30 μg / ml, or the compound is present at a concentration of 0.01 to 10 μg / ml, or the compound is present at a concentration of 0.01 μg / Or the compound is present at a concentration of 5 μg / ml to 10 μg / ml, or the compound is present at a concentration of 0.1 μg / ml to 5 μg / ml, or the compound is present at a concentration of 0.1 μg / / Ml, < / RTI > The compound is present at a concentration of 0.1 to 10 μg / ml, or the compound is present at a concentration of 0.1 to 15 μg / ml, or the compound is present at a concentration of 0.1 μg / ml to 20 μg / Or the compound is present at a concentration of from 0.1 μg / ml to 30 μg / ml, or the compound is present at a concentration of from 1 μg / ml to 5 μg / ml, or the compound is present at a concentration of from 1 μg / / Ml, or the compound is present at a concentration of 1 / / ml to 10 / / ml, or the compound is present at a concentration of 1 / / ml to 20 / / ml, or the compound is present at a concentration of 1 ㎍ / Or the compound is present at a concentration of from 3 μg / ml to 5 μg / ml, or the compound is present at a concentration of from 3 μg / ml to 7.5 μg / ml, or the compound Is present at a concentration of 3 [mu] g / ml to 10 [mu] g / ml, or the compound is present at a concentration of 3 [mu] g / Alternatively, the compound may be present at a concentration of 3 μg / ml to 20 μg / ml, or the compound may be present at a concentration of 3 μg / ml to 30 μg / ml or the compound may be present at a concentration of 4 μg / Or the compound is present at a concentration of from 4 μg / ml to 7.5 μg / ml, or the compound is present at a concentration of from 4 μg / ml to 10 μg / ml, or the compound is present at a concentration of from 4 μg / Or the compound is present at a concentration of from 4 μg / ml to 20 μg / ml, or the compound is present at a concentration of from 4 μg / ml to 30 μg / ml, or the compound is present at a concentration of 4 The compound is present at a concentration of from 5 μg / ml to 8 μg / ml, or the compound is present at a concentration of from 5 μg / ml to 9 μg / ml, The compound is present at a concentration of 5 [mu] g / ml to 10 [mu] g / ml, or the compound is present at a concentration of 5 [mu] g / Alternatively, the compound may be present at a concentration of 5 μg / ml to 20 μg / ml, or the compound may be present at a concentration of 5 μg / ml to 30 μg / ml or the compound may be present at a concentration of 7 μg / Or the compound is present at a concentration of from 7 / / ml to 9 / / ml, or the compound is present at a concentration of from 7 / / ml to 10 / / ml, or the compound is present at a concentration of from 7 / / Or the compound is present at a concentration of 7 / / ml to 20 / / ml, or the compound is present at a concentration of 7 / / ㎖ to 30 / / ml.

The present invention relates to a method of treating cancer or treating cancer, cancer growth, cancer infiltration, cell infiltration, cancer cell infiltration, The invention provides the use of a compound or method for inhibiting cell adhesion, cell adhesion, cell cycle, cancer migration, metastasis or growth, wherein the invention provides a compound of the invention, or a pharmaceutically acceptable salt thereof, Wherein the compound is present in a concentration from 0.008 μM to 80 μM or the compound is present in a concentration from 0.01 μM to 60 μM or the compound is present in a concentration from 0.01 μM to 50 μM Or the compound is present in a concentration of 0.01 μM to 40 μM or the compound is present in a concentration of 0.01 μM to 30 μM or the compound is present in a concentration of 0.01 μM to 20 μM or the compound is present in a concentration of 0.01 μM to 10 μM , Or the compound is present at a concentration of 5 μM to 10 μM, or the compound is present at a concentration of 0.1 μM to 5 μM Or the compound is present in a concentration of 0.1 μM to 7.5 μM or the compound is present in a concentration of 0.1 μM to 10 μM or the compound is present in a concentration of 0.1 μM to 15 μM or the compound is present in a concentration of 0.1 μM to 20 μM Or the compound is present at a concentration of 0.1 μM to 30 μM or the compound is present at a concentration of 0.1 μM to 40 μM or the compound is present at a concentration of 0.1 μM to 50 μM, Or the compound is present in a concentration of 0.1 [mu] M to 80 [mu] M, or the compound is present in a concentration of 1 [mu] M to 5 [mu] M, or the compound is present in a concentration of 1 [ 1 μM to 10 μM, or the compound is present at a concentration of 1 μM to 15 μM, or the compound is present at a concentration of 1 μM to 20 μM, The compound is present at a concentration of 1 [mu] M to 30 [mu] M, or the compound is present at a concentration of 1 [mu] M to 40 [mu] M, or the compound is present at a concentration of 1 [ , The compound is present in a concentration of 1 [mu] M to 80 [mu] M, or the compound is present in a concentration of 3 [mu] M to 5 [mu] M, or the compound is present in a concentration of 3 [mu] M to 7.5 [mu] M, , The compound is present at a concentration of 3 μM to 15 μM or the compound is present at a concentration of 3 μM to 20 μM or the compound is present at a concentration of 3 μM to 30 μM or the compound is present at a concentration of 3 μM to 40 μM, The compound is present at a concentration of 3 [mu] M to 50 [mu] M, or the compound is present at a concentration of 3 [mu] M to 60 [mu] M, Or the compound is present at a concentration of 5 μM to 8 μM or the compound is present at a concentration of 5 μM to 10 μM or the compound is present at a concentration of 5 μM to 15 μM or the compound is present at a concentration of 5 μM to 20 μM Or the compound is present at a concentration of 5 μM to 30 μM or the compound is present at a concentration of 5 μM to 40 μM or the compound is present at a concentration of 5 μM to 50 μM or the compound is present at a concentration of 5 μM to 60 μM , The compound is present at a concentration of 5 μM to 80 μM or the compound is present at a concentration of 7 μM to 8 μM or the compound is present at a concentration of 7 μM to 10 μM or the compound is present at a concentration of 7 μM to 15 μM, The compound is present at a concentration of 7 μM to 20 μM, or the compound is present at a concentration of 7 μM to 30 μM, or the compound is present at a concentration of 7 μM To 40 μM, or the compound is present at a concentration of 7 μM to 50 μM, or the compound is present at a concentration of 7 μM to 60 μM, or the compound is present at a concentration of 7 μM to 80 μM.

While the invention will be better understood with reference to the following examples, those skilled in the art will readily appreciate that certain experiments are illustrative only and are not intended to limit the invention as described herein, But is limited by the claims.

Throughout the present invention, various references or publications are cited. The entire contents of such references or publications are incorporated herein by reference for the purpose of more fully describing the art to which this invention pertains.

The terms "including", "containing", or "transitional term", which is synonymous with "comprising", are inclusive or open-ended, and additional non- Or method steps are not excluded.

Example  One

10 mg , 20 mg , 30 mg Of active compound < RTI ID = 0.0 >

The active compound, cellulose, and some corn starch were mixed and granulated with 10% corn starch paste. The resulting granulate was sieved, dried and combined with the remaining corn starch and magnesium stearate. The resulting granulate was then pressurized with tablets containing 1, 5, 10, 20, 30 mg of active ingredient per tablet, respectively.

Example 2

Intravenous  Solution preparation

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

1-1O [mu] g of active compound

Sodium citrate 5-50 mg

Citric acid 1-15 mg

Sodium chloride 1-8 mg

A sufficient amount to 1 ml of water for injection (USP)

Using this amount, the active compound was dissolved in a prepared solution of sodium chloride, citric acid, and sodium citrate in water for injection at room temperature.

Example  3

Intravenous Dot  Products ( intravenous drip preparation )

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

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

Treatment of a number of standard chlorinating reagents, including phosphoric acid, phosphorus oxychloride, phosphorus trichloride and thionyl chloride, forms the diglyoyl chloride. Oxalyl chloride forms a 2: 1 ratio of angeloyl chloride to tigloyl chloride. Treatment of potassium salt in diethyl ether with oxalyl chloride and catalytic amount of DMF for 2 h at 0 C forms pure angeloyl chloride.

Acid hydrolysis of the following compounds:

a) Zonifolia (Y),

또는 화학명: 3-O-[β-D-갈락토피라노실 (1→2)]-α-L-아라비노푸라노시 (1→3)-β-D-글루쿠로노피라노실-21,22-0-디안겔로일-3β,15α,16α,21β,22α,28-헥사하이드록시올레안-12-엔;

Or the chemical name: 3-O- [beta -D-galactopyranosyl (1 → 2)] - α-L-arabinofuranosyl (1 → 3) 22-0-dianergoyl-3β, 15α, 16α, 21β, 22α, 28-hexahydroxy olean-12-ene;

c) Zonifolia (Y2),

또는 화학명: 3-O-[β-D-글루코피라노실-(1→2)]-α-L-아라비노푸라노시 (1→3)-β-D-글루쿠로노피라노실-21,22-0-디안겔로일-3β,15α,16α,21β,22α,24β,28-헵타하이드록시올레안-12-엔;

Or the chemical name: 3-O- [beta -D-glucopyranosyl- (1 → 2)] - α-L-arabinofuranosyl (1 → 3) 22-0-dianergoyl-3β, 15α, 16α, 21β, 22α, 24β, 28-heptahydroxy olean-12-ene;

d) Zonifolia (Y8),

또는 화학명: 3-0-[β-D-글루코피라노실 (1→2)]-α-L-아라비노푸라노실 (1→3)-β-글루쿠로노피라노실-21,22-0-디안겔로일-3β,16α,21β,22α,24β,28-헥사하이드록시올레안-12-엔;

Or the chemical name: 3-0- [beta -D-glucopyranosyl (1 → 2)] - α-L-arabinofuranosyl (1 → 3) - β- glucuronopyranosyl- - dianergoyl-3β, 16α, 21β, 22α, 24β, 28-hexahydroxy olean-12-ene;

f) Zonifolia (Y10),

또는 화학명: 3-0-[β-갈락토피라노실 (1→2)]-α-아라비노푸라노실 (1→3)-β-글루쿠로노피라노실-21,22-0-디안겔로일-3β,16α,21β,22α,28-펜타하이드록시올레안-12-엔.

Or a chemical name: 3-0- [? - galactopyranosyl (1? 2)] -? - arabinofuranosyl (1? 3) -? - glucuronopyranosyl-21,22-0- Roile-3 [beta], 16 [alpha], 21 [beta], 22 [alpha], 28-pentahydroxy oleucan-12-

j) Structure (M10)

m) Structure (bES):

After acid hydrolysis of the compound, a separate or purified or synthetic compound is formed having a structure (ACH) selected from the following structures:

The composition comprises a bioactive compound from a native plant or from a plant. The program is based on the purification method and biological assay of the present application, including the MTT assay (see International Application No. PCT / US05 / 31900, filed September 7, 2005, November 28, 2005 U.S. Serial No. 11/289142, filed on May 17, 2005, and U.S. Serial No. 11/131551, filed May 17, 2005, and PCT / US2008 / 002086, 1188-ALA-PCT , 12 / 344,682, 1020-B1-US, filed December 29, 2008, the contents of which are incorporated herein by reference. Details of the analysis of gene expression of ES2 cells after Y-treatment by microarrays, data analysis methods and Western blots in PCT / US2008 / 002086, 1188-ALA-PCT filed on February 15, 2008, and 2010 Cell infiltration test method of International Application No. PCT / US2010 / 0042240, filed July 16, 2006, the contents of which are incorporated herein by reference).

Hemolytic assay ( haemolytic assay )

Red blood cells (RBC) were isolated from human blood (EDTA whole blood, random collection). 50 [mu] l of 10% RBC suspension (in PBS) was added to 2 ml of sample solution in PBS (concentration range from 0.1 [mu] g / ml to 400 [mu] g / ml). The mixture was briefly vortexed and allowed to saturate at room temperature for 60 minutes. The mixture was spun at 3K for 10 minutes and the relative amount of dissolved hemoglobin in the supernatant was measured at 540 nm. The synthetic compounds of the present invention were tested in this way.

Acid hydrolysis of saponin

15 mg of Zonifolia-Y was dissolved in 1 ml of methanol. 1 ml of 2N HCl was then added. The mixture was refluxed in a water bath at 80 ° C for 5 hours. The solution was then neutralized by the addition of 2 ml of 1N NaOH (final pH 4-6). The aglycon was subsequently extracted with 3 ml x 2 of ethyl acetate. The extracts were combined and collected. Additional separation of aglycon (ACH-Y) was achieved by HPLC with isocratic elution of 80-100% acetonitrile. The experiment was repeated with compounds Z4, Y10, Y2, Y8, Y7, Y0, X, M10 and ESCIN (bES) to give respectively the following compounds: ACH-Z4, ACH-Y10, ACH-Y2, ACH- -Y7, ACH-Y0, ACH-X, ACH-E, ACH-Z5, ACH-M10 and ACH-bES [see Experiments in International Application No. PCT / US2010 / 0042240, filed July 1, 2010 Method, the contents of which are incorporated herein by reference).

Removal of acyl groups by alkaline hydrolysis

20 mg of Zonifolia-Y was dissolved in 0.5 ml of 1N NaOH. The solution was incubated in a water bath at 80 캜 for 4 hours. It was cooled to room temperature and then neutralized with 0.5 ml 1N HCl (pH adjusted to about 3). The mixture was extracted three times with 2 ml of 1-butanol. The butanol fraction was collected and lyophilized. The hydrolyzed saponin was further purified by HPLC on a C-18 column eluting with 25% acetonitrile.

The compounds AKOH-Y and AKOH-M10 do not exhibit the ability to inhibit cancer growth, cancer invasion, cell invasion or cancer cell invasion.

Core compound

Core compounds or pentacyclic triterpenes, hydroxylated triterpenes were obtained by acid and alkali hydrolysis of saponins from natural sources. The pentacyclic triterpene can also be obtained by a synthetic method. The method for synthesizing the core compound is as follows:

Beta-Escin, Compound Y, Y10, Y2, Y8, Y7, YO, 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 DEG C for 5 hours. The solution was neutralized with NaOH. The hydrolyzed product was extracted with ethyl acetate, which was subsequently removed by evaporation. (E4A) was accomplished by FPLC chromatography on a C18 column with a balance of 70% acetonitrile / TFA at a flow rate of 1 ml / min. Core compound.

The core compound does not exhibit the ability to inhibit cancer growth, cancer invasion, or cell adhesion.

Structure of core compound:

, 또한 bES-코어, E IV A, ES4A, E4A 또는 (E)라 명명됨;

, Also called bES-core, E IV A, ES4A, E4A or (E);

, 또한 ES V, E5A 또는 (F)라 명명됨;

, Also referred to as ES V, E5A or (F);

Wherein R 1, R 2, R 5, R 8 are OH; R3 is OH, is H or a member, R4, R10 is CH ₃ or CH ₂ OH, and; R9, R11, R12, R13, R14, R15 is CH _3;

Wherein R 1, R 2, R 5, R 8, R 17, R 18 are OH; R3 is OH, H or absent; R9, R11, R12, R13, R14, R15 is CH _3.

Typical numbering from 1 to 30 carbon positions of pentacyclic triterpene.

, 또는 E4A 또는 (E)라 명명됨, 상기 식에서, R1, R2, R5, R8, R17, R18은 OH이며; R9, R11, R12, R13, R14, R15는 CH3이다.

, Or E4A or (E), wherein R1, R2, R5, R8, R17, R18 are OH; R9, R11, R12, R13, R14 and R15 are CH3.

The present invention provides a method for synthesizing a novel active compound. A method of attaching a functional group to a core compound that does not include but is not limited to (A), (B), (C), (D), (E) The core compound can be reacted with a compound selected from the group consisting of tigloyl chloride, angeloyl chloride, acetyl chloride, crotonoyl chloride, 3,3-dimethyl acryloyl chloride, semenoyl chloride, cinnamoyl chloride, pentenoyl chloride, hexanoyl chloride, benzoyl chloride 1, 2, 5, 10, 30, 1, 2, or 18 hours at 0, 25, or 75 C with acyl chloride including, but not limited to, Hour, 2 days or 3 days. At the end of the reaction, it was added 5 ml of 2N HCl or 1M NaHC0 ₃ in the reaction mixture. Thereafter, the solution was extracted three times with 10 ml of ethyl acetate, then evaporated under vacuum and at 45 캜 and lyophilized. The reaction product was dissolved in 80% acetonitrile-0.005% trifluoroacetic acid. The active esterification product was purified by HPLC. The activity of the acyl chloride, the post-reaction solution, the individual fractions, and the individual compounds was tested to perform MTT activity. The core compound is synthetic, semisynthetic or is derived from a natural source. The core compounds include terpenes, isoprene, triterpenes, and hydroxylated triterpenes.

At a different reaction time of 5 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 2 days, or 3 days at 0, 25, The MTT activity of acylation was studied. The core compound E4A is reacted with a compound selected from the group consisting of tigloyl chloride, angeloyl chloride, acetyl chloride, crotonoyl chloride, 3,3-dimethyl acryloyl chloride, semenoyl chloride, cinnamoyl chloride, pentanoyl chloride, hexanoyl chloride, The HPLC profile of the esterification product of benzoyl chloride or ethyl butyryl chloride has been shown to change the composition when the reaction time or temperature is changed (cf. FIGS. 1 to 21).

Peaks, fractions and compounds are selected according to the activity and peak change of the time study. The choice of HPLC fraction for separation depends on the cytotoxic activity of the reaction product obtained at a particular time. Compounds with strong activity to weak activity were selected and isolated. The anticancer sex hormones are composed of bone (U20S), lung (H460), bladder (HTB-9), ovary (ES2), colon (HCT116), pancreas (Capan), ovary (OVCAR3) (MT) of the cervix (HeLa), leukemia (K562), brain (T98G), liver (HepG2), and kidney (A498).

Esterification of the core compound to tigloyl chloride and separation of the compound by HPLC provides the following compounds:

Esterification of core compound E4A with angeloyl chloride and separation of the compound by HPLC provides the following compounds:

Esterification of core compound E4A with (3,3-dimethyl acryloyl chloride) senesioyl chloride and separation of the compound by HPLC provides the following compounds:

Sen = Senesi oil

Esterification of core compound E4A with 4-pentenoyl chloride and separation of the compound by HPLC provides the following compounds:

Pen = 4-pentenoyl

Esterification of core compound E4A with hexanoyl chloride and separation of the compound by HPLC provides the following compounds:

Hex = hexanoyl

Esterification of the core compound E4A with 2-ethylbutyryl chloride and separation of the compound by HPLC provides the following compounds:

Eth = 2-ethylbutyryl

Esterification of core compound E4A with acetyl chloride (H) and separation of the compound by HPLC provides the following compounds:

Acy = acetyl

Esterification of core compound E4A with crotonyl chloride and separation of the compound by HPLC provides the following compounds:

Cro = crotone

Esterification of core compound E4A with cinnamoyl chloride and separation of the compound by HPLC provides the following compounds:

Cin = Cinnamoile

Esterification of core compound E4A with benzoyl chloride and separation of the compound by HPLC provides the following compounds:

Ben = Benzoyl

Esterification of Core Compound E4A with Senecionyl Chloride and Isolation of Compounds by HPLC provides the following compounds:

Esterification of E4A-Tig-N with crotonoyl chloride and separation of the compound by HPLC provides the following compounds:

Esterification of E4A-Tig-N with acetyl chloride and separation of the compound by HPLC provides the following compounds:

Esterification of E4A-Tig-N with 4-pentenoyl chloride and separation of the compound by HPLC provides the following compounds:

Esterification of E4A-Tig-N with hexanoyl chloride and separation of the compound by HPLC affords the following compounds:

Esterification of E4A-Tig-N with cinnamoyl chloride and separation of the compound by HPLC provides the following compounds:

Esterification of E4A-Tig-N with an Angeloyl chloride and separation of the compound by HPLC provides the following compounds:

Esterification of E4A-Tig-N with 2-ethylbutyryl chloride and separation of the compound by HPLC affords the following compounds:

(A), (B), (C), (D), (E) Esterification with acyl chlorides including chloride, 3,3-dimethyl acryloyl chloride, semeny oil chloride, cinnamoyl chloride, pentenoyl chloride, hexanoyl chloride, benzoyl chloride or ethyl butyryl chloride. The compound changes its composition when the reaction time or temperature changes. Peaks, fractions and compounds are selected according to the activity of the time study and the change in peak. Compounds with strong activity to weak activity were selected and isolated. The anticancer activity is expressed in bone (U20S), lung (H460), bladder (HTB-9), ovary (ES2), colon (HCT116), pancreas (Capan), ovary (OVCAR3) (MT) of the cervix (HeLa), leukemia (K562), brain (T98G), liver (HepG2), and kidney (A498). The active esterification product was purified by HPLC. The mixture and the reaction products of the individual compounds were tested for MTT cell activity assays. The details of the method are the same as Experiment 3 of the present invention. The second esterification of the compound can be selected from the above experimental results to form a novel active compound. The partially esterified compound was selected from these experiments to perform the second esterification or to repeat the third esterification with the different acyl chloride to produce the novel active compound by the experiment in the present invention.

The method comprises the steps of 1) dissolving the core compound or triterpenic core, hydroxylated triterpene core in pyridine, 2) adding acyl chloride, and 3) mixing the mixture at different temperatures for 5 seconds, 1 minute, 2 minutes, 5 minutes , 10 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 2 days or 3 days, and 4) at the end of the reaction, an acid or a weak base aqueous solution or water is added to the reaction mixture, 5) the solution is then extracted with ethyl acetate and lyophilized, 6) the reaction product is dissolved in acetonitrile containing trifluoroacetic acid or DMSO, 7) the reaction product of the mixture and the individual fractions is subjected to MTT cytotoxicity assays 8) HPLC fractionation for separation is chosen according to the cytotoxic activity of the reaction product obtained at the particular reaction time, 10) the active esterification product is purified by HPLC, 11) the products are combined and 12) the product Wherein the core compound is a terpene, isoprene, or triterpene core or a hydroxylated triterpene core, the core compound is dissolved in pyridine, and the acyl chloride is selected from the group consisting of tigloyl chloride, angeloyl chloride, acetyl chloride, Tosyl chloride, 3,3-dimethyl acryloyl chloride, semenioyl chloride, cinnamoyl chloride, pentenoyl chloride, hexanoyl chloride, benzoyl chloride and ethyl butyryl chloride; The reaction time for the mixture is 5 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 2 days or 3 days, or a 75 ℃ temperature, the base comprising an acid or NaHC0 ₃ containing HCl is added to the reaction mixture, the solution is extracted three times after with ethyl acetate and freeze-drying, the reaction product is 80% acetonitrile-0.005% The HPLC fraction for separation is dissolved in trifluoroacetic acid or DMSO and the reaction is performed at 5 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 2 days, or 3 days Is selected based on the cytotoxic activity of the reaction product obtained over time. In one embodiment, the reaction time can be at least 3 days. In one embodiment, the experiment can be performed at 0 DEG C or less. In one embodiment, the experiment can be performed at < RTI ID = 0.0 > 75 C < / RTI >

The antitumor activities of Tig-R compounds were as follows: IC50 of bone (U20S) 4.5 ㎍ / ㎖, lung (H460) 4.8 ㎍ / ㎖, bladder (HTB- 9) 2.5 ㎍ / (SK-Mel-5) was 5.1 ㎍ / ㎖ in the ovary (OVCAR3), 5.8 in the prostate (DU145) , Brain (T98G), brain (T98G), brain (T98G), and brain (T98G) were administered at a dose of 5 μg / 8 ㎍ / ㎖ for leukemia (K562), 2 ㎍ / ㎖ for leukemia (K562) and 5 ㎍ / ㎖ for cervix (HeLa).

The antitumor activity of the Tig-V compound was evaluated in the same manner as in Example 1. The antitumor activity of the Tig-V compound was evaluated in the following manner: IC50 of bone (U20S) was 7 μg / ml, lung (H460) was 6.8 μg / ml, bladder (HTB- (SK-Mel-5), the prostate (DU145) was 4 ㎍ / ㎖, and the skin (SK-Mel-5) (T98G), 12 μg / ml of liver (HepG2), 9 μg / ml of breast (MCF-7), 4 μg / ) Was 14 ㎍ / ㎖, leukemia (K562) was 4 ㎍ / ㎖, and cervix (HeLa) was 7 ㎍ / ㎖.

The antitumor activity of the Tig-N compound was evaluated in the following manner: IC50 of bone (U20S) was 15 占 퐂 / ml, lung (H460) was 13 占 퐂 / ml, bladder (HTB- (SK-Mel-5), ovary (OVCAR3) and prostate (DU145) were administered at a dose of 15 μg / ml, pancreas 8 μg / (A498), 11 (㎍ / ㎖) for breast (MCF-7), 18 ㎍ / ㎖ for liver (HepG2) T98G) is 19 ㎍ / ㎖), leukemia (K562) is 6 ㎍ / ㎖, and cervix (HeLa) is 15 ㎍ / ㎖.

The antitumor activities of Tig-Q compounds were 20 ㎍ / ㎖ for the bone (U20S), 18 ㎍ / ㎖ for the lung (H460), 10 ㎍ / ㎖ for the bladder (HTB- (SK-Mel-5), the prostate (DU145) and the skin (SK-Mel-5) were administered at a dose of 20 μg / (A498), 13 (㎍ / ㎖) for breast (MCF-7), 24 ㎍ / ㎖ for liver (HepG2) T98G) is 29 ㎍ / ㎖), leukemia (K562) is 6 ㎍ / ㎖, and cervix (HeLa) is 20 ㎍ / ㎖.

The antitumor activities of Tig-T compounds were 20 ㎍ / ㎖ for the bone (U20S), 21 ㎍ / ㎖ for the lung (H460), 12 ㎍ / ㎖ for the bladder (HTB- (SK-Mel-5), the prostate (DU145) and the skin (SK-Mel-5) were administered at a dose of 10 μg / ml, 23 μg / ml of colon (HCT116), 10 μg / ml of pancreas (Capan), 25 μg / ml of OVCAR3 (T98G), 22 ㎍ / ㎖, 20 ㎍ / ㎖ for liver (HepG2), 13 ㎍ / ㎖ for mouth (KB), 15 ㎍ / ㎖ for kidney ) Was 26 ㎍ / ㎖, leukemia (K562) was 9 ㎍ / ㎖, and cervix (HeLa) was 18 ㎍ / ㎖.

The antitumor activities of Tig-S compounds were as follows: IC50 of bone (U20S) was 5.2 ㎍ / ㎖, lung (H460) was 5.6 ㎍ / ㎖, bladder (HTB- (SK-Mel-5), the prostate (DU145) and the skin (SK-Mel-5) (A498), 4.8 ㎍ / ㎖ in kidney (A498), 6.3 ㎍ / ㎖ in breast (MCF-7), 8.5 ㎍ / ㎖ in liver (HepG2) ), Leukemia (K562) is 4.3 ㎍ / ㎖, and cervix (HeLa) is 7 ㎍ / ㎖.

The antitumor activities of Tig-U compounds were as follows: IC50 of bone (U20S) was 23 ㎍ / ㎖, lung (H460) was 19 ㎍ / ㎖, bladder (HTB- (SK-Mel-5), ovary (OVCAR3), 27 ㎍ / ㎖, prostate (DU145) and 15 ㎍ / (A498), 18 ㎍ / ㎖, breast (MCF-7) 25 ㎍ / ㎖, liver (HepG2) 23 ㎍ / ㎖, brain (1 ㎍ / T98G) was 22 ㎍ / ㎖, leukemia (K562) was 10 ㎍ / ㎖, and cervix (HeLa) was 17 ㎍ / ㎖.

The present invention provides a compound of formula (2A) for the treatment of cancer, inhibiting cancer growth, inhibiting cancer invasion, inhibiting cancer metastasis, regulating cell adhesion and inhibiting cell adhesion, The use of a compound for the manufacture of a medicament, a method, or a medicament for a medicament selected from the group consisting of:

Wherein R 1, R 2, R 3, R 4, R 5, R 8, R 9, R 10, R 11, R 12, R 13, R 14 and R 15 are independently selected from the group consisting of hydrogen, hydroxyl, methyl, O-angeloyl, O- O-acetyl, O-crotonoyl, 0-3,3-dimethyl acryloyl, O-cinnamoyl, O-pentenoyl, O-hexanoyl, O-benzoyl, , O-alkanoyl, O-alkanoyl, O-alkenoyl, O-benzoylalkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl CH20-thienyl, CH2O-acetyl, CH2O-acetyl, CH2O-, and the like, which are optionally substituted with one or more substituents selected from the group consisting of O-alkyl, O-aryl, O-acyl, O-heterocyclyl, O- CH20-O-alkyl, O-alkyl, O-alkyl, O-alkyl, O-alkanoyl, O-alkanoyl, O-alkenoyl, O-benzoylalkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O- , O- acyl, O- heterocyclic, O- heteroaryl, O- alkenyl-carbonyl, alkane, is independently selected from the group consisting of an alkene and the sugar moiety or a derivative thereof; Structure (2A) is selected from the group consisting of O-angeloyl, O-tigloyl, O-semenooxy, O-acetyl, O-crotonoyl, O-3,3-dimethyl acryloyl, O-cinnamoyl, O O-alkanoyl, O-benzoylalkyl substituted O-benzoylalkyl, O-benzoylalkyl, O-benzoylalkyl, O-benzoyl, O- At least two groups selected from O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocyclic, O-heteroaryl, O-alkenylcarbonyl Include; R 1 and R 2 are independently selected from the group consisting of O-angeloyl, O-thioglooyl, O-semenooxy, O-acetyl, O-crotonoyl, O-3,3-dimethyl acryloyl, O- Alkanoyl, O-alkanoyl, O-benzoylalkyl substituted O-alkanoyl, O-benzoylalkyl, O-alkanoyl, O-benzoylalkyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocyclic, O-heteroaryl, O-alkenylcarbonyl; In one embodiment, R < 1 > and R < 2 > In one embodiment, R4 and R10 are independently selected from the group consisting of CH2O-angeloyl, CH2O-thiogloyl, CH2O-senesy oil, CH2O-acetyl, CH2O- crotonoyl, CH2O3,3-dimethyl acryloyl, Cinnamoyl, CH2O-pentenoyl, CH2O-hexanoyl, CH2O-benzoyl, or CH2O-ethylbutyryl. In one embodiment, R 3 and R 8 are hydrogen or hydroxyl. In one embodiment, R9, R11, R12, R13, R14, R15 are independently bonded to methyl. In one embodiment, R4 is CH3, CHO, CH2R6 or COR6 wherein R6 is selected from the group consisting of hydroxyl, O-angeloyl, O-thioglyoyl, O- O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkanoyl, O-alkenoyl, O-benzoylalkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O- , O-alkenylcarbonyl, and derivatives thereof; In one embodiment, R < 3 > is H or OH; In one embodiment, R8 is H or OH and in one embodiment, R16 is H, CH3, OH or R4 and R16 together are -CH2-X-, CH (OH) -X- or C X-, wherein -X- can be O or NH or S; When C12-13 of ring 3 of triterpene has a double bond, R16 is a member. In one embodiment, R10 is CH3, CHO, or CH2R6 wherein R6 is selected from the group consisting of hydroxyl, O-angeloyl, O-thioglyoyl, O- O-benzoyl, O-benzoyl, O-benzoyl, O-benzoyl, O-benzoyl, O-ethylbutyryl, O- Alkanoyl, O-alkenoyl, O-benzoylalkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocyclic, O -Heteroaryl, O-alkenylcarbonyl, and derivatives thereof.

In one embodiment, R5 is a hydrogen, hydroxyl, heterocyclic or O-sugar moiety (s) wherein the sugar moiety (s) is selected from the group consisting of glucose, galactose, rhamnose, arabinose, xylose, fucose, , Maltose, gulose, gulose, idose, ricinose, mannose, psicose, ribose, sorbose, tagatose, talose, fructose, aldonic acid, glucuronic acid, galacturonic acid and derivatives or combinations thereof Lt; / RTI > R9, R10, R11, R12, R13, R14 and R15 are independently selected from the group consisting of CH3, CH2OH, CHO, COOH, COO-alkyl, COO-aryl, COO- heterocyclic, COO- CH2O-heteroaryl, an alkyl group, a hydroxyl, an acetyl group; R4 and R16 form a divalent radical of the formula CH2O, CH (OR7) O, or COOR7 where R7 is hydrogen, alkyl, angeloyl, tigloyl, senesinoyl, dibenzoyl, benzoyl, alkanoyl, Alkenoyl, benzoylalkyl substituted alkanoyl, aryl, acyl, heterocyclic, heteroaryl, and derivatives thereof; At least two of R 1, R 2 and R 6 are selected from the group consisting of O-angeloyl, O-tigloyl, O-semenoxy, O-acetyl, O- O-alkanoyl, O-benzoylalkyl substituted O < RTI ID = 0.0 > -Alkanoyl, O-aryl, O-acyl, O-heterocyclic, O-heteroaryl, and derivatives thereof; At least one of R 1, R 2 and R 4 is selected from the group consisting of geloyl, acetyl, tigloyl, senesy oil, crotonoyl, 3,3-dimethyl acryloyl, cinnamoyl, pentenoyl, hexanoyl, A sugar moiety having at least two groups selected from the group consisting of acyl, acyl, acyl, heterocycle, heteroaryl, and derivatives thereof; R4 is CH2R6 wherein R6 is selected from the group consisting of hydroxyl, O-angeloyl, O-thioglooyl, O-semenooxy, O-acetyl, O-crotonoyl, 0-3,3-dimethyl acryloyl, O-cinnamoyl, O-pentenoyl, O-hexanoyl, O-ethyl butyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O- O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocyclic, O-heteroaryl, O-alkenylcarbonyl and derivatives thereof Lt; / RTI > R5 is a sugar moiety (s) selected from the following sugars and aldulonic acids: glucose, galactose, rhamnose, arabinose, xylose, fucose, alos, altrose, gulose, idose, ricosus, mannose, Wherein R5 is selected from the group consisting of hydroxyl, O-angeloyl, O-thioglooyl, thiomorpholinyl, thiomorpholinyl, , O-semenoxy, O-acetyl, O-crotonoyl, 0-3,3-dimethyl acryloyl, O-cinnamoyl, O-pentenoyl, O-hexanoyl, O- O-alkanoyl, O-alkanoyl, O-alkenoyl, O-benzoylalkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituent O-aryl, O-acyl, O-heterocyclic, O-heteroaryl, O-alkenylcarbonyl and derivatives thereof. In one embodiment, R 1, R 2, R 3, R 4, R 5, R 8, R 9, R 10, R 11, R 12, R 13, R 14 or R 15 comprise one or more sugar moieties. In one embodiment, R1, R2, R3, R4, R5, R8, R9, R10, R11, R12, R13, R14 or R15 comprise at least one acid. In one embodiment, at least one, or two, or three, or four of R 1, R 2, R 3, R 4, R 5, R 8, R 9, R 10, R 11, R 12, R 13, R 14 and R 15 are hydroxyl. In one embodiment, at least 2 or 3 or 4 or 5 or 6 or 7 of R 1, R 2, R 3, R 4, R 5, R 8, R 9, R 10, R 11, R 12, R 13, R 14 and R 15 is O- O-cinnoyoyl, O-acetyl, O-crotonoyl, 0-3,3-dimethyl acryloyl, O-cinnamoyl, O-pentenoyl, O- O-alkanoyl, O-alkanoyl, O-alkanoyl, O-benzoylalkyl substituted O-alkanoyl, O-alkanoyl, O-benzoyl, O-benzoyl, O-ethylbutyryl, O-alkyl, O-dibenzoyl, O-alkenylcarbonyl, alkanes, alkenes, and derivatives thereof, wherein the phenyl ring is optionally substituted with one or more substituents selected from the group consisting of phenyl, heteroaryl, heteroaryl, heteroaryl, heteroaryl, Wherein at least one or two or three or more of R 1, R 2, R 3, R 4, R 5, R 8 and R 10 are bonded to the triterpene directly or by linking moiety (s) 4 or 5 or 6 or 7 are O-angeloyl, O-tigloyl, O-senecio O-acetyl, O-crotonoyl, 0-3,3-dimethyl acryloyl, O-cinnamoyl, O-pentenoyl, O-hexanoyl, O-benzoyl, Alkanoyl, O-alkanoyl, O-alkenoyl, O-benzoylalkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O- CH20-thienyl, CH20-seceneoxy, CH20-acetyl, CH20-crotono, Benzoyl, CH20-ethylbutyryl, CH3, CH2OH, O-alkyl, O-dibenzoyl, O-alkanoyl, O-alkenoyl, O-benzoylalkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O- O-heterocyclyl, O-heteroaryl, O-alkenylcarbonyl, and derivatives thereof. And the group is bonded directly or by connection portion (s) to the triterpene. In one embodiment, the cancer is selected from the group consisting of breast cancer, leukocyte cancer, liver cancer, ovarian cancer, bladder cancer, prostate cancer, skin cancer, bone cancer, brain cancer, leukemia cancer, lung cancer, colon cancer, Kidney cancer, cervical cancer, esophageal cancer, testicular cancer, spleen cancer, kidney cancer, lymphatic cancer, pancreatic cancer, stomach cancer and thyroid cancer; The cell may be a breast cell, a white blood cell, a liver cell, an ovary cell, a bladder cell, a prostate cell, a skin cell, a bone cell, a brain cell, a leukemia cell, a lung cell, a colon cell, a CNS cell, Cells, esophageal cells, testicular cells, spleen cells, kidney cells, lymphatic cells, pancreatic cells, gastric cells and thyroid cells. In one embodiment, the compound is selected from the following structures:

R13, R14 and R15 are independently selected from the group consisting of CH3, CH20H, hydrogen, hydroxyl, methyl, O-angeloyl, O- O-cyclodecanyl, 0-3-dimethyl acryloyl, O-cinnamoyl, O-pentenoyl, O-hexanoyl, O-benzoyl, O- O-alkanoyl, O-alkanoyl, O-benzoylalkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl O-aryl, O-acyl, O-heterocyclic, O-heteroaryl, O-alkenylcarbonyl, CH2O-angeloyl, CH2O-tigloyl, CH2O- CH2O-alkyl, CH2O-alkyl, < RTI ID = 0.0 > CH2O- < / RTI > Benzoyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoylalkyl substituted O-alkanoyl, CH2O-alkanoyl substituted phenyl, CH2O- Hwandoen phenyl, CH2O- aryl, acyl CH2O-, CH2O- heterocyclic, CH2O- heteroaryl, CH2O- alkenyl-carbonyl, alkanes, independently selected from the group of alkenes and sugar moiety, or derivatives thereof;

One or two or three or four of R 1, R 2, R 3, R 4, R 5, R 8 and R 10 is O-angeloyl, O-thioglyoyl, O- O-benzoyl, O-benzoyl, O-benzoyl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoylalkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocyclic , CH2-O-heteroaryl, O-alkenylcarbonyl, CH2O-angeloyl, CH2O-tungoyl, CH2O-senesy oil, CH2O- acetyl, CH2O- crotonoyl, CH2O- CH2O-cinnamoyl, CH2O-pentenoyl, CH2O-hexanoyl, CH2O-benzoyl, CH2O-ethylbutyryl; R9, R11, R12, R13, R14, R15 are independently bonded to CH3; R10 is selected from the group consisting of O-angeloyl, O-tigloyl, O-semenooxy, O-acetyl, O-crotonoyl, 0-3,3-dimethyl acryloyl, O-cinnamoyl, O-alkanoyl, O-benzoylalkyl substituted O-alkanoyl, O-benzoylalkyl, O-benzoyl, O-benzoyl, O-ethylbutyryl, O-alkyl, O-dibenzoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocyclic, O-heteroaryl, O-alkenylcarbonyl, CH20-cinnamoyl, CH20-acetyl, CH20-crotonoyl, CH20-3,3-dimethyl acryloyl, CH20-cinnamoyl, CH20-pentenoyl, CH20-hexanoyl, CH20-ethylbutyryl, CH20-alkyl, CH20-dibenzoyl, CH20-benzoyl, CH20-alkanoyl, CH20-alkenoyl, CH20-benzoylalkyl substituted O-alkanoyl, CH20- Alkanoyl substituted phenyl, CH20-aryl, CH2O-acyl, CH2O-heterocycle, CH2O-heteroaryl, CH2O- alkenylcarbonyl and Or R4 and R10 are selected from the group consisting of O-angeloyl, O-tigloyl, O-senesy oil, O-acetyl, O-crotonoyl, 0-3,3-dimethyl acryloyl, O- O-pentenoyl, O-hexanoyl, O-benzoyl, O-ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O- O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocyclic, O-heteroaryl, O-alkenylcarbonyl, CH20-thienyl, CH20-semenooxy, CH20-acetyl, CH20-crotone, CH20-3,3-dimethylacryloyl, CH20-cinnamoyl, CH20-pentenoyl, CH20-benzoyl, CH20-ethylbutyryl, CH20-alkyl, CH20-dibenzoyl, CH20-benzoyl, CH20-alkanoyl, CH20-alkenoyl, CH20-benzoylalkyl substituted O-alkanoyl, Phenyl, CH20-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocycle, CH2O-heteroaryl, CH2 Independently, is 0-alkenylcarbonyl; R3 is OH or H or absent; R1, R2, R3, R5, R8 are OH or H or absent; R9, R11, R12, R13, R14, and R15 are CH3; R1, R2, R5, R8 are OH; R3 is OH, H or absent; R4, R10 are CH20 < / RTI > R9, R11, R12, R13, R14, R15 are CH3; R1, R2, R5, R8 are OH or O-Tigloyl; R3 is OH, H, or member; R4 and R10 are < RTI ID = 0.0 > CH20 < / RTI > R9, R11, R12, R13, R14, R15 are CH3; Benzoyl, ethylbutyryl, alkyl, dibenzoyl, benzoyl, cyclopentyl, cyclohexyl, cyclohexyl, cyclohexyl, cyclohexyl, cyclohexyl, Alkanoyl, alkanoyl, benzoylalkyl substituted alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocyclic, heteroaryl, and alkenylcarbonyl Lt; / RTI > are interchangeable. These may be the same group or a combination thereof. Substitution, removal and / or addition of any group in the compounds described above by other group (s) will be apparent to those skilled in the art based on the teachings of the present invention. In other embodiments, the substitution, removal and / or addition of the group (s) in the compounds of the present invention do not substantially affect the biological function of the compound.

In one embodiment, the compound is selected from the following structures:

A composition comprising an effective amount of a compound selected from the above formula or a salt, ester, metabolite or derivative thereof may be used as a medicament for blocking invasion, migration and metastasis of cancer cells, inhibiting tumor or cancer cell growth, Wherein the cancer is selected from the group consisting of breast cancer, leukocyte cancer, liver cancer, ovarian cancer, bladder cancer, prostate cancer, skin cancer, bone cancer, brain cancer, leukemia cancer, lung cancer, colon cancer, , Cervical cancer, esophageal cancer, testicular cancer, splenic cancer, kidney cancer, lymphatic cancer, pancreatic cancer, stomach cancer and thyroid cancer.

The present invention relates to a method for treating cancer, inhibiting a virus, preventing brain aging, improving memory, and improving brain function; Enuresis, frequent urination, urinary incontinence; Dementia, Alzheimer's disease, autism, brain trauma, Parkinson's disease or other disorders caused by brain dysfunction; Arthritis, rheumatism, poor blood circulation, arteriosclerosis, Raynaud's syndrome, angina, heart disease, coronary heart disease, headache, dizziness, kidney disease; For treating cerebrovascular disease; For inhibiting NF-kappa B activation; Treating brain edema, severe acute respiratory syndrome, viral respiratory disease, chronic venous insufficiency, hypertension, chronic venous disease, o edema, inflammation, hemorrhoids, peripheral edema formation, varicose veins, flu, post-traumatic edema and postoperative pain for; For inhibiting thrombosis; To inhibit ethanol absorption; To lower blood sugar; There is provided a composition comprising a compound provided herein for modulating adrenoceptor corticotropin and corticosterone levels. The present invention relates to a pharmaceutical composition for preventing or treating an anti-MS, anti-aneurysm, anti-asthma, anti-edema, inflammation, anti-Bradykin, hematocrit, anti headache, Anti-inflammatory agents, anti-inflammatory agents, anti-inflammatory agents, anti-inflammatory agents, anti-inflammatory agents, anti-inflammatory agents, antiinflammatory agents, anti-inflammatory agents, antiinflammatory agents, antiinflammatory agents, antihypertensive agents, antihistamines, antihypertratics, Anti-varicose, anti-dizziness, cancer suppression, corticosteroid source, diuretic, fungicide, hemolyzing agent, hyaluronidase inhibition, lymphatic material, sodium diuretic, insecticide, pituitary stimulant, thymus tissue dissolution, vascular protection, , Adhesion of cancer cells or regulation of angiogenesis, anti-parasites; An increase in the expression of genes, ANGPT2, DDIT3, LIF and NFKB1 Z, and a composition for the manufacture of an adjuvant composition and a treatment for a venous incision.

Alkenyl means an unsaturated linear or branched structure having one or more double bonds and the formula R2C = CR2, and combinations thereof. Examples of alkenyl groups include vinyl, propenyl, isopropenyl, butenyl, s- and t-butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, and hexadienyl.

Aryl is a functional group of an organic molecule derived from an aromatic compound such as benzene, a 6- to 14-membered carbocyclic aromatic ring system containing 1-3 benzene rings. When two or more aromatic rings are present, 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 substituents independently selected from halogen, alkyl or alkoxy.

Acyl is a functional group that can be obtained from an organic acid by removal of a carboxyl group. Acyl groups may be described using the formula -COR wherein a double bond is present between carbon and oxygen. The name of the acyl group is usually terminated as -O-, examples of which include polymethyl, acetyl, propionyl, butyryl and benzoyl. Benzoyl is one of the acyl, C6H5COR derived from benzoic acid by removal of the carboxyl.

A heterocyclic compound is a compound containing a heterocyclic ring referred to as a non-aromatic ring having from one to four heteroatoms, wherein said ring is a three- or four-membered ring containing 0 to 4 heteroatoms, aryl and heteroaryl, The heterocyclic compound is selected from the group consisting of pyrrolidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, imidazolinyl, thiomorpholinyl, and the like. .

Heterocyclyl groups are derived from heteroarenes by removing hydrogen atoms from any ring atom.

Alkanyl is a generic name for the organic functional group RCO-, where R is hydrogen or an alkyl group. Examples of alkanoyl are acetyl, propionyl, butyryl, isobutyryl, pentanoyl and hexanoyl.

Alkenoyl is alkenylcarbonyl, wherein alkenyl is as defined above. Examples are pentenoyl (thiogloyl) and hexenoyl (angeloyl).

Alkyl is a radical containing only carbon and oxygen atoms arranged in a chain, branched, cyclic or bicyclic structure, or combinations thereof, having from 1 to 18 carbon atoms. Examples are methyl, ethyl, propyl, isopropyl, butyryl, s- and t-butyl, pentyl, hexyl, heptyl, octyl, nonyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, cyclopropyl, But are not limited to, butyl, cyclobutyl, and cyclohexyl.

Benzoylalkyl substituted alkanoyl refers to a linear or branched alkanoyl substituted with at least one benzoyl and at least one alkyl, wherein the benzoyl is attached to a linear or branched alkyl. An example of a benzoylalkyl substituted alkanoyl is benzoylmethylisobutanoyl.

The sugar moiety is a segment of a molecule comprising at least one sugar or derivative thereof or an allylonic acid thereof.

Isobutyryl is a synonym for 2-methylpropanoyl.

(Y) Y3, Y and Y3 are the same compounds.

YM and (ACH-Y) are the same compounds.

The linking moiety is a substructure or atomic group linking a functional group to a core compound. For example, angeloyl is linked to the triterpene core by sugar moieties.

, 안겔로일 클로라이드

, 티글로일 클로라이드

, 세네시오일 클로라이드

, 크로토노일 클로라이드

, O-3,3-디메틸아크릴로일 클로라이드

, 신나모일 클로라이드

, 펜테노일 클로라이드

, 헥사노일 클로라이드

, 벤조일 클로라이드

, 에틸부티릴 클로라이드

를 포함한다.The building blocks used in the present invention are selected from the group consisting of triterpenes, hydroxylated triterpenes, acetyl, angeloyl, thiogloyl, senesy oil, crotonoyl, 0-3,3-dimethyl acryloyl, cinnamoyl, Alkanoyl, alkanoyl, benzoylalkyl-substituted alkanoyl, alkanoyl-substituted phenyl, alkenoyl-substituted alkanoyl, alkanoyl, alkanoyl, Substituted phenyl, aryl, acyl, heterocyclic, heteroaryl, alkenylcarbonyl, acetyl chloride, angeloyl chloride, thiogloyl chloride, semenioyl chloride, crotonoyl chloride, 0-3,3- Toluenesulfonyl chloride, toluenesulfonyl chloride, toluenesulfonyl chloride, toluenesulfonyl chloride, toluenesulfonyl chloride, benzylsulfonyl chloride,

, Angeloyl chloride

, Tigloyl chloride

, Senesioyl chloride

, Crotonoyl chloride

, O-3,3-dimethyl acryloyl chloride

, Cinnamoyl chloride

, Pentenoyl chloride

, Hexanoyl chloride

, Benzoyl chloride

, Ethyl butyryl chloride

.

In the experiments described, the concentration of drug inhibiting 15% cell-growth or less (i. E., 85% or more of the control) compared to the non-drug control (DMSO) is considered a non-cytotoxic concentration. In one embodiment, the concentration of the drug that inhibits 10% cell-growth or less (i. E., 90% or more of the control) compared to the non-drug control (DMSO) is considered a non-cytotoxic concentration. In one embodiment, the concentration of the drug that inhibits 5% cell-growth or less (i. E., 95% or more of the control) compared to the non-drug control (DMSO) is considered a non-cytotoxic concentration. In one embodiment, the concentration of the drug that inhibits 20% cell-growth or less (i.e., 80% or more of the control) compared to the non-drug control (DMSO) is considered a non-cytotoxic concentration. In one embodiment, the concentration of the drug that inhibits 25% cell-growth or less (i. E., 75% or more of the control) compared to the non-drug control (DMSO) is considered a non-cytotoxic concentration. In one embodiment, the concentration of the drug that inhibits 30% cell-growth or less (i. E., 70% or more of the control) compared to the non-drug control (DMSO) is considered a non-cytotoxic concentration. In one embodiment, the concentration of the drug that inhibits 45% cell-growth or less (i. E., 55% or more of the control) compared to the non-drug control (DMSO) is considered a non-cytotoxic concentration.

A triterpene compound or a compound selected from the present invention can be administered to a subject in need thereof to treat the subject, which can prevent cancer, provide side effects to the subject, inhibit the onset or proliferation of cancer, Killing tumor cells, or inhibiting cancer cell infiltration. In one embodiment, the compound inhibits the activation of nuclear factor-kB, wherein it inhibits ocalization or binds DNA. In one embodiment, the compound induces apoptosis of cancer cells.

Tables 1 to 12, the effect of Y and YM on gene expression (PCT / US2008 / 002086, filed February 15, 2008, Tables 1 to 12 of 1188-ALA-PCT, incorporated herein by reference) Table 13 to Table 19, Effects of Y and YM on gene expression (PCT / US2009 / 034115 filed February 15, 2008, Tables 13-19 of 1188-D-PCT are incorporated herein by reference).

Measurement of gene expression by real-time PCR method ( Brilliant QPCR , Agilent Technologies) : Real-time polymerase chain reaction further confirms the results obtained from microarray analysis. Real-time PCT results (described below) confirm that compounds Y3 and YM increase the expression of the genes (ANGPT2, DDIT3, LIF and NFKB1Z), see PCT / US09 / 34115 ≪ / RTI > are incorporated herein by reference.

Saponins are partially hydrolyzed into a mixture of products which can be separated by HPLC. Partial hydrolysis of saponins can also be achieved by enzymes. Glycosidases catalyze the hydrolysis of glycosidic linkages. Galactosidase is an enzyme that catalyzes the hydrolysis of galactoside. Glucosidase is an enzyme that breaks down the cucuron from saponin. Examples of other enzymes include silanase, lactase, amylase, kittinase, sucrase, maltase, and neuraminidase.

The sugar moiety of triterpenoid saponin (e.g., Zonifolia Y) can be removed by hydrolysis. A synthetic compound of ACH-Y is obtained. ACH-Y is a triterpene having an acyl group without a sugar moiety. The acyl group of saponin (e.g., Zonifolia Y) can be removed by alkaline hydrolysis. Synthetic compound AKOH-Y can be obtained. AKOH-Y is a pentacyclic triterpene having a sugar moiety. Pentacyclic triterpenes can be obtained by acid and alkali hydrolysis of saponins from natural sources. Pentacyclic triterpenes can be obtained by synthetic methods (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 having sugar moieties can also be obtained by synthesis (Pie et al., Synthesis of L-arabinopyranose containing hederagenin saponins, Tetrahedron 61 (2005) 4347-4362). Acylation is the step of adding an acyl group to the compound. The Friedel-Crafts reaction is an example of such a process. The active compound can be obtained by acylating pentacyclic triterpene, or hydroxylated triterpene. In one embodiment, the acylations C24, C28, C21, and C22 of the pentacyclic triterpene, or hydroxylated triterpene, inhibit cancer growth, cancer invasion, cell invasion, cancer cell invasion, cell adhesion or cell cycle ≪ / RTI > In one embodiment, the acyl group (s) may be present at C3. In one embodiment, the sugar moiety is at C21, 22, or 28, wherein the sugar moiety is combined with two acyl groups. In one embodiment, the acylation of compounds (A), (B), (C), (D), (F), (G), and (H) is selected from the group consisting of cancer infiltration, cell infiltration or cancer cell infiltration; Cancer metastasis; Or to inhibit cancer growth. The building blocks in the present invention are used to synthesize active saponins.

The acylation of the compound (G) and the anthroyl or tigloyl group provides the following compounds:

Wherein R1, R2, R5, R8 are OH or O-angeloyl; R3 is OH, H or O-angeloyl; R4, R10 are CH3, CH2OH or CH20 angelyl; R3 is OH, H or O-angeloyl; R9, R11, R12, R13, R14, R15 are CH3; R1, R2, R5, R8 are OH or O-Tigloyl; R3 is OH, H or O-Tigloyl; R <4>, R <10> is CH3, CH20H or CH20 tigloyl; R9, R11, R12, R13, R14, R15 are CH3; Wherein the compound inhibits cancer growth, cancer invasion, cell invasion or cancer cell invasion.

The compound (G) is reacted with a compound represented by the general formula (I): wherein R 1 and R 2 are independently selected from the group consisting of an alkylene group, an alkylene group, an alkylene group, an alkenyl group, Alkanoyl, alkanoyl, benzoylalkyl substituted O-alkanoyl, alkanoyl substituted phenyl, alkenoyl substituted phenyl, aryl, acyl, heterocyclic, heteroaryl, CH20-alkenylcarbonyl, alkane , Acylation of an alkene provides compound (K) wherein R1, R2, R5 and R8 are independently selected from the group consisting of OH, O-angeloyl, O-thioglyoyl, O- O-benzoyl, O-ethylbutyryl, O-alkyl, O-dibenzoyl, O-dibenzoyl, O- O-alkanoyl, O-alkenoyl, O-benzoylalkyl substituted O-alkanoyl, O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-hetero O-heteroaryl, O-alkenylcarbonyl; R4 and R10 are independently selected from the group consisting of CH3, CH2OH, CH2O-angeloyl, CH2O-thiogloyl, CH2O-senesy oil, CH2O- acetyl, CH2O- crotonoyl, CH2O3,3-dimethyl acryloyl, CH2O-benzoyl, CH2O-alkanoyl, CH2O-alkenoyl, CH2O-benzoylalkyl substituents such as CH2O-pentenoyl, CH2O-hexanoyl, CH2O-benzoyl, CH2O- Alkanoyl substituted phenyl, CH2O-alkenoyl substituted phenyl, CH2O-aryl, CH2O-acyl, CH2O-heterocycle, CH2O-heteroaryl, CH2O-alkenylcarbonyl, alkane, Alkene; R3 is a member selected from the group consisting of OH, H, O-angeloyl, O-tigloyl, O-semenooxy, O- acetyl, O- O-pentanoyl, O-hexanoyl, O-benzoyl, O-ethylbutyryl, O-alkyl, O-dibenzoyl, O-benzoyl, O-alkanoyl, O-alkenoyl, O-benzoylalkyl O-alkanoyl substituted phenyl, O-alkenoyl substituted phenyl, O-aryl, O-acyl, O-heterocyclic, O-heteroaryl, O-alkenylcarbonyl; R9, R11, R12, R13, R14, R15 are CH3; Wherein the compound inhibits cancer growth, cancer invasion, cell invasion or cancer cell invasion; The compound is for use as a mediator or inhibitor of adhesion proteins or angiopoietin; The use of compounds as mediators of the secretion, expression, or synthesis of adherent proteins reduces fibronectin to inhibit cell adhesion, cell adhesion or cellular circulation, where attachment proteins include fibronectin, integrin family, myosin, , Collagen, laminin, polyglycan, carderine, heparin, tenascin, CD ₅₄ , and CAM; The compounds are used to target adhesion molecules for adhesion therapy and for therapeutic purposes.

Applicants further describe that targeting of adhesive molecules for anti-adherence therapy and therapy is a new direction for the development of drugs. Some examples of anti-adhesive drugs in clinical trials are Efalizumab, Odulimomab, Alicaforsen, Aselizumab, etc., which alter the target adhesion protein [ Reference, TEXT BOOK, Adhesion Molecules: Function and Inhibition, (Reference 2), edited by Klaus Ley page 289-291, 297]

Adhesion molecules in inflammatory disease (cf. 4), Abstract, line 7-8 "Blockade of expression of CAM has emerged as a new therapeutic target in inflammatory diseases". Applicants' invention is an adhesion therapy which is a novel use of compounds as an agent or an inhibitor of adhesion proteins and angiopoietin. This inhibits excessive adhesion and inhibits cell adhesion. In the present invention, Applicants have used compounds selected from Structure (2A) as mediators or inhibitors of adhesion proteins and angiopoietin for antiadhesive therapy and for modulation of cell attachment and cell adhesion.

Experiment details

Herb extraction, analysis of extracted components by HPLC, determination of the cell-growth activity achieved by Johnny Polyya Y from cells derived from different human organs using MTT assay, purification of viable components from plant extracts, purification by FPLC Experimental details of fractionation of plant extracts, isolation of component Y by preparative HPLC, determination of chemical structure, cell experiments and animal studies are described in PCT / US05 / 31900, US Serial No. 11/289142, US Serial No. 10/906303, US Serial No. 11/131551 and US Serial No. 11/683198 (filed March 7, 2007), PCT / US 2007/077273 (filed August 30, 2007) No. 60 / 890,380 (filed February 16, 2007), US Serial No. 60 / 947,705 (filed July 3, 2007), PCT / US2008 / 002086, 1188-ALA-PCT Filed on February 15, 2009), the contents of which are incorporated herein by reference. Experiments 1 to 23 in PCT / US2008 / 002086, 1188-ALA-PCT filed on February 15, 2008 are incorporated herein by reference.

Experiment 1: Removal of 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 a water bath at 80 ° C for 5 hours. Afterwards, the solution was neutralized by adding 2 ml of 1 N NaOH (to a final pH of 4-6) and the aglycon was subsequently extracted with 3 ml x 2 of ethyl acetate. The extracts were combined and brought together. Additional separation of aglycon (sugar-removed saponin) was achieved by HPLC with isocratic elution of 80-100% acetonitrile.

Experiment 2: Elimination of acyl groups by alkali hydrolysis

Method: 20 mg of saponin was dissolved in 0.5 ml of 1N NaOH. The solution was incubated in a water bath at 80 캜 for 4 hours. It was cooled to room temperature and then neutralized with 0.5 ml 1N HCl (pH adjusted to about 3). The mixture was extracted three times with 2 ml of 1-butanol. The butanol fraction was collected and lyophilized. The hydrolyzed saponin further purified by HPLC on a C-18 column was eluted with 25% acetonitrile.

Experiment 3: In esterification  by Tritterpen  Addition of acyl group

Method: In a 50 ml tube, 40 mg of triterpene cores (fraction IV) were dissolved in 1 ml of pyridine. 0.2 ml of acyl chloride (tgyloyl chloride, angeloyl chloride, acetyl chloride, crotonoyl chloride, 3,3-dimethyl acryloyl chloride (senesioyl chloride), cinnamoyl chloride, pentenoyl chloride, hexanoyl Chloride, benzoyl chloride or ethyl butyryl chloride). The mixture was stirred for 5 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 2 days or 3 days at 0 캜, 25 캜 or 75 캜. At the end of the reaction, it was added 5 ml of 2N HCl or 1M NaHC0 ₃ in the reaction mixture. The solution was then extracted three times with 10 ml ethyl acetate, then evaporated under vacuum and at 45 &lt; 0 &gt; C and lyophilized. The reaction product was dissolved in 80% acetonitrile-trifluoroacetic acid or DMSO and separated by HPLC. The HPLC fraction for separation was chosen according to the cytotoxic activity of the reaction products obtained at the particular reaction time. The active esterification product was purified by HPLC. The mixtures and the reaction products of the individual compounds were tested by MTT cytotoxicity assays (cf. FIGS. 1 to 12).

Experiment 4: E4A Manufacturing

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 DEG C for 5 hours.

4. The solution was neutralized with NaOH.

5. The hydrolyzed product was extracted with ethyl acetate and then removed by evaporation.

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

Experiment 5: E4A of Tiglillo  Chloride Esterification

1. 50 mg of E4A in 1 ml pyridine was gently stirred in a 50 ml tube. Esterification was carried out at 25 [deg.] C by adding 200 [mu] l of thiogloyl chloride.

2. After stirring for 1 minute, immediately add 5 ml 2N HCl.

3. Stir for 1 hour and stir overnight at room temperature.

4. The esterification product was extracted with 10 ml ethyl acetate.

5. Ethyl acetate was evaporated.

6. Samples were dissolved in 1 ml DMSO.

7. The reaction product was fractionated by HPLC.

8. Samples were combined.

Experiment 6: HPLC Of E4A - Tig  Isolation of active compounds

1. Column: ZORBAX ODS 9.4 x 250 mm, 5 탆

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

3. Chromatographic conditions: a) elution: solvents A to B in 80 min; Then, for 70 minutes, the solvent B; Flow rate: 1 ml / min, c) Monitor OD: 207 nm.

Experiment 7: MTT  Experiment

cell. (Breast), HCT116 (colon), HepG2 (liver), U20S (bones), HTB-9 (bladder), HeLa-S3 (cervix), DU145 (prostate), H460 ), T98G (brain), SK-MEL-5 (skin) and OVCAR 3, ES2 (ovary), pancreas (Capan), mouth (KB), kidney (A498).

MTT assay. The MTT assay procedure follows the method described by the modified document [Carmichael et al. (1987)]. Cells were seeded in 96-well plates for 24 hours before drug treatment. Subsequently, the cells were exposed to the drug for 48, 72, or 96 hours. After drug-treatment, MTT (0.5 mg / mL) was added to the culture and incubated for 1 hour. Formazan (reduced product of tetrazolium by viable cells) was formed, dissolved in DMSO and OD at 490 nm and measured by ELISA reader. The MTT level of cells before drug-treatment was also measured (TO). The cell-growth rate (% G) was calculated according to the formula:% G = (TD-T0 / TC-T0) x 100 (1), where TC or TD represents the OD reading of the drug-treated cells. Cytotoxicity (LC) expressed as% of control when TO> TD was calculated as:% LC = (TD-TO / TO) x 100 (2).

Experiment 8: E4A - Tig -R chemical synthesis, separation and characterization

Chemical synthesis of E4A-Tig-R: 1. Preparation of E4A; 2. Esterification of E4A with thiogloyl chloride; 3. Isolation of E4A-Tig-R by HPLC

Measurement of cytotoxic activity: 1. MTT assay

Determination of chemical structure: 1. NMR analysis; 2. Mass spectrum analysis

Reference 23-30

Reference Table 1

compound E4A - Tig -R

24,28-0- Tiglillo -3β, 16α, 21β, 22α, 24β, 28- Hexahydroxy oleane -12- yen

Experiment 9: E4A - Tig Chemical synthesis, separation and characterization of -N

Chemical synthesis of E4A-Tig-R: 1. Preparation of E4A; 2. Esterification of E4A with thiogloyl chloride; 3. Isolation of E4A-Tig-N by HPLC

Measurement of cytotoxic activity: 1. MTT assay

Determination of chemical structure: 1. NMR analysis; 2. Mass spectrum analysis

Experiment 10: E4A - Tig Chemical synthesis, separation and characterization of -Q

Chemical synthesis of E4A-Tig-R: 1. Preparation of E4A; 2. Esterification of E4A with thiogloyl chloride; 3. Isolation of E4A-Tig-Q by HPLC

Measurement of cytotoxic activity: 1. MTT assay

Determination of chemical structure: 1. NMR analysis; 2. Mass spectrum analysis

Experiment 11: E4A - Tig Chemical synthesis, separation and characterization of -V

Chemical synthesis of E4A-Tig-V: 1. Preparation of E4A; 2. Esterification of E4A with thiogloyl chloride; 3. Isolation of E4A-Tig-V by HPLC

Measurement of cytotoxic activity: 1. MTT assay

Determination of chemical structure: 1. NMR analysis; 2. Mass spectrum analysis

Experiment 12: E4A - Tig -T chemical synthesis, separation and characterization

Chemical synthesis of E4A-Tig-T: 1. Preparation of E4A; 2. Esterification of E4A with thiogloyl chloride; 3. Isolation of E4A-Tig-T by HPLC

Measurement of cytotoxic activity: 1. MTT assay

Determination of chemical structure: 1. NMR analysis; 2. Mass spectrum analysis

Experiment 13: E4A - Tig -U chemical synthesis, separation and characterization

Chemical synthesis of E4A-Tig-S: 1. Preparation of E4A; 2. Esterification of E4A with thiogloyl chloride; 3. Isolation of E4A-Tig-S by HPLC

Measurement of cytotoxic activity: 1. MTT assay

Determination of chemical structure: 1. NMR analysis; 2. Mass spectrum analysis

Experiment 14: E4A - Tig -S chemical synthesis, separation and characterization

Chemical synthesis of E4A-Tig-S: 1. Preparation of E4A; 2. Esterification of E4A with thiogloyl chloride; 3. Isolation of E4A-Tig-S by HPLC

Measurement of cytotoxic activity: 1. MTT assay

Determination of chemical structure: 1. NMR analysis; 2. Mass spectrum analysis

Experiment 15: Esterification of acetyl, angeloyl, thiogloyl, senesy oil, crotyl, cinnamoyl, pentenoyl, hexanoyl, ethylbutyryl with the method used in Experiment 8 provided the following compounds .

Experiment 16: E4A - Tig Of -N Senesy oil  Chloride Esterification

Chemical synthesis of E4A-Tig-Sen-1: 1. Esterification of E4A-Tig-N with senesioyl chloride; 3. Isolation of E4A-Tig-Sen-1 by HPLC

Measurement of cytotoxic activity: 1. MTT assay

Determination of chemical structure: 1. NMR analysis; 2. Mass spectrum analysis

Experiment 17: A mixture of E4A-Tig-N angeloyl chloride, acetyl chloride, crotonoyl chloride, 3,3-dimethyl acryloyl chloride, semenoyl chloride, cinnamoyl chloride, pentenoyl chloride, hexanoyl chloride, Esterification with chloride or ethyl butyryl chloride; Separation by HPLC; Determination of cytotoxic activity: Determination of the chemical structure by the method of Experiment 8 provided the following compounds:

Experiment 18: Suppression of cell adhesion

Methods and Results. ES2 or Hey8A cells are selected from the group consisting of E4A-Tig-R, E4A-Tig-V, E4A-Tig-S, E4A-Tig-N, E4A-Tig- / Ml &lt; / RTI &gt; of the compound. The cultures were incubated for 5 hours. Cells attached by trypanization were removed from the flask and counted. Compared to the non-drug control, 80 + 4% ES2 cells and 60 + 4% Hey8A cells were identified to be attached to the flask under these conditions. At 5 [mu] g / ml of the above compound, at least 90% of the nonadherent cells are alive, measured by trypan blue exclusion assay and by their ability to reattach to the flask when plating into media without the test compound. However, in the case of the compound tested at 10 μg / ml, less than 40% of cells were attached to the flask, and many of these were dead cells. These experiments indicate that the compound tested inhibits the cell attachment process.

Experiment 19: Fibronectin  Secretory experiment

Western blot was applied in the present invention as a method for detecting a specific protein in cells and untreated cells treated with a compound of the present invention wherein the cell is a bladder, a cervix, a prostate, a lung, a breast, a leukemia, 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 T25 flasks and grown for 24 hours before drug-treatment.

Drug-treatment: Cell cultures were treated with 2.5 [mu] l DMSO (as control) [D]; Or 10, 20, 30, 40, 80 ug / ml of the tested compound.

After 24 hours, aliquots of the culture medium were taken for fibronectin measurement (western blotting method).

Cell viability was measured by MTT assay at 24 hours. The culture was replaced with RPMI medium (5 ml) containing MTT and incubated for 1 hour. The formation of formazan was dissolved in 10 ml of DMSO and the OD at 570 nm was measured (MTT unit).

Western Blot: The spent culture medium was mixed with SDS sample buffer, and boiled for 3 minutes before loading on SDS gel. Samples were applied to 6-10% SDS gels and electrophoresis was performed at 100 volts for 2 hours. The proteins were transferred to the nitrocellulose membrane by electrophoresis. Nitrocellulose blots were incubated with the first and second antibodies (AP conjugated, Promega S3721). Immuno-band immuno-band) was developed with BCIP / NBT color development system.

Measurement of Western Band Intensity: The band of Western blot-image was captured with a digital camera, and the intensity of the band was determined using "Image J" software.

The results show that compounds of E4A-Tig-R, E4A-Tig-V, E4A-Tig-S, E4A-Tig-N, E4A- (20 to 40%) of fibronectin secretion in breast, leukemia, colon, liver, bone, brain, skin, ovary, pancreas, mouth (KB) and kidney.

Table 1

table. ¹³ C and ¹ H NMR data for E4A-Tig-R (DMSO-d ₆ ) ^a

Table 2:

table. ¹³ C and ¹ H NMR data for E4A-Tig-V (DMSO-d ₆ ) ^a

Claims (17)

delete delete delete delete delete delete delete delete Wherein the compound is selected from the group consisting of:
a) a separate or purified or synthesized compound having the structure:

, Or the chemical name: 24,28-O-Tigloyl-3 ?, 16 ?, 21 ?, 22?, 24?, 28-
b) a separate or purified or synthesized compound having the structure:

, Or the chemical name: 24-O-tigloyl-3 ?, 16 ?, 21 ?, 22 ?, 24?, 28- hexahydroxy olean-
c) a separate or purified or synthesized compound having the structure:

, Or the chemical name: 22,28-O-Tigloyl-3 ?, 16 ?, 21 ?, 22 ?, 24 ?, 28-
d) a separate or purified or synthesized compound having the structure:

, Or the chemical name: 21,24,28-O-Tigloyl-3 ?, 16 ?, 21 ?, 22 ?, 24 ?, 28-hexahydroxy olean-
e) a separate or purified or synthesized compound having the structure:

, Or the chemical name: 22,24,28-O-Tigloyl-3 ?, 16 ?, 21 ?, 22 ?, 24 ?, 28-hexahydroxy olean-
f) a separate or purified or synthetic compound having the structure:

, Or the chemical name: 3,21,28-O-Tigloyl-3 ?, 16 ?, 21 ?, 22 ?, 24 ?, 28-hexahydroxy olean-
g) a separate or purified or synthesized compound having the structure:

, Or the chemical name: 21,24-O-Tigloyl-3β, 6α, 21β, 22α, 24β, 28-hexahydroxy olean-12-ene. A composition for treating cancer comprising a compound according to claim 9 and a pharmaceutically acceptable carrier or diluent. 10. The method according to claim 9, wherein the compound is for treating cancer, inhibiting cancer growth, inhibiting cancer infiltration, inhibiting cancer metastasis, regulating cell adhesion, or controlling cell adhesion, Cancer, leukemia cancer, lung cancer, colon cancer, CNS cancer, melanoma cancer, renal cancer, cervical cancer, esophageal cancer, breast cancer, liver cancer, liver cancer, liver cancer, ovarian cancer, bladder cancer, prostate cancer, skin cancer, Testicular cancer, spleen cancer, kidney cancer, lymphatic cancer, pancreatic cancer, gastric cancer, and thyroid cancer; The present invention relates to a method for treating or preventing cancer in a mammal such as a breast, a white blood cell, a liver cell, an ovarian cell, a bladder cell, a prostate cell, a skin cell, a bone cell, a brain cell, a leukemia cell, A cell, an esophageal cell, a testicular cell, a spleen cell, a kidney cell, a lymphatic cell, a pancreatic cell, a stomach cell and a thyroid cell. 10. The method of claim 9, wherein the cancer is treated, cancer growth, cancer invasion, cell invasion, cancer cell invasion; Inhibiting cell adhesion, cell adhesion, cell cycle, inhibiting virus, preventing brain cerebralization, improving memory, improving brain function, nocturia, frequent urination, urinary incontinence; Or a pharmaceutically acceptable salt thereof, for the treatment of diseases caused by dementia, Alzheimer's disease, autism, brain trauma, Parkinson's disease or brain dysfunction, or for treating arthritis, rheumatism, poor blood circulation, arteriosclerosis, Raynaud's syndrome, angina, heart disease, coronary heart disease, , Dizziness, kidney disease; Or inhibiting NF-kappaB activation or inhibiting the activation of NF-kappa B activation or a pharmaceutically acceptable salt thereof in the treatment of cerebral edema, severe acute respiratory syndrome, viral respiratory disease, chronic venous insufficiency, hypertension, chronic venous disease, edema, inflammation, For the treatment of sexually transmitted diseases, influenza, post-traumatic edema and post-operative boogie, to inhibit thrombosis, to inhibit ethanol absorption, to lower blood glucose levels, to control adrenocorticotropin and corticosterone levels, Aneurysms, Anti-asthma, Anti-edema, Anti-inflammatory, Anti-Bradykin, Hemorrhage, Hemorrhage, Anti Headache, Antiepileptic, Antiepileptic, Antiepileptic, Antiepileptic, Antiepileptic, Flu Remedy, Fracture , Anti-gingivitis, anti-hematoma, antiseptic, antihistamine, anti-hydratritic, anti-mensteritis, antioxidant, anti-periodontal disease, anti-phlebitis, anti pleatitis, hoarseness treatment, anti-rhinitis, anti-tonsillitis, anti-ulcer, , Cancer suppression , A corticosteroid source, a diuretic, a fungicide, a hemolytic agent, a hyaluronidase inhibitor, a lymphatic substance, a sodium diuretic, an insecticide, a pituitary stimulant, a thymic tissue dissolution, a blood vessel protection, , For anti-parasites; Gene, ANGPT2, DDIT3, LIF, and NFKB1Z, for the manufacture of an adjuvant composition or for the treatment of a venous incision. 10. The compound according to claim 9, wherein the compound is for the treatment of adherence or cancer. A composition comprising an effective amount of a compound of claim 9 as a medicament for the treatment of cancer. 10. A compound according to claim 9, wherein the compound can be obtained by the following method:
1. dissolving the core compound or triterpene, hydroxylated triterpene core in pyridine,
2. Addition of acyl chloride,
3. The mixture was stirred at different temperatures for 5 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 2 days or 3 days,
4. At the end of the reaction, an aqueous solution of an acid or base, or water is added to the reaction mixture,
5. Thereafter, the solution is extracted with ethyl acetate and lyophilized,
6. dissolve the reaction product in acetonitrile containing trifluoroacetic acid or DMSO,
7. The reaction products of the mixture and the individual fractions were treated with MTT cytotoxicity assays,
8. HPLC fractionation for separation is selected according to the cytotoxic activity of the reaction product obtained at a particular reaction time,
9. Purify the active esterification product by HPLC,
10. Combining the products,
11. Test the above products. 16. The method of claim 15 wherein the core compound is terpene, isoprene, or triterpene core, the core compound is hydroxylated, the core compound is soluble in pyridine, the acyl chloride is selected from the group consisting of tigloyl chloride, angeloyl chloride, acetyl chloride Is selected from the group consisting of crotonoyl chloride, 3,3-dimethyl acryloyl chloride, semenoyl chloride, cinnamoyl chloride, pentenoyl chloride, hexanoyl chloride, benzoyl chloride and ethyl butyryl chloride, The reaction time is 5 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 2 days or 3 days, and 75 ℃ temperature, the acid HCl or the base is a weak base or a NaHC0 ₃ is added to the reaction mixture, and the solution after being extracted three times with ethyl acetate and freeze-dried, The product is dissolved in 80% acetonitrile-0.005% trifluoroacetic acid or DMSO and the HPLC fraction for separation is eluted at 5 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 2 days, or 3 days of reaction time. delete

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