NOVEL BETULINIC ACID DERIVATIVES HAVING ANTIANGIOGENIC ACTIVITY, PROCESSES FOR PRODUCING SUCH DERIVATIVES AND THEIR USE FOR TREATING TUMOR ASSOCIATED ANGIOGENESIS

Abstract: The invention relates to novel betulinic acid derivatives having antiangiogenic activity, processes for producing such derivatives and their use for treating tumor associated angiogenesis.

Description: NOVEL BETULINIC ACID DERIVATIVES HAVING ANTIANGIOGENIC ACTIVITY, PROCESSES FOR PRODUCING SUCH DERIVATIVES AND THEIR USE FOR TREATING TUMOR ASSOCIATED ANGIOGENESIS. Field of Invention This invention relates to novel betulinic acid derivatives and a composition containing betulinic acid derivatives, processes for preparation of such betulinic acid derivatives and method of treatment of tumor and such proliferative disease using these derivatives. This invention also relates to the use of the novel betulinic acid derivatives for inhibiting and/or preventing tumor associated angiogenesis, more specifically angiogenesis associated with prostate, lung, ovary and colon cancers. Background of the Invention Betulinic acid is a pentacyclic triterpene. It can derived from several natural (botanical) sources. It can also be chemically derived from betulin, a substance found in abundance in the outer bark of white birch trees (Betula alba). Betulinic acid has been found to selectively kill human melanoma cells (Nature Medicine, Vol.l(10),1995, WO 96/29068). The cytotoxic potential of betulinic acid was tested using three human melanoma cell lines, Mel-1, Mel-2, and Mel-4. The growth of all the cell lines was inhibited significantly by treatment with betulinic acid. The effectiveness of betulinic acid against melanoma cancer cells was also tested using athymic mice. It seems to work by inducing apoptosis in cancer cells. The anti-cancer activity of betulinic acid and some of its derivatives has also been demonstrated using mouse sarcoma 180 cells implanted s.c. in nude mouse (JP 87,301,580), inhibition of growth of p388 lymphocytic leukemia cells in vitro (Choi.Y-H et al., Planta Medica Vol.XLVII,51 1-513,1988) and inhibiting growth of cancer cells, particularly by inhibiting omithine decarboxylase (Yasukawa, K et al, Oncology 48:72-76, 1991 ; WO 95/04526). Recently, the applicants reported anti-leukemia and anti-iymphoma activity and anti- prostate, anti-lung and anti-ovarian cancer activity of betulinic acid and its derivatives with ED 50 values less than 4.0 μg ml. (US Application Number 09/040,856 filed on March 18, 1998 and US Application Number 09/251 ,309 filed on February 17, 1999). Further, antiangiogenic activity of betulinic acid and its derivatives was recently reported by the applicants in US Application number 09/166,809 filed on October 06, 1998 wherein betulinic acid and its derivatives were shown to inhibit the formation of tube-like-structures (TLS) of endothelial cells when grown on Matrigel coated surface. The endothelial cell anti-proliferative activity along with anti-TLS activity was shown to suggest the anti-angiogenic activity of betulinic acid derivatives. Anderson et al (WO 95/04526) disclose that for certain cancers to spread throughout a patients' body, a process termed metastasis, cell-cell adhesion must take place. Specifically, cancer cells must migrate from their site of origin and gain access to blood vessel to facilitate colonization at distant sites. Certain cancer cells are known to adhere to E-Selectin via E-Selectin ligands on their cell surface and this event is one component of the metastasis process. Betulinic acid and its derivatives interfere with Selectin binding. Betulinic acid inhibited P-Selectin binding to 2,3, sLex, a chemical known to bind to P-Selectin, with an IC 5 o of 125 uM. Also it inhibited P- Selectin binding to HL-60 cells in a dose-dependent way with an IC 50 of 0.75 mM. Betulinic acid and derivatives also significantly interfere with the binding to colon cancer cells, LS174T to E-Selectin. Dasgupta et al (WO 96/29068) disclosed a method and composition for inhibiting tumor growth using the active compound betulinic acid. The invention provides a method and composition for inhibiting tumor growth and, particularly, for inhibiting growth of melanoma using a natural product derived compound. The invention also provides a treatment method using betulinic acid to prevent growth or spread of cancer cells, wherein betulinic acid is applied in a topical preparation. Pezzuto et al (US Patent 5,869,535) disclose method and composition for probes inhibiting tumor growth using betulinic acid or a derivative thereof. Betulinic acid has been isolated from stem bark of Ziziphus mauritiana, by mediating a selective cytotoxic profile against human melanoma in a subject panel of human cancer cell lines, conducting a bioassay directed fractionation based on the profile of bioactivity using cultured human melanoma cells (MEL-2) as the monitor, and betulinic acid has been obtained therefrom as the active compound. The resulting betulinic acid can be used to inhibit tumor growth, or can be converted to a derivative to prevent which prevents or inhibits tumor growth. The invention also provides a treatment method using betulinic acid to prevent the growth or spread of cancerous cells, wherein betulinic acid or derivatives thereof is applied in a topical preparation. Betulinic acid was found to inhibit in vitro growth of MEL-2 cells. However, none of the other cell lines tested [A431 (squamous cells), BC-1 (breast), COL-2 (colon), HT1080 (sarcoma), KB (human oral epidermoid carcinoma), LNCaP (prostate), LU-1 (lung), U373 (glioma) and ZR-75-1 (breast)] were affected by betulinic acid (ie., ED 50 values of greater than 20 μg/ml). Lee et al (WO 96/39033) disclose betulinic acid and dihydrobetulinic acid acyl derivative to have potent anti-HTV activity. The C -hydroxy, Cι -carboxylic acid and C 2 o-exomethylene groups have been modified. Anti-HTV assays indicate potent anti- HTV activity of betulinic acid and dihydrobetulinic acid derivatives in acutely infected H9 lymphocytes with EC 50 values of less than 1.7 x 10 "5 μM respectively. Objects of the invention The invention provides a method of treating angiogenesis by administering a pharmaceutically effective dosage of betulinic acid derivatives. This invention also provides for novel betulinic acid derivatives and compositions containing them with pharmaceutically acceptable additives, diluents, carriers and excipients with or without betulinic acid. Another object of the invention relates to providing novel betulinic acid derivatives, which are used for inhibiting angiogenesis. Another object of the invention is to provide a compound and compositions for treating, inhibiting and/or preventing angiogenesis using a natural product-derived compound and its derivatives. Another object of the invention is to provide a treatment method using betulinic acid derivatives to inhibit angiogenesis, wherein the derivatives are administered systemically. Yet another object of the invention is to overcome the problem of high toxicity associated with standard antiangiogenic chemotherapeutic agents by using a natural product-derived compound, e.g., betulinic acid or its derivatives. Still another object of the invention is to overcome the problem of insufficient availability associated with synthetic antiangiogenic anticancer agents by using readily available semisynthetic derivatives of betulinic acid. Another object of the invention is to overcome the problem of high costs of synthetic antiangiogenic agents by utilizing the readily available natural product derived compound, e.g. betulinic acid and its derivatives which is expected to be less expensive than other chemotherapeutic drugs. These and other objects of the present invention will become apparent from the description of the invention disclosed below, which descriptions are intended to limit neither the spirit or scope of the invention but are only offered as illustrations of the preferred embodiments of the invention. Summary of the invention The above objects and others have been achieved by providing novel betulinic acid derivatives of formulae 1 and 2 which are described in the present description. Detailed Description of Invention The present invention provides a pharmaceutical composition useful for preventing / inhibiting angiogenesis. Betulinic acid derivatives inhibit endothelial cell proliferation and exhibiting high endothelial cell specificity thereby specifically targeting endothelial cells. The derivatives also inhibit the formation of tube-like-structures (TLS) of endothelial cells when grown on Matrigel coated surface. The endothelial cell anti-proliferative activity along with anti-TLS activity very strongly suggests the anti-angiogenic activity of betulinic acid derivatives. The method compπses admmistei ing a theiapeutically effective dose of betulinic acid deuvatives eithei alone oi in a pharmaceutical composition containing the compounds • j 35 so as to kill, inhibit oi pi event the multiplication of tumoi associated endothelial cells In a pi efened embodiment, phaimaceutically acceptable cai nei s, diluents, excipients a d/oi solvents aie used with betulinic acid/oi its deuvatives The method of tieatment of the pi esent invention may be particulai ly useful in inhibiting 1 40 The novel deuvatives of betulinic acid have a basic skeleton of betulinic acid as shown herebelow in Figure 1 1 55 wheiem R, Ri R 2 , R 3 , RA, R. R 6 and R 7 independently oi in combination lepresent the following gioups 1 60 1 6 5 R 2 is H and R, is OH, OCO(CH )„Crl 3 (wheie n = 0 to 14), OCOC(CH 3 OCO(CH 2 )„X (where n = l to 7. X = H. Cl, Bi , F), OCOCH 2 C 6 H n X [n = 2 to 4, X = H, Cl, Br, F, I, CN, N0 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 , CHC1 2 or C n H 2n+1 (n=l to 7)], OS0 2 (CH 2 ) n X (where n = 1 to 7, X = H or Cl), 0SO 2 ONH 2 , OCOCeH-X [n = 0 to 4, X = H, Cl, Br, F, I, CN, N0 2 , NH 2 , CF 3 , OH, OCH 3 , OC 2 H 5 , CHC1 2 or 1 70 C n H 2n+1 (n = 1 to 7)], NH 2 , NH(CH 2 ) n OR [(n = 2 to 4), R = H or COCH 3 ], NHR, N(R) 2 [where R = CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 ], NHCeH-X, NHCH 2 C6H n X (where n = 2 to 4), NHCH 2 C,oH n X (n = 2 to 7) [X = H,C1, Br, F, I, CHC1 2 , CN, CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+1 (n = 1 to 7)], RCH 2 NOH (R = H,CH 3 ,C 2 H 5 ,C 3 H 7 ,C 4 H 9 ), NHOR (R = H, COCH 3 , COC 6 H n X, OCH 2 C 6 H„X, OC 6 H n X) [n = 2 to 4, X = Cl, Br, F, I, CF 3 , 1 75 CHC1 2 , CN, N0 2 , CH 3 , NH 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+ .(n = 1 to 7)], N=CHC 6 H n X (where n = 2 to 4), N=CHCι 0 H n X (n = 2 to 6)[X = H, Cl, Br, F, I, CF 3 , CN, N0 2 , NH 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+1 (n = 1 to 3)], OCO(CH 2 ) n NH 2 (n = 1 to 8), NHCO(CH 2 ) n X (X = H,C1 or Br, n = 1 to 4), NHCOCβH π X, NHCOCι 0 H„X (n = 2 to 6), NHCOCH 2 C 6 H n X (n = 2 to 4), NHCOCH 2 C 10 H π X (n = 2 to 6)[X = Cl, Br, F, I, 1 80 CF 3 , CN, N0 2 , NH 2; OH, OCH 3 , OC 2 H 5 , CHC1 2 or C„H 2n+ ,(n = 1 to 7)], NHCOCβHjCOOH, NHCOC 6 H n (COOH)X [ where n = 2 or 3, X = H, Cl, Br, F, N0 2 or NH 2 ), OCOC^COOH, OCOC6H n (COOH)X (where n = 2 or 3, X = H, Cl, Br, F, NO 2 or NH 2 ), OCOCHRRi, (R = H, CH 3 or Ph; R, = OH, Cl, Br or OCOCH 3 ), NHNHCeH n X (n = 2 to 4), NHNHCH(OH)C 6 H n X (n = 2 to 4), NHNHC 10 H„X (n = 2 8 5 to 6), NHNHCH(OH)CιoH„X (n = 2 to 6)[X = Cl, Br, F, I, OH, OCH 3 , OC 2 H 5 , NO 2 , NH 2 , CHC1 2 , CF 3 or C n H 2n+1 (n = 1 to 7)], OCOCH = C(R) 2 (R is H, CH 3 or C 2 H 5 ), O- CO-CH=CH-COOH, 0-CO-C(Br)=CHCOOH, OCOCH 2 C(R) 2 COOH (R = H or [R = NH 2 , NHC 6 H n X (n = 2 to 4), NHC, 0 H n X (n = 2 to 6), NHCO(CH 2 ) n X (n = 1 to 16)[X = H, Cl, F, Br], NHCOC 6 H n X, NHCOCH 2 C 6 H n X (n = 2 to 4), NHCOC 10 H n X (n = 2 to 6), N=CHC 6 H n X (n = 2 to 4), N=CHC 10 H n X (n = 2 to 6), NHCH 2 C 6 H n X (n = ! 95 2 to 4), NHCH 2 C 10 H n X (n = 2 to 6)[X = H, Cl, Br, F, I, CN, N0 2 , NH 2 , CF 3 , CHC1 2 , OCH 3 , OC 2 H 5 or C n H 2n+ i (n = 1 to 7), NHS0 2 (CH 2 ) n X (n = 1 to 7), NHS0 2 C 6 H n X (n = 2 to 4)[X = H, Cl, Br, F, CH 3 , N0 or NH 2 ] , R 2 and R 3 together are O, NNHC 6 H n X, NNHCOC 6 H n X (n = 2 to 4), NNHC, 0 H n X (n 00 = 2 t0 6) > NNHCOCιoH n X (n = 2 to 6), NC 6 H n X (n = 2 to 4), NC, 0 H„X (n = 2 to 6)[X = H, Cl, Br, F, I, CN, N0 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+ ι (ή = 1 to 7)], NNHC 6 H n BrX [(n = 2 or 3), X = F, Cl, N0 , NH 2 , OCH 3 , OC 2 H 5 , C n H 2n+ ι (n = 1 to 7)], NOS0 3 H, N-OX, NHOX [X being H, CH 3 , C 2 H 5 , COCH 3 , S0 2 C 6 H4CH 3 , COCgH n X, CβHnX, CH 2 C 6 H n X [(n = 2 to 4)X = H,C1, Br, F, I, CN, N0 2 , NH 2 , OH, 205 OCH 3 , OC 2 H 5 , C n H 2n+ ι ( n = 1 to 7], CF 3 or CHC1 2 ], NNHR [R is CH 3 , C 2 H 5 , C 2 EL 4 OY, Y = H, alkyl, phenyl, benzyl or its substituted derivative with Cl, Br, F, I, N0 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+1 (n = 1 to 7)], R 7 is O and P is H, OH , Cl, N 3 , NH 2 , OR (R = CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 ), 2 1 0 0(CH 2 ) n COY (n = 1 to3)[Y = OH, OCH 3 , OC 2 H 5 , Cl, CN, N 3 , NH 2 ], OCH 2 CH 2 OY [Y = H, CH 3 , C 2 H 5 , COCH 3 ], OCOCH=C(R) 2 (R = H, CH 3 or C 2 H 5 ), OCO(CH 2 ) π X (n = 1 to 16), (X = H, Cl, F or Br), OCOCeH n X ( n = 0 to 4), OCOCH 2 C 6 H n X (n = 2 to 4)[X = H, Cl, Br, F, I, CN, N0 2 , CF 3 , CHC1 2 , OH, OCH 3; OC 2 H 5 or C n H 2n+ , (n = 1 to 7)], NH(CH 2 ) n CH 3 (n = 0 to 9), NH(CH 2 ) n COOH (n = 1 to 8), OCH 2 CHO, 1 5 OCH 2 CH=NOX, OCH 2 CH 2 NHOX[X = H, CH 3 , S0 2 C6H,CH 3 , OCOCH 3 , OCOC 6 H 5 , phenyl or benzyl substituted derivatives], OCH 2 CH=NNHC 6 H n X, OCH.CH.NHNHCeH n X (n = 2 to 4), OCH 2 CH=NNHCιoH n X (n = 2 to 6), OCH 2 CH CH 2 NHNHCιoH„X [X = H, Cl, Br, F, I, CN, CF 3 , CHC1 2 , N0 2 , NH 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+1 (n = 1 to 7)], 0CH 2 CH 2 N(R) 2 (R = H, CH 3 , C 2 H 5 , C 3 H 7 , 20 C" 1 * 0 ' c eH5, C6H 5 CH 2 or its substituted derivative e.g.: Cl, Br, CN, F, I, N0 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+ ι (n = 1 to 7)], P M is H and R 7 is NOH, NHOR, N-OR [R is H, CH 3 , C 2 H 5 , S0 2 C 6 H 4 CH 3 , COCH 3 , CH 2 C 6 H n X, COCeHπX (n = 2 to 4), X = Cl, Br, F, I, CN, N0 2 , NH 2 , OH, OCH 3 , 25 OC 2 H5, CF 3 , CHCI 2 or C n H 2n+ , (n = 1 to 7) ], RCH 2 NOH (R = H, CH 3 or C 2 H 5 ), NH 2 , NHS0 2 (CH 2 ) n X (n = 1 to 7), NHSO 2 C 6 H n X (n = 2 to 5)[X = H, Cl, Br, CH 3 , N0 2 or NH 2 ], (NR) 2 ( R is H, CH 3 , C 2 H S , C 3 H 7 , C 4 H 9 , Phenyl or Benzyl or its substituted derivative), N=CHC 6 H n X, NHCH 2 C 6 H n X (n = 2 to 4), N=CHC, 0 H n X, NHCH 2 C|oH n [X (n = 2 to 6) X = H, Cl, Br, F, I, CN, N0 2 , NH 2 , CF 3 , CHC1 2 , OH, 30 OCH 3 , OC 2 H 5 or C„H 2n+1 ( n = 1 to 7)], NNHC 6 H n X, NHNHC 6 H n X, NHNHCH(OH)C 6 H n X, NNHCOC 6 H n X (n = 2 to 4), NNHC 10 H n X, NNHCOC, 0 H n X, NHNHCioHnX, NHNHCH(OH)C, 0 HnX [where n = 2 to 6, X = H, Cl, Br, F, I, CN, N0 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , C n H 2π+1 (n = 1 to 7)], NHCOR [R is CH 3 , CH 2 C1, CHC1 2 , CCI 3 , C 2 H 5 , C 2 H_,C1, C 3 H 7 , C 3 H 6 OH, C 3 H 6 C1, C 6 H 5 , C 6 H n X, CH 2 C 6 H n X, 235 COCH 2 C 6 H n X (n = 2 to 4), C 10 H n X, CH 2 C,oH n X, COCH 2 C 10 H„X (n = 2 to 6), X = Cl, Br, CN, F, I, N0 2 , NH 2 , CF 3 , OH, OCH 3 , OC 2 H 5 , CHC1 2 or C n H 2n+ ι (n = 1 to 7)], R 5 is H or Br, R* is CH 3 , CH 2 Br, CH 2 OR [R is CO(CH 2 ) n X, (n = 1 to 7; X = H, Cl, Br or F), CHO, CHNOY, CH 2 NHOY, [Y = H, CH 3 , C 2 H 5 , S0 2 C6H 5 , SO Z C O H J O ;, 240 CH 2 C 6 H n X, CeH n X (n = 2 to 4), X = H, Cl, Br, F, I, CN, N0 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , 0C 2 H 5 , C n H 2n+ , (n = 1 to 7)], RCH 2 NOH [where R is H, CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 ], CH 2 NH 2 , CH 2 NHR or CH 2 N(R) 2 [R is CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , CβHs, C 6 H n X or CH 2 C 6 H n X, COCH 2 C6H n X ( n = 2 to 4), CH 2 C, 0 H n X, COCH 2 C 10 H n X (n = 2 to 6)[X = H, Cl, Br, F, CN, I, N0 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , CF 3 , CHC1 2 or C n H 2n+1 (n 245 = 1 to 7)], COOH, COC1, CONHR (R is alkyl or aryl substituted group), CO-OCOR (R is alkyl or aryl substituted group), COCH 2 COR (R is OH, OCH 3 , OC 2 H 5 , NH 2 or Cl), COCH 2 CH 2 OR [R is H, CO(CH 2 )„X (n = 1 to 16), COC 6 H„X, COCH 2 C 6 H„X, (n = 2 to 4, X = H, Cl, Br, CN, F, I, N0 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+1 (n = 1 to 7)], COO(CH 2 ) n H (n = 1 to 5), COO(CH 2 ) n COY (n = 1 to 5, Y = 250 OH, OCH 3 , OC 2 H 5 , Cl or Br), CH=NC 6 H I1 X (n = 2 to 4), CH=NC 10 H„X (n = 2 to 6), CH=NNHC 6 H„X, CH=NNHCOC 6 H n X (n = 2 to 4), CH=NNHC 10 H n X, CH=NNHCOCιoH n X (n = 2 to 6), CH.NHNHCβH n X (n = 2 to 4), CH 2 NHNHC,oH π X (n = 2 to 6), CH 2 NHNHCH(OH)C6H n X (n = 2 to 4), CH 2 NHNHCH(OH)C 10 H„X (n = 2 to 6) [where X = H, Cl, Br, F, I, CN, CF 3 , N0 2 , NH 2 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or 255 C n H 2n+ ι (n = 1 to 7)], R 5 and Rό together is O, OH, 0(CH 2 ) n X (n = 1 to 6, X = H, Cl or Br), OCOC δ H n X, OCOCH 2 C6H-X [n = 2 to 5, X = Cl, Br, F, I, CN, N0 2 , NH 2 , CF 3 , OH, OCH 3 , OC 2 H 5 or C n H 2n+ ι (n = 1 to 7)], 0(CH 2 ) n COOH (n = 1 to 3), NOR, NHOR (R = H, CH 3 , 260 2 H s, C 3 H 7 , COCH3, COC 6 H 5 , phenyl or benzyl substituted derivatives), NH 2 , (NR) 2 (R = H, CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , CeHnX, CH 2 C 6 H n X; n = 2 to 5, X = Cl, Br, F, I, CF 3 , CN, N0 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , C π H 2n+1 (n = 1 to 7)], NHCO(CH 2 ) n X [n = 1 to 16, X = Cl or Br], NHCOC 6 H n X, NHCOCH 2 C 6 H π X (n = 2 to 4), NHCOC, 0 H n X, NHCOCH 2 CιoH„X (n = 2 to 6) (X = Cl, Br, F, I, CN, CF 3 , N0 2 , NH 2 , OH, OCH 3 , 65 OC 2 H5, C n H 2n+ ι (n = 1 to 7)], N=CHC 6 H n X (n = 2 to 4), N=CHC, 0 H n X (n = 2 to 6), NHCH 2 C 6 H„X (n = 2 to 5), NHCH 2 C 10 H n X (n = 2 to 6), NNHC 6 H n X, NC 6 H n X, NHC 6 H n X (n - 2 to 4), NC 10 H n X, NHC 10 H n X, NNHC, 0 H n X (n = 2 to 6), NNHCOC 6 H n X (n = 2 to 4), NNHCOC, 0 H n X (n = 2 to 6), NR [R = C 6 H n X (n = 2 to 5), CioH π X (n = 2 to 7)[X = H, Cl, Br, Cl, F, I, CN, N0 2 , NH 2 , CF 3 , CHC1 2 , OCH 3 , 270 OC 2 H 5 , OH or C n H 2n+ , (n = 1 to 7)]. Preparation of Betulinic acid derivatives. The following procedures are either used alone or in combination to produce the 275 derivatives of the present invention. Example 1 Preparation of 3 — o-acyl derivatives Method 1 : Substrate in organic base is treated with suitable anhydride at room 280 temperature for approximately 4-16 hours. Examples of anhydrides that can be used in this process are represented by general formula (RCH 2 C0 2 )0 wherein R=H, CH 3 , C 2 Hi, etc. The reaction was worked by evaporation of the reaction mixture, addition of water and extraction with an organic solvent. The organic layer was dried over anhydrous sodium sulfate, evaporated and residue crystallized to yield the 285 corresponding pure 3-0 acyl derivatives respectively. Examples of organic bases that can be used in this method are TEA, pyrdine and DMPA. Method II: Substrate in halogenated organic solvent was treated with suitable acyl chloride as in Method 1. The reaction was worked up as described in Method I to 290 yield the corresponding 3-o-acy' derivatives in the pure form. Examples of acyl chlorides that can be used are RCH 2 (CH 2 ) n , COCI wherein R=H, Cl or F Br and n=0 to 9 or RCH 2 (CH) „ XCOCI wherein R=H, X=OH, OCOCH 3 and n=l . The halogenated solvent may be selected from CC1 4 CH 2 Cl 2 , C 6 H 5 CH 3 or the like. 295 Example 2 Preparation of 3-oxo-derivatives. The substrate was dissolved in an organic solvent and conventional oxidizing agent was added under normal reaction conditions. The reaction was worked up as described in Method I to yield the corresponding 3-oxo derivatives in the pure form. 300 Example of oxidizing agents that can be used are Cr0 3 /Py; Cr0 3 /H 2 S0 ; Cr0 3 ./AcOH or the like. The normal reaction condition is stirring the substrate with oxidizing agent at from 0°C to room temperature for a few hours. The org&iTicTsδl err. ήla^-be selected from acetone, CH 2 C1 2 , AcOH, mixtures thereof or the like. Example 3 Preparation of 2, 20,29-tribromo 3-oxo derivative A -3-oxo betulinic acid derivative prepared according to the process of Example 3 was dissolved in halogenated organic solvent. To this was added dropwise liquid bromine dissolved in the same solvent and the temperature was maintained between 0-10°C. The reaction mixture was brought to room temperature and stirred for a few hours. The reaction was worked up as described in Method I of Example 2. The organic layer was washed with 5-10% aqueous alkaline solution and evaporated. The crystallized product yielded pure 2, 20, 29-tribromo-3-oxo derivatives. Examples of halogenated solvents that can be used are CC1 4 ,CH 2 C1 , CHC1 3 and the like; Examples of aqueous alkaline solution that can be used are bicarbonate or carbonate of an alkali metal in water, and the like. 3-Oxo-derivative of betulinic acid, dihydrobetulinic acid or their derivatives can be used in the processes of Examples 4, 5, 8,10 and 14. Example 4 Preparation of 3-oximino derivative A 3-oxo derivative is mixed in an alcoholic solvent such a methanol, ethanol, propanol and the like. To this was added hydroxylamine, phenyl hydroxylamine or benzyl hydroxylamine or its substituted derivatives and sodium acetate. The mixture was refluxed for a few hours. The reaction mixture was evaporated to dryness. The reaction was worked up as described in Method I of Example 2 and yielded crude-3-oximino derivative which crystallized to yield the corresponding P ure 3-oximino derivative. Example 5 Preparation of phenylhydrazone of 3-oxo derivative 335 Phenylhydrazine or its phenyl substituted analogs or a salt thereof, and sodium acetate were added to 3-oxo derivative dissolved in alcoholic solvent such as methanol, ethanol, propanol and the like, and was refluxed for about four hours. The reaction was worked up as described in Method I of Example 2 to yield the corresponding pure phenylhydrazone derivative in pure form. 340 Example 6 Preparation of 17 and /or 20-carboxyalkyl carboxylate 345 To the substrate dissolved in dry dimenthylformamide, sodium hydride was added and the mixture was stirred at room temperature for about two hours. A suitable haloalkyl carboxyester was added to the above reaction mixtures and the mixture was stirred at room temperature for 16-20 hours. The reaction was worked up as described in Method I of Example 2 to yield pure 17 and/or 20-carboxyalky 350 carboxylate derivative. Examples of haloalkyl carboxy esters that can be used are chloro or bromo derivative of methyl or ethyl acetate, or chloro or bromo derivative of propionate and the like. Example 7 355 Preparation of 17 and/or 20-carboxyalkyl carboxylic acid 17 and or 20-carboxyalkyl carboxylate was dissolved in an alcoholic solvent such as methanol, ethanol, propanol or the like to which a hydroxide such as sodium or potassium hydroxide or the like was added. The mixture was warmed to 40-50° C for 60 2-4 hours. The reaction was worked up as described in Method 1 of Example 2 to yield pure 17 or 20 -carboxyalkyl carboxylic acid derivative. Example 8 Preparation of 2-bromo-3-oxo-derivative: 355 3-oxo-dihydrobetulinic acid derivative was dissolved in halogenated organic solvent such as CCl 4 ,CH 2 ,Cl 2 , CHC1 3 or the like. Liquid bromine dissolved in the same solvent was added dropwise while maintaining the temperature between 0-10°C. The reaction mixture was brought to room temperature and maintained for a few hours. The mixture was worked up in the usual manner, the organic layer was washed with 370 5-10% aqueous alkaline solution followed by water. Evaporation and crystallization yielded pure 2-Bromo-3-oxo derivatives. Examples of aqueous alkaline solution that can be used are bicarbonate or carbonate of an alkali metal in water, and the like. 375 Example 9 Preparation of 20, 29-dibromo derivative: Betulinic acid or its derivative (except 3-oxo-betulinic acid or its derivatives) was dissolved in halogenated organic solvent. To this liquid bromine dissolved in the 380 same solvent was added dropwise and temperature maintained between 0-10°C. The reaction mixture was brought to room temperature and stirred for few hours. The reaction mixture was worked up as described in Method I of Example 2. The organic layer was washed in 5-10% aqueous alkaline solution and evaporated. The crystallized product yield pure 20,29-dibromo derivative. 385 Examples of halogenated solvents that can be used are CC1 ,CH 2 C1 2 ,CHC1 3 and the like. Example 10 3gθ Preparation of 3-amino derivatives a] 3-oximino derivative is dissolved in glacial acetic acid and shaken under hydrogen atmosphere (60-70psi) in presence of platinum oxide catalyst for several hours. Reaction mixture is filtered, mother liquor evaporated under g5 vacuum to remove glacial acetic acid and the residue worked up in the usual manner to yield the corresponding 3-amino derivative. b] 3 oxo-derivative is dissolved in methanol added ammonium sulphate and sodium borodhyride and reflected for 2-4 hrs. Reaction mixture evaporated to dryness, added water, filtered the solid and crystallized to give 3-amino 400 derivatives. Example 11 Preparation of 3-o- benzoyl derivatives 405 Substrate in organic base is treated with suitable benzoyl chloride for approximately 6-16 hours at an ambient temperature. Examples of benzoyl chloride that can be used are represented by general formula R n (Ar)CoCl wherein n = 1 to 3, R = H, Cl, Br, F, CF3 and Ar = C6H5, C6H4, C6H3 or C6H2. The reaction was worked up by addition of water and extraction with organic solvent. The organic layer was dried over 1 0 anhydrous sodium sulphate, evaporated and residue crystallized to yield pure 3-o- benzoyl derivatives respectively. Examples of organic bases that can be used are pyridine, piperidine. Example 12 1 5 Peparation of 3-o- mesylate derivatives Substrate is dissolved in halogenated solvent and added methane sulphonyl chloride slowly to it at 5-10 C. Stirred the mixture at an ambient temperature for 2-4 hours. Worked up the reaction mixture by washing the organic layer with water. Organic 20 layer dried over anhydrous sulfate, filtered, evaporated to dryness to get a residue which was crystallized from acetonitrile to yield pure 3-o- mesylate derivative. Example 13 Preparation of 3 -phenyl hydrazino or its phenyl substituted derivative 25 3-phenylhydrazone or its phenyl substituted derivative of betulinic acid or dihydrobetulinic acid is dissolved in glacial acetic acid and shaken under hydrogen atmosphere (50-70- psi) in presence of platinum sponge catalyst for 3-5 hours. Reaction mixture was filtered, mother liquor evaporated under vacuum to remove 430 glacial acetic acid and the residue crystallized from alcoholic solvent to yield pure 3- phenyl hydrazino or its phenyl substituted derivative. Alcoholic solvents used are methanol, ethanol or iso propanol. Example 14 435 Preparation of 3-N-Hydroxyethyl derivative 3-oxo-derivative is dissolved in absolute alcoholic solvent such as methanol / ethanol and to it added 15-20% alcoholic hydrochloric acid and 2-aminoethanol and stirred at room temperature for 30 - 60 minutes. To this added sodium cyanoborohydride and 440 further stirred at room temperature for approximately 72 hours. Worked up by adding water followed by filtration of solid to yield crude product, which was crystallized from alcohol to yield pure 3-N-hydroxyethyl derivative. Example 15 445 Preparation of 3-N-Benzylidene derivative 3-amino derivative is dissolved in alcoholic solvent, such as methanol / ethanol and to it added benzaldehyde or substituted benzaldehyde derivative in presence or absence of alkali carbonate, such as sodium or potassium carbonate. The mixture was stirred for few hours at ambient temperature to 50 C approximately. The reaction mixture 450 was worked up by removing alcohol under vacuum and addition of water. The aqueous layer either filtered or extracted with halogenated organic solvent, followed by evaporation yielded 3-N-benzylidene derivative. Example 16 455 30μl of ECV304 cell suspension (50 x 10 4 cells/ml in RPMI 1640 containing 10% FBS) followed by 150μl of medium was added to the wells of a 96-well tissue culture plate (Nunc, Denmark) and incubated (37°C, 5% C0 2 ) overnight. 20μl of the betulinic acid derivative to be tested was then added at concentrations ranging from 0.5ug/ml to 4 ug/ml. Each concentration was plated in triplicates. 20μl of medium 460 alone was added to control wells. After 72 hours of incubation an MTT assay (Mosmann, 1983) was performed and percentage inhibition in proliferation of treated cells was calculated with respect to control cells. The cytotoxicity assays for tumor cells have been described in detail in our application No. 09/040,856 filed in US on March 18, 1998. Table I shows the ED 50 455 values against ECV304 and four different tumor cell lines and the endothelial cell specificities of seventeen potent derivatives. Table - 1 ED50 Concentration (μg/ml) of drug that causes 470 50%>Cytotoxicity, where Percent Cytotoxicity = [ 1 - O.D. Treated O.D. control] * 100 ECS ratio = ED 5 0 Tumor cell growth / ED 50 Endothelial cell growth. ECS less than 10 Low ECS ECS between 10 and 20 = Moderate ECS 475 ECS greater than 20 = High ECS We predict that the 'high' and 'moderate' ECS compounds specifically target endothelial cells and can be grouped under potent anti-angiogenic compounds while 'low ' ECS compounds would supplement their already reported cytotoxic activity against tumor cells. 480 Example 17 Several derivatives of betulinic acid were prepared by making substitutions and/or structural changes at C 3 , C| , C 20 , and/or C 2 positions of betulinic acid as described in the examples. The derivatives were characterized on the basis of spectral data. 485 Table II refers to the structures of Figure 2 wherein R to R 4 which are clearly indicated including lists the structures of forty derivatives. Figure 2 wherein R to R are shown herebelow: TABLE II 500 Dihydrobetulinic acid Example 18 Matrigel (70 μl) was placed into each well of a 96-well culture plate at 4°C and was 505 allowed to polymerize by incubation at 37°C for 30 min. ECV304 (1 x 10 4 ) cells were seeded on the Matrigel in 200 μl DMEM containing 10% FBS. Betulinic acid and derivatives to be tested were added to marked wells at non-cytotoxic concentrations and incubated at 37 C for 24 - 48 hours. The absence of cytotoxicity of betulinic acid and its derivatives on ECV304 cells at the above time points was 51 0 confirmed by suitable controls. Five different phase-contrast microscopic fields (4X) were viewed and total tube area of the tube-like-structures (TLS) measured using Video Pro 32 Image Analysis system. Percent reduction in total tube area was given as the mean of the data from five fields. Percent inhibition of TLS was calculated with reference to Controls(no drug). Table - III Example 19 A suitable formulation of betulinic acid derivatives was prepared as follows. The derivatives were solubilized in a minimum volume of methanol. The derivatives may also be solubilized in isopropyl alcohol, dimethylformamide, dimethylsulfoxide or any other suitable solvent. Substituted beta-cyclodextrin, such as 2-hydroxypropyl beta-cyclodextrin, sulfobutyl ether beta-cyclodextrin was separately dissolved in water to a concentration of approximately 50 to 1000 mg per ml, preferably 250 to 7 0 mg per ml. The solubilized betulinic acid derivative was added in small aliquots to the derivatized beta cyclodextrin solution and sonicated at low temperature until a clear solution developed. The organic solvent was then removed by rotary evaporation and the final solution filtered to give a sterile product. The resulting solution was lyophilized. Systemic administration refers to oral, rectal, nasal, transdermal and parentral (i.e., intramuscular, intraperitoneal, subcutaneous or intravenous). In accordance with good clinical practice, it is preferred to administer the composition at a dose that will produce antiangiogenic effects without causing undue harmful side effects. The composition may be administered either alone or as a mixture with other therapeutic agents. Pharmaceutical compositions which provide from about 10 mg to 1000 mg of the composition per unit dose are preferred as tablets, lozenges, capsules, powders, aqueous or oily suspensions, syrups, elixirs, implants or aqueous solutions by any conventional method. The nature of pharmaceutical composition employed will, of course, depend on the desired route of administration. The human dosage of the composition is in the range of 1.0 to 200 mg/kg/day and the preferred range is 1.0 to 50 mg/kg/day. One embodiment of the invention relates to a method of using novel betulinic acid derivatives or a combination thereof to treat a patient with tumor associated angiogenesis by administering a pharmaceutically effective dosage of said betulinic acid derivative or its combination to the patient. The patient of the invention can be human, mammal or other animal. The ED 50 value of betulinic acid derivatives against human umbilical vein endothelial cells is 0.35 to 4.0 μg/ml. The endothelial cell specificity of betulinic acid derivatives for prostate cancer is 1.04 to 21.1. As regards the endothelial cell specificity of betulinic acid derivatives for lung cancer is 0.43 to >10. However, regarding the endothelial cell specificity of betulinic acid derivatives for ovarian cancer is 0.54 to 7.0. With regard to the endothelial cell specificity of betulinic acid derivatives for colon cancer is 0.87 to 14.3. The betulinic acid derivative is administered to the patient in a pharmaceutically acceptable additive, carrier, diluent, solvent, filler, lubricant, excipient, binder or stabilizer. Preferably, the betulinic acid derivative is administered in the form of a tablet, lozenge, capsule, powder, aqueous or oily suspension, syrup, elixir, implant or aqueous solution and the betulinic acid derivative or derivatives or combination thereof is administered to the patient systemically.

Claims: 570 Claims I . Novel betulinic acid derivatives of formula wherein R. R| , R 2 , R 3 . R4, R 5 Rg and R7 independently or in combination represent the following groups : 585 R is H ; R, is H, Br, Cl, F or I ; 5 g 0 R 2 is H and R 3 is OH, OCO(CH 2 )„CH 3 (where n = 0 to 14), OCOC(CH 3 ) 3 , OCO(CH 2 )„X (where n = 1 to 7, X = H, Cl, Br, F), OCOCH 2 C 6 H n X [n = 2 to 4, X = H, Cl, Br, F, I. CN, N0 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 , CHC1 2 or C n H 2π+ ι (n=l to 7)], OS0 2 (CH 2 )„X (where n = 1 to 7, X = H or Cl), OS0 2 ONH 2 , OCOC 6 H„X [n = 0 to 4, X = H, Cl, Br, F, I, CN, NO,, NH 2 , CF 3 , OH, OCH 3 , OC 2 H 5 , CHC1 2 or 595 C n H 2n+ ι(n = 1 to 7)], NH 2 , NH(CH 2 )„OR [(n = 2 to 4), R = H or COCH 3 ], NHR, N(R) 2 [where R = CH 3 , C 2 H 5 . C3H7, C4H9], NHC 6 H n X, NHCH 2 C 6 H„X (where n = 2 to 4), NHCH 2 C 1 0H-X (n = 2 to 7) [X = H,C1, Br, F, I. CHC1 2 , CN, CF 3 . CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n +ι (n = 1 to 7)], RCH 2 NOH (R = H,CH 3 ,C 2 H5,C3H 7 ,C 4 H 9 ), NHOR (R = H, COCH 3 , COC 6 H„X, OCH 2 C 6 H n X, OC 6 H„X) [n = 2 to 4, X = Cl, Br, F, I, CF 3 , 600 CHC1 2 , CN, N0 2 , Ch . NH 2 , OH, OCH 3 . OC 2 H 5 or C n H 2 „-ι(n = 1 to 7)], N=CHC 6 H„X (where 11 - 2 to 4). N=CHC 10 H„X (n = 2 to 6)[X = H, Cl, Br, F, I, CF 3 , CN, N0 2 , NH 2 , OH, OCH3, OC 2 H, or C„H 2π+ 1 (n = 1 to 3)], OCO(CH 2 ) n NH 2 (n = 1 to 8), NHC0(CI 1 2 )„X (X = H,C1 or Br, n = 1 to 4), NHCOC 6 H n X, NHCOC i 0 H n X (n = 2 to 6), NHCOCH 2 C 6 H n X (n = 2 to 4), NHCOCH 2 C, 0 H n X (n = 2 to 6)[X = Cl, Br, F, I, 605 CF 3 , CN, N0 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , CHC1 2 or C n H 2n +ι(n = 1 to 7)], NHCOCβfLCOOH, NHCOC 6 H n (COOH)X [ where n = 2 or 3, X = H, Cl, Br, F, N0 2 or NH 2 ), OCOC 6 H 4 COOH, OCOC 6 H n (COOH)X (where n = 2 or 3, X = H, Cl, Br, F, N0 2 or NH 2 ), OCOCHRR,, (R = H, CH 3 or Ph; R, = OH, Cl, Br or OCOCH 3 ), NHNHC 6 H n X (n = 2 to 4), NHNHCH(OH)C 6 H n X (n = 2 to 4), NHNHCι 0 H n X (n = 2 6 1 0 to 6), NHNHCH(OH)C,oH n X (n = 2 to 6)[X = Cl, Br, F, I, OH, OCH 3 , OC 2 H 5 , N0 2 , NH 2 , CHC1 2 , CF 3 or C n H 2n+1 (n = 1 to 7)], OCOCH = C(R) 2 (R is H, CH 3 or C 2 H 5 ), O- CO-CH=CH-COOH, 0-CO-C(Br)=CHCOOH, OCOCH 2 C(R) 2 COOH (R = H or CH 3 ), OCO(CH 2 ) n COOH (n 61 5 [R = NH 2 , NHC 6 H n X (n = 2 to 4), NHC l0 H n X (n = 2 to 6), NHCO(CH 2 ) n X (n = 1 to 16)[X = H, Cl, F, Br], NHCOC 6 H n X, NHCOCH 2 C 6 H n X (n = 2 to 4), NHCOCι 0 H n X (n = 2 to 6), N=CHC 6 H n X (n = 2 to 4), N=CHCι 0 H n X (n = 2 to 6), NHCH 2 C 6 H n X (n = 2 to 4), NHCH 2 Ci 0 H n X (n = 2 to 6)[X = H, Cl, Br, F, I, CN, N0 2 , NH 2 , CF 3 , CHC1 2 , 620 0CH 3> OC 2 H 5 or C n H 2n+ , (n = 1 to 7), NHS0 2 (CH 2 ) n X (n = 1 to 7), NHSOzCβHnX (n = 2 to 4)[X = H, Cl, Br, F, CH 3 , N0 2 or NH 2 ]., R 2 and R 3 together are O, NNHC 6 H n X, NNHCOCeH-X (n = 2 to 4), NNHC 10 H n X (n = 2 to 6), NNHCOC, 0 H n X (n = 2 to 6), NC δ H-X (n = 2 to 4), NC,oH n X (n = 2 to 6)[X 625 = H, Cl, Br, F, I, CN, N0 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+ ι (n = 1 to 7)], NNHC 6 H n BrX [(n = 2 or 3), X = F, Cl, N0 2 , NH 2 , OCH 3 , OC 2 H 5 , C n H 2n+ ι (n = 1 to 7)], NOS0 3 H, N-OX, NHOX [X being H, CH 3 , C 2 H 5 , COCH 3 , S0 2 C 6 H 4 CH 3 , COC 6 H n X, C 6 H n X, CH 2 C6H n X [(n = 2 to 4)X = H,C1, Br, F, I, CN, N0 2 , NH 2 , OH, OCH 3 , OC2H5, C n H 2n+ ι ( n = 1 to 7], CF 3 or CHC1 2 ], NNHR [R is CH 3 , C 2 H 5 , 30 C 2 H OY, Y = H, alkyl, phenyl, benzyl or its substituted derivative with Cl, Br, F, I, N0 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C„H 2n+ ι (n = 1 to 7)], R 7 is O and R4 is H, OH , Cl, N 3 , NH 2 , OR (R = CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 ), 0(CH 2 ) n COY (n = 1 to3)[Y = OH, OCH 3 , OC 2 H 5 , Cl, CN, N 3 , NH 2 ], OCH 2 CH 2 OY 35 [Y = H, CH 3 . C 2 H 5 , COCH3], OCOCH=C(R) 2 (R = H, CH 3 or C 2 H 5 ), OCO(CH 2 ) n X (n = 1 to 16), (X = H, Cl, F or Br), OCOC 6 H n X ( n = 0 to 4), OCOCH 2 C 6 H n X (n = 2 to 4)[X = H, Cl, Br, F, I, CN, N0 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+ ι (n = 1 to 7)], NH(CH 2 )„CH 3 (n = 0 to 9), NH(CH 2 ) π COOH (n = 1 to 8), OCH 2 CHO, OCH 2 CH=NOX, OCH 2 CH 2 NHOX[X = H, CH 3 , SO2C6H4CH3, OCOCH 3 , OCOC6H5, 640 phenyl or benzyl substituted derivatives], OCH 2 CH 2 NHNHC 6 H n X (n = 2 to 4), OCH 2 CH=NNHCιoH n X (n = 2 to 6), OCH 2 CH 2 CH 2 NHNHC| 0 H n X [X = H, Cl, Br, F, I, CN, CF 3 , CHC1 2 , N0 2 , NH 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+ ι (n = 1 to 7)], OCH 2 CH 2 N(R) 2 (R = H, CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , C 6 H 5 , C 6 H 5 CH 2 or its substituted derivative e.g.: Cl, Br, CN, F, I, N0 2 , NH 2 , 645 CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+ ι (n = 1 to 7)], R 4 is H and R 7 is NOH, NHOR, N-OR [R is H, CH 3 , C 2 H 5 , S0 2 C 6 i CU 3 , COCH 3 , CH 2 C6H n X, COC δ H n X (n = 2 to 4), X = Cl, Br, F, I, CN, N0 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , CF 3 , CHC1 2 or C n H 2n+1 (n = 1 to 7) ], RCH 2 NOH (R = H, CH 3 or C 2 H 5 ), NH 2 , 650 NHS0 2 (CH 2 ) n X (n = 1 to 7), NHS0 2 C 6 H n X (n = 2 to 5)[X = H, Cl, Br, CH 3 , N0 2 or NH 2 ], (NR) 2 ( R is H, CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , Phenyl or Benzyl or its substituted derivative), N=CHC 6 H n X, NHCH 2 C 6 H π X (n = 2 to 4), N=CHC 10 H n X, NHCH 2 Ci 0 H n [X (n = 2 to 6) X = H, Cl, Br, F, I, CN, NO 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+1 ( n = 1 to 7)], NNHC 6 H n X, NHNHCβH-X, 655 NHNHCH(OH)C6H n X, NNHCOCgHπX (n = 2 to 4), NNHCi 0 H n X, NNHCOC 10 H n X, NHNHdoHnX, NHNHCH(OH)C I0 HnX [where n = 2 to 6, X = H, Cl, Br, F, I, CN, N0 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , C n H 2n+ ι (n = 1 to 7)], NHCOR [R is CH 3 , CH 2 C1, CHCI 2 , CC1 3 , C2H5, C2ΪL4CI, C 3 H 7 , C 3 H 6 OH, C 3 H 6 C1, C ό Hs, C 6 H n X, CH 2 C 6 H n X, COCH 2 C 6 H n X (n = 2 to 4), Cι 0 H n X, CH 2 CιoH n X, COCH 2 Cι 0 H n X (n = 2 to 6), X = Cl, 660 Br > CN > F, I, N0 2 , NH 2 , CF 3 , OH, OCH 3 , OC 2 H 5 , CHC1 2 or C n H 2n+1 (n = 1 to 7)], Rs is H or Br, Ro is CH 3 , CH 2 Br, CH 2 OR [R is CO(CH 2 ) n X, (n = 1 to 7; X = H, Cl, Br or F), CHO, CHNOY, CH 2 NHOY, [Y = H, CH 3 , C 2 H 5 , S0 2 C 6 H 5 , S0 2 C6H,CH 3 , CH 2 C 6 H n X, 665 C 6 H n X (n = 2 to 4). X = H, Cl, Br, F, I, CN, N0 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC2H5, C n H 2n+ ι (n = 1 to 7)], RCH2NOH [where R is H, CH 3 , C 2 H 5 , C 3 H 7 , C H 9 ], CH 2 NH 2 , CH 2 NHR or CH 2 N(R) 2 [R is CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , Cβϊ s, ^X or CH 2 C 6 H n X, COCH.CβH n X ( n = 2 to 4), CH 2 C, 0 H n X, COCH 2 C 10 H n X (n = 2 to 6)[X = H, Cl, Br, F, CN, I, N0 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , CF 3 , CHC1 2 or C n H 2n+ ι (n = 1 to 670 7 )] > COOH, COCl, CONHR (R is alkyl or aryl substituted group), CO-OCOR (R is alkyl or aryl substituted group), COCH 2 COR (R is OH, OCH 3 , OC 2 H 5 , NH 2 or Cl), COCH 2 CH 2 OR [R is H, CO(CH 2 )„X (n = 1 to 16), COC 5 H n X, COCH 2 C 6 H n X, (n = 2 to 4, X = H, Cl, Br, CN, F, I, N0 2 , NH 2 , CF 3 , CHC1 2 , OH,OCH 3 , OC 2 H 5 or C n H 2n+ ι (n = 1 to 7)], COO(CH 2 ) n H (n = 1 to 5), COO(CH 2 ) n COY (n = 1 to 5, Y = OH, OCH 3 , 675 OC 2 H 5 , Cl or Br), CH=NC 6 H„X (n = 2 to 4), CH=NC 10 H n X (n = 2 to 6), CH=NNHC 6 H n X, CH=NNHCOC 6 H n X (n = 2 to 4), CH=NNHCι 0 H n X, CH=NNHCOC,oH n X (n = 2 to 6), CH 2 NHNHC 6 H n X (n = 2 to 4), CH 2 NHNHC I0 HπX (n = 2 to 6), CH 2 NHNHCH(OH)C 6 H n X (n = 2 to 4), CH 2 NHNHCH(OH)C, 0 H n X (n = 2 to 6) [where X = H, Cl, Br, F, I, CN, CF 3 , N0 2 , NH 2 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or 680 C n H 2n+ ι (n = 1 to 7)], R 5 and R^ together is O, OH, 0(CH 2 ) n X (n = 1 to 6, X = H, Cl or Br), OCOC 6 H n X, OCOCH 2 C 6 H n X [n = 2 to 5, X = Cl, Br, F, I, CN, N0 2 , NH 2 , CF 3 , OH, OCH 3 , OC 2 H 5 or C n H 2 „ + ι (n = 1 to 7)], 0(CH 2 )„COOH (n = 1 to 3), NOR, NHOR (R = H, CH 3 , 685 C 2 H 5 , C3H7, COCH3, COC 6 H 5 , phenyl or benzyl substituted derivatives), NH 2 , (NR) 2 (R = H, CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , C 6 H n X, CH 2 C 6 H n X; n = 2 to 5, X = Cl, Br, F, I, CF 3 , CN, N0 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , C n H 2n+ , (n = 1 to 7)], NHCO(CH 2 ) n X [n = 1 to 16, X = Cl or Br], NHCOCeH-X, NHCOCH 2 C 6 H n X (n = 2 to 4), NHCOCιoH n X, NHCOCH 2 C,oH n X (n = 2 to 6) (X = Cl, Br, F, I, CN, CF 3 , N0 2 , NH 2 , OH, OCH 3 , 690 OC 2 H 5 ; C n H 2n+ ι (n = 1 to 7)], (n = 2 to 6), NHCH.CβH-X (n = 2 to 5), NHCH 2 C 10 H n X (n = 2 to 6), NNHCeH-X, NCβH n X, NHC δ H π X (n = 2 to 4), NC 10 H„X, NHC 10 H n X, NNHC 10 H n X (n = 2 to 6), NNHCOC 6 H π X (n = 2 to 4), NNHCOCι 0 H n X (n = 2 to 6), NR [R = C 6 H n X (n = 2 to 5), C, 0 H n X (n = 2 to 7)[X = H, Cl, Br, Cl, F, I, CN, N0 2 , NH 2 , CF 3 , CHC1 2 , OCH 3 , 695 OC 2 H 5 , OH or C„H 2n+ , (n = 1 to 7)] . 2. Novel betulinci acid derivatives as claimed in claim 1 having formula 2 wherein R = H R, = H or Br R 2 = O, NNHC 6 H 5 , NHNHCβ H 5 , NHNHC 6 H 4 (CH 3 )(4), OCOC 6 H 3 F 2 (3,5), OCOC 6 71 0 H 3 F 2 (2,4), OCOC 6 H 4 (CF 3 )(3), OCOC 6 H 4 (CF 3 )(2), N=CHC 6 H 3 F 2 (3,4), OH, OCOC 6 H 4 (C 5 H, ,)(4), OCOCH 2 C 6 H 3 (OCH 3 ) 2 (2,5), N=CHC 6 H 3 F 2 (2,4), OCOC^C, H 15 )(4), OCOC 6 F 5 , NOH, OCOC 6 H 3 (OCH 3 ) 2 (3,4), OCOC 6 H 3 (OCH 3 ) 2 (2,4), OCOCCIF 2 , OCOC 6 H 4 (C 6 H 5 )(4), OCOCH(CI)C 6 H 5 , OCO(CH 2 ) 3 COOH, OCOC 6 H 4 CI(4), OCOC 6 H 4 (CHCl 2 )(3),- OCOC 6 H 2 (COOH)(2)CI 2 (3,6), OCOCH(CI)CH 3 7 T 5 NNHCOC 6 EL,OH(2) or NOCH 2 C 6 H 4 N0 2 (4). R3 = H, - CH 2 CH 2 COOCH 3 -COCH =CH or 3-deoxydihydro betulinic acid. R 4 = CH 2 CCH 3 -CH(CH 3 ) 2 or BrCH 2 C(Br)CH 3 , 3. A process for preparing 3-O-acyl betulinic acid derivative of betulinic acid as 720 claimed in claim 1 wherein said process comprising (i) treating betulinic acid or a betulinic acid derivative with an anhydride or acyl chloride at room temperature for 4 to 16 hours; (ii) adding water to the solution of step (i), and (iii) extracting with an organic solvent to obtain an organic layer, said 3-o-acyl betulinic acid derivative being present in the organic layer. 725 4. A process for preparing a 3-oxo-derivative of betulinic acid as claimed in claim 1 wherein said process comprising dissolving a betulinic acid or a betulinic acid derivative in an organic solvent and adding an oxidising agent to obtain the 3-oxo- derivative. 730 5. A process for preparing a 2-bromo-3-oxo-dihydrobetulinic acid derivative as claimed in claim 1 wherein said process comprising dissolving 3-oxo- dihydrobetulinic acid or its derivative in a halogenated organic solvent, adding at a temperature between 0-10°C liquid bromine dissolved in the halogenated organic 735 solvent and heating the mixture to room temperature to obtain 2-bromo-3-oxo derivative. 6. A process for preparing a 3-oximino-betulinic acid derivative as claimed in claim 1 wherein said process comprising adding an alcoholic solvent to a 3-oxo- 740 betulinic acid derivative, adding hydroxylamine, phenyl hydroxylamine, benzyl hydroxlamine or a salt thereof and sodium acetate, refluxing the mixture to obtain 3- oximino betulinic acid derivative. 7. A process for preparing phenylhydrazone of 3-oxo-betulinic acid derivative as 745 claimed in claim 1 wherein said process comprising dissolving an 3-oxo-betulinic acid derivative in an alcoholic solvent adding phenylhydrazine or its phenyl substituted analog or salt thereof, and sodium acetate, refluxing the reaction mixture and obtaining the phenylhydrazone derivate. 750 8. A process for preparing 17-carboxyalkyl carboxylate betulinic acid derivative as claimed in claim 1 wherein said process comprising dissolving betulinic acid or a derivative of betulinic acid in dry dimethyl formamide, adding sodium hydride, stirring at room temperature, adding haloalkylcarboxyester and obtaining 17- carboxylalkyl carboxylate betulinic acid derivative. 755 9. A process for preparing 17-carboxyalkylcarboxylic acid of betulinic acid as claimed in claim 1 wherein said process comprising dissolving a 17-carboxyalkyl carboxylate of betulinic acid derivative in an alcoholic solvent, adding a hydroxide, heating the reaction mixure to 40-50°C for 2-4 hours and obtaining 17-carboxyalkyl 760 carboxylic acid betulinic acid derivative. 10. A process for preparing a 2, 20, 29-tribromo betulinic acid derivative as claimed in claim 1 wherein said process comprising dissolving an 3-oxo-betulinic acid or its derivative in a halogenated organic solvent, adding liquid bromine 765 dissolved in the organic solvent, maintaining the reaction mixture at a temperature 0- 10°C, bringing the reaction mixture to room temperature and obtaining 2,20,29- tribromo betulinic acid derivative. 1 1. A process for preparing a 20, 29 dibromo betulinic acid derivative as claimed 770 in claim 1 wherein said process comprising dissolving betulinic acid or its derivative in a halogenated organic solvent with the proviso that the derivative is not 3-oxo- betulinic acid or its derivative, adding liquid bromine dissolved in the organic solvent, maintaining the reaction mixture at a temperature 0-10°C, raising the temperature of the reaction mixture to room temperature and obtaining 20, 29 77 tribromo betulinic acid derivative. 12. A process for preparing 20-carboxyalkyl carboxylate betulinic acid as claimed in claim 1 wherein said process comprising dissolving betulinic acid or a derivative of betulinc acid in dry dimethyl formamide, adding sodium hydride, stirring at room 780 temperature, adding haloalkylcarboxyester and obtaining 20-carboxyalkyl carboxylate betulinic acid derivative. 13. A process for preparing 20-carboxyalkylcarboxylic acid of betulinic acid as claimed in claim 1 wherein said process comprising dissolving a 20-carboxyalkyl 785 carboxylate of betulinic acid derivative in an alcoholic solvent, adding a hydroxide , heating the reaction mixture to 40-50°C for 2-4 hours and obtaining 20-carboxylkyl carboxylic acid betulinic acid derivative. 14. A process for preparing 3-amino derivative of betulinic acid as claimed in 7gθ claim 1 wherein said process comprising (a) dissolving 3-oximino derivative of betulinic acid in glacial acetic acid in the presence of platinum oxide catalyst, (b) filtering the mixture of step (a), and (c) evaporating the filterate of step (b) to obtain the amino derivative of betulinic acid; a] issolving 3-oxo derivative of betulinic acid in methanol (b) is adding sodium 795 borohydride and ammonium sulphate and refluxing; and c] readion mixture evaporated to dryness and adding water to yield 3-amino derivatie. 15. A process for preparing 3-0-benzoyl betulinic acid derivative, as claimed in 800 claim 1 wherein said process said process comprising a) mixing betulinic acid or a betulinic acid derivative with a benzoylchloride or its phenyl substituted derivative at room temperature for 6-10 hrs; b) adding water to the solution of step (a), and 805 c ) extracting with an organic solvent to obtain an organic layer, said 3-0- benzoylbetulinic acid derivative being present in the organic layer. 16. A process for preparing 3-0-mesylate betulinic acid derivative as claimed in claim 1 wherein said process comprising: 8i o a) dissolving betulinic acid derivative in halogenated solvent; b) treating the mixture of step (a) with methane sulfonyl chloride at 0 -10°C; c) stirring the mixture at ambient temperature for 2-4 hrs; d) adding water to the mixture of the step (c) and separating organic layer; and e) drying the organic layer over anhydrous sodium sulfate, evaporation and 81 5 crystallizing the residue using nitrile solvent to yield pure 3-0 mesylate derivative. 17. A process for preparing 3-phenyl hydrazino or its phenyl substituted derivative of betulinic acid as claimed in claim 1 wherein said process comprising:. 820 a) dissolving 3-phenyl hydrazone or its phenyl substituted derivative of betulinic acid glacial acetic acid in the presence of platinum sponge catalyst under hydrogen atmosphere (50-70 psi) for 3-5 hours; b] filtering mixture of step (a); • »*__ 825 °] evaporating the filtrate of step (b), and crystallizing to obtain 3- phenylhydrazino betulinic acid derivative. 18. A process for preparing 3-N-hydroxyethyl derivative of betulinic acid as claimed in claim 1 said process comprising dissolving 3- oxo~ 830 derivative in absolute alcoholic solvent such as methanol / ethanol, adding 15-20% alcoholic hydrochloric acid and 2-aminoethanol in the above solution and stirred at room temperature for 30 - 60 minutes, thereafter adding sodium cyanoborohydride and further stirred at room temperature for approximately 72 hours and finally worked up by adding water followed by filtration of solid to yield crude product, which was 835 crystallized from alcohol to yield pure 3-N-hydroxyethyl derivative. 1 9. A process for preparing 3-N-Benzylidene derivative of betulinic acid as claimed in claim 1 said process comprising dissolving 3-amino derivative in alcoholic solvent such as methanol/ethanol and to it added benzaldehyde or 840 substituted benzaldehyde derivative in presence or absence of alkali carbonate, such as sodium or potassium carbonate, the above mixture was stirred for few hours at ambient temperature to about 50 C, and finally the reaction mixture was worked up by removing alcohol under vacuum and addition of water, the aqueous layer either filtered or extracted with halogenated organic solvent, followed by evaporation 845 yielded 3-N-benzylidene derivative. 20. A method of using novel betulinic acid derivatives or a combination thereof to treat a patient with tumor associated angiogenesis by administering a pharmaceutically effective dosage of said betulinic acid derivative or its combination 850 t0 tne patient. 21. A method as claimed in Claim 20 wherein said patient is human, mammal or other animal. 855 22. A method as claimed in Claim 20 wherein ED50 value of betulinic acid derivatives against human umbilical vein endothelial cells is 0.35 to 4.0 μg/ml. 23 A method as claimed in Claim 20 wherein the endothelial cell specificity of betulinic acid derivatives for prostate cancer is 1.04 to 21.1. 860 24 A method as claimed in Claim 20 wherein the endothelial cell specificity of betulinic acid derivatives for lung cancer is 0.43 to >10. 25. A method as claimed in Claim 20 wherein the endothelial cell specificity of 865 betulinic acid derivatives for ovarian cancer is 0.54 to 7.0. 26. A method as claimed in Claim 20 wherein the endothelial cell specificity of betulinic acid derivatives for colon cancer is 0.87 to 14.3. 870 27. A method as claimed in claim 20 wherein the betulinic acid derivative is administered to the patient in a pharmaceutically acceptable additive, carrier, diluent, solvent, filler, lubricant, excipient, binder or stabilizer. 28. A method as claimed in claim 20 wherein the betulinic acid derivative is administered in the form of a tablet, lozenge, capsule, powder, aqueous or oily 875 suspension, syrup, elixir, implant or aqueous solution. 29- A method as claimed in claim 20 wherein the dosage for human is in the range of 1 .0 to 200 mg/kg/day. 880 30. A method as claimed in claim 20 wherein the betulinic acid derivative or derivatives or combination thereof is administered to the patient systemically. 885 890 895 900 905 AMENDED CLAIMS [received by the International Bureau on 30 September 2000 (30.09.00); original claims 1-30 replaced by new claims 1-31 (13 pages)] Novel betulinic acid derivatives of formula 1 In which R, Rj, R 2 , R 3 , R_t, R5, Re and R independently or in combination represent the following groups and the provisios (a) to (j) have been incorporated to exclude groups in relevant places: Wherein R is H ; Wherein Ri is H, Br, Cl, F or I ; Wherein R 2 is H and R 3 is OH, OCO(CH 2 )„CH 3 (where n = 0 to 14), OCOC(CH 3 ) 3 , OCO(CH 2 ) n X p (where n = 1 to 7, p = 1, X = H, Cl, Br, F), OCOCH . CβH-X p [n = 2 to 4, p = 1 to 3, X = H, Cl, Br, F, I, CN, NO 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 , CHC1 2 or C n H 2n+1 (n=l to 7)], OSO 2 (CH 2 )„X p (where n = 1 to 1, p = 1, X = H or Cl), OSO 2 ONH 2 , OCOCeH-Xp [n = 0 to 4, p = 1 to 5, X = H, Cl, Br, F, I, CN, NO 2 , NH 2 , CF 3 , OH, OCH 3 , OC 2 H 5 , CHC1 2 or C„H 2n+1 (n = 1 to 7)], NH 2 , NH(CH 2 ) n OR [(n = 2 to 4), R = H or COCH3], NHR, N(R) 2 [where R = CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 ], NHCβH-X p , NHCH 2 C 6 H n X P (where n = 2 to 4, p = 1 to 3), NHCH 2 Cι 0 H n X p (n = 2 to 6, p = 1 to 5) [X = H,C1, Br, F, I, CHC1 2 , CN, CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+I (n = 1 to 7)], RCH 2 NOH (R = H,CH 3 ,C 2 H 5 ,C 3 H 7 ,C 4 H 9 ), NHOR [R = H, COCH 3 , CO H-X p , CH 2 C 6 H n X p , C 6 H„Xp, n = 2 to 4, p = 1 to 3, X = Cl, Br, F, I, CF 3 , CHC1 2 , CN, NO 2 , CH 3 , NH 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+ ι(n = 1 to 7)], N = CHC 6 H n X p (where n = 2 to 4, p = 1 to 3), N = CHCιoH n X p (n = 2 to 6, p = 1 to 5) [X = H, Cl, Br, F, I, CF 3 , CN, NO 2 , NH 2 , OH, OCH 3 , OC2H 5 or C n H 2n+ ι (n = 1 to 3)], OCO(CH 2 ) n NH 2 (n = 1 to 8), NHCO(CH 2 ) n Xp (X = H,C1 or Br, n = 1 to 4, p = 1), NHCOC 6 H n X p , NHCOCH 2 C 6 H n X p (n = 2 to 4, p = 1 to 3),NHCOC 10 H n X p , NHCOCH 2 CιoH n Xp (n = 2 to 6, p = 1 to 5) [X = Cl, Br, F, I, CF 3 , CN, NO 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , CHC1 2 or C n H 2n+ ι(n = 1 to 7)], NHCOC 6 H 4 COOH, NHCOC 6 H π (COOH)X p , OCOC 6 H 4 COOH, OCOC 6 H n (COOH)X p [ where n = 2 or 3, p = 1 or 2, X = H, Cl, Br, F, NO 2 or NH 2 ), OCOCHRRj, (R = H, CH 3 or Ph; Ri = OH, Cl, Br or OCOCH 3 ), NHNHC6H n X p , NHNHCH(OH)C 6 H n X p (n = 2 to 4, p = 1 to 3), NHNHCioHnXp NHNHCH(OH)C 10 H n X p (n = 2 to 6, p = 1 to 5)[X = Cl, Br, F, I, OH, OCH 3 , OC 2 H 5 , NO 2 , NH 2 , CHC1 2 , CF 3 or C n H 2n+ ι (n = 1 to 1)], OCOCH = C(R) 2 (R is H, CH 3 or C 2 H 5 ), O-CO-CH=CH-COOH, O-CO-C(Br)=CHCOOH, OCOCH 2 C(R) 2 COOH (R = H or CH 3 ), OCO(CH 2 ) n COOH (n = 0 to 3), — o, *-&s - , -OOCCH(OH)CH(Ph)R R = NH 2 , NHCβH π Xp (n = 2 to 4, p = 1 to 3), NHC 10 H„Xp (n = 2 to 6, p = 1 to 5), NHCO(CH 2 ) n X p (n = 1 to 16, p = 1)(X = H, Cl, F, Br), NHCOQjH n X p , NHCOCH 2 C 6 H„X p , N=CHC 6 H π X p , NHCHzQjH n Xp , NHSO 2 C 6 H n X p (n = 2 to 4, p = 1 to 3), NHCO oHnXp, N=CHCιoH n X p , NHCH 2 CιoH n X p (n = 2 to 6, p = 1 to 5)[X = H, Cl, Br, F, I, CN, NO 2 , NH 2 , CF 3 , CHC1 2 , OCH 3 , OC 2 H 5 or C n H 2n+ 1 (n = 1 to 7)], NHSO 2 (CH 2 ) n X p [n = 1 to 7, p = 1 , X = H, Cl, Br, F, CH 3 , NO 2 or NH 2 ], with a provisio that the following groups are excluded: a) R = Ri = R 2 = H; R 3 = OH, OCO(CH 2 )nCH 3(n __ 0) , OCO(CH 2 ) n X p [n = l,p = 1; X = H, Cl, Br, F],OSO 2 (CH 2 ) n X p (n = 1, p = 1 and X = H),OCOC 6 H-X p [n= 2 to 4, p = 1 to 3; X=H, Cl, Br, F, I, OCH 3 , NO 2 , C n H 2n + 1 (π= l to 7)]; NH 2 , NHR, N(R) 2 [R = CH 3 , C 2 H 5 , C 3 H 7 , C4H9], NH C ό H n X p (n=4, p = 1 to 3; X=H, Cl, Br, F or I); NHCO(CH 2 ) n X p [n= l, p = 1; X = H, Cl or Br]; NHCOC ό H- X p [ n = 2 to 4, p = 1 to 3; X = H, Cl, Br, F, I, NO 2 , OCH 3 , C n H 2n + l (n = 1)]; R 4 = OH; R 7 = 0; R 5 = H, R δ = CH 3 ; R 5 and R* together is = CH 2 . b) R = Ri = R 2 =H; R 3 =OCOCH 2 C(R) 2 COOH (R= H or CH 3 ), NH 2 , NHCO(CH 2 ) n X p (X = H, n = 1, p = 1), OCO(CH 2 ) n COOH (n=2 and 3); R4=OH, OR (R = H, CH 3 , C 2 H 5 , C 3 H 7 , C4H9, OCH 2 COOCH 3 ); R 7 = O; R 5 =H, Re=CH 3 ; R 5 and R δ together is = CH 2j Wherein R 2 and R 3 together are O, NNHC 6 H„X P , NNHCOC δ H n X p (n = 2 to 4, p = 1 to 3), NNHC 10 H n X p , NNHCOC 10 H„X(n = 2 to 6, p = 1 to 5), NC 6 H n X p (n = 2 to 4, p = 1 to 3), NC 10 H n X p (n = 2 to 6, p = 1 to 5) [X = H, Cl, Br, F, I, CN, NO 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+ ι (n = 1 to 7)], NNHC 6 H n BrX p [n = 2 or 3, p = 1 or 2, X = F, Cl, NO 2 , NH 2 , OCH 3 , OC 2 H 5 , C n H 2n+ ι (n = 1 to 7)], NOSO 3 H, N-OX p , NHOX p [p = 1, X = H, CH 3 , C 2 H 5 ,COCH3,SO 2 C 6 H 4 CH 3 ],COC6HnXp, C6HπX p ,CH 2 C6H n X p [n = 2 to 4, p = 1 to 3, X = H, Cl, Br, F, I, CN, NO 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , C n H 2n+ ι (n = 1 to 7), CF 3 or CHC1 2 ], NNHR [R is CH 3 , C 2 H 5 , C 2 H4OY, Y = H, alkyl, phenyl, benzyl or its substituted derivative with Cl, Br, F, I, NO 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C rt H 2n+ ι (n = 1 to 7)], with a provisio that the following groups are excluded: c) R=H; R] = H, or Br; R 2 and R 3 = 0; Rj = OH; R 7 = 0; R 5 = H and Ra = CH 3 ; R 5 and Re together is = CH 2 and R = R, = H; R 2 and R 3 together is NOMe, NOH; R 7 = 0 and ^ = OH; R 5 and R δ together is = CH 2 and R 5 = H and R δ = CH 3; R = H; d) Rj = H, Br, Cl, F or I; R 2 and R 3 together is NNH C 6 H n X p [n = 4, p = 1 , X = H; n = 2, p = 3, X = Cl; n = 4, p = 1, X=OCH 3 or OH; n = 3, p = 2, X = Br and OCH 3 ]; R 4 = OH, Cl, N 3 , NH 2 , OCH 3 , NH (CH 2 ) n CH 3 (n = 0 to 9), 0(CH 2 ) n COY [n = 1 to 3; Y = OH, OCH 3 ,OC 2 H 5 , Cl]; OCH 2 CH 2 OY [Y = H, OCOCH 3 , OC 2 H 3 ]; R 7 = 0; R 5 = H and R δ = CH 3 Wherein R 7 is O and R 4 is H, OH , Cl, N 3 , NH 2 , OR (R = CH 3 , C 2 H 5 , C 3 H 7 , C^y, O(CH 2 ) n COY (n = 1 to3)[Y = OH, OCH 3 , OC 2 H 5 , Cl, CN, N 3 , NH 2 ], OCH 2 CH 2 OY [Y = H, CH 3 , C 2 H 5 , COCH 3 ], OCOCH=C(R) 2 (R = H, CH 3 or C 2 H 5 ), OCO(CH 2 ) n X p (n = 1 to 16, p = 1, X = H, Cl, F or Br), OCOCδH n X P , OCOCH 2 CδH n X p (n = 2 to 4, p = 1 to 3) [X = H, Cl, Br, F, I, CN, NO 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+ ι (n = 1 to 7)], NH(CH 2 )„CH 3 (n = 0 to 9), NH(CH 2 )„COOH (n = 1 to 8), OCH 2 CHO, OCH 2 CH=NOX p , OCH 2 CH 2 NHOX p [p = 1, X = H, CH 3 , OCOCH 3 , OCOC δ H 5 , phenyl or benzyl substituted derivatives], OCH 2 CH=NNHC6H n X p , OCH 2 CH 2 NHNHC δ H n X p (n = 2 to 4, p = 1 to 3), OCH 2 CH=NNHC 10 H„X P , OCH 2 CH 2 CH 2 NHNHC 10 H n X p ( = 2 to 6, p = 1 to 5), X = H, Cl, Br, F, I, CN, CF 3 , CHC1 2 , NO 2 , NH 2 , OH, OCH 3 , OC 2 H 5 or C„H 2 „ +1 (n = 1 to 7)], OCH 2 CH 2 N(R) 2 (R = H, CH 3 , C 2 H 5 , C 3 H 7 , C H9, CδH 5 , C δ H 5 CH 2 or its substituted derivative with a proviso that the following groups are excluded: e) R = R, = R 2 = H; R 3 = OH, OCO(CH 2 ) n CH 3 (n = 0), OCOCH 2 C(R) 2 COOH (R = H or CH 3 ) or OCOC δ H n X p (n = 4, p = 1, X = H); R 7 = O; t = OCOC δ H n X p [n = 2, p = 3, X =OCH 3 ], NH (CH 2 )„ CH 3 (n=0 to 9), OR (R=CH 3 , C 2 H 5 , C 3 H 7 , H 9 ), O(CH 2 ) n COY[n = 1,Y = OH or OCH 3 ]; R 5 = H, R δ = CH 3 and R 5 and Rδ together is = CH 2J Wherein R 4 is H and R 7 is NOH, NHOR, N-OR [R is H, CH 3 , C 2 H 5 , SO 2 C δ H 4 CH 3 , COCH 3 , CH 2 C δ H n Xp, COC δ H n X p [n = 2 to 4, p = 1 to 3, X = Cl, Br, F, I, CN, NO 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , CF 3 , CHC1 2 or C n H 2n+ 1 (n = 1 to 7) ], RCH 2 NOH (R = H, CH 3 or C 2 H 5 ), NH 2 , NHSO 2 (CH 2 ) n X p (n = 1 to 7, p = 1), NHSO 2 C δ H n X p (n = 2 to 5, p = 0 to 3)[X = H, Cl, Br, CH 3 , NO 2 or NH 2 ], (NR) 2 ( R is H, CH 3 , C 2 H 5 , C 3 H 7 , C4H 9 , Phenyl or Benzyl or its substituted derivative), N=CHC δ H n X p , NHCH 2 C δ H n X p (n = 2 to 4, p = 1 to 3), N=CHCioH„Xp, NHCH 2 C 10 H n X p (n =2 to 6, p = 1 to 5) [X = H, Cl, Br, F, I, CN, NO 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+ 1 (n = 1 to 7)], NNHC δ H„X p , NHNHCeH n Xp, NHNHCH(OH)C δ H n X p , NNHCOC δ H n X p (n = 2 to 4, p = 1 to 3), NNHC 10 H n X p , NNHCOC 10 H n Xp, NHNHC 10 HnXp, NHNHCH(OH)C 10 HnX p (n = 2 to 6, p = 1 to 5) [X = H, Cl, Br, F, I, CN, NO 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , C n H 2n+ ι (n = 1 to 7)], NHCOR [R is CH 3 , CH 2 C1, CHC1 2 , CC1 3 , C 2 H 5 , C2H CI, C 3 H 7 , C 3 H 6 OH, C 3 H 6 C1, CδH 5 , C δ H n X p , CH 2 C 6 H n X p , COCH 2 CδH n X p (n = 2 to 4, p = 1 to 3), C 10 H n X P , CH 2 C 10 H n X p , COCH 2 C 10 H n Xp (n = 2 to 6, p = 1 to 5][X = H, Cl, Br, CN, F, I, NO 2 , NH 2 , CF 3 , OH, OCH 3 , OC 2 H 5 , CHC1 2 or C n H 2n+ i (n = 1 to 7)], with a proviso that the following groups are excluded: f) R = Rj = R 2 = H; R 3 = OH; R 4 = H; R 7 = N-OR [R = H, CH 3 or C 2 H 5 , COCH3, COC 6 H n X p (n = 2, p = 3, X = OCH 3 ; n = 4, p = 1, X = H, Cl, Br, F, I, NO 2 , OCH 3 and C n H 2n + 1 (n = 1)] R 5 =H, R δ =CH 3; R 5 and R δ together is =CH 2 f g) R = Rj = R 2 = H. R 3 = NH 2 or NHCO(CH 2 )„X p (n = 1, p = 1, X = H); R 7 = NH 2 or NHCOR (R = CH 3 ); t = H, OH or OR(R = CH 3 ); R 5 and Rδ together is = CH 2 - h) R = R t = H; R 2 and R 3 together is O, N-OX p (p = 1, X = H); R4 = H, OH or OCH 3 ; R 7 = NH 2 or NHCOR (R = CH 3 ); R 5 and R δ together is = CH 2 j Wherein R 5 is H or Br, Wherein Rg is CH 3 , CH 2 Br, CH 2 OR [R is CO(CH 2 )„X p , (n = 1 to 7; p = 1, X = H, Cl, Br or F), CHO, CHNOY, CH 2 NHOY, [Y = H, CH 3 , C 2 H 5 , SO 2 CδH 5 , SO2C 6 H4CH 3 , CH 2 C δ H n X p , C δ H„X p (n = 2 to 4, p = 1 to 3) [X = H, Cl, Br, F, I, CN, NO 2 , NH 2 , CF 3 , CHC1 2 , OH, OCH 3 , OC 2 H 5 , C n H 2n+1 (n = 1 to 7)], RCH 2 NOH [where R is H, CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 ], CH 2 NH 2 , CH 2 NHR or CH 2 N(R) 2 [R is CH 3 , C 2 H 5 , C 3 H 7 , C4H 9 , C δ H 5 , C δ H n X p or CH 2 C 6 H„X P , COCH 2 C δ H n X p ( n = 2 to 4, p = 1 to 3), CH 2 C 10 H n X p , COCH 2 CιoH n X p (n = 2 to 6, p = 1 to 5)[X = H, Cl, Br, F, CN, I, NO 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , CF 3 , CHC1 2 or C„H 2n+ 1 (n = 1 to 7)], COOH, COCl, CONHR (R is alkyl or aryl substituted group), CO-OCOR (R is alkyl or aryl substituted group), COCH 2 COR (R is OH, OCH 3 , OC 2 H 5 , NH 2 or Cl), COCH 2 CH 2 OR [R is H, CO(CH 2 ) n X P (n = 1 to 16, p = 1), COC 6 H n X p , COCH 2 C δ H n X p , CH = NC δ H n X p , CH = NNHC δ H n X p , CH = NNHCOC 6 H n Xp, CH 2 NHNHC δ H n X p , CH 2 NHNHCH(OH)C 6 H n X p , (n = 2 to 4, p = 1 to 3), CH = NC 10 H n X p , CH = NNHC 10 H n X p , CH = NNHCOC 10 H n X p , CH 2 NHNHCιoH n X p , CH 2 NHNHCH(OH)CιoH„X p (n = 2 to 6, p = 1 to 5) [where X = H, Cl, Br, F, I, CN, CF 3 , NO 2 , NH 2 , CHC1 2 , OH, OCH 3 , OC 2 H 5 or C n H 2n+1 (n = 1 to 7)], COO(CH 2 ) n H (n = 1 to 5), COO(CH 2 )„COY (n = 1 to 5, Y = OH, OCH 3 , OC 2 H 5 , Cl or Br), with a proviso that the following groups are excluded : (i) R = Ri = H, R and R 3 together is O, NOH; R 7 = O, R = OH, NH(CH 2 )„CH (n = 0 to 9), R 5 = H, R6 =CH 2 OR [R =CO(CH 2 ) n X p , n = 1, p = 1 , X = H], CH 2 NH 2 , CH 2 NHR (R = CH 3 , C 2 H 5 , C 3 H 7l C 4 H 9 ) Wherein R 5 and Rs together is =CH 2 O, OH, O(CH 2 )„X p (n = 1 to 6, p = 1, X = H, Cl or Br), OCOC 6 H n X p , OCOCH 2 C 6 H n X p [n = 2 to 4, p = 1 to 3, X = H, Cl, Br, F, I, CN, NO 2 , NH 2 , CF 3 , OH, OCH3, OC 2 H 5 or C n H 2n+1 (n = 1 to 7)], O(CH )„COOH (n = 1 to 3), NOR, NHOR (R = H, CH 3 , C 2 H 5 , C 3 H 7 , COCH 3 , COC 6 H 5 , phenyl or benzyl substituted derivatives), NH 2 , (NR) 2 (R = H, CH 3 , C 2 H 5 ,C3H 7 ,C 4 H 9 ,C 6 H n X p , CH 2 C 6 H n X p ; n = 2 to 4, p = 1 to 3,X = Cl, Br, F, I, CF 3 , CN, NO 2 , NH 2 , OH, OCH 3 , OC 2 H 5 , C n H 2n+ ι (n = 1 to 7)], NHCO(CH 2 ) n X p [n = 1 to 16, p = 1, X = Cl or Br], NHCOC 6 H n X p , NHCOCH 2 C 6 H n X p , N=CHC 6 H n X, NHC 6 H n Xp, NHCH 2 C 6 H„X p , NNHC 6 H n X p , NC 6 H n X p , NNHCOC 6 H n X p (n = 2 to 4, p = 1 to 3), NHCOCι 0 H n Xp, NHCOCH 2 Cι 0 H„X p , N=CHC 10 H n X p , NHCH 2 CioH„X p , NCioH„Xp, NHC 10 H n X p , NNHC 10 H n X p , NNHCOC 10 H n X p (n = 2 to 6, p = 1 to 5), NR [R = C 6 H n X p (n = 2 to 5, p = 0 to 3), Cι 0 H n X p (n = 2 to 7, p = 0 to 5) [X = H,C1, Br, F, I, CN, NO 2 , NH 2 , CF 3 ,CHCl 2 ,OCH 3 , OC 2 H 5 ,OH or C n H 2n+ ι (n = 1 to 7)] with a proviso that the following groups are excluded: j) R = Ri = R 2 = H; R 3 = OH; R4 = OH; R 7 = O; R 5 and Rs together is O,OH, OCOCH 3 ; O(CH 2 ) n X p (n = 1 to 6, p = 1; X = H), OCOC 6 H n X p [n = 2 to 4, p = 1 to 3, X = H, Cl, Br, F, I, NO 2 , OCH 3 and C„H 2n+ ι (n = 1)]; NOR [R=H, CH 3 , C 2 H 5 , C 3 H 7 , COCH 3 and COC_H 5 ]; NHR [R = CH 3 , C 2 H 5 , C 3 H 7 , C^, C 6 H n X p , COC 6 H n X p (n = 4, p = 1), X = H, Cl, Br, F, I, NO 2 , OCH 3 , C n H 2n+1 (n = D] 2. Novel betulinic acid derivatives as claimed in claim 1 having formula 2 Fig . 2 wherein R = H R 2 = O, NNHC 6 H 5 , NHNHC 6 H 5 , NHNHC 6 H4(CH 3 )(4), OCOCδH 3 F 2 (3,5), OCOC 6 H 3 F 2 (2,4), OCOC 6 H 4 (CF 3 )(3), OCOC 6 H4(CF 3 )(2), N=CHC 6 H 3 F 2 (3,4), OH, OCOC 6 I_ 4 (C_ H n )(4), OCOCH 2 C δ H 3 (OCH 3 ) 2 (2,5), OCOC δ H 4 (C 7 H 15 )(4), OCOC 6 F 5 , NOH, OCOC δ H 3 (OCH 3 ) 2 (3,4), OCOC δ H 3 (OCH 3 ) 2 (2,4), OCOCCIF 2 , OCOC 6 H 4 (C 6 H 5 )(4), OCOCH(CDC δ H 5 , OCO(CH 2 ) 3 COOH, OCOC 6 H4CI(4), OCOC 6 H 4 (CHCI 2 )(3) ,-OCOC δ H 2 (COOH)(2)CI 2 (3 ,6) , OCOCH(CI)CH 3 ; NNHCOC δ H 4 θH(2) and/or NOCH 2 C 6 H 4 NO 2 (4); R 3 = H, - CH 2 CH 2 COOCH 3 ,-COCH =CH 2 or 3-deoxydihydro betulinic acid. R 4 = CH 2 CCH 3 , -CH(CH 3 ) 2 or BrCH 2 C(Br)CH 3 , with a provisio that the following compounds are excluded: R = R ! = H; R 2 = OCOC 6 H 4 Cl, or OCO(CH 2 ) 3 COOH; R 3 = H; R4 = -CH 2 =C-CH 3 or -CH(CH 3 ) 2 . 3. A process for the preparation of 3-O-acyl betulinic acid derivatives said process comprising the steps of: (i) treating betulinic acid or its derivative with an anhydride or acyl chloride at room temperature for 4 to 16 hours; (ii) adding water to the solution of step (i), and (iii) extracting with an organic solvent to obtain an organic layer containing said 3-O-acyl betulinic acid derivatives; and (iv) separation of the derivatives by conventional methods. 4. A process for the preparation of 3-oxo-derivatives of betulinic acid said process comprising the steps of dissolving betulinic acid or its derivative in an organic solvent and adding an oxidising agent to obtain the 3-oxo- derivative. 5. A process for the preparation of 2-bromo-3-oxo-dihydrobetulinic acid derivatives, said process comprising the steps of dissolving 3-oxo- dihydrobetulinic acid or its derivative in a halogenated organic solvent, adding at a temperature between 0-10°C liquid bromine dissolved in the halogenated organic solvent and heating the reaction mixture to room temperature to obtain 2-bromo-3-oxo derivative. 6. A process for the preparation of 3-oximino-betulinic acid derivatives, said process comprising the steps of adding an alcoholic solvent to a 3-oxo- betulinic acid derivative, adding hydroxylamine, phenyl hydroxylamine, benzyl hydroxylamine or a salt thereof and sodium acetate, refluxing the mixture to obtain 3-oximino betulinic acid derivative. 7. A process for the preparation of phenylhydrazone derivatives of betulinic acid, said process comprising the steps of dissolving a 3-oxo-betulinic acid derivative in an alcoholic solvent adding phenylhydrazine or its phenyl substituted analog or salt thereof, and sodium acetate, refluxing the reaction mixture to obtain phenylhydrazone derivate. 8. A process for the preparation of 17-carboxyalkyl carboxylate betulinic acid derivatives said process comprising the steps of dissolving betulinic acid or its derivatives in dry dimethyl formamide, adding sodium hydride, stirring at room temperature, adding haloalkylcarboxyester to obtain 17- carboxylalkyl carboxylate betulinic acid derivative. 9. A process for the preparation of 17-carboxy alky lcarboxy lie acid derivatives of betulinic acid said process comprising dissolving a 17-carboxyalkyl carboxylate of betulinic acid derivative in an alcoholic solvent, adding a hydroxide, heating the reaction mixture to 40-50°C for 2-4 hours and obtaining 17-carboxyalkyl carboxylic acid betulinic acid derivative. 10. A process for the preparation of 2, 20, 29-tribromo betulinic acid derivatives, said process comprising steps of dissolving 3-oxo-betulinic acid or its derivative in a halogenated organic solvent, adding liquid bromine dissolved in the organic solvent, maintaining the reaction mixture at a temperature 0-10°C, bringing the reaction mixture to room temperature and obtaining 2,20,29-tribromo betulinic acid derivative. 11. A process for the preparation of 20, 29 dibromo betulinic acid derivative, said process comprising the steps of dissolving betulinic acid or its derivative in a halogenated organic solvent, said derivative not being 3- oxo-betulinic acid or its derivative, adding liquid bromine dissolved in the organic solvent, maintaining the reaction mixture at a temperature 0-10°C, raising the temperature of the reaction mixture to room temperature and obtaining 20, 29 tribromo betulinic acid derivative. 12. A process for the preparation of 20-carboxyalkyl carboxylate betulinic acid, said process comprising the steps of dissolving betulinic acid or a derivative of betulinc acid in dry dimethyl formamide, adding sodium hydride, stirring at room temperature, adding haloalkylcarboxyester and obtaining 20-carboxyalkyl carboxylate betulinic acid derivative. 13. A process for the preparation of 20-carboxyalkylcarboxylic acid of betulinic acid, said process comprising the steps of dissolving a 20- carboxyalkyl carboxylate of betulinic acid derivative in an alcoholic solvent, adding a hydroxide , heating the reaction mixture to 40-50°C for 2- 4 hours and obtaining 20-carboxylkyl carboxylic acid betulinic acid derivative. 14. A process for the preparation of 3-amino derivative of betulinic acid, said process comprising the steps of (a) dissolving 3-oximino derivative of betulmic acid in glacial acetic acid in the presence of platinum oxide catalyst, (b) filtering the mixture of step (a), and (c) evaporating the filtrate of step (b) to obtain the amino derivative of betulinic acid; 15. A process for the preparation of 3-amino derivative of betulinic acid, said process comprising the steps of: (a) dissolving 3-oxo derivative of betulinic acid in methanol, adding sodium borohydride and ammonium sulphate and refluxing; and (b) evaporating the reaction mixture to dryness and adding water to obtain 3-amino derivative. 16. A process for the preparation of 3-0-benzoyl betulinic acid derivative, said process comprising the steps of: a) mixing betulinic acid or a betulinic acid derivative with a benzoylchloride or its phenyl substituted derivative at room temperature for 6-10 hrs; b) adding water to the solution of step (a), and c) extracting with an organic solvent to obtain an organic layer, said 3- O-benzoylbetulinic acid derivative being present in the organic layer. 17. A process for the preparation of 3-0-mesylate betulinic acid derivative, said process comprising the steps of: a) dissolving betulinic acid derivative in halogenated solvent; b) treating the mixture of step (a) with methane sulfonyl chloride at 0 - 10°C; c) stirring the mixture at ambient temperature for 2-4 hrs; d) adding water to the mixture of the step (c) and separating organic layer; and e) drying the organic layer over anhydrous sodium sulfate, evaporation and crystallizing the residue using nitrile solvent to yield pure 3-0 mesylate derivative. 18. A process for the preparation of 3-phenyl hydrazino or its phenyl substituted derivative of betulinic acid, said process comprising the steps of: a) dissolving 3-phenyl hydrazone or its phenyl substituted derivative of betulinic acid glacial acetic acid in the presence of platinum sponge catalyst under hydrogen atmosphere (50-70 psi) for 3-5 hours; b) filtering mixture of step (a); and c) evaporating the filtrate of step (b), and obtaining 3-phenylhydrazino betulinic acid derivative by crystallization. 19. A process for the preparation of 3-N-hydroxyethyl derivatives of betulinic acid, said process comprising the steps of dissolving 3-oxo-derivative in absolute alcoholic solvent such as methanol/ethanol, adding 15-20% alcoholic hydrochloric acid and 2-aminoethanol stirring at room temperature for 30 - 60 minutes, adding sodium cyanoborohydride and further stirring at room temperature for approximately 72 hours and finally working up by adding water followed by filtration of the solid to obtain crude product, which is crystallized from alcohol to yield pure 3-N- hydroxyethyl derivative. 20. A process for the preparation of 3-N-Benzylidene derivative of betulinic acid, said process comprising the steps of dissolving 3-amino derivative in alcoholic solvent such as methanol/ethanol, adding benzaldehyde or substituted benzaldehyde derivative in presence or absence of alkali carbonate, such as sodium or potassium carbonate, stirring mixture for few hours at ambient temperature to about 50°C, and finally working up the reaction mixture by removing alcohol under vacuum and adding water, filtering the aqueous layer extracting with halogenated organic solvent, followed by evaporation to yield 3-N-benzylidene derivative. 21. Use of novel betulinic acid derivatives or a combination thereof as claimed in claim 1 to manufacture a composition useful in the treatment of a patient with tumor associated angiogenesis. 22. Use as claimed in claim 21 wherein said patient is human, mammal or other animal. 23. Use as claimed in claim 21 wherein ED 50 value of betulinic acid derivatives against human umbilical vein endothelial cells is 0.35 to 4.0 μg/ml. 24. Use as claimed in claim 21 wherein the endothelial cell specificity of betulinic acid derivatives for prostate cancer is 1.04 to 21.1. 25. Use as claimed in claim 21 wherein the endothelial cell specificity of betulinic acid derivatives for lung cancer is 0.43 to >10. 26. Use as claimed in claim 21 wherein the endothelial cell specificity of betulinic acid derivatives for ovarian cancer is 0.54 to 7.0. 27. Use as claimed in claim 21 wherein the endothelial cell specificity of betulinic acid derivatives for colon cancer is 0.87 to 14.3. 28. Use as claimed in claim 21 wherein the betulmic acid derivative is administered to the patient in a pharmaceutically acceptable add tive, carrier, diluent, solvent, filler, lubricant, excipient, binder or stabilizer. 29. Use as claimed in claim 21 wherein the betulinic acid derivative is administered in the form of a tablet, lozenge, capsule, powder, aqueous or oily suspension, syrup, elixir, implant or aqueous solution. 30. Use as claimed in claim 21 wherein the dosage for human is in the range of 1.0 to 200 mg/kg/day. 31. Use as claimed in claim 21 wherein the betulinic acid derivative or derivatives or combination thereof is administered to the patient systemically. STATEMENT UNDER ATICLE 19 1. The Examiner has cited eleven references on pages 2 & 3 of the Search Report under the category "X". The applicant submits that references D 1 to Dl l do not disclose the compounds claimed in the instant application. The applicant has amended claims 1 and 2 to eliminate the compounds found in the cited references Dl to Dll by adding provisos (a) to (j) in claim 1. Comments on the respective documents follow: The documents at Dl and D2 are WO 98/51294 A2 and WO 98/51293 Al respectively, disclosing betulinic acid derivatives and dihydro betulinic acid derivatives obtained by effecting modifications at C-3, C-17 and C-20 positions respectively. The compounds so obtained have been eliminated by addition of provisos (a), ( b), (c), (f) and (j) to claim 1. D3 is WO 96/39033 Al, a document disclosing betulinic acid derivatives obtained by effecting modifications of the C-3 hydroxy, C-17 carboxylic acid and C-20 exomethylene groups. The citation also discloses succinyl derivatives. These compounds have been eliminated by virtue of proviso (b) added to claim 1 Documents D4 and D5 are WO 93/10142 Al and EP 0542622 Al respectively. In addition to these documents, US patent 5,468,888 being the equivalent of these documents has also been considered. These documents disclose lupane derivatives obtained by modification at C-3, C-7 and C-20 positions of betulinic acid. Proviso (i) of claim 1 excludes the lupane derivatives disclosed by this reference. Document D6 is GB 1415601 A (BIOREX), a document disclosing dihydrobetulinic acid derivatives obtained by effecting modification at C-3 and C-17 positions. These derivatives disclosed have been eliminated by the addition of proviso (b) to claim 1 D7 is a publication by H1KOSHI et al disclosing derivatives of betulinic and dihydro betulinic derivatives. Proviso (a) added to claim 1 eliminates the compounds found in this citation. D8 is a publication by JAGADEESH et al, 1998, disclosing triterpene derivatives of betulinic acid. These derivatives have been eliminated by addition of proviso (a) to claim 1. D9 is a publication by FUJIOKA et al, 1994, disclosing triterpenenoids derivatives of betulinic acid. These have been eliminated by the addition of proviso (j) to claim 1. D10 is a publication by PRADHAN et al, 1993, disclosing 2-bromo- ketotritepenoids of lupanone and methyldihydrobetulonate with lithium aluminium hydride and sodium borohydride. These compounds have been eliminated by the addition of proviso (c) to claim 1. Dll is a publication by WRZECIONO et al, 1972, disclosing triterpene nitrogenous derivatives (VI) and 3- and 28-aminolupane derivatives (III). These compounds have been eliminated by the addition of provisos (g) and (h) to claim 1 In addition to the references cited by the International Searching Authority, in the Search Report, the Applicant has found the following relevant references and accordingly has distinguished the compounds claimed in the present invention of the instant application from the references by amending claim 1. (i) J S Lee "cytotoxic constituents from the Forsythiae fructus against L1210 and HL60 cells", Page 58; XP002139933, Abstract & YAKHAK HOECHI, Vol. 40, No. 4, 1996, pages 462-467. The citation discloses 3β-acetyl betulinic acid and betulinic acid obtained by modification at C-3 position. Proviso (a) added to claim 1 eliminates these compounds. (ii) Miles D H, et al "Tumor inhibitors I: Preliminary investigation of antitumor activity of Sarracenia Flava", Journal of Pharmaceutical Sciences, US, American Pharmaceutical Association, Washington, Vol. 3, No. 4, 1 April 1974 (1974-04-01), Pages 613-615, XP000611702 ISSN : 0022-3549 discloses betulinic acid which is eliminated by proviso (a) added to claim 1. (iii) Chemical Abstracts, Vol. 35, No.12, 20 June 1941 (1941-06-20) Columbus, Ohio, US; Abstract No. 3997, Riiti Kawaguti et al : "The constituents of Zyzyphus vulgaris Lamark var. spinosus Bunge" discloses betulinic acid and dihydrobetulinic acid which have been eliminated by proviso (a) added to claim 1. (iv) Kim D S H et al: "Synthesis of betulinic acid derivatives with activity against human melanoma" Bioorganic & Medicinal Chemistry Letters, GB, Oxford, Vol. 8, No. 13, 7 July 1998 (1998- 07-07), Pages 1707-1712, XP004137114 ISSN : 0960-894X. This reference discloses compounds obtained by effecting modifications at C-3, C-20 and C-28 positions of betulinic acid. These compounds have been eliminated by provisos (b) and (c) added to claim 1. (v) US 6048847 (Ramadoss et al) discloses betulinic acid and its derivatives which are eliminated by adding proviso (d) to claim 1.

Inventors: AHMED SIDDIQUI MOHAMMAD JAMSHE, JAGGI MANU, RAJENDRAN PRAVEEN, RAMADOSS SUNDER, AHMED SIDDIQUI, MOHAMMAD, JAMSHED, JAGGI, MANU, RAJENDRAN, PRAVEEN, RAMADOSS, SUNDER
Applicants: AHMED SIDDIQUI MOHAMMAD JAMSHE, DABUR RES FOUNDATION, JAGGI MANU, RAJENDRAN PRAVEEN, RAMADOSS SUNDER, AHMED SIDDIQUI, MOHAMMAD, JAMSHED, DABUR RESEARCH FOUNDATION, JAGGI, MANU, RAJENDRAN, PRAVEEN, RAMADOSS, SUNDER
Application year: 1999
Main classification: C07J63
IPC codes: A61K31, A61P35, A61P43, C07J63