US 2002001 0208A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2002/0010208A1 Shashoua et al. (43) Pub. Date: Jan. 24, 2002 (54) DHA-PHARMACEUTICAL AGENT Related U.S. Application Data CONJUGATES OF TAXANES (63) Continuation of application No. 09/135,291, filed on (76) Inventors: Victor Shashoua, Brookline, MA (US); Aug. 17, 1998, now abandoned, which is a continu Charles Swindell, Merion, PA (US); ation of application No. 08/651,312, filed on May 22, Nigel Webb, Bryn Mawr, PA (US); 1996, now Pat. No. 5,795,909. Matthews Bradley, Layton, PA (US) Publication Classification Correspondence Address: Edward R. Gates, Esq. (51) Int. Cl." ............................................ A61K 31/337 Wolf, Greenfield & Sacks, P.C. (52) U.S. Cl. .............................................................. 514/449 600 Atlantic Avenue Boston, MA 02210 (US) (57) ABSTRACT The invention provides conjugates of cis-docosahexaenoic (21) Appl. No.: 09/846,838 acid and pharmaceutical agents useful in treating noncentral nervous System conditions. Methods for Selectively target 22) Filled: Mayy 1, 2001 ingg pharmaceuticalp agents9. to desired tissues are pprovided. Patent Application Publication Jan. 24, 2002 Sheet 1 of 14 US 2002/0010208A1 1 OO 5 O -5OO - 1 OO-9 -8 -7 -6 -5 -4 LOG-10 OF SAMPLE CONCENTRATION (MOLAR) CCRF-CEM-o- SR ----- RPM-8226----- K-562- - -A - - HL-60 (TB) -g- - MOLT4: ... O Fig. 1 1 OO 5 O -5OO -1 O O -8 -7 -6 -5 -4 -- 9 LOGo OF SAMPLE CONCENTRATION (MOLAR) A549/ATCC-o-NS326. NCEKVX 28. --Q-- NCI-H322M-...-a---Eidsf8::... NC-H522--O-- HOP-62---Fig. 2 a-- NC-H460.-------- Patent Application Publication Jan. 24, 2002 Sheet 2 of 14 US 2002/0010208A1 COLON CANCER 1 OO - T - S. O 50 CD li O ()LL O - Z 5-501. -100-9 -8 -7 -6 -5 -4 LOGo OF SAMPLE CONCENTRATION (MOLAR) COLO 205-O- HCT-116--Q-- HCT-15- - -a- - - KM12- - e -- SW-62O---4--- HT29.....O...... Fig. 3 1 OO CNS CANCER 5 O -5OO -1 O O -8 -7 -6 -5 - 9 LOGo OF SAMPLE CONCENTRATION (MOLAR) SF-268-O-- SF-295 --K)-- SF-539 - - -A- - - SNB-75 -- 0 - - U251 - -e- - - SNB-19 . ] . Fig. 4 Patent Application Publication Jan. 24, 2002 Sheet 3 of 14 US 2002/0010208A1 MELANOMA 1 OO -8 -7 -6 -5 -4 LOGO OF SAMPLE CONCENTRATION (MOLAR) LOX IMV. -o- UACC-257- - - - SK-MEL-2- . O' SK-MEL-5 - - e - - M14 --- a-- MALME-3M--(- - Fig.UACC-62- 5 : A' -- 1 OO ARAN CANCE 5 O -5OO - 1 OO 9 -8 .7 -6 -5 -4 LOGo OF SAMPLE CONCENTRATION (MOLAR) GROW1 -O- OVCAR-3--O-- OVCAR-4--A- - OCWAR-8 - - - - SK-OV-3 - - - - - OVCAR-5 ro-r Fig. 6 Patent Application Publication Jan. 24, 2002. Sheet 4 of 14 US 2002/0010208A1 OO 5 O O -9 LOG10 OF SAMPLE CONCENTRATION (MOLAR) 786-O-O- A498- -g- - ACHN- - -A- - - CAK-1: ... RX 393-- e - - K-10---0--- UO-31 - - -A- Fig. 7 1 OO PROSTRATE CANCER O O LOGo OF SAMPLE CONCENTRATION (MOLAR) PC-3-O- Fig. 8 Patent Application Publication Jan. 24, 2002 Sheet 5 of 14 US 2002/0010208A1 1 OO BREAST CANCER 5 O -5 OO -100 -8 -7 -6 -5 -4 LOGo OF SAMPLE CONCENTRATION (MOLAR) MCF7 -O- MDA-N- - - - - - HS 578...-O.... MDA-MB-435 - -e- - MDA-MB-231/ - - A - - T-47D-...-a-...- MCF 7ADR-RE-i-o- Fig.BT-549-a- 9 1 OO LEUKEMA 5 O -5 OO -1 OO -9 -8 -7 -6 -5 -4 CCRF-CEM--o-LOGo OF SAMPLE CONCENTRATIONSR- - - - - - (MOLAR) RPM-8226--e--- K-562 - - -A - - HL-60 (TB) -g- -Fig. MOLT-4 10 to . Patent Application Publication Jan. 24, 2002 Sheet 6 of 14 US 2002/0010208A1 1 OO 5 O -5OO -100-9 -8 -7 -6 -5 -4 LOGo OF SAMPLE CONCENTRATION (MOLAR) A549/ATCC-O- EKVX --Q-- NCI-H322M----a--- NCI-H226-- - - NC-H23----- HOP-92.......... NC-H522--O-- HOP-62----Fig. 11 NC-H460-a--- 1 OO 5 O O -5 -100 9 -8 -7 -6 -5 -4 LOGO OF SAMPLE CONCENTRATION (MOLAR) COLO 205-O- HCT-116--Q-- HCT-15---a--- KM12-- e - -Fig. SW-620 12 - - - - - - HT29. Patent Application Publication Jan. 24, 2002 Sheet 7 of 14 US 2002/0010208A1 1 OO 5 O -5OO -1 O O-9 -8 -7 -6 -5 -4 LOGo OF SAMPLE CONCENTRATION (MOLAR) SF-268-O- SF-295 --K)-- SF-539 - - - A - - - SNB-75 -- 9 - - Fig.251 - 139 - SNB-19-0- 1 OO 5 O -5OO - 1 O O 9 -8 -7 -6 - -4 LOG-10 OF SAMPLE CONCENTRATION (MOLAR) LOX IMV. --O- UACC-257- --- SK-MEL-2..... 0-..... SK-MEL-5 - -e-- M14 --- a-- MALME-3M--Q- - UACC-62 - - - -a - Fig. 14 Patent Application Publication Jan. 24, 2002. Sheet 8 of 14 US 2002/0010208A1 OVARAN CANCER 1 OO Cys's S. 5 O -5 OO - 100 a -9 -8 -7 -6 -5 -4 LOGo OF SAMPLE CONCENTRATION (MOLAR) GROW 1 -O- OVCAR-3--K) -- OWCAR-4--A- - OCVAR-8 - - - - SK-OV3Fig. - - - 15 - - OVCAR-5 “d OO RENAL CANCEF 5 O -5 OO -1 O O -9 -8 -7 -6 -5 -, LOGo OF SAMPLE CONCENTRATION (MOLAR) 786-O-O-, - A498--K) - ACHN- - -A--- CAK-1: . RXF 393-- 9 - - TK-10---0-- UO-31 - A - Fig. 16 Patent Application Publication Jan. 24, 2002 Sheet 9 of 14 US 2002/0010208A1 1 OO PROSTRATE CANCER -- O - 1 OO -9 -8 -7 -6 -5 -4 LOGo OF SAMPLE CONCENTRATION (MOLAR) PC-3-O Fig. 17 1 OO 5 O -5 OO -100 -8 -7 -6 -5 -4 LOG10 OF SAMPLE CONCENTRATION (MOLAR) MCF 7 -O- MDA-N- -0-- HS 578T....... MDA-MB-435- -o- - MDA-MB-231/ --A-- T-47D-...- MCF 7/AOR-RE--K)-- BT-549 -...-A-...- Fig. 18 Patent Application Publication Jan. 24, 2002. Sheet 10 of 14 US 2002/0010208A1 EUKEMA 1 O -9 -8 -7 -6 LOGo OF SAMPLE CONCENTRATION (MOLAR) CCRF-CEM-O- MOLT-4---A- - - K-562 - (- - Fig.f SR 19 . .O. NON-SMALL CELL LUNG CANCER C - - -- -6 LOGo OF SAMPLE CONCENTRATION (MOLAR) A549/ATCC-o-, NC.-H.322M-...-0.-. NC-H226 .......O...... NC-H23---- HOP-62 --- a--- NC-H522 - - -si-...- EKWX-- C -- NC-H46O -- a--- Fig. 20 Patent Application Publication Jan. 24, 2002 Sheet 11 of 14 US 2002/0010208A1 COLON CANCER l H S. O 1. CD 1 O l CD H l O 1. I - O -9 -8 -7 -6 LOGO OF SAMPLE CONCENTRATION (MOLAR) COLOHT29- 205-O- - - - city'sKM12---0--- HCT-15 soro so e - ea as a HCC-2995- - C)-- SW-620-...--A-...-Fig. 21 CNS CANCER 1 OO - - - - - H s O 1. CD l O 9- 2. il O 1. L Cl - 1 OO- 1 O -9 -8 -7 -6 LOGo OF SAMPLE CONCENTRATION (MOLAR) SF-268-O-- SF-295 - -g- - SF-539 - - - a- - - SNB-75 -- 0 - - U251 - '-8- - - SNB-19.- ... O. Fig. 22 Patent Application Publication Jan. 24, 2002 Sheet 12 of 14 US 2002/0010208A1 MELANOMA 1 O -9 -8 -7 -6 LOGO OF SAMPLE CONCENTRATION (MOLAR) LOX MV -o- SK-MEL-5- - - - SK-MEL-2. --O-.... SK-MEL-28 - () - M14 --- a-- UACC-62 -...-a- " - MALME-3M--Q-- Fig.UACC-257- 23 - - A - OWARIAN CANCER LOGO OF SAMPLE CONCENTRATION (MOLAR) IGROV1 -o- OVCAR-3--O--OVCAR-4--a-- OCVAR-8 - - - - SK-OV-3-...-0.-Fig. 24 : OWCAR-5"O" Patent Application Publication Jan. 24, 2002 Sheet 13 of 14 US 2002/0010208A1 Her S. O 1. CD L O L CD CC - 2. 5-50Y - 1 OO- 10 -9 -8 .7 -6 LOGO OF SAMPLE CONCENTRATION (MOLAR) 786-O-O- . A498--KX- - ACHN- - - A - - - CAK-1: .... O RXF 393 - - 3 - - SN12CFig. -- (- - - 25 TK-1 O - A - UO-31 - ... a PROSTRATE CANCER o -9 -8 -7 -6 LOGo OF SAMPLE CONCENTRATION (MOLAR) PC-3-O- DU-145-Fig. -O-- 26 Patent Application Publication Jan. 24, 2002 Sheet 14 of 14 US 2002/0010208A1 BREAS CANCER E S. O 1. CD ll O CD 2. O 1. l - O g - -7 -6 LOG10 OF SAMPLE CONCENTRATION (MOLAR) MCF 7 -O- MDA-N---4--. - HS 578T. O.... MDA-MB-435- -e- - MDA-MB-231/ -- a-- -47D---a-...- MCF 71ADR-RE--&- B-549-...-A-...- Fig. 27 US 2002/0010208A1 Jan. 24, 2002 DHA-PHARMACEUTICAL AGENT CONJUGATES 0006 Taxol has the following formula: OF TAXANES BACKGROUND OF THE INVENTION AcO O OH 0001 Improving drug selectivity for target tissue is an established goal in the medical arts. In general, it is desirable H O to deliver a drug Selectively to its target, So that dosage and, X. consequently, Side effects can be reduced. This is particu Y Yo Y Y Ph 3 2 O i larly the case for toxic agents Such as anti-cancer agents HO because achieving therapeutic doses effective for treating the H PhCO2 A.C cancer is often limited by the toxic side effects of the anti-cancer agent on normal, healthy tissue. The problems relating to lack of drug Selectivity can be exemplified by 0007 Taxanes have the basic three ring structure (A. B TaxolcE). and C), Substituted or unsubstituted. 0002 Taxole (paclitaxel) was first isolated in 1971 from 0008 Taxol's carbons are numbered conventionally as the bark of Taxus brevifolia and was approved in 1992 by the follows: US Food and Drug Administration for treatment of meta Static Ovarian cancer and later for breast cancer. Its mecha nism of action is believed to involve promoting formation and hyperStabilization of microtubules, thereby preventing the disassembly of microtubules necessary for completion of cell division. It also has been reported that Taxol induces expression of cytokines, affects the activity of kinases and blockS processes essential for metastasis, in as yet unchar acterized mechanisms of action.